poster abstracts
Transcription
poster abstracts
Monday 22 June – Poster session Rhizosphere Microbiome 5 Branching out: towards a trait-based understanding of fungal ecology in the rhizosphere Carlos A. Aguilar-Trigueros*1, Stefan Hempel1, Jeff R. Powell2, Ian C Anderson3, Janis Antonovics4, Joana Bergmann1, Timothy R Cavagnaro5, Baodong Cheng6, Miranda M Hart7, John Klironomos7, Jana S Petermann1, Erik Verbruggen8, Stavros D Veresoglou1, Matthias C Rillig1 1 Freie Universität Berlin, Germany, 2Univeristy of Western Sydney, Australia, 3University of Western Sydney, Australia, 4Univeristy of Virginia, USA, 5University of Adelaide, Australia, 6Chinese Academy of Science, China, 7University of British Columbia, Canada, 8University of Antwerp, Netherlands Rhizosphere fungal ecology lags behind in the use of traits (i.e. phenotypic characteristics) to understand ecological phenomena. We argue this is a missed opportunity and that the selection and systematic collection of trait data throughout the fungal kingdom will reap major benefits in ecological and evolutionary understanding of fungi. To develop our argument, we first employ plant trait examples to show the power of trait-based approaches in understanding ecological phenomena such as identifying species allocation resource patterns, inferring community assembly and understanding diversity-ecosystem functioning relationships. Second, we discuss ecologically relevant traits in fungi (focusing on rootassociated fungi) that could be used to answer such ecological phenomena and can be measured on a large proportion of the fungal kingdom. Third, we identify major challenges and opportunities for widespread, coordinated collection and sharing of fungal trait data, for which we solicit input from the community of researchers. The view that we propose has the potential to allow mycologists to contribute considerably more influential studies in the area of fungal ecology and evolution, as has been demonstrated by comparable earlier efforts by plant ecologists. This represent a change of paradigm, from community profiling efforts through massive sequencing tools, to a more mechanistic understanding of fungal ecology in the rhizosphere. 6 The relationship between strigolactones, symbiotic fungi and drought tolerance in rice Beatriz Andreo Jimenez*1, Carolien Ruyter-Spira1, Philippe Vandenkoornhuyse2, Amandine Le Van2, Marie Duhamel2, Harro Bouwmeester1 1 Wageningen University, Netherlands, 2Rennes University, France Abiotic stresses, such as drought, are the primary causes of yield reduction in rice. One of the responses of plants to cope with drought is optimization of their root system architecture. Root architecture is controlled by the joint interaction of several plant hormones. Recently, it has been shown that strigolactones also play an important role in this process. Strigolactones are also known to stimulate the beneficial symbiosis with arburscular mycorrhizal (AM) fungi, which is also of importance during periods of water limitation because these fungi provide plants not only with nutrients but also with water. 1 Rhizosphere Microbiome 1 Monday 22 June – Poster session One of the objectives of our study is to monitor how a group of rice lines that differ in drought tolerance respond to drought with respect to strigolactone levels and endophytic fungi/AMF community structure. For this purpose we selected two well-known rice indica varieties, which historically have been selected under different cultivation managements: Apo (non-flooded conditions, drought tolerant) and IR64 (flooded conditions, drought sensitive). Interestingly, strigolactones levels in roots from the drought tolerant variety are higher when compared to the drought sensitive variety. This difference correlates with their tillering behavior, being lowest for Apo. Furthermore, SSU rRNA gene amplicons mass sequencing was used to analyze the entire endophytic and AM fungal community associated with these two varieties under field conditions (plants were grown in an upland field in the Philippines). In both rice genotypes, drought clearly affects endophyte community structure. These preliminary results may suggest a role for strigolactones in plant adaptation to water deficit through their effect on AMF symbiosis and plant architecture. The hypothesis that wet cultivation practices have led to more drought sensitive plants, having a lower endophytic and AM fungal association, higher tiller number and lower strigolactone levels will further be explored in a larger set of rice ecotypes. 7 Rhizobacterial community structure in Mahikeng rhizospheric soil and associated plant growth promoting potential Olubukola Oluranti Babalola* North-West University, South Africa Denaturing gradient gel electrophoresis (DGGE) profiles of Mahikeng soil can indicate dominant soil bacterial types and Plant Growth Promoting Rhizo-Bacteria (PGPR) can stimulate the growth of the host plant. The aforementioned were examined in relation to nine rhizospheric soils. Rhizobacteria with PGPR traits were selected for use in pot experiments on tomato and spinach. The rhizobacterial isolates tested were found to produce ammonia; several of them produced indole acetic acid (IAA; 38%) and hydrogen cyanide (HCN; 38%). Also exhibited are 1aminocyclopropane-1-carboxylate (ACC) deaminase activity (48%), phosphate solubilisation (48%) and antifungal activity (21%) against test pathogen Fusarium solani. All the HCNproducing bacteria belong to the genus Bacillus. B. amyloliquefaciens indicated high cyanogenic potential compared to other strains. The treatment of both crops with the bacterial inoculants promoted plant growth in terms of increased shoot length at P<0.05. B. amyloliquefaciens MR16 had significantly higher growth at P<0.05 compared to the uninoculated control treatment. DNA revealed some percentage identity with yet uncultured Bacillus sp. (94%), Rubrobacter sp. (90%), Rhizobiales bacterium (95%), and soil bacterium (87%) besides the culturable B. megaterium (97%) and Cohnella sp. (84%). PGPR can be used to make reliable and accessible products such as biofertilizers for farmers. Metagenomics holds the promise to reveal several important questions regarding the unculturable fraction of the rhizosphere community. 2 Rhizosphere Microbiome 1 Monday 22 June – Poster session 8 Comparative analysis of autochthonous and zymogenous bacteria in Mahikeng Agricultural Ecosystem Olubukola Oluranti Babalola*, Keokeditswe Raven Motsewabangwe North-West University, South Africa Soil is a complex biological system and it is difficult to determine the composition of microbial communities within it. Autochthonous and zymogenous soil bacteria collected in Mahikeng, South Africa were investigated. Compounded nutrient agar and soil extract agar used for isolation had glucose amendments of 0.0 to 2.0 g. Media containing 0.0 to 0.01 g of glucose were considered as having poor carbon substrate amendment. Media that contained glucose at a mass of 0.25 to 2.00 g were considered as having rich carbon substrate amendment. Soil analysis revealed between 6.5 and 7.0 average pH, 20.25 kg/acre of nitrogen, 14.50 kg/acre of phosphorus and 75 kg/acre of potassium. A great variety of bacterial species were isolated from poor media including species such as Agrobacterium tumefaciens, Planomicrobium sp, Cronobacter turicensis, Enterobacter hormaechei, Bacillus pumilis as well as B. subtilis. Bacillus species isolated from rich media were B. subtilis, a bacterial species that functions as a fungicide in the roots of plants. B. mojavensis, a unique group of bacteria that form endophytic associations with plants for the prevention of diseases, B. pumilis and, B. aryabhattai, species that colonise plant root systems and portray antibacterial and antifungal properties, Arthrobacter globiformis and A. ramosus species has the ability to degrade soil pesticides as well as pollutants such as hexavalent chromium as well as Rhizobium sp. bacteria that colonize plant cells within root nodules where they convert atmospheric nitrogen to ammonia and then provide organic nitrogenous compounds to the plant. 9 Microbes From Inner Space: I. Comparison of biological soil suppression potential against invertebrate pests across ten New Zealand pastoral soils Nigel Bell*1, Katharine Adam1, Damien Fleetwood2, Gabriela Burch3, Richard Johnson3, Alison Popay3, Faith Mtandavari3, Rhys Jones3, Vanessa Cave3 1 AgResearch Ltd, New Zealand, 2Biotelliga Ltd, New Zealand, 3AgResearch, New Zealand White clover (Trifolium repens) is the key legume component of New Zealand pastoral agriculture due to the high quality feed and nitrogen inputs it provides. There are a number of invertebrate pests which constrain white clover growth in New Zealand soils and our work is investigating rhizosphere or endosphere associated microbial controls for these pests. The degree of suppressiveness of ten soils from across New Zealand to added Meloidogyne hapla nematodes and Costelytra zealandica scarab larvae was measured in untreated soil and, in a second experiment, a comparison of plant growth between untreated and irradiated soil carried out on five of those soils using Pyronota sp. as the scarab larvae. As expected, of the ten soils most showed no suppressive activity against these pests but, promisingly, two showed activity against M. hapla and two against C. zealandica larvae. Suppression was expressed as significantly reduced galling/ mm root by nematodes and significantly reduced survival of scarab larvae compared to the other soils. Soil irradiation reduced white clover shoot growth by ca 60–70% in two soils, demonstrating the importance of the microbial flora in these soils. In a further two soils there was no significant effect of irradiation on plant 3 Rhizosphere Microbiome 1 Monday 22 June – Poster session growth, while in one soil there was a ca 16× increase in plant growth after irradiation, likely due to pathogen release as evidenced by a significant reduction in root rotting. Lack of consistent changes in soil macronutrients and pH post-irradiation suggest these were only partially responsible for plant responses. Rhizosphere and endosphere microbes responsible for pest suppressive and plant growth responses are being elucidated via next-gen sequencing and results will be detailed in an aligned presentation (Johnson et al, this volume), as will potential for beneficial microbes to be carried endophytically in white clover seed (Monk et al, this volume). 10 Release of secondary plant metabolites in soil as mediated by arbuscular mycorrhizal fungi in response to Fusarium oxysporum Fernando Monroy1, Benedetta Cangelosi1, Paolo Curir1, Valeria Bianciotto2, Roberto Borriello*2 1 Consiglio per la Ricerca e l'Analisi dell'Economia Agraria (CRA-FSO), Italy, 2Institute of Plant Protection (IPP-CNR), Italy The production of root exudates containing defensive compounds able to inhibit soilborne microbial pathogens has been proposed as one of the mechanisms involved in plant protection by arbuscular mycorrhizal fungi (AMF). However, there is little evidence of the induced release of such compounds in the soil and of their direct or indirect effects on pathogenic microorganisms. We carried out a full-factorial experiment under greenhouse conditions to check for the release of secondary metabolites by two cultivars of Ranunculus asiaticus in soils inoculated with a mix of three AMF species (Rhizophagus intraradices, Funelliformis mosseae, Glomus sp.) and the vascular wilt fungus Fusarium oxysporum f. sp. ranunculi. Chemical analyses of the rhizosphere soil revealed the presence of significant amounts of three secondary metabolites, namely syringic acid, ferulic acid and isorutin, released only by the mycorrhized plants challenged with F. oxysporum. Although the production of these compounds has been previously recorded in mycorrhized plants, we found that none of them had a direct inhibitory effect on F. oxysporum growth. Additional inoculation tests carried out with syringic acid showed a persistence of less than one day for this compound in non-sterilized soil, and the formation of syringic-derived decomposition products. These results suggest that rhizosphere microorganisms are biochemically involved in the defense response induced by AMF to F. oxysporum. 11 Characterization of the microbiome involved in the Nitrogen cycle from Kavango soils of Namibia using qPCR- and omics- based methods Claudia Sofía Burbano*, Thomas Hurek, Barbara Reinhold-Hurek Department of Microbe-Plant Interactions, University of Bremen, Germany Nitrogen is the biggest limiting factor in plant productivity in the savanna ecosystems of southern Africa. It resides in the soil organic matter and its microbial mineralization is critical for plant nutrition and soil productivity. To learn about the relation between soil management practices and the microbial nitrogen cycle in the savanna soils, we are studying 4 Rhizosphere Microbiome 1 Monday 22 June – Poster session key transformations processes of it on soils from the Kavango region of Namibia. Different land use strategies are included from two types of soils. Quantitative analyses on key functional marker genes for nitrification, denitrification and nitrogen fixation were done by qPCR and were targeted at the DNA (microbial population) and RNA (actual transcription activity) level. Microorganisms involved in those processes were present in all type of soils. At the functional level nitrification from Archaea was the most prominent process. Transcripts from nitrogen fixation were not detected in most soils studied. To characterize the structure and activity of the soil microbiome without a bias introduced by PCR we applied metagenomic and metatranscriptomic approaches to selected soils from the different land uses. We obtained five metagenomes and two metatranscriptomes using Illumina (MiSeq) paired-end sequencing technology. We obtained ~3 and ~15 Million sequences for all the metagenomes and metatranscriptomes, respectively. Our initial metagenome taxonomic analysis showed that bacterial sequences are predominant in all soils. They are affiliated to 16 phyla with Actinobacteria and Proteobacteria representing the more prevalent ones. Archaeal sequences were also present in the soils but in low abundances. The taxonomic analysis of the metatranscriptomes together with all the functional annotation will also be presented. The omics- and qPCR-based approach will give a better understanding of how the nitrogen cycling microbial communities behave under different land uses and will give a clue on how they contribute to the agricultural productivity of Namibian savanna soils. 12 Characterization of beneficial microorganisms isolated from the rhizosphere of saffron Imane Chamkhi*, Jamal Aurag, Laila Sbabou Mohammed V University, Faculty of Sciences, Morocco Saffron or Crocus sativus L., is a local product or terroir product with high added value, which is included in the national Green Morocco Plan (PMV= Plan Maroc Vert) with a fixed goal to increase three times its production by 2020. The production of this spice is not exceeding presently 3 tons per year and still rivalled in the international market. This production is limited to the region of Taliouine-Taznakht (area <600 ha) in the south of Morocco. Furthermore, saffron fields show a decline in production from the 6th year of use and require regular renewal. The project aims the conception of microbial biofertilizers for saffron to improve plants yield and quality. A first year experiments was carried and 90 microorganisms were isolated from rhizosphere sampled from Taliouine-Taznakht, on specific media. Isolates were screened for their plants growth promoting abilities (phosphate solubilisation, auxin and siderophores production). The isolates were identified using the 16S rDNA gene sequencing. Six genera were identified (Rahnella, Pseudomonas, Variovorax, Delftia, Bacillus and Luteibacter). 5 Rhizosphere Microbiome 1 Monday 22 June – Poster session The best three isolates, with higher levels of biological activities, were selected and used for the conception of saffron biofertilizers. In situ and ex situ trials (field and greenhouse conditions) are ongoing. 13 The function of the root cap in shaping the microbiome of Arabidopsis rhizosphere Xu Cheng*, Rene Geurts, Carolien Franken, Ben Scheres, Ton Bisseling Wageningen University, Netherlands Microbial communities play a pivotal role in the functioning of plants by influencing their physiology and development. In the rhizosphere, plant roots release low molecular weight compounds (amino acids, organic acids and sugars), mucilage and proteins. These rhizodeposits are a major driving force in the regulation of microbial diversity and activity on plant roots, and also affect plant to modulate the rhizosphere microbiome for selecting beneficial microorganisms. However, to what extend the root cap contributes to this root exudate is not clear. As part of the maturation of root cap cells they detach from the root after which they stay alive for some time in the soil. The availability of Arabidopsis mutants that make fewer or immature root cap cells provides good opportunities to determine the role of the root cap in shaping the microbiome. Previous studies showed that a loss of function mutation in FEZ results in a reduced number of columella and lateral root cap layers. SOMBRERO (SMB) BEARSKIN1 (BRN1) and BRN2 regulate the cellular maturation of root cap cells. Lateral root cap cells fail to detach from the root in smb-3 mutant, columella cells fail to detach in brn1-1/brn2-1 double mutants and cells fail to mature in all parts of the root cap in the smb-3/brn1-1/brn2-1 mutant. To reveal the function of root cap in shaping plant rhizospheric microbiome, in this study, we grew Arabidopsis Columbia wild type and smb-3, fez-2, brn1-1/brn2-1 and smb-3/brn1-1/brn2-1 mutants under controlled condition. Total genomic DNA was isolated from the rhizosphere and microbial compositions were analyzed by using 16S rDNA amplicom sequencing with Miseq250 platform. The role of the root cap on shaping the microbiome will be discussed. 14 The influence of protist grazing on corn rhizodeposition and recalcitrant litter decomposition Fionn Clissmann*1, Sebastian Löppmann2, Anna Gunina2, Johanna Pausch2, Yakov Kuzyakov2, Robert Köller3, Michael Bonlowski1 1 University of Cologne, Germany, 2University of Göttingen, Germany, 3Forschungszentrum Jülich, Germany Plants provide carbon (C) to the C limited microbial community by means of root exudates and thereby increase decomposition and N release from recalcitrant litter in soil, the so called plant `priming` effect. Protists have also been found to increase N release from the microbial community, through the ingestion and destruction of bacterial cells and excretion of ammonia. This boosts plant growth, increasing exudate release, leading to further microbial decomposition and N release, the so called `microbial loop`. 6 Rhizosphere Microbiome 1 Monday 22 June – Poster session To investigate the contribution of protists to the plant `priming` effect we isotopicaly labelled both C channels, organic litter (13C, 15N) and root exudates (14C) in corn (Zea mays)-planted microcosms and duplicated all treatments with an added model protist (Acanthamoebae castellani). Plants were transferred into soil after initial fluxes of easily available C had subsided from the parent soil and mixed 13C, 15N recalcitrant litter (Lolium perenne root material) allowing for a elucidation of root exudation induced increases in microbial decomposition and N release. CO2 released from the soil was trapped throughout the experimental duration and plants were pulse labelled with 14C three days before harvest. 14C content in the shoot, root and microbial biomass and CO2, phospholipid fatty acid and microbial biomass 13C content, total DNA and enzymatic activity were analysed. First results include a higher 14C activity detected in the microbial biomass in the presence of protozoa and increased DNA content in litter amended and planted soil in the presence of protozoa.This indicates that the added protozoa significantly boosted plant C allocation to the soil, potentially speeding the decomposition of recalcitrant soil litter. 15 Impact of benzoxazinoids in root exudates of maize on interactions with rhizosphere bacteria and arbuscular mycorrhizal fungi T.E. Anne Cotton*1, Steve Rolfe1, Duncan Cameron1, Pierre Pétriacq1, Georg Jander2, Matthias Erb3, Jurriaan Ton1 1 University of Sheffield, United Kingdom, 2Boyce Thompson Institute for Plant Research, United States, 3University of Bern, Switzerland Interactions between plant roots and soil microbes are important determinants of plant growth and health. Plants can modify their rhizosphere by the exudation of primary and secondary metabolites. Benzoxazinoids (BXs) are tryptophan-derived secondary metabolites of maize and other grasses that contribute to aboveground resistance against pests and diseases. When exuded from roots, BXs can act as belowground recruitment signals for beneficial soil microbes, such as Pseudomonas putida, and root-feeding pests, such as the corn rootworm (Diabrotica virgifera). Plant growth promoting rhizobacteria and arbuscular mycorrhizal (AM) fungi have also been reported to alter BX levels in roots. Hence, BXs are emerging as important regulators of belowground plant-biotic interactions. However, little is known about the exact mechanisms by which BXs are secreted by roots, their degradation pathways in the rhizosphere, and their impacts on rhizosphere microbial communities. The European research consortium BENZEX will address these shortfalls in our current knowledge. Within this consortium, we will use maize plants with mutations in three BX biosynthesis genes (Bx1, Bx2 and Bx6), in conjunction with molecular methods, such as high throughput sequencing, to examine the effects of BXs on agricultural rhizosphere bacterial communities. In addition, we will examine the impact of AM fungi on BX exudation as a possible driver of mycorrhizosphere development and biocontrol of belowground pests and examine the impact of BXs on mycorrhizal development. Our results will both enhance our fundamental understanding of soil-microbial interactions and provide valuable insights into the potential application of soil microbes in control strategies for sustainable agricultural production. 7 Rhizosphere Microbiome 1 Monday 22 June – Poster session 16 Combined effects of compost and arbuscular mycorrhizal inoculation on soil fertility, microbial community structure and plant growth Vincenza Cozzolino*1, Vincenza Cozzolino1, Hiarhi Monda1, Vincenzo Di Meo1, Riccardo Spaccini2, Alessandro Piccolo1 1 University of Naples Federico II, Italy, 2University of Naples, Italy Increasing the sustainability of cropping systems requires management strategies that involve the beneficial activity of rhizosphere microorganisms and the recycling of organic wastes. Limited information are available on the combined application of arbuscular mycorrhizal fungi (AMF) inoculants and composted organic wastes on soil microbial community structure and their effects on crop productivity and soil fertility. Therefore, a field study was conducted to evaluate the effects of commercial inoculation (CI) and the application of composted municipal organic wastes on physical-chemical and biological parameters. Three fertilizer treatments were compared: 1. inorganic fertilizer (NPK), 2. compost (CM), 3. half inorganic fertilizer dose plus compost (HCM). Maize plants in all treatments were inoculated (CI+) or uninoculated (CI-). The effects on maize growth, soil physical-chemical properties (soil aggregation, N, C total and available P), mycorrhizal fungi growth (root colonization, C16:1ω5 phospholipid-PLFA and neutral-NLFA fatty acid content) and soil microbial community structure (PLFA profiles) were determined in soil and plant samples. CI+ maize treatments increased AMF root colonization, yield and P uptake in NPK and CM treatments, in respect to CI- plots, although the largest yield were obtained with NPK. On the contrary, application of compost reduced AMF growth as indicated by the decline of root colonization and NLFA C16: 1ω5 content. Compost increased total PLFA and altered soil microbial community structure, by decreasing Gram-positive to Gram-negative bacteria ratio and enhancing fungal PLFA, as compared to NPK. In combination with CI treatments, our compost favored soil macroaggregates formation and enhanced total C, N and available in both CM and HCM treatments, by improving structural and soil fertility parameters, whereas its related yields were not comparable to NPK treatments and depressed AMF growth. Alteration of soil microbial community promoted activity of an antagonistic microflora, thereby hindering the relationship between AMF and plants and compromising plant growth. 17 The rhizosphere microbiome of seagrasses and their role in sulfur processes Catarina Cúcio*1, Aschwin Engelen2, Gerard Muyzer1 1 Institute for Biodiversity and Ecosystem Dynamics, University of Amsterdam, Netherlands, 2Centre of Marine Sciences, University of Algarve, Portugal Seagrass meadows grow in coastal, biogeochemically active sediments. These productive ecosystems are distributed from tropical to temperate areas and are dominated by seagrasses, a group of marine angiosperms that is often threatened by the presence of hydrogen sulfide, a phytotoxic gas produced by belowground bacterial communities. The seagrass rhizosphere is still poorly described, however the occurrence of die-off events caused by high levels of sulfide stresses the need to understand these plants and bacterial communities present in their rhizosphere – rhizobiome. Using Next Generation Sequencing of 8 Rhizosphere Microbiome 1 Monday 22 June – Poster session 16S rDNA amplicons, we described the bacterial communities of three North Atlantic seagrasses, Zostera marina, Z. noltii and Cymodocea nodosa. Comparative analysis revealed that the rhizobiome of seagrasses is strongly influenced by the plant and significantly differs from the communities present in their surrounding environment (i.e., bulk sediment and seawater). However, we found that the rhizobiomes of the different seagrass species did not vary at a local scale, but were significantly different between plants from different geographical locations. The core rhizobiome of seagrasses is mainly composed of OTUs related to bacteria known for being involved in the sulfur cycle (sulfate reduction and sulfur oxidation), and for their ability to fix nitrogen. Moreover, our results point to a niche differentiation of sulfur bacteria, in which sulfide oxidation is performed by different taxa. These results are a strong indication of the importance of sulfur bacteria in seagrass ecology. 18 The selection on sugarcane rhizosphere upon the Pseudomonas community Lucas Dantas Lopes*, Michele de Cássia Pereira e Silva, Luana Bresciani, Fernando Dini Andreote University of Sao Paulo, Brazil The knowledge on interactions between plants and key microbial groups is essential for a better exploration of rhizosphere interactions though an efficient and sustainable plant production system. Here we targeted the selection exerted by sugarcane (an important crop for sugar and bioethanol production) rhizosphere upon the community of Pseudomonas spp., a well-known bacterial group involved in plant growth promotion. Six randomly selected plants were sampled in a cultivation field, and bulk soil was contrasted with rhizosphere (manually separated by selecting soil highly adhered to the maximum of 1 mm or the roots vicinity). The responsiveness of Pseudomonas spp. to the sugarcane rhizosphere was determined by culture independent analyses, where quantitation results (copies of the 16S rRNA gene per g of soil) showed that Pseudomonas is significantly enriched in the rhizosphere (p < 0.01), shifting from 2.5x104 in the bulk soil to 8.7x105 in the rhizosphere. A NMDS constructed by using the OTU table, from a sequence-based analysis of the 16S rDNA performed by Illumina, indicated a clear distinction between the structures of Pseudomonas spp. communities in bulk soil and rhizosphere. We gained insights on the intra-genus variations by cultivation of 34 isolates of Pseudomonas in Pseudomonas Agar Base medium (21 from bulk soil and 13 from rhizosphere). The 16SrDNA sequencing revealed the presence of three groups; two in both environments (P. fluorescens/P. koreensis and P. plecoglossicida/P. monteilii), while the group affiliated to P. putida was exclusively found in the rhizosphere. BOX-PCR analysis indicated a deeper variation on genome contents of these isolates, clustering 7 genotypes (some exclusive in soils or rhizosphere composing 23 BOX profiles). Our final target will be to determine whether these variations are commonly observed among the Pseudomonas cells in soils, or if the genomic re-arrangements are important on rhizosphere-succeeded isolates. 9 Rhizosphere Microbiome 1 Monday 22 June – Poster session 19 Microbial eco-compatible strategies for improving wheat quality traits and rhizospheric soil sustainability (MIC-CERES) Cindy Dasilva*1, Nathalia Arias Rojas1, Daniel Garçia Seco de Herrera1, Eddy Ngonkeu2, Valentina Fiorilli3, Florence Wisniewski-Dyé4, Diégane Diouf5, Candida Vannini6, Valeria Terzi7, Jacques Le Gouis8, Aboubacry Kane5, Madiama Cisse9, Marcella Bracale6, Paola Bonfante3, Ralf Koebnik1, Lionel Moulin1 1 IRD, UMR Interactions Plant Microorganisms Environment, France, 2Institute of Agricultural Research for Development & University Yaoundé, Cameroon, 3University of Torino, Dpt. Life Science and Systems Biology, Italy, 4University Claude Bernard Lyon I, UMR-CNRS 5557 Microbial Ecology Laboratory, France, 5University Cheikh Anta Diop, Common laboratory of Microbiology (IRD/UCAD/ISRA), Senegal, 6University of Insubria, Dpt. Biotechnology and Life Science, Italy, 7CRA-GPG, Genomics Research Centre, Italy, 8INRA/UBP, UMR 1095, Génétique, Diversité et Ecophysiologie des Céréales, France, 9Senegal Institute of Agricultural Research, Common laboratory of Microbiology (IRD/UCAD/ISRA), Senegal Wheat (Triticum aestivum L. subsp. aestivum) is one of the most important sources for food, animal fodder and industrial raw materials. However these past years, world-wide wheat production has not met demand, largely due to the adverse effects of climate change, soil salinization, and limited water availability. Here, we present the project MIC-CERES (20142016) funded by Agropolis (France) and Cariplo (Italy) Fondations, with seven groups from four countries (France, Italy, Senegal, and Cameroon), and in collaboration with the International Maize and Wheat Improvement Center (CIMMYT) and INRA (France). The goal of this project is to characterize the response of wheat to colonisation by arbuscular mycorrhizal fungi and symbiotic beneficial bacteria, with a view to using them as natural biofertilizers and bioprotectors in integrated strategies for wheat cultivation. Our main objectives are: 1) determine structure, diversity and activity of the microbiome associated to the roots of wheat by a barcoding approach of ribosomal markers with Illumina MiSeq, coupled with a culturable approach to isolate plant growth-promoting bacteria. Several varieties of wheat are tested on soils with different characteristics (from four countries) in order to understand the impact of both soil origin and plant variety on the microbiota associated to wheat roots; and 2) characterize the molecular wheat responses to beneficial and harmful microbes by both transcriptomic and proteomic analysis to better understand the wheat-microbes interactions. Wheat will be challenged with a mycorhizal fungi, Azospirillum brasilense, Burkholderia graminis or Xanthomonas translucens in single or multiple inoculations. We will present the first data on the culturable approach on the wheat plant-growth promoting bacteria. 20 Effect of biochar amendment on the rhizosphere microbiome of lettuce and strawberry Jane Debode*, Caroline De Tender, Pieter Cremelie, Bart Vandecasteele, Martine Maes Institute for Agricultural and Fisheries Research (ILVO), Plant Sciences Unit, Belgium Biochar, a solid coproduct of biomass pyrolysis, can be used as soil amendment. It thereby has potential to help mitigate climate change, as it permanently sequesters carbon from the atmosphere and it may be beneficial for agricultural crops. We investigate the effects of 10 Rhizosphere Microbiome 1 Monday 22 June – Poster session biochar soil amendment on plant health and rhizosphere microbiology (FP7-Fertiplus project). For 2-3 months, lettuce was grown in field soil and strawberry was grown in peat, each amended with 0, 1 or 3% biochar produced from holm oak. Changes in chemical soil/substrate and plant properties were measured. The associated changes in the rhizosphere microbiome were studied using phospholipid fatty acid (PLFA) analysis and amplicon sequencing of the bacterial V3-V4 region of the 16SrDNA. Biochar amendment had a different effect on the lettuce-soil versus the strawberry-peat rhizosphere. For the lettuce-soil rhizosphere, PLFA analysis showed that the 3% biochar application increased the population of arbuscular mycorrhizal fungi, whereas no significant shifts could be observed in the bacterial microbiome based on both PLFA and 16S analysis. For the strawberry-peat rhizosphere, PLFA analysis showed a significant clustering of the 3% biochar application as compared to the 0% and 1% applications. This was confirmed by the 16S analysis, which dedicated the shift to the reduction in relative abundance of the Proteobacteria, mainly Burkholderia species. This microbial information was combined with plant and substrate properties, showing a relationship between strawberry rhizobiome, biochar application rate, plant growth, plant resistance to Botrytis cinerea and substrate moisture and water-soluble phosphorus content. In sum, PLFA analysis and 16S amplicon sequencing showed that biochar incorporation can have an influence on the rhizosphere microbiome, associated with induced plant growth and health. This influence was more pronounced in the peat substrate than in soil, which is a more diverse and complex environment and is thus probably less sensitive to disturbance. 21 Mollicutes-related endobacteria in basal fungi: genetic determinants of the fungal microbiome Alessandro Desirò*1, Martin Bidartondo2, Paola Bonfante3, Gregory Bonito1 1 Department of Plant, Soil, Microbial Sciences, Michigan State University, USA, 2Imperial College London and Royal Botanic Gardens, Kew, United Kingdom, 3Department of Life Sciences and Systems Biology, University of Turin, Italy Fungi are often associated with microorganisms such as bacteria, which colonize their outer surface or, in some fungal taxa, live in their cytoplasm as endobacteria. An example of endobacterium-fungus interaction is represented by arbuscular mycorrhizal fungi (Glomeromycota), which are important component of the root and rhizosphere microbiome. Two types of endobacteria are known in arbuscular mycorrhizal fungi: a β-proteobacterium called Candidatus Glomeribacter gigasporarum, and a coccoid bacterium which represents an undescribed taxon of Mollicutes-related endobacteria. Interestingly, the two bacterial populations have also been simultaneously detected in some Glomeromycota spores hosting a newly described fungal microbiota. However, some evidence indicates that other fungi belonging to Mucoromycotina contain endobacteria, e.g. an endobacterium closely related to Candidatus Glomeribacter gigasporarum has been identified in Mortierella elongata, while endobacteria similar to Mollicutes-related endobacteria have been morphologically detected in Endogone flammicorona. 11 Rhizosphere Microbiome 1 Monday 22 June – Poster session Here we investigate i) bryophytes-associated fungi, and ii) Endogone fruiting bodies in order to identify their endobacteria. Molecular analyses revealed the presence of Mollicutes-related endobacteria both in bryophytes-associated fungi and Endogone fruiting bodies. The partial sequencing of the 16S rRNA gene on the MiSeq platform shed light on the variability of Mollicutes-related endobacteria dwelling in fruiting bodies. Phylogenetic reconstructions placed the Mollicutes-related endobacteria sequences retrieved from Endogone with the ones from Glomeromycota, forming, however, a separate new clade. These results confirm previous morphological studies on Endogone and allow to identify Mollicutes-related endobacteria in bryophytes-associated fungi and Endogone fruiting bodies. The presence of exclusive Mollicutes-related endobacteria phylotypes, both in Glomeromycota and Endogone, suggests that they may be useful markers of fungal presence. The presence of Mollicutes-related endobacteria in a group of basal fungi, Endogone, close to, but potentially more ancient than arbuscular mycorrhizal fungi, opens new questions concerning the relationships between Mucoromycotina and Glomeromycota, and the origin and evolution of this little known group of endobacteria. 22 Landscape to rhizosphere: large and small-scale drivers of Pseudomonas community structure in New Zealand pastoral soils Bryony Dignam*1, Leo Condron2, Maureen O'Callaghan3, George Kowalchuk4, Jos Raaijmakers5, Steve Wakelin3 1 AgResearch Ltd and Bio-Protection Research Centre, Lincoln University, New Zealand, 2Bio-Protection Research Centre, Lincoln University, New Zealand, 3AgResearch Ltd, New Zealand, 4Utrecht University, Netherlands, 5Netherlands Institute of Ecology, Netherlands Pseudomonas species are ubiquitous in soils and contribute towards suppression of various soil-borne phytopathogens. This study aims to identify opportunities to manage Pseudomonas communities as ‘biological resources’ for sustainable agricultural systems. Sequential investigations were performed across spatial scales to identify edaphic, environmental and agricultural management factors affecting the composition of Pseudomonas communities in New Zealand pastoral soils. A country-wide survey of 50 soils showed that farm management practises were more dominant than geographic or environmental properties, with farm intensification and soil properties (notably organic matter quality (C:P ratio)) associated with Pseudomonas community structure. Soil and plant-specific influences were experimentally tested among ‘brown’ and ‘recent’ soils, each planted to ryegrass, clover and kale. No direct influence of soil order was evident. However there were differences in community structure among individual soils (P=0.001), driven by soil pH and E.C., and to a lesser extent, among plant species. Thus, physicochemical properties were stronger drivers of Pseudomonas communities than plantbased selection. To determine the influence of selection at the soil-root interface, a single soil was planted to three pasture plant species and Pseudomonas communities profiled in the bulk soil, 12 Rhizosphere Microbiome 1 Monday 22 June – Poster session rhizosphere and rhizoplane compartments. There were significant differences between plant species (P=0.005), but more strongly among compartments (P=0.001). Surprisingly, all bulk and rhizosphere communities were similar; the majority of variation among Pseudomonas communities occurred at the rhizoplane (P=0.001). Ryegrass rhizoplane communities were highly dissimilar to those of the other two plant species. Our findings show that soil properties are stronger drivers of Pseudomonas communities than plant-based selective pressures at larger spatial scales. However, at the small spatial scales of the root-soil interface plant influences were observed especially in the rhizoplane. Ongoing work is determining whether ‘disease suppressive’ functional components of the Pseudomonas communities (i.e. functional genes) are driven by similar factors to the phylogenetic component. 23 Cultivation of bacteria from the sugarcane rhizosphere and the role of roots exudates on its development Danielle Gonçalves dos Santos, Simone Raposo Cotta, Fernando Dini Andreote* University of São Paulo, Brazil The sugarcane (S. officinarum) is a perennial gramineous used for bioethanol production, a renewable energy source. Studies related to the improvement of cultivation conditions are of great importance. It is known that the rhizosphere is an environment that hosts an intimate interaction between plants and their respective microbiomes, being it mediated by the roots exudates. This study aimed to understand the effect of plant exudation on composition and behavior of microbiome through cultivation-dependent methodologies. Soil and rhizosphere samples from a sugarcane field were used for bacteria cultivation, followed by the genetic (BOX-PCR and partial sequencing of the 16S rRNA gene) and metabolic (BIOLOG®) characterization of isolates. Results demonstrated higher numbers of culturable bacteria in rhizosphere when compared to soil samples, with the prevalent affiliation of isolates to the phylum Proteobacteria (especially with the classes Gammaproteobacteria and Betaproteobacteria). The BOX-PCR results showed a great genetic diversity, even when isolates affiliated to the same taxa are compared. In counterpart, the analysis of the 16S rRNA gene sequence indicated that several isolates preserve high similarities in the ribosomal gene. The metabolic profiling results corroborated with the BOX-PCR data, which isolates highly correlated in the taxonomical analyses presented distinct capacities to use the carbon sources that were tested. At the end, this metabolic diversity was evidenced by the distinct behavior of isolates belonging to the same genera, but isolated from soils or rhizosphere samples, when cultivated in the presence of roots exudates. In general, this study demonstrated that sugarcane plants can influence the behavior of bacterial communities present in soils. It can also be indicated the individuality of components of the rhizosphere microbiome, with distinct behavior regardless to the taxonomical affiliation of isolates. 13 Rhizosphere Microbiome 1 Monday 22 June – Poster session 24 Long-term fertilization influenced the dynamics of rhizosphere microorganisms in maize Ivica Djalovic*1, Yinglong Chen2, Nastasija Mrkovacki1, Dragana Bjelic1 1 Institute of Field and Vegetable Crops, Serbia, 2School of Earth and Environment, and UWA Institute of Agriculture, University of Western Australia, Australia The aim of this study was to investigate dynamics of microorganisms in the rhizosphere of maize in long-term monoculture in dependance of inoculation with azotobacter. The study treatments were: 1. Ø (control treatment without mineral or organic fertilizers); 2. Ø (control traetment with inoculation); 3. NPK (fertilized with mineral fertilizer); 4. NPK + AC (fertilized with mineral fertilizer with inoculation); 5. NPK + M (fertilized with manure and mineral fertilizer); 6. NPK + M + AC (fertilized with manure and mineral fertilizer with inoculation); 7. NPK + CR (plowing crop residues - maize and mineral fertilizer); 8. NPK+ CR + AC (plowing crop residues – maize and mineral fertilizer with inoculation). Inoculation was done with liquid culture of Azotobacter chroococcum (mixture of three strains). Soil samples were taken for microbiological analysis at three dates (20th May, 7th June and 2nd August). The application of differetiated fertilization and the succesive stages of plant development found a reflection in the changes of the number of the studied soil microorganisms. At the first sampling period significantly higher total number was obtained in treatments 2, 4 and 5 in relation to the other treatments. At the second period treatment 6 had the largest total microbial number, and significantly higher number compared to treatments 1, 5 and 7 was achieved in treatments 2, 3, 4 and 8. At the third period only treatments 5 and 6 had significantly higher total number compared to other treatments. The total number of microorganisms decreased during the growing season, except at the second sampling period in NPK + M + AC treatment. At the first period of sampling the highest total microbial number was recorded, while the lowest values were obtained at the third period. NPK + M + AC treatment positively affected the total number of microorganisms. 25 Factors determining the composition of rhizosphere and root-inhabiting bacterial communities of Arabis alpina Nina Dombrowski*1, Klaus Schläppi2, George Coupland1, Paul Schulze-Lefert1 1 Max Planck Institute for Plant Breeding Research, Germany, 2Agroscope, Switzerland Plants growing in soil associate with a plethora of microorganisms, including nematodes, fungi and bacteria. Bacteria can colonize the rhizosphere and roots of plants and influence plant growth via hormone modulation, nutrient mobilization and suppression of defense responses. However, little is known about the structure and functions of bacterial consortia living in association with plant roots. To address this gap, we used the arctic-alpine plant Arabis alpina as a model to unravel factors influencing community structure. To determine the influence of plant residence time in soil and plant development on community composition, A. alpina wild-type and pep1 mutant plants were grown under controlled environmental conditions for seven months. The pep1 mutant does not require a vernalization treatment to flower and allows a direct comparison of a flowering and nonflowering plant at the same time point. This enabled us to disentangle the influence of plant residence time in soil and plant development on community structure. Profiling of the 16S rRNA gene by Illumina sequencing identified a distinctive taxonomic structure of bacterial 14 Rhizosphere Microbiome 1 Monday 22 June – Poster session communities within the soil, rhizosphere and root compartments. Additionally, the taxonomic composition of bacterial communities was only partially stable, for example Bacteroidetes were outcompeted over time. Notably, plant developmental stage did not influence community structure. To validate these observations and to assess the importance of soil type and environmental conditions, we harvested A. alpina roots at a natural site in the French Alps and profiled bacterial communities. We could confirm the absence of an effect of plant developmental stage on community assembly. In addition, we found that soil type and environmental conditions induce pronounced shifts in rhizosphere and root-inhabiting bacterial communities. Surprisingly, ~30% of those communities were stable across soil types and environmental conditions, allowing us to identify a shared bacterial microbiota, which might contribute bacterial functions for plant growth. 26 Intercropping is an effective way to control fusarium wilt of faba bean and improve rhizosphere microbe diversity Yan Dong*1, Li Tang1, Yi Zheng2 1 Yunnan Agricultural University, China, 2Southwest Forestry University, China Fusarium wilt of faba bean is one of the most widespread and destructive diseases that reduce faba bean yield and quality. As a soil-borne fungal disease, fusarium wilt is difficult to control with conventional chemicals in field production. Intercropping has been practiced for a long history in crop production in China, and widely accepted as a sustainable practice due to its yield advantage and disease suppression. Field experiment was conducted to examine the effects of wheat and faba bean intercropping on faba bean yield, rhizosphere microecology and occurrence of faba bean fusarium wilt. Results showed that aboveground dry weight,grain yield and 100-seed weight of faba bean significantly increased by 99.11%, 44.29% and 12.17%, fusarium wilt incidence and disease index of faba bean significantly decreased by 20% and 30.4% respectively as intercropped with wheat. The Biolog analysis with ECO plate could interpret the metabolic functions diversity of rhizosphere microbial community, results showed that intercropping significantly increased the average well color development (AWCD) of faba bean, the AWCD value of intercropped faba bean increased by 82.7% in comparison with that of monocropped. Functional diversity index of Shannon index (H) and substrate richness (S) in Biolog ECO microplates were significantly higher with intercropping than that with monocropping, substrate richness of intercropped faba bean increased by 30.3%. Intercropping changed the microbial community diversity, the activity of invertase, urease and catalase in rhizosphere of faba bean. It is suggested that wheat and faba bean intercropping could improve rhizosphere soil micro eco-environment and control the soilborne disease caused by faba bean continuous cultivation. 15 Rhizosphere Microbiome 1 Monday 22 June – Poster session 27 Influence of primextra on rhizosphere microorganisms of cowpea Fatuyi Ekundayo*, Adebanke Adedeji Federal University of Technology, Akure, Nigeria Fungicides are known to either be stimulatory or inhibitory to microorganisms that are essential for plant growth. This present investigation was therefore aimed at determining the effects of primextra at four different concentrations on the rhizosphere microorganisms of cowpea. A field experiment was carried out at the back of Crop, Soil and Pest Management Department, Federal University of Technology, Akure, Ondo State, Nigeria for a period of 30 days. Cowpea seeds were planted and after 14 days, four different concentrations (0.00, 0.50, 1.00, 3.00) of primextra were added. After experimental trial, rhizosphere microorganisms were isolated using Wakes A and B agar. The ability of the isolated microorganisms to solubilise tricalcium phosphate was evaluated on Pivoskaya’s medium. The microbial isolates were also screened for phosphatases by standard techniques. There was decrease in the bacterial and fungal counts with increase in the concentration of primextra. The bacterial isolates obtained from primextra treated soils include Bacillus subtilis, B. cereus, Klebsiella pneumoniae, Proteus mirabilis, Escherichia coli, Serratia marcescens, Staphylococcus aureus and Micrococcus luteus while the fungal isolates were Aspergillus niger, Penicillium italicum. The isolated microorganisms were able to solubilize tricalcium phosphate in varying degrees.The concentration of solubilized tricalcium phosphate ranged from 97.87 to 559.47µ/mg. Bacillus subtilis, B. cereus, Klebsiella pneumoniae, P. mirabilis, Serratia marcescens had high phosphatase activity while E. coli, S. aureus low phosphatise activity. The microorganisms with high phosphatase activity could be inoculated into the rhizosphere soil of cowpea in primextra treated soil. 28 Effects of bio-effectors on the growth and rhizosphere microbiome of tomato plants grown in low phosphorus soil Namis Eltlbany*1, Guoch Ding2, Mohamed Baklawa1, Kornelia Smalla1 1 Julius Kühn-Institut, Federal Research Centre for Cultivated Plants (JKI), Institute for Epidemiology and Pathogen Diagnostics, Braunschweig, Germany, 2College of Resources and Environmental Science, China Agricultural University, China Bio-effectors are a viable microorganism which directly or indirectly affect plant performance and can contribute to a reduced fertilizer and pesticide use in crop production based on biological mechanisms interfering with soil-plant-microbe interactions. Low phosphorus availability limits plant growth in many soils and is a common constraint to agricultural productivity. Greenhouse experiment was conducted aiming to study the effect of four bioeffectors (B1: Trichoderma harzianum T-22; B2: Pseudomonas sp.; B3: Bacillus amyloliquefaciens FB01, B4: Pseudomonas jessenii RU47) on the growth of tomato plants in a phosphorus limited soil and their effects on the indigenous rhizosphere bacterial community compared to non-inoculated plants (B0). At each sampling time the rhizosphere competence and colonization patterns of the bio-effectors were monitored in rhizosphere samples using colony forming units counts. Effects on the bacterial community composition were determined using denaturing gradient gel electrophoresis (DGGE) and pyrosequencing of 16S rRNA gene fragments amplified from total bulk and rhizosphere community DNA. All bacterial bio-effectors had a good rhizocompetence and promoted the growth of tomato 16 Rhizosphere Microbiome 1 Monday 22 June – Poster session with B3 and B4 showing the best activity. The UPGMA analysis for DGGE and amplicon sequences showed significant differences between the rhizosphere bacterial community composition of B0 and the inoculated samples. Amplicon sequencing allowed us not only to reveal bacterial genera with significantly increased or decreased relative abundance in the tomato rhizosphere compared to the bulk soil dominant genera enriched in the rhizosphere but also responders to the inoculation. B4 became a dominant population and caused strong but transient bacterial community changes. At the last sampling time a significantly increased relative abundance of Bacteroidetes and Betaproteobacteria was observed for all bio-effector treatments and for Lysobacter for all bacterial inoculants. Our data showed that inoculants cause major but often transient shifts in the bacterial community which might also contribute to the PGP effects observed. 29 Exudates of arbuscular mycorrhizal hyphae trigger phytin mineralization and P turnover in the hyphosphere Fei Wang*, Ning Shi, Rongfeng Jiang, Fusuo Zhang, Gu Feng China Agricultural University, China The aim of this study was to elucidate a direct pathway for the translocation of photoassimilated carbon from maize plants to extraradical mycelium-associated phosphate solubilizing bacteria (PSB) in the hyphosphere, and in turn, these PSB mediate the mineralization and turnover of organic P. The flux of photosynthesized C from maize to bacteria associated with extraradical hyphae of AM fungus was traced using stable isotope probing (SIP). The microbes in the hyphosphere soil receiving 13C from maize were assessed through T-RFLP analysis with stable isotope probing (DNA-SIP). The bacteria actively assimilating C derived from pulse-labeled maize plants were Massilia aurea (Oxalobacteraceae), Streptomyces spp. (Streptomycetaceae) and Pseudomonas alcaligenes (Pseudomonadaceae). In turn, PSB interacted with AM fungus in the hyphosphere chamber to mediate a marked decline in the organic P concentration and an increase in the microbial biomass phosphorus concentration in hyphosphere soil but did not contribute to the uptake of P by maize. These results provide the first in situ demonstration of the pathway underlying the carbon flux from plants to the AM mycelium-associated PSB and that the PSB rewards the acceleration of phytin mineralization and turnover with carbohydrates. 30 Structure of fungal and bacterial communities in a boreal forest podzol and their response to nitrogen fertilisation Roger Finlay*, Srisailam Marupakula, Juan Santos-González, Shahid Mahmood SLU, Uppsala BioCenter, Sweden Symbiotic ectomycorrhizal fungi form diverse communities in boreal forests, mobilising nutrients from organic polymers with different degrees of recalcitrance and from mineral substrates that are weathered at different rates. These fungi may also sequester photosynthetically-derived carbon in the soil, but the different roles of individual taxa and their location within the soil profile are still poorly understood. Structure and activity of soil microbial communities are influenced by both atmospheric nitrogen deposition and 17 Rhizosphere Microbiome 1 Monday 22 June – Poster session applications of fertiliser, but detailed knowledge of the community dynamics of these responses is still lacking. In this study we investigated fungal and bacterial community structure in different horizons of an 80 year old pine forest at Lamborn, Sweden, using highthroughput sequencing. Samples were taken in three replicate plots of two treatments. Half the plots were unfertilised, the other plots had been fertilised with 150 kg N ha-1 16 months prior to sampling. Ten replicate cores were taken from each plot, divided into O, E and B horizons and pooled prior to DNA extraction, PCR and pyrosequencing of both soil and root fungi. Sequences were quality filtered and clustered using the bioinformatics pipeline SCATA for fungi and the RDP pipeline for bacteria. A total of 807 fungal OTUs N>2 reads (496 OTUs with N>5 reads) was found in the soil samples with corresponding figures of 546 and 350 OTUs in the root samples. Fungal community structure in soil differed significantly between soil horizons but not between N treatments. Total numbers of OTUs declined significantly down the soil profile and were higher for soil DNA in N+ treatments in the O horizon than in the corresponding N- treatment. The horizon effect on OTUs from roots was not significant but there was a significant decrease in the number of root-associated OTUs in response to N. 31 Structure and successional changes in communities of bacteria, archaea and fungi colonising granitic rock surfaces in a boreal forest Roger Finlay*, Shahid Mahmood SLU, Uppsala BioCenter, Sweden In boreal forest ecosystems, bedrock outcrops and boulders are generally covered by morlayer soil, with distinct mycelial mats at the interface between the rock surface and mor-layer. Using such rocks as model systems, we are studying microbial diversity and patterns of colonisation in relation to biogeochemical weathering processes that lead to mobilisation of mineral nutrients essential for plant growth and ecosystem function. The nutrients released from the rocks are utilised by the microbial community, taken up directly by roots or transported to roots via mycelia of ectomycorrhizal fungi forming symbiotic associations with tree roots. We hypothesise that both microbial diversity and abundance will be greater on rocks receiving higher amounts of recently fixed photosynthetic carbon (to produce weathering agents such as organic acids, or siderophores) compared to bare rocks. To test this we sampled rock surfaces that were either: 1) bare or colonised only by lichens, 2) colonised by mosses or 3) colonised by tree roots and fungal mycelia. We analysed communities of bacteria, archaea and fungi using high-throughput 454-pyrosequencing. Significant differences were found between fungal and bacterial communities colonising the three rock-types. Lichen forming fungi were dominant on bare rocks while rocks with tree roots were dominated by ectomycorrhizal fungi and also exhibited higher fungal diversity than the bare rocks or rocks with mosses. A decline in abundance of lichens from bare to ectomycorrhizal rocks suggests that lichens may have facilitated the colonisation of mosses and subsequently ectomycorrhizal mycelia and tree roots during the course of boreal ecosystem development. Rocks with mosses and ectomycorrhizal roots had higher numbers of bacterial sequences than the bare rocks. Extremely low abundance of archaea on rocks suggests that they probably have little or no direct role in biological weathering. However crenarchaeota appear to have a specific association with rocks colonised by mosses. 18 Rhizosphere Microbiome 1 Monday 22 June – Poster session 32 Phytophthora diversity in Bavarian oak forests Carmen Morales Rodríguez, Wolfgang Oßwald, Frank Fleischman* Technische Universität München, Germany Phytophthora species are considered to be devastating pathogens of many herbaceous and woody plants. Up to now, about 140 species or taxa has been described, most of them cause soil born root diseases. However, recent studies in different forest ecosystems using next generation sequencing techniques reviled an unexpected high diversity of Phytophthora species in various soils without plants showing severe disease symptoms, questioning the ecological relevance of Phytophthora species as pathogens in their natural habitats. In Bavarian (South Germany) oak forest a decline symptom of mature trees can be observed. As cause for this dieback a complex of edaphic and climatic factors, root pathogens mainly of the genera Phytophthora and Armillaria, mildew leaf disease and most of all herbivory is discussed. The most frequently isolated Phytophthora species in central European oak forest (Quercus robur or Quercus petrea) are P. plurivora and P. quercina, and their occurrence can be correlated with above ground dieback symptoms. In a two year survey using MiSeq-ITS1amplicon sequencing we characterize the Phytophthora microbiome of oak roots in 15 symptomatic and asymptomatic oak stand, to elucidate the possible contribution of Phytophthora in this decline complex. The first year results will be presented. 33 Investigation on the interactions between soil nutrient availability, rhizosphere microbiome, as well as plant growth and development – a pot experiment Davide Francioli*, Elke Schulz, Witoon Purahong, François Buscot, Thomas Reitz Helmholtz Centre for Environmental Research - UFZ, Germany The composition and functioning of the rhizosphere microbiome is directly affected by the nutrient content of the soil. Besides this, the availability of nutrients also strongly influences the structure and performance of plant communities, which in addition affects the microbial community structure and accompanying microbial traits due to difference in the quantity and/or the quality of rhizodeposition. The composition and functioning of the rhizosphere microbiome is, in turn, one of the major driving factors in plant-soil feedback interactions and directly influences plant growth and development. In order to determine the role of soil microorganisms for plant nutrient acquisition at different nutrient levels we set up a pot experiment. The soil for this study was taken from a long-term agricultural experiment, where various fertilization regimes were consistently applied for the last 113 years. The soil microbial communities, related to four different fertilization regimes (no fertilization, mineral fertilization, organic fertilization, and combined mineral and organic fertilization), were analysed revealing significant differences in their structure and activity. For the pot experiment the four soils were gamma sterilized, and subsequently cross re-inoculated in a factorial design with the four distinct microbial communities of the studied soils. In the pots common grassland species were sown including the grasses Poa annua and Bromus mollis and the herbs Viola arvensis and Matricaria inodora. We examined several phenotypic traits of the plants and we performed fingerprinting analysis of the soil microbial community at several time points of the experiment to identify the microbial key actors and processes determining plant growth and development under the different nutrient levels. The results 19 Rhizosphere Microbiome 1 Monday 22 June – Poster session indicated the soil nutrient content as the main driver in plant growth. In addition, notable shifts in microbial community composition were observed within the rhizosphere microbiome at specific plant growth stages and/or different soil nutrient levels. 34 A novel isolation procedure to isolate bacteria from Penicillium bilaii hyphae Behnoush Ghodsalavi*, Ole Nybroe, Stefan Olsson, Mette Haubjerg Nicolaisen University of Copenhagen, Denmark Deficiency of plant-available forms of phosphorus limit plant production in many soils. Plants are generally assisted by microbial associations, where fungi have major importance for phosphate solubilization and may function as biofertilizers. For mycorrhiza fungi, which may also improve plant growth, it is known that their hyphae are colonized by bacteria, which include helper bacteria with positive impact on fungal growth and performance. The nonmycorrhiza fungi Penicillium bilaii has been shown to increase plant-available phosphorus, and thereby improve plant growth. However, it remains unknown if comparable relationships exist between P. bilaii and its hyphae associated bacteria. The objective of the current study was to characterize bacteria associating with P. bilaii hyphae in a close-to-natural soil system. We initially established a novel microcosm system to mimic the natural habitat for fungal-bacterial interaction in the soil. Hyphal growth of P. bilaii was established on glass cover slides that were placed in mesh bags and transferred to soil. After incubation for 8 days, the presence of hyphae-associated bacteria was confirmed by SYBR Green staining and fluorescence microscopy. Quantification of culturable bacteria from washed, colonized cover slips versus un-colonized controls showed more than 102 fold difference supporting the notion that bacteria isolated from colonized cover slips were indeed hyphae-associated. A strain collection of 100 hyphae associated isolates was established. By universally primedPCR finger printing, the isolates could be assigned to 25 different groups. Representative isolates were subjected to 16S rRNA gene sequencing, which revealed that the hyphaassociated bacteria primarily belonged to the genera Bacillus and Pseudomonas. Future studies will test if growth and activity of P. bilaii can be improved by hyphae associated helper bacteria in order to improve plant growth. 35 Effects of inoculation with Serratia proteamaculans S4 on structure and allocation of plant-derived C in root and rhizosphere communities of oilseed rape Konstantia Gkarmiri*, Shahid Mahmood, Sadhna Alström, Nils Högberg, Roger Finlay Swedish University of Agricultural Sciences, Sweden The rhizosphere is an active and dynamic niche where plant-derived carbon supports microbial growth. Rhizosphere communities can be source of high biodiversity but there are few studies of how this may be affected by applications of biological control agents. Serratia spp. have demonstrated plant growth promoting and antagonistic effects, and should compete more successfully than indigenous communities for plant-derived carbon and alter C allocation patterns to root and rhizospheric microbial communities. A greenhouse study, using field soil, was performed to identify bacterial and fungal communities actively 20 Rhizosphere Microbiome 1 Monday 22 June – Poster session assimilating plant-derived C in the root and rhizosphere of oilseed rape plants following inoculation with Serratia bacteria and the fungal phytopathogen Verticillium longisporum. Oilseed rape seedlings were inoculated with Serratia proteamaculans S4 and were sown in soil artificially inoculated with Verticillium. Factorial combinations of Serratia and Verticillium inoculation were used, as well as additional plant-free soil treatments. Plants were grown for four weeks. Continuous labelling for 6 hours was initiated 4 weeks after the seeds were sown in pots. The first harvest took place 24h prior to labelling (SIP t0). Following 13CO2 injection, 4 time points were decided for harvesting. No morphological differences or difference in shoot or root dry weight was observed between the treatments. However total 13C enrichment in rhizospheric microorganisms was different between the different treatments suggesting a treatment effect on carbon allocation below-ground, either in the level of microorganisms or due to increased root exudation because of stress. The community structure associated with the roots and rhizosphere was examined using 454 pyrosequencing and RNA-based stable isotope probing (SIP) of 13C-labelled 16S rRNA fragments was used to identify taxa actively involved in the assimilation of root-derived carbon. The main focus of this study was to improve our understanding of the relationship between plants, indigenous microbial communities and externally applied biocontrol bacteria. 36 Comparative root microbiome studies of a Verticillum longisporum resistant and susceptible rapeseed line Stefanie Glaeser*, Christian Obermeier, Ebru Cevik, Nima Haghighi, Rod Snowdon, Peter Kämpfer Justus-Liebig-University Giessen, Germany Agricultural plants harbor a high diversity of microbes that colonizing endophytic compartments of roots. Among those are several antagonistic bacteria, which protect plants against root pathogenic fungi. Root exudation pattern and other so far unknown plantderived factors strongly affect the root colonization by these specifically adapted microbes. We investigated bacterial root-endophytic communities of two contrasting oilseed rape double haploid lines from a cross of a susceptible and a resistant parent. One of the rapeseed lines is resistant, the other susceptible to Verticillium longisporum infection. We compared the microbiome of both lines after two weeks of growth in soil and sand to determine if specific root microbiomes can be linked to resistance or susceptibility against Verticillium. Roots were harvested (a pool of 6 plants per treatment), the rhizosphere was removed and roots further purified by sonication and washing. Genomic DNA was extracted from roots to compare endophytic bacterial communities by 16S rRNA gene amplicon Illumina sequencing. Bacterial communities showed higher similarities between the two rapeseed lines grown either in soil or sand than for the same rapeseed lines after growth in sand and soil. However, specific endophytic bacterial communities were developed for both rapeseed lines. In summary 37 bacterial phyla or candidates phyla were detected. The most abundant phyla in all samples were the Proteobacteria, with 60-62% relative abundance if rapeseed lines were grown in soil and >54% if grown in sand. Second and third most abundant phyla after growth in soil were Actinobacteria (6 to 9%) and Bacteroidetes (7 to 10%). A strong rapeseed line 21 Rhizosphere Microbiome 1 Monday 22 June – Poster session specific effect was determined after growth in sand with an unexpected high abundance of Planctomycetales (20%) in the Verticillium resistant rapeseed line. Our data gave a first insight in rapeseed line specific microbiomes which may contribute to the obtained resistance against Verticillium. 37 Influence of rhizosphere, root and P soil on fungal and bacterial communities associated with maize genotypes with contrasting P use efficiency Eliane Gomes*1, Ubiraci Lana1, Bangzhou Zhang2, Christiane Oliveira1, Lauro Guimarães1, James Tiedje2 1 Embrapa Maize and Sorghum, Brazil, 2Michigan State University, USA Maize (Zea mays L.) represents one of the main economic crops for food and energy in the world, and its associated microbial communities have been intensively investigated using different approaches. However, low-resolution profiling methods often make it difficult to understand the complicated microbial communities. Using Illumina MiSeq high-throughput sequencing, we analyzed the bacterial and fungal communities from washed roots (endosphere) and from the rhizosphere soil both originated from the same root sample of two maize genotypes with contrasting phosphorus (P) use efficiency cultivated in two different P soil concentrations. A total of 2.588.087 and 2.516.453 reads were identified from 16S-V4 (bacteria) and ITS (fungi) rRNA gene regions, respectively, and all taxonomic classifications were assigned using the naïve Bayesian algorithm developed for the RDP Classifier. We observed higher Chao 1 richness and Shannon–Weaver diversity indices in the rhizosphere than inside the roots, suggesting that endosphere has a specific microbial community. The predominant bacteria in rhizosphere were from the phyla Proteobacteria (63.9%), Actinobacteria (8.4%), and Bacteroidetes (6.7%), while the main phyla inside the roots were Proteobacteria (50.5%), Tenericutes (15.8%) and Bacteroides (13.4%). The soil P level also influenced the bacterial community distribution since members of Burkholderia and Ralstonia genera were enriched in the low P rhizosphere while Pseudomonas and Leclercia were predominant in the high P rhizosphere soils. Considering the fungal communities, Ascomycota (85.7%) was the dominant phylum, followed by Basidiomycota (9.5%) and Glomeromycota (3.0%) in the rhizosphere and endosphere in both P conditions. Glomeromycota phylum (mycorrhizal fungi) was represented at 1% in the rhizosphere and at 5% in the endosphere, and was especially abundant in the low P soil. In general, we conclude that rather than maize genotypes, the overall patterns of soil microbial diversity was influenced by the rhizosphere and endosphere habitats followed by the soil P concentration. 38 Microbial community dynamics of growing media in soilless culture systems Oliver Grunert*1, Emma Hernandezsanabria1, Marie-Christine Van Labeke2, Dirk Reheul2, Maaike Perneel3, Nico Boon4 1 Laboratory of Microbial Ecology and Technology (LabMET), Ghent University, Belgium, 2Department of Plant Production, Ghent University, Coupure Links 653, Belgium, 3Peltracom, Belgium, 4Laboratory of Microbial Ecology and Technology (LabMET), Ghent University, Coupure Links 653, Belgium 22 Rhizosphere Microbiome 1 Monday 22 June – Poster session The hairy roots syndrome caused by Agrobacterium rhizogenes is an increasing problem in greenhouse horticulture in Spain, France, Germany, UK, Ireland, Poland, Canada, USA, Mexico, the Netherlands and Belgium. Current control measures are limited to removal of excessive leaves from the plants and the plastic cover from the slabs. There is a lack of effective control strategies and the search for alternative strategies also has been stimulated by public concerns about the adverse effects of biocides on the environment and human health. Natural disease-suppressive soils and probably horticultural growing media and its microbial community can effectively protect plants against soilborne pathogens. In this study, we compared the physicochemical and microbial community characteristics of an mineral and organic horticultural growing medium in relation to the presence of the plant pathogen Agrobacterium rhizogenes. We aimed to identify how microbial and environmental interactions influenced the development and spread of this disease in a soilless cultivation system. In this study we hypothesised that there is a fundamental difference of a young (mineral) and an old (organic) growing medium-associated microbial community in its general suppressiveness against Agrobacterium rhizogenes causing the hairy roots syndrome and resulting according to the definition of Baker and Cook in little or no visible damage to the plant, although the pathogen may persist in the growing medium. Multivariate statistical analysis performed to assess the characteristics of each growing medium revealed the key variables impacting the microbial community, such as potassium and ammonia concentrations. High throughput sequencing analysis of the bacterial abundance of the communities present in organic substrate showed significant interactions among Methylophilaceae and Actinobacteridae with A. rhizogenes. Our results clarified the complex bacterial relationships in horticultural growing media. Knowledge regarding these relationships may be used to develop strategies to control the hairy roots syndrome. 39 Microbes in root-soil interfaces: colonization patterns in oxic/anoxic microenvironments of wetland rice Hannes Schmidt*1, Thilo Eickhorst2 1 University of Vienna, Department of Microbiology and Ecosystem Science, Division of Microbial Ecology, Austria, 2University of Bremen, Soil Microbial Ecology, Germany In paddy soils, the presence of numerous electron acceptors and root-derived substances in oxic/anoxic micro-environments such as the rhizosphere may attract, stimulate, or inhibit soil microorganisms. The intensity of influence of the plant on microbes and vice versa is likely to be greatest in close vicinity to the root surface and may decrease with distance from the rhizoplane. It is therefore essential to detect sites of microbial rhizosphere colonization, and thus to identify potential areas of microbe-plant interaction. A 16S rRNA-targeted approach including PCR-based fingerprinting and CARD-FISH was applied to identify, localize, quantify, and visualize microbial cells associated with roots (rhizoplane, rhizosphere) and selected soil compartments (bulk soil, oxidized layer). With emphasis on microbial rhizoplane communities, the spatial distribution of archaea, bacteria, and relevant microbial populations was investigated. The growth stage of rice and the distance to the root surface strongly influenced microbial colonization. Patchy distribution patterns of microbes observed at early stages turned into 23 Rhizosphere Microbiome 1 Monday 22 June – Poster session more uniform colonization with increasing plant age. Accordingly, the highest colonization densities shifted from the tip to more mature regions of rice roots and were found at flowering stage. β-Proteobacteria became predominant in the oxygenated rhizosphere that were affiliated with lithoautotrophic iron-oxidizing bacteria. They were further visualized to densely colonize areas of pronounced iron coatings indicating an active role in the biotic formation of Fe-plaque on rice roots. The co-localization of oxygen-sensitive methanogenic archaea with oxygen-dependent methanotrophic bacteria on the rhizoplane indicated the formation of micro-sites of contrasting oxic/anoxic conditions within short distance. Beneficial effects of a close association with root surfaces were indicated by proportionally higher numbers of methane-oxidizing bacteria on the rhizoplane compared to the rhizosphere. The presented data provides essential quantitative and spatio-temporal information on microbial rhizosphere colonization and thus may ideally complement studies on microbial diversity as analyzed via high-throughput sequencing. 40 Taxonomic and functional diversity of the microbiome of agricultural plants and its relation to the rhizosphere effect Julie Hernandez-Salmeron*1, Eduardo Valencia1, Gabriel Moreno-Hagelsieb2, Gustavo Santoyo1 1 Universidad Michoacana de San Nicolas de Hidalgo, Mexico, 2Wilfrid Laurier University, Canada Plants rhizosphere is the microenvironment that connects free life microorganisms in soil with the plant, through the flow of rhizodeposits by the roots. Each plant species has a differential effect on the way they select or set a microbiome in its particular rhizosphere, whose main function is to promote plant growth and phyto-pathogen antagonism. In this study, we hypothesized that after growing different plant species in the same soil with the same repertoire of microorganisms they will diferencially select and promote a microbiome specific to the plant genotype, we selected plants of alimentary importance for our country: wheat, maize, sorghum and beans. The in vitro experiment was conducted under controlled conditions of temperature and light in a growth chamber using the same soil type where the plants were grown, so that the only variable was attributed to the plant genotype. Rhizospheric and bulk soil was removed after four weeks of plants growth, from which DNA was extracted and metagenomic bacterial diversity was initially screened using DGGE, these results showed a clear variation in the diversity of microorganisms in each plant species. Therefore, metagenomic DNA was purified and sent to pyrosequencing to analyze genetic functional diversity in each plant rhizosphere. We expect to obtain sequences in the following months and complete the in silico analysis of metagenomes by using MEGAN to show the results in this event. Diversity of culturable microorganisms was also analyzed by RAPD fingerprinting, where the highest haplotype percentage was found in sorghum plants, followed by wheat, and corn rhizosphere. This study would allow us to deepen more about the factors that influence the structure of bacterial communities in the rhizosphere and infer ecological functions relevant during plantmicroorganism interaction. 24 Rhizosphere Microbiome 1 Monday 22 June – Poster session 41 Activity profiling of extra-radicular enzymes in urban trees Josef Valentin Herrmann*, Ullrich Gilge Bavarian State Institute for Viticulture and Horticulture, Germany Urban trees grow in a stressful, unnatural environment. Limited tree pits restrict their rooting zone, degraded, compacted soil disturbs water and air balance. Multiple pollutants as well as frequent mechanical damage affect the tree vitality. Additionally, climate change has worsened these demanding conditions. Under such adverse circumstances mycorrhizal associations play an even more significant role for plant health. To select urban trees for the future, a long-term project was started in 2010. Different tree species were planted at three sites with differing climate conditions. Representing different mycorrhizal preferences, some species (Carpinus betulus, Fraxinus pennsylvanica ’Summit’, Magnolia kobus, Ostrya carpinifolia, Parrotia persica, Quercus cerris, Tilia tomentosa ’Brabant’) were chosen to study extraradicular enzymes relevant to tree nutrition and connected to mycorrhization. At planting half of the trees were inoculated with a commercial mycorrhizaproduct (INOQ). Since 2011 root sampling has been performed twice a year. To measure the activity of 8 hydrolytic and oxidative enzymes (cellulose and hemicellulose degrading xylosidase, glucoronidase, cellobiohydrolase, β-glucoronidase, chitin degrading Nacetylglucosaminidase, phosphate mobilizing phosphomonoesterase, protein degrading leucinaminopeptidase) a rapid and sensitive fluorometric microplate multiple enzymatic test was performed on single root tips. In addition, each root tip was microanalyzed for mycorrhizal colonization. The data revealed a high diversity of activity profiles. The pattern showed a tree species specificity may be representing the species specific mycorrhization. In addition, a seasonality could be detected with higher activities in spring compared to summer and autumn. A difference in enzymatic profiles between inoculated and non-inoculated trees could not be observed. Comparing mycorrhizal and non mycorrhizal root tips significant (Mann-Whitney Rank Sum Test) alterations were detected with some enzymes displaying higher and some lower activities. Correlation analysis to substrate soil, soil condition and planting site are under way. 42 Impact of microbial species loss on rhizosphere microbiome assembly Gera Hol*, Wietse de Boer, Wim van der Putten Netherlands Institute of Ecology, Netherlands Plant growth may respond positively or negatively to the soil microbial community, depending on the balance between pathogens and mutualists. Growing plants repeatedly in the same soil may result in increase of pathogens (negative plant soil feedback) or increase of mutualists (positive plant-soil feedback). While the concept of plant-soil feedback is well- known, there is a gap in the knowledge of microbial community development during feedback, in particular with regards to the role of less abundant species. Loss of microbes is expected to affect plant growth when the balance between pathogens and mutualists is 25 Rhizosphere Microbiome 1 Monday 22 June – Poster session disturbed. Here we tested to what extent loss of rare microbes will affect the development of plant-soil feedback. Inoculation of sterilized soils with serial diluted suspensions from two soil origins and subsequent incubation resulted in soils with similar microbial biomass but differences in microbial community. Pyrosequencing of 16S rRNA was used to track how dilution and incubation affected microbial community composition directly after incubation and after one or two cycles of plant growth. Dilution decreased species richness and similarity as compared to the original field soil. Growing plants on the inoculated soils once or twice increased similarity of the bacterial community to that of the field soil. While the most diluted communities showed the least overlap with the original field soil, the similarity between replicates was high. Also the similarity between microbial communities after one and two growing cycles was highest in the most diluted community. On basis of the sequence data and literature we identified potential pathogens and mutualists of the host plant Triticum aestivum and followed how those bacteria species reacted to a second growing cycle with the same host plant. We conclude that loss of species will make microbial community dynamics more predictable. 43 Hybrid rice stimulates greater abundance of AOB relative to AOA in the rhizosphere Qaiser Hussain*1, Genxing Pan2 1 Pir Mehr Ali Shah Arid Agriculture University, Pakistan, 2Institute of Resource, Ecosystem and Environment of Agriculture, Nanjing Agricultural University, China It is well documented that rice paddy soils have activities of both AOB and AOA, but impact of hybrid rice on the ecology of ammonia oxidizers (AOB & AOA) is still poorly understood under field condition. We hypothesized that hybrid rice with the trait of vigorous root system releases more oxygen that can influence the structure and abundance of AOB and AOA in the rhizophere. A field experiment was initiated to assess the structure and abundance of ammonia oxidizers in the rhizosphere of hybrid and conventional rice cultivars at three growing stages. Rhizosphere samples were collected at selected growth stages for each rice cultivar: 45 days after planting (tillering stage-S1, 81 days after planting (grain filling stage-S2) and 107 days after planting (ripening stage-S3). PCR-DGGE and real-time PCR approaches were used to evaluate the structures and relative abundances of ammonia oxidizers (AOA and AOB) by targeting a functional gene fragment coding ammonia monooxygenase (amoA). Moreover, important band retrieved from DGGE gel were analyzed using sequencing and phylogenetic analysis. Hybrid rice promoted 53%, 30% and 23% higher rhizosphere amoA (AOB) gene abundance relative to conventional cultivar at S1, S2 and S3 stages, respectively. Though, rice cultivars showed no considerable effect on the amoA (AOA) gene abundance. Principal component analyses (PCA) of PCR-DGGE profile revealed a profound shift of AOB community structure between two cultivars across particular growing stages. However, rice cultivars did not significantly influence the structure of archaeal-amoA (AOA) in rhizosphere showing the higher stability of AOA communities. Diversity of AOA was limited to uncultured Crenarchaeote, whereas AOB analyses revealed several distinct operational taxonomic units 26 Rhizosphere Microbiome 1 Monday 22 June – Poster session markedly resemble to Nitrosospira sp. in the rhizosphere of hybrid cultivar compared to conventional cultivar. Hybrid rice can select from a particular group of AOB in the rhizosphere relative to AOA throughout growth periods. 44 Effects of different organic fertilizers on soil rhizospheric microbial functional diversity of greenhouse chrysanthemum Rong Jiang*1, Zhi Xu2, Li Tang2 1 College of Resources and Environmental Science,Yunnan Agricultural University, China, 2College of Resources and Environmental Science, Yunnan Agricultural University, China Organic fertilization can affect microbial community in rhizosphere soils which is thought to be responsible for biological processes that are necessary for maintaining a healthy soil. However, its efficacy of different organic fertilizers on soil microbial community structure in highly intensive flower industry under greenhouse condition had little been elucidated.Field experiment was conducted to study the effects of applying different organic fertilizers and their combination with 20% reduced chemical fertilizers on the microbes in rhizosphere soil of chrysanthemum. As compared with conventional chemical fertilization (100% chemical fertilizers, CF), applying bio-organic fertilizer in combination with 20% reduced chemical fertilization (BIO) significantly increased the number of bacterial by 101.2%, actinomycetes by 84.3% and the total microbial number by 100.1% in the rhizospheric soil, and decreased the fungi number by 64.2%, while applying refined organic fertilizer in combination with 20% reduced chemical fertilization (OF) only decreased the fungi number by 30.4%. As compared with CF, the BIO treatment increased the rhizospheric microbial diversity of Shannon index (H) and McIntash index (U) of chrysanthemum by 8.6% and 21.6%, respectively, and the OF treatment showed no significant difference. OF and BIO could significantly increased the carbohydrates utilization ability by 13.3% and 15.8%, the phenolic carbon source utilization ability increased by 13.0% and 36.5%,repectively. The BIO also increased the capacity of amino acids significantly. The principal component analysis (PCA) demonstrated that, BIO significantly affected the soil microbial community structure, mainly depending on the utilization of carbohydrates, amino acids and carboxylic acids. Under 20% chemical fertilizers reduction during the full growth stage, applying bio-organic fertilizer (BIO) increased the stem length and stem diameter significantly, showed the increasing of the flower diameter and stalk length, yield and dry matter, improved the flower quality of chrysanthemum. 45 Plant growth promoting rhizobacterial efficacy in cowpea (Vigna unguiculata (L.) walp.) Kannan Kanthaiah*, Velu Rajesh Kannan Bharathidasan University, India Plant growth promoting rhizobacteria (PGPR) are known to influence the growth of the plant by various plant growth promoting activities such as Indole Acetic Acid (IAA) production, phosphate solubilization activity, ammonia production (NH3), nitrogen fixation, siderophore production and hydrogen cyanide production (HCN). In search for the potential PGPR from 27 Rhizosphere Microbiome 1 Monday 22 June – Poster session the Rhizosphere soil of cowpea, a total of 16 rhizobacteria (B1-B6 & D1-D9) were isolated. By comparing the results two potential rhizobacteria (B6 & D4) have been selected for the further evaluation of Plant Growth Promoting (PGP) activity. These isolates biochemically characterized, B6 and D4 confirmed as Pseudomonas and Bacillus by 16SrRNA gene sequencing. Their ability of IAA production ranging from (95 and 220.5 µg/ml) respectively, they showed positive results to ammonia, nitrogen fixation, hydrogen cyanide production and siderophore production. Their phosphate solubilizing ability ranging from 16 mm and their solubilization efficiency (SE) ranging from 200 and 155 and their Solubilization index (SI) 3.0 and 2.5 respectively. In vitro and in vivo nursery field studies revealed that 100% seed germination rate using Bacillus safensis and 92% for Pseudomonas aeruginosa showing promosing results. The study showed the characters of benefits to use these organisms as biofertilizers for sustainable agriculture to improve crop yield. 46 Pseudomonas species in the secretion of auxin Nazanin Khakipour*1, Kazem Khavazi2, Abdolreza Akhgar3 1 Islamic Azad University- Savadkooh branch, Iran, 2Soil and Water Institute, Iran, 3Vali-e Asr university of Rafsanjan, Iran P.putida and P.fluorescence stimulate plant growth and crop yield can be increased. One of the important characteristics of fluorescent Pseudomonads bacteria produce organic compounds in concentrations too low to be able to control physiological processes. Secretion of growth regulators as one of the most important mechanisms of plant growth is active. The purpose of this study was to compare different strains of bacteria P.putida and P.fluorescence secrete a variety of auxin and it was under the same conditions. For this purpose, 23strains P.fluorescence and 27 strains P. putida randomly of Microbial bank was prepared. The IAA strains P.fluorescens of zero to 31.6milligrams per liter respectively. Of the 23 strains, 7 strains did not produce IAA. The amount of IAM at P.fluorescens strains was from zero to 16.12 mg per liter respectively. Of the 23 strains did not produce 2 strain IAM. The ILA also from zero to 7.2 milligrams per liter respectively. IBA did not generate any of the strains studied P.fluorescens. Among different strains P.fluorescens this study the prevalence of strains capable of producing IAA, IAM, ILA and IBA 8,15,10 and zero, respectively. The IAA strains P.putida of zero to 24.8 milligrams per liter .7 strains of 27 strains did not produced IAA . The amount of IAM at P.putida strains was from zero to a maximum of 17.2 milligrams per liter respectively. Of the 27 strains did not produce 6 strain IAM. The ILA also from zero to 10 milligrams per liter respectively. Of the 27 strains, 8 strains did not produce ILA. P.putida strains studied did not generate any of the IBA. P.putida among different strains of the study, the prevalence of strains capable of producing IAA, IAM, ILA and IBA 11, 13, 12 and zero, respectively. 28 Rhizosphere Microbiome 1 Monday 22 June – Poster session 47 Microbial diversity analysis of the rhizosphere of tomato cultivars that are resistant or susceptible to bacterial wilt Seon-Woo Lee1, Hyun-Gi Kong*1, Min-Jung Kwak2, Eun Joo Jung1, Ju Yeon Song2, Jihyun Kim2 1 Dong-A University, South Korea, 2Yonsei University, South Korea Bacterial wilt caused by Ralstonia solanacearum is lethal wilt resulting in significant yield loss on many Solanacea plants. Bacterial wilt resistance in several resistant plants is controlled by quantitative trait loci and also by rhizosphere conditions probably by microbial composition. In this study, we investigated the microbial community structure of tomato rhizosphere using two different tomato cultivars, Hawaii7996 as a bacterial wilt resistant cultivar and Moneymaker as a bacterial wilt susceptible cultivar. We cultivated tomato cultivars under greenhouse condition with field soils at Dong-A Agricultural experimental station. Microbial communities of tomato rhizosphere of both cultivars were analyzed by amplifying 16S rRNA genes from those rhizosphere soil and pyrosequencing of the amplified clones through GSFLX 454 sequencer. Microbial community structure was analyzed using MG-RAST system. The most abundant microbial phylum was proteobacteria both in bulk soil and in tomato rhizosphere soil. Compared to bulk soils, the proportions of the phyla such as Bacteroidetes and Firmicutes were significantly increased in tomato rhizosphere, while that of phylum Acidobacteria was decreased in tomato rhizosphere. Especially, bacterial members in class Flavobacteria were highly abundant in tomato rhizosphere. Comparison of microbial communities of tomato rhizosphere revealed that rhizosphere microbiome between Hawaii7996 and Moneymaker was distinguishable. The tomato rhizosphere showed the similar pattern of microbial diversity for two consecutive years. Our result suggested that the unique microbial community forms in tomato rhizosphere in a cultivar specific manner. 48 Selection of rhizobial and rhizospheric isolates associated to chickpea (Cicer arietinum) for promoting its culture in region dedicated for grain culture Saadia Laabas*, Zineb faiza Boukhatem, Tsaki Hassini, Abdelkader Bekki University of Oran 1 Ahmed Benbella, Algeria, Algeria Consumption of chickpea (Cicer arietinum L.) holds a special place in Algerian culture but its production, however, is far from to be sufficient for the population needs. It has been attempted to introduce this legume in wilaya of Tissemsilt (Algeria), in rotation with cereal crops. This study aims to establish the best microbiological approach to improve the legume performance. Strains isolated from Cicer arietinum L. rhizospheric soil from five selected sites were screened for their plant growth promoting (PGPR) potential under controlled conditions, for IAA production and P solubilization ability. In a second step, were selected native rhizobial strains with high nitrogen-fixing potential. Then the single effect of PGPR isolates was compared to their combination with rhizobial inoculant on plant growth, under greenhouse conditions. 29 Rhizosphere Microbiome 1 Monday 22 June – Poster session 49 Effect of phosphorus fertilization on bacterial microbiome in microsites of ryegrass rhizosphere Lorena Lagos*1, Oscar Navarrete2, Fumito Maruyama3, María de la Luz Mora1, Milko Jorquera1 1 Universidad de La Frontera. Center of Plant, Soil Interaction and Natural Resources Biotechnology, Chile, 2Universidad de La Frontera, Chile, 3Kyoto University. Section of Microbiology,Graduate School of Medicine, Japan The structure and diversity of microbial communities along the rhizosphere differ in their composition, activity and abundance according to diverse biotic and abiotic factors, such as root exudates, plant species, type of soil, agronomic practices like fertilization. In the present report we analyzed the effect of P fertilization on rhizobacterial composition in microsites of Lolium perenne grown in a volcanic soil (Andisol) characterized by low-P available. Lolium perenne grown in rhizotrons were separately fertilized with inorganic (phosphate; KH2PO4; 300 mg P kg-1 of soil) and organic (phytate; C6H18O24P6 and H2O; 2.5 mM (825 mg phytate kg-1 of soil) P sources. Total DNA samples were extracted from two microsites of the rhizosphere (root tip and mature zone). Control without fertilization and bulk soil were also analyzed. The bacteria DNA sequences were analyzed by pyrosequencing of 16S ribosomal RNA (rRNA) genes followed for a silico analysis by MOTHUR. Our results revealed according to the alpha diversity analysis that in all rhizosphere samples there was a higher relative abundance of the Proteobacteria (45%; alpha- and Betaproteobacteria) followed by Actinobacteria (17%), Cloroflexi (12%) and Acidobacteria (11%). In contrast, in bulk soil more relative abundance groups were Proteobacteria (68%; alpha- and gammaproteobacteria) and Firmicutes (21%, Bacilli). On the other hand, beta diversity analyses indicated difference in ocurrence (presence/absence) based on index of Jaccard for microsites of the rhizosphere and bulk soil. Moreover, Yue and Clayton theta index indicated similarity for bacterial community of microsites of the rhizosphere. The analyses were evaluated at level of species and classes 0.03 and 0.10 OTUs, respectively. The present study showed that P fertilization induced changes in the composition of bacterial communities at microsite level in the rhizosphere of Lolium perenne. 50 Host-based microbial recruitment at the aquatic rhizosphere Yong Jian Lee*, Sanjay Swarup National University of Singapore, Singapore The aquatic rhizosphere is a region around the roots of aquatic plants. Many studies focusing on terrestrial rhizosphere have led to a good understanding of the interactions between the roots, its exudates and its associated rhizobacteria. The rhizosphere of free-floating roots, however, is a different habitat that poses several additional challenges, including rapid diffusion rates of signals and nutrient molecules, which are further influenced by the hydrodynamic forces. These can lead to rapid diffusion and complicates the studying of diffusible factors from both plant and/or rhizobacterial origins. These plant systems are being increasingly used for self purification of water bodies to provide sustainable solution. A better understanding of these processes will help in improving their performance for ecological engineering of freshwater systems. The same principles can also be used to improve the yield of hydroponic cultures. 30 Rhizosphere Microbiome 1 Monday 22 June – Poster session We have begun to unravel the complexity of rhizobacterial communities associated with aquatic plants. Using fluorescence in-situ hybridization (FISH) and Illumina Miseq Next Generation Sequencing of 16S amplicons and metagenomic DNA, we investigated the rootassociated microbial community of established P. amaryllifolius grown in four different water bodies. The community structure of rhizobacteria from plants grown in these water bodies are highly similar. The top three phyla belonged to Proteobacteria, Bacteriocedes and Actinobacteria, as validated by FISH analyses. This suggests that the rhizosphere have an innate ability to attract and recruit rhizobacterial communities, possibly through the metabolic compounds secreted through root exudation. The selection pressure through plant host is higher compared to environmental pressures that are different between the two water sources. In comparison with the terrestrial rhizosphere, the aquatic rhizosphere microbiome seems more specialised and has a high influence by the host. We are using these findings to further understand the role of microbes in the performance of freshwater aquatic plants. 51 Motility in atrazine-degrading Arthrobacter bacteria and its possible ecological role Chengyun Li*1, Dmitry Bazhanov1, Hongmei Li2, Jishun Li1, Hetong Yang2 1 Key Laboratory for Applied Microbiology of Shandong Province, Biotechnology Center of Shandong Academy of Sciences, China, 2Biology Institute of Shandong Academy of Sciences, China Bacteria of the genus Arthrobacter are known as ubiquitous and most frequently isolated soil inhabitants. It is believed that the prevalence of Arthrobacter spp. is due to their resistance to stressful conditions. No behavioral mechanisms contributing to the survival of Arthrobacter bacteria in soil were so far known. Based on the results of ERIC-PCR genotyping and phylogenetic analysis of directly isolated atrazine degraders, we found that Arthrobacter spp. prevailed in the root-associated communities of atrazine-degrading bacteria both in industrial and agricultural soils. Genetically similar A. ureafaciens bacteria were predominant atrazine degraders in the maize rhizosphere at geographically distant agricultural sites. Regardless their geographical origin, atrazine-degrading A. ureafaciens strains were motile, spread along fungal hyphae and colonized plant roots after seed inoculation. By characterizing more than 100 atrazinedegrading strains, we revealed that active motility was exhibited by representatives of various phylogenetic groups of the genus Arthrobacter. Some strains spread in semisolid agar and on the surface of soft agar, suggesting various types of motility. Our data suggest that active motility is widely distributed among Arthrobacter bacteria and can play an important role in their rhizosphere competence and soil survival. Some atrazine-degrading strains we have isolated can be considered promising model bacteria for studying root colonization by Arthrobacter spp. Mechanisms of the active motility possessed by Arthrobacter bacteria and their role in the root colonization will be further investigated. 31 Rhizosphere Microbiome 1 Monday 22 June – Poster session 52 Effect of Bio-organic fertilizer on bacterial wilt and rhizospheric soil microbes of flue-cured tobacco Zhang Yunwei, Tang Li*, Xu Zhi, Li Yanhong, Song Jianqun Yunnan Agricultural University, China Two years field experiments were carried out to study the effects of refined organic fertilizer and bio-organic fertilizer, both combined with 20% reduced chemical fertilizer on flue-cured tobacco yield and quality, resistance to bacterial wilt and soil microbes. The effects of different organic fertilizers on soil microbial quantity, pathogen number, and microbial functional diversity were studied by means of traditional enumeration methods and BIOLOG technology. Under reduction of 20% chemical fertilizers, the effects of applying organic fertilizer (OF) on tobacco yield and quality was not significantly, but applying bio-organic fertilizer (BIO) significantly improved tobacco yield, output value and the proportion of high grade tobacco leaves by 12%, 25% and 27%, respectively. Compared with conventional chemical fertilization (CF, 100% NPK chemical fertilizers), both treatments applying either organic fertilizer (OF), or bio-organic fertilizer (BIO), decreased the bacterial wilt, increased the bacterial number and improved the microbial activity significantly. Compared with CF, the disease incidence and the disease index of bacterial wilt by BIO treatment were reduced 20%~32.1% and 36.2%~40.9%. Compared with OF, BIO treatment decreased the disease incidence of bacterial wilt by 11.1%~13.7% and the disease index of BIO were significantly reduced by 20.9%~31.1%, respectively. BIO had 70.5%~171.7% higher bacteria counts and 2~4 times higher actnomyces counts than OF. Compared with OF, The fungi number and pathogen number of BIO treatment were decreased by 64% ~74.3% and 55.8%~66.7%, respectively. Biolog ECO results showed that, AWCD data which indicate the soil microbial activity varied significantly in order from highest to lowest of BIO>OF>CF. The same trend (BIO>OF>CF) was found when functional diversity was expressed as Shannon’s diversity index (H), McIntosh’s index and Simpson’s index. Compared with CF, the utilization rate of carboxylic acids, amino acids and phenols of BIO were increased by 30%~32%, 27.2%~39.7% and 77.2%~117.8%, respectively. 53 Rhizosphere bacteria constitute a principal influential force on the outcome of cotton replanted diseases Xiaogang Li*, Xingxiang Wang Institute of Soil Science, Chinese Academy of Sciences, China Cotton yield and quality are seriously compromised by consecutive cropping of cotton in the primary cotton regions of China. The primary hypothesis of this study was that the rhizobacterial community would constitute a principal influential force for governing cotton replant disease. To assess this possibility, plant growth, disease resistance, and the effect of root exudates on the development of Fusarium oxysporum and on rhizobacterial community composition were evaluated. The experiment was conducted in two different replanted soils, 32 Rhizosphere Microbiome 1 Monday 22 June – Poster session one in use for four years and the other in use for 15 years with a fallow soil as the control. The replanted soil significantly influenced plant growth and rhizosphere-secreting characteristics with a lower resistance to Fusarium wilt. Also, a bioassay revealed a loss of spore germination and sporulation of F. oxysporum in cotton root exudates from replanted soils compared to those from the control soil, and MiSeq sequencing of 16S rRNA gene amplicons demonstrated clear variations in the rhizobacterial community of cotton grown in replanted soils and in the control soil. Specifically, the Gammaproteobacteria and Betaproteobacteria, including Xanthomonadaceae, Comamonadaceae, Oxalobacteraceae, and Caulobacteraceae known to comprise strains with plant-beneficial and disease-suppressive properties, were identified as the most dynamic taxa associated with cotton health in the control soil, while fewer bacterial taxa (Flavobacteriaceae, Streptomycetaceae, Sphingomonadaceae) were associated with cotton-replanted soils, possibly representing greater cotton root damage in replanted soil. Collectively, these results suggest that the complex phenomenon of cotton replant disease may not simply be ascribed to accumulations of pathogens or a single group, but is most likely associated with the rhizobacterial community recruited by cotton for its protection against pathogen infections. 54 The consecutive monoculture problems of Pseudostellaria heterophylla and its control strategy Wenxiong Lin*, Linkun Wu, Hongmiao Wu, Jun Chen, Xianjin Qin, Yongpo Zhao, Juanying Wang Fujian Agriculture and Forestry University, China The consecutive monoculture problems are one of the big issues in modern agriculture. In this study, Pseudostellaria heterophylla, an important medicinal plant was used as experimental material. Based on our previous study, the changes in the rhizosphere microbial community under consecutive monoculture were studied through the approaches of modern rhizosphere biology. The results showed that P. heterophylla consecutive monoculture altered the rhizospheric microbial composition with fewer beneficial microorganisms and more pathogenic microbes. The qRT-PCR analysis confirmed that beneficial microbes including the Pseudomonas, Bacillus subtilis, etc. decreased with the increasing years of monoculture, the reverse was true in case of the pathogenic microbes such as Salmonella sp., Fusarium oxysporum, etc.. Further study found the mixtures of phenolic acids in the same ratio as determined in P. heterophylla root exudates could significantly promote the growth of the specific pathogenic fungi but inhibit the beneficial microbes. Some pathogenic microbes, such as Salmonella sp. showed a chemotaxis response to several root exudates. The sequent test showed these pathogens could cause serious wilt disease on P. heterophylla seedlings and led to the up-regulation of the genes involving in calcium signaling transduction system and energy production, which explains in part the underlying mechanism of declines in yield and quality of plants in response to consecutive monoculture stress. Accordingly, different control strategies were used, suggesting that the specific microbial fertilizers and some cropping sequences such as paddy-upland rotation patterns could effectively remediate the imbalanced soil microbial community, in turn leading to increased yield of replanted P. heterophylla. It is therefore concluded that the plant-microbe interactions play important roles in consecutive monoculture problems of P. heterophylla, and to develop a specific 33 Rhizosphere Microbiome 1 Monday 22 June – Poster session biofertilizer with a good combination of beneficial microbes to the disease-conducive soil is an efficient way to overcome the soil-borne disease of replanted P. heterophylla. 55 Characterization of bacterial communities from halophytes growing in Khewra Salt Ranges by culture-dependent and metagenomic approaches Salma Mukhtar, Samina Mehnaz, Kauser Malik* Forman Christian College (A Chartered University), Pakistan The microbial diversity in the rhizosphere of halophytes can provide information regarding various mechanisms enabling the plants to survive under such an extreme condition of abiotic stress. Only 1-3% of the soil microbial population is culturable. DNA-based methods have been developed to have an insight into the remaining fraction of soil microflora. This study, investigated the distribution of culturable and non culturable bacteria in the rhizosphere of halophytes. Culturable bacterial isolates were characterized morphologically, biochemically and through sequencing of 16S rRNA gene. These were identified as strains of different species of Bacillus, Pseudomonas, Staphylococcus, Micrococcus, Planococcus, Vibrio, Burkholderia, Klebsiella, and Kocuria. Based on salt tolerance, fifty five strains were selected. Plasmid curing of isolates was done to study the effect of plasmid conferring salt tolerance. These plasmids were isolated, transferred into E. coli and growth response of original and transformed E. coli strains was compared at 1.5 to 4 M concentration of NaCl. Almost all of these, showed optimum growth at 1- 3.5 M NaCl. Cured isolates did not grow in halophilic medium but grew well on LB medium. Most of the transformed E. coli strains grew up to 3M NaCl concentration. Culture independent diversity was studied by PCR and sequencing of 16S rRNA gene. Metagenomic analysis from rhizosphere showed that 40% bacteria were uncultured and unclassified. Firmicutes, Proteobacteria, Acidobacteria, Bacteriodetes, Plantomycetes, Cyanobacteria, Thermotogae and Choroflexi were predominant groups. Results revealed a wide diversity of culturable and non-culturable halophilic bacteria in the rhizosphere of halophytes. 56 Metagenomic analysis of the rhizosphere microbiome of the common bean resistant to Fusarium oxysporum Lucas William Mendes*1, Rodrigo Mendes2, Siu Mui Tsai1 1 University of Sao Paulo, Brazil, 2EMBRAPA Environment, Brazil The rhizosphere microbiome plays a key role in the functioning of the host plant, influencing its physiology and development. It has been suggested that plants use mechanisms present in the rhizosphere microbiome to fend off infections, such as fungal diseases. This work aimed to assess the microbial community inhabiting the common bean rhizosphere in order to identify potential groups related to the suppression of the soil-borne pathogen Fusarium oxysporum. Therefore, using shotgun metagenomic sequencing (Illumina Miseq), we investigated the phylogenetic and potential functional diversity of microbial communities colonizing the rhizosphere of four cultivars of common bean with different levels of 34 Rhizosphere Microbiome 1 Monday 22 June – Poster session resistance to the fungus, ranging from high susceptibility to resistant. Quantitative PCR of total bacteria in rhizosphere samples showed an increase of 16S rRNA copy number with the increase of resistance to the fungus. Mesocosms experiments, including four common bean cultivars cultivated in Amazonian Dark Earth and three replicates, were conducted in greenhouse conditions and we obtained over than 12 million metagenomic sequences. The overall microbial diversity did not present significant variations across common bean cultivars. From the classified sequences, 97.4% were affiliated to Bacteria and 1.48% to Archaea. Proteobacteria represented the most abundant phyla (41.7%), followed by Actinobacteria (29.4%), Firmicutes (5.9%) and Acidobacteria (4.1%). The microbial communities structure were different between bulk soil and rhizosphere samples. Comparing all bean cultivars, the resistant one showed an overrepresentation of the phyla Spirochaetes, Nitrospirae and Euryarchaeota. The resistant bean cultivar presented high number of sequences affiliated to the genus Bacillus. Interestingly, the resistant and moderately resistant cultivars, presented high proportion of sequences related to bacteriocin, a narrow spectrum antibiotics. Preliminary analysis showed that different common bean cultivars could select differential microbial groups in the rhizosphere environment. Further analysis will search for bacterial groups potentially related to the fungal antagonism. 57 Improving the recovery outcomes of the critically endangered Wollemi pine: is success determined by soil microbes? Jessica Mowle*1, Ian Anderson1, Brajesh Singh1, Steve Clarke2, Catherine Offord3, Jeff Powell1 1 Hawkesbury Institute for the Environment, University of Western Sydney, Australia, 2University of Western Sydney, Australia, 3Science and Conservation, Royal Botanic Gardens and Domain Trust, Australia Wollemi pine (Wollemia nobilis W. Jones, K. Hill & J. Allen) is a monotypic species, of which fewer than 100 trees are known in the wild. The Wollemi pine Recovery Team has proposed translocation as a conservation strategy to establish 'back‐ups' to the wild population; however, knowledge regarding the environmental/ biotic requirements of individuals planted in new environments is limited. One of the most important limitations to the introduction or reintroduction of tree species is the presence of suitable microbial partners. Plants in novel environments will encounter fewer co-evolved mutualists. Wollemi pine grows on shallow soils of poor nutrient status and high acidity and is likely to be highly dependent on mycorrhizal fungi, which have been observed associated with the roots of Wollemi pine, and bacteria that contribute to nutrient cycling. A plant-soil feedback experiment was undertaken to estimate the effect of soil microbial communities on seedling growth and survival, under semi-realistic conditions. We found that microbial communities associated with soils under Wollemi pine in the wild differed from those under neighbouring species. We also found that Wollemi pine seedlings were slightly larger at 5 months when interacting with their own microbes than with microbes associated with these neighbours, suggesting that generalist pathogens and/or a lack of host-specific beneficial associations may be an important factor limiting Wollemi pine recruitment in the wild. 35 Rhizosphere Microbiome 1 Monday 22 June – Poster session 58 Bioinoculants versus chemical fertilizers: Assessment of rhizospheric microbial communities in pigeonpea (Cajanuns cajan) under field condition Shilpi Sharma*1, Richa Sharma2, Preeti Chopra3, Vijay Pooniya4, Virendra Swarup Bisaria2, Karivaradharajan Swarnalakshmi3 1 Indian Institute of Technology Delhi, India, 2Dept of Biochem Engg and Biotechnology, IIT Delhi, India, 3Division of Microbiology, Indian Agricultural Research Institute, India, 4Division of Agronomy, Indian Agricultural Research Institute, India In today’s agriculture for environmental sustainability there is an increased emphasis on employing eco-friendly agricultural amendments like bioinoculants. While there are various reports on their performance with respect to enhancement of crop’s growth and yield under field conditions, a largely ignored aspect has been their impact on rhizospheric microflora. Application of plant growth promoting microbes in numbers larger than what are naturally present in the rhizosphere is bound to exert, at least a transient, effect on the resident microflora, thereby affecting crucial soil processes. Hence, the present study was undertaken to assess the effects (target and non–target) of plant growth promoting bioinoculants, viz. Bacillus sp., Pseudomonas sp. and Azotobacter sp. (individually and in different combinations) in pigeon pea. The effects of bioinoculants were compared with recommended dose of chemical fertilizers. Together with measuring plant growth parameters and soil nutrient status, cultivation–dependent (enumeration on specific media) and –independent [qPCR and denaturing gradient gel electrophoresis (DGGE)] tools were employed to characterise the structural and functional diversity of rhizospheric microflora. The triple inoculation proved to be the best compared to other treatments of bioinoculants in terms of promotion of plant's growth, with results comparable to that of chemical fertilizers. The same trend was also observed with respect to nitrogen, phosphorous and potassium uptake at vegetative stage. A marked rhizosphere effect could be observed for all treatments as compared to uninoculated control. Plant’s growth stage exerted a more pronounced impact on culturable microbial diversity compared to the treatments. However, both qPCR and DGGE revealed the non-target effects of the bioinoculants on abundance of 16S rRNA gene and genes involved in various steps of nitrogen cycle. With the triple inoculation’s efficiency being comparable to that of chemical fertilizers, and no detrimental effect observed on rhizospheric microflora, the combination can be termed as ecologically "safe” for agricultural application. 36 Rhizosphere Microbiome 1 Monday 22 June – Poster session Signaling 61 The interkingdom volatile signal indole promotes root development by interfering with auxin signalling Aurélien Bailly*1, Ulrike Groenhagen2, Stefan Schulz2, Markus Geisler3, Leo Eberl4, Laure Weisskopf1 1 Agroscope, Switzerland, 2Technische Universität Braunschweig, Germany, 3University of Fribourg, Switzerland, 4University of Zurich, Switzerland The plant rhizosphere attracts and hosts complex microbial communities in a dynamic environment where rapid and targeted communication between soil microbes and plants is an important asset for the survival of both partners. Bacteria produce volatile organic compounds (VOCs) that mediate communication with plants. Although some bacterial VOCs that promote plant growth have been identified, their underlying mechanism of action is unknown. Here we demonstrate that indole, which was identified using a screen for Arabidopsis growth promotion by VOCs from soil-borne bacteria, is a potent plant-growth modulator. Beyond indole-to-auxin conversion, its prominent role in increasing the plant secondary root network is mediated by interfering with the auxin signalling machinery. Using auxin reporter lines and classic auxin physiological and transport assays we show that the indole-derived signal invades the plant body, reaches zones of auxin activity and acts in a polar auxin transport-dependent bimodal mechanism to trigger differential cellular auxin responses. Our results suggest that indole, alongside its importance as a bacterial signal molecule, can serve as a remote messenger to manipulate plant growth and development. Therefore VOCs-mediated shifts in the plant hormonal balance may represent an effective mechanism by which bacteria directly modify their ecological niche to their advantage. Better understanding of inter-kingdom volatile communications in soil may prove useful for designing cost-effective sustainable agricultural strategies. 62 Fluorescent pseudomonad injectisomes and manipulation of plant defenses: biocontrol versus pathogenic rhizosphere agents Dorian Bergeau*1, Xavier Latour2, Sylvie Mazurier3, Marie-Laure Follet-Gueye4, Abderrakib Zahid4, Jean-Claude Mollet4, Nicole Orange5, Marc Feuilloley5, Philippe Lemanceau3, Maïté Vicré-Gibouin4, Xavier Latour5 1 Normandie Université, Laboratoire de Microbiologie Signaux et Microenvironnement EA 4312 & IRIB, France, 2Normandy University (University of Rouen), France, 3INRA Dijon, UMR 1347 Agroécologie, France, 4Normandie Université, Laboratoire de Glycobiologie et Matrice Extracellulaire Végétale - EA 4358, France, 5Normandie Université, Laboratoire de Microbiologie Signaux et Microenvironnement - EA 4312, France Fluorescent pseudomonads are called ‘rhizobacteria’ since their density and activities are strongly stimulated and most elevated in the plant rhizosphere. They have a broad capability to adapt to fluctuating environments, for example by exchanging signaling molecules with the host. Among them, Pseudomonas syringae pathovars are major microbial pathogens that can cause diseases on economically important plants. During the parasitic phase, P. syringae 37 Signaling Monday 22 June – Poster session forms a type-III secretion system (T3SS) i.e. a long and flexible pilus involved in cell-to-cell communication. This structure is essential for the injection of multiple effector proteins into the plant cell, to suppress plant innate immune defenses, to manipulate hormone signaling and to elicit cell death. In contrast, numerous P. fluorescens strains are mentioned to improve plant health and are considered as biocontrol agents of soilborne diseases. They synthesize secondary metabolites, which are implied in antagonistic activity against various phytopathogens, affect the plant physiology by producing growth substances, and elicit defense reactions of the host plant (induced systemic resistance). Recently, several plant P. fluorescens isolates were found to carry T3SS genes although their role is unknown. De facto, the structure of P. fluorescens T3SS and its potential role in virulence or in beneficial interaction with eukaryotic hosts are also ignored. The objective of this work is to investigate the T3SS morphology by transmission electron microscopy and the plant immune response induced by a biocontrol P. fluorescens strain in comparison with a phytopathogenic P. syringae strain. Induction of T3SS genes by exudates analogous were also compared for these strains. 63 Nitric oxide is involved in phosphate deficiency-induced stomatal closure in maize Lingyun Cheng*, Ying Wang, Jianbo Shen China Agricultural University, China Phosphorus is an essential element required for plant growth, and P deficiency could decrease plant water use. Nitric oxide (NO) is emerging as an important messenger molecule involved in many important physiological processes in plants. We hypothesized that NO is involved in phosphate deficiency-induced stomatal closure in maize. In this work, we studied the effect of NO donor sodium nitroprusside (SNP) and NO scavengers methylene blue (MB) on the response of stomatal behavior of maize (Zea mays L.) grown at different phosphate treatments. In contrast to phosphate-sufficient treatment, long-term phosphate starvation and short-term deprivation treatments strikingly induced the NO accumulation in lateral root tips of maize, as wells as the reduction of stomatal conductance and stomatal aperture of leaves. After short-term phosphate deprivation, detached maize leaves that were pretreated with MB restored the stomatal conductance and stomatal aperture to the level found in phosphate-sufficient leaves. Coincidently, exogenous supply of SNP to phosphate-sufficient leaves was able to induce a 50% closure of stomata in leaves. In parallel experiments, applying SNP or MB to the root medium regulated the stomatal behavior in a similar way as applying to the detached leaves. Taken together, these results suggest that NO might confer the stomatal closure in response to phosphate starvation. 38 Signaling Monday 22 June – Poster session 64 Indole acetic acid biosynthesis and nitric oxide metabolism crosstalk in Azospirillum brasilense SM Vatsala Koul*, Alok Adholeya, Mandira Kochar The Energy and Resources Institute, India Plant growth promoting (PGP) rhizobacteria, Azospirillum brasilense SM releases phytohormone-Indole-3-acetic acid (IAA) and other plant growth regulators into the rhizosphere which enhances plant development. A crosstalk between IAA and gasotransmitter, Nitric oxide (NO) is speculated which may further benefit the plants. To corroborate this, mutant strains over-expressing essential NO metabolism genes i.e. nitrous oxide reductase (nosZ), nitrous oxide reductase regulator (nosR) and nitric oxide reductase (norB) were generated. The production of NO was established by fluorescence assay using NO-specific probe: 4,5- diamino-fluorescein diacetate (DAF-2DA) and NO scavenger: 2-(4carboxyphenyl)-4,4,5,5,-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO). Increased release of NO was observed by all the mutants in the presence of Tryptophan (IAA substrate) and IAA. Quantitative IAA estimation suggested that nosR and norB influences regulation of IAA biosynthesis in A. brasilense SM. The NO quencher, inhibitor and donor reduced or blocked IAA biosynthesis in wild type and mutants, emphasizing a common regulatory role of these molecules in IAA biosynthesis. Surface colonization of strain SM on sorghum roots was established by electron microscopy and improved plant development was observed with the mutants, possibly mediated by increased NO and IAA levels. Expression studies by qPCR revealed that apart from Arginine, Tryptophan is able to induce expression of NO metabolism genes. IAA biosynthesis gene, indole-3-pyruvate decarboxylase (ipdC) was influenced positively by Tryptophan but negatively by Arginine. These results impress some shared signalling mechanism or potential crosstalk involving IAA and NO in strain SM. 65 Plant activating proteins from microbes (PAPM) as plant health determinants Sivakumar Uthandi, Tamilarasi Mani* Tamil Nadu Agricultural University, India In order to feed the estimated global population of 9 billion in the year 2050, agricultural yields will have to increase by 70 to 100% which can be achieved by improved understanding of plant-microbe interactions to secure nutrients, deter pathogens and resist environmental stress. While plants recognize and respond to environmental signals, including pathogen infection, an incompatible interaction will not only restrict pathogen proliferation by causing hypersensitive response (HR) but also leads to a systemic acquired resistance (SAR) that prevent subsequent infections by other pathogens. HR responses are exerted by secreted protein from the Gram negative plant pathogenic bacteria through type III secretory system which acts as an elicitor of plant defence. The hrp (hypersensitive response & pathogenicity) genes encode elicitor proteins (harpin like proteins) and induce resistance in a variety of plants against a broad array of pathogens and enhancing plant growth.In the present study, Xanthomonas axonopodis pv. dieffenbachiae which cause blight disease in ornamental plants was cultured in Hrp inducing media. The cell-free extracellular proteins upon concentration were infiltrated onto tobacco leaves and the response was monitored until visible necrosis like HR. Further, Hydrogen peroxide (H2O2) detected at the site of infiltration suggesting the activation of signal transduction for the production of reactive oxygen species (ROS) 39 Signaling Monday 22 June – Poster session scavenging enzymes. Consequently, ROS scavenging enzymes including super oxide dismutase, polyphenol oxidase and catalases were measured. In addition, callose deposition on leaves suggests that HR was mediated by the applied proteins. To identify the proteins responsible for HR, the secretome of X. axonopodis pv. dieffenbachiae under apoplast fluid based induction was analysed and a unique protein having molecular weight around 26 kDa was detected. Further confirmation of an indicator of SAR signalling pathway, the nonexpresser of pathogenicity related1 (npr1) gene regulation and over expression of gene responsible for PAPM are underway. 66 Investigating mechanisms for vermicompost promotion of tomato growth in organic production systems Juana Munoz Ucros* Cornell University, United States One of the main limitations of conventional farming is nutrient management, where crops tend to be nutrient limited and animal production operations result in excess nutrients. In this proposed work we intend to understand the mechanisms by which vermicompost (VC) of dairy farm waste can provide plant growth promotion to horticultural crops as an alternative to synthetic fertilizers, optimizing food production and nutrient recycling while reducing waste. Tomato transplants will be grown in the greenhouse amended with VC that has been autoclaved, non-autoclaved, and autoclaved plus re-inoculated with the microbial community found naturally in the VC. Plants will be monitored weekly for changes in germination and growth parameters, and plant gene expression. Soil samples will be collected to evaluate soil exoenzyme activity, available N, and microbial community composition. The results of this work will contribute to the understanding of the mechanisms by which VC amendments promote plant growth. 67 Root exudate- induced influences on surface properties and plant infectivity of slow and fast growing strains nodulating Glycine max.L Satish Naik*, David Biate, Annu Kumari, Lakkineni Vital, Kiran K Reddy, Rajesh Kumar, Kannapalli Annapurna Indian Agricultural Research Institute, India Root exudates have been implicated as playing an important role in mediating recognition and specificity in the Rhizobium–legume nitrogen-fixing symbiosis. To consolidate this hypothesis, we characterized the root exudate profiles of five soybean genotypes for organic acids, amino acids and flavonoids. High nodulating soybean genotypes showed propanoic acid, benzenedicarxylic acid, D-threo-Pentonic acid –methyl glutaconic acid where as the low nodulating genotypes showed distinct presence of octadecanoic acid. Among amino acids high amounts of glutamic acid and aspartic acids were found in high and low nodulating genotypes respectively. We also studied the influence of these exudates on the surface properties, protein profiles and plant infectivity of slow and fast growing rhizobial strains nodulating soybean. Soybean root exudates were collected from plants grown under hydroponic conditions for 7 days under 16h and 8h day/night cycle, concentrated 10 fold, 40 Signaling Monday 22 June – Poster session filter sterilized and tested on the slow growing Bradyrhizobium strain KAS1 and the fast growing Rhizobium strain DS-1. Root exudates increased the growth yield of the two strains in the presence of a carbon source but the strains were unable to utilize root exudates as sole carbon or energy source. Exposure to root exudates altered Congo-red dye binding, which indicated changes in the bacterial surface properties at the fatty acid level. Fourier transform infrared spectroscopy (FTIR) confirmed fatty acid changes and revealed further carbohydrate changes. Root exudates –induced changes in surface components of the strains may contribute to successful root colonization and nodule formation with subsequent plant growth promotion. 68 EcfE, a master regulator of pea root attachment and colonization of Rhizobium leguminosarum bv viciae 3841 Vinoy Ramachandran*1, Eduardo Balsanelli2, Alison East2, Andrew McNally2, Karunakaran Ramakrishnan2, Philip Poole1 1 University of Oxford, United Kingdom, 2John Innes Centre, United Kingdom Attachment and colonization of legume roots by rhizobia is one of the very early and critical steps in symbiosis. Legumes tightly regulate attachment and colonization by very specific time-dependent signals and the ability to perceive and respond to such signals by rhizobia is a requirement for successful colonization. From our earlier rhizosphere transcriptomics studies, we identified a three-gene operon coding for lipoprotein (LppE), ECF (EcfE) and anti-ECF (AsfE) sigma factors to be highly and specifically transcribed during root colonization and weakly in lab culture, only when supplemented with phenylalanine. Current work, attempts to characterize the role of these genes in root attachment and colonization by taking advantage of cutting-edge lux luminescence assays, confocal imaging, RNA-seq and ChIP-seq. In-planta root colonization assays with wild-type harboring lppE promoter lux fusion showed activation of lppE operon specifically on pea root elongation zone, emphasizing an important role of this operon in early stages of Rhizobia-legume interaction. Further in-vitro induction studies of lppE promoter lux fusion by phenylalanine in different mutant background demonstrated that EcfE regulates this operon. In-vitro root attachment assay showed that, lppE mutant were severely attenuated in root attachment whereas asfE (EcfE over-expressing strain) mutant showed a hyper-attachment phenotype. In contrast, in-planta colonization assay showed that lppE and ecfE mutants were able to colonize pea roots as wild-type whereas asfE mutant failed to colonize. Together these results emphasize the necessity of temporal regulation of specific genes in the attachment regulon to allow rhizobia to spread and colonize. To identify the further genes regulated by EcfE, we performed microarray analysis of asfE mutant, which showed elevated expression of several EcfE target genes, unraveling a complex hierarchical regulatory network governing pea root attachment and colonization. Together, this suggests that EcfE is a master regulator of pea root attachment and colonization. 41 Signaling Monday 22 June – Poster session 69 Influence of type VI secretion system (T6SS) on rhizosphere competence of a rhizobacterium Alain Sarniguet*, Muriel Marchi, Charline Lecomte, Morgane Boutin, Anne-Yvonne Guillerm-Erckelboudt, Lionel Lebreton, Kévin Gazengel, Stéphanie Daval, Alain Sarniguet INRA, France Rhizosphere competence of rhizobacteria is governed by multiple mechanisms enabling surface colonization. Protein secretion systems such as type VI secretion system (T6SS) may be involved in such rhizobacterial life traits. The role of one T6SS in the ability of the rhizobacterial Pseudomonas fluorescens Pf29Arp strain to colonize different environments was investigated. A deletion mutant of the T6SS-IV cluster was obtained and compared to the wild type. This ΔT6SS-IV mutant displayed distinct colony morphology and cell rearrangement, suggesting a role of T6SS-VI genes in phenotype variability. However, the ΔT6SS-IV mutant was a better colonizer of wheat rhizosphere than the wild type strain whereas the deletion did not affect the bacterial survival in non-sterile bulk soil. Some of the T6SS-IV genes are involved in colony morphology and in rhizosphere colonization abilities of the P. fluorescens Pf29Arp. 70 Early root responses to beneficial rhizobacteria and elicitors Ioannis Stringlis*, Christos Zamioudis, Corné Pieterse Utrecht University, Plant-Microbe Interactions, Netherlands To defend themselves against pathogens, plants have evolved a sophisticated immune system that recognizes conserved components of microbial origin, collectively referred to as Microbe-Associated Molecular Patterns (MAMPs). Although, innate immune signaling in leaves has been studied in depth, information available on MAMP-triggered immunity in the roots is limited. Root colonization by selected strains of non-pathogenic rhizobacteria triggers an induced systemic resistance (ISR) in diverse plant species that is effective against a broad range of pathogens. Here, by employing Affymetrix ATH1 whole-genome transcriptional profiling and Gene Ontology (GO) analysis, we demonstrate that the transcriptome of Arabidopsis roots two days after bacterization by the reference strain Pseudomonas fluorescens WCS417 is associated with immune responses commonly activated upon perception of MAMPs, including metabolic and cellular defense responses. Aiming to further characterize early events upon perception of microbial signals and dissect signaling mechanisms underpinning MAMP-triggered immunity locally in roots, we treated Arabidopsis roots with fungal and bacterial elicitors and P. fluorescens WCS417. Application of the wellstudied elicitors flg22 and chitin resulted in a transient upregulation of the MAMP-responsive genes MYB51 and CYP71A12, indicating that roots actively respond to immune elicitors of bacterial and fungal origin. However, P. fluorescens WCS417 caused weak upregulation of these genes that returned earlier back to basal levels, suggesting that initial host immune responses become suppressed, possibly to accommodate colonization of the beneficial. Amongst various hormonal marker-genes that we have tested for their responsiveness to immune-elicitors, we observed induction of ethylene and auxin responsive genes, pointing to a role of ethylene-mediated signaling and auxin distribution in MAMP-triggered immunity in roots. An RNA-sequencing experiment following this initial findings will provide us with a 42 Signaling Monday 22 June – Poster session better understanding on the molecular mechanisms and the timing of the responses involved in early steps of root-microbes interactions. 71 Expanding the repertoire of mRNA targets controlled by the post-transcriptional Gac/Rsm cascade in Pseudomonas protegens CHA0 Patricio Sobrero, Claudio Valverde* Universidad Nacional de Quilmes, Argentina The rhizospheric strain Pseudomonas protegens CHA0 regulates the expression of several traits involved in biocontrol of phytopathogenic fungi through a well characterized network that includes an extracellular unknown autoinducer-like signal, the GacS/A two component system, two RNA-binding proteins (e.g., RsmA/E) and three small non-coding regulatory RNAs (RsmX, RsmY and RsmZ). The Gac/Rsm pathway orchestrates the induction of cyanide and antibiotics production, and of several extracellular hydrolytic enzymes. Based on structural and sequence requirements of the interacting elements of the cascade (proteins and RNAs) it is anticipated that expression of novel uncharacterized genes/operons may be subject to regulation by the Gac/Rsm system, and thus contribute to the ecological fitness and biocontrol activity of strain CHA0. In order to get a broader insight into the regulon of the Gac/Rsm network, we carried out an in silico prediction of novel Gac/Rsm targets by inspecting the genome sequence of P. protegens strain CHA0 for the presence of RsmEbinding sites near or at their cognate ribosome binding site within the untranslated region (UTR) of all annotated genes. The output list was seized according to local secondary structure folding and prediction of correct RNA-RsmE residue contacts, and by conservation of the putative binding site among related pseudomonads. Among a set of 45 identified putative novel Gac/Rsm targets, a subset of candidates was experimentally tested by means of translational ‘lacZ reporter fusions in the genetic background of Gac/Rsm mutant strains. We present evidences of translational regulation by Gac/Rsm of the following mRNA genes: 1) PFLCHA0_c21910, encoding a LuxR-like transcriptional factor that is physically associated with a cyclic lipopeptide biosynthetic operon in different pseudomonads; 2) dprA (PFLCHA0_c00250), encoding a putative DNA protecting protein; 3) pksP (PFLCHA0_c02690), encoding a putative acyl carrier protein. Our results broaden the repertoire of genes/operons under the control of the post-transcriptional Gac/Rsm system. 72 Root cell type-specific gene expression in response to plant growth-promoting rhizobacteria Eline H. Verbon*1, Christos Zamioudis1, Louisa M. Liberman2, Philip N. Benfey2, Corné M.J. Pieterse1 1 Utrecht University, Netherlands, 2Duke University, USA Plant growth-promoting rhizobacteria are known to promote growth and to induce systemic resistance against pathogen attack in a wide variety of crop species, indicating that these bacteria can be used as natural fertilizers and pesticides. To maximize the agricultural potential of the bacteria, a detailed understanding of the molecular mechanisms underlying both growth promotion and systemic resistance is essential. In the shoot, the plant 43 Signaling Monday 22 June – Poster session endogenous jasmonic acid and ethylene defense signaling pathways are generally activated to induce systemic resistance. In the root no signaling pathways are known that translate root colonization into a signal that primes the shoot defense responses. Previous experiments have shown that the transcription factor MYB72 is upregulated in the root upon colonization by beneficial Pseudomonas spp. and is essential for the induction of systemic resistance. The upregulation of MYB72 depends on an increase in photosynthesis in the shoot. Interestingly, MYB72 is also involved in the iron deficiency response. Our aim is to uncover root specific gene regulatory networks activated upon root colonization and to determine the role of MYB72 in these networks. We used fluorescent activated cell sorting to enrich for vascular, endodermal, cortical or epidermal cell populations of fluorescently tagged roots grown in either sterile or colonized conditions. Next, we sequenced the RNA libraries of each of these cell types. Analysis of these transcriptomes will allow us to investigate the molecular details of the early root responses elicited upon colonization by beneficial rhizobacteria. These responses will subsequently be tested to determine whether they are involved in the induction of systemic resistance or promotion of plant growth. 73 When signaling gets smelly: bacterial volatiles and their effects on rhizosphere inhabitants Laure Weisskopf* Agroscope, Switzerland In addition to soluble molecules, bacteria emit a wide range of volatile organic compounds, which have raised increasing interest over the last decade. An overview of our recent work on volatile-mediated effects of bacteria on plants, fungi, oomycetes and bacteria themselves will be presented. Particular emphasis will be made on the protective effect of bacterial volatiles against disease-causing organisms such as the late blight causing agent Phytophthora infestans. Detailed profiling of the volatiles emitted by a collection of newly isolated rhizosphere bacterial strains revealed high chemical diversity even within closely related taxa. These volatile signals had target-specific effects and some of them, mostly long chain alkenes and sulphur compounds, induced drastic growth inhibition and physiological perturbations in the disease-causing agents. In bacteria-bacteria interactions, we have shown that bacteria are able to discriminate between their own volatiles and those of other, distantly related species. Future challenges, such as the need to investigate volatile emission in the natural environment of the bacterial strains, or to elucidate the cues triggering volatile emission in rhizosphere bacteria, will be highlighted. The potential of applying these volatile signals or their bacterial producers for sustainable crop enhancement and protection in the field will be discussed. 44 Signaling Monday 22 June – Poster session 74 Hijacking common mycorrhizal networks for defense signal transfer and underground interplant communication Song Yuanyuan*1, Wen Xiong Lin1, Suzanne W. Simard2, Ren Sen Zeng1 1 Fujian Agriculture and Forestry University, China, 2Department of Forest and Conservation Sciences, University of British Columbia, Canada How plants can communicate each other is an intriguing scientific question. Although increasing evidences show existence of plant-plant communication, the vast majority of the studies conducted so far mainly focus on induced volatile-mediated communication. Mycorrhizae are ubiquitous plant-fungus symbiosis in land ecosystems. Common mycorrhizal networks (CMNs) link multiple plants together in ecosystems, and ecological significance of plant-to-plant carbon and other nutrient movement through CMNs is well addressed. We hypothesized that plants can hijack CMNs as an underground conduit for transferring induced defense signals and interplant communication. We established CMN between two tomato plants in pots with mycorrhizal fungus Funneliformis mosseae, challenged a ‘donor’ plant with either a caterpillar or a pathogen, and investigated defense responses and resistance in neighbouring CMN-connected ‘receiver’ plants. After CMN establishment caterpillar infestation or pathogen infection on the ‘donor’ plant led to increased resistance and activities of putative defensive enzymes, induction of defense-related genes and activation of jasmonate (JA) pathway in the ‘receiver’ plant. However, use of a JA biosynthesis defective mutant spr2 as ‘donor’ plants resulted in no induction of defense responses and no change in resistance in ‘receiver’ plants, suggesting that JA signaling is required for CMNmediated interplant communication. Our results suggest that plants are able to hijack CMNs for induced defense signal transfer and interplant defense communication. 45 Signaling Monday 22 June – Poster session Rhizosphere and Climate Change 75 Above- and belowground plant responses to long- and short-term frost manipulations Gesche Blume-Werry*1, Juergen Kreyling2, Hjalmar Laudon3, Ann Milbau1 1 Climate Impacts Research Centre, Umeå University, Sweden, 2University Greifswald, Germany, 3SLU Umeå, Sweden Boreal forests make up one third of the world’s forested areas and store 30% of the global terrestrial carbon pool. They are shaped by distinct seasonality with a persistent snow cover during wintertime. However, with decreasing extent and duration of snow cover with climate change, soils will experience more severe frost which has been shown to affect soil carbon and nutrient fluxes. Vegetation responses, in particular root mortality, have been proposed as possible mechanisms. We directly compared short- (1 year) and long-term (11 years) effects of an absent snow cover, and thereof resulting severe air and soil frost, on above- and belowground growth, phenology and biomass of vegetation in a Norway Spruce forest in northern Sweden. In the long-term treatment vascular plants had reduced shoot growth, their cover had decreased by 93% and root biomass had decreased by 40% compared to the control. In the short-term manipulation, the same effects were seen as trends, yet were less substantial and thus not significant. However, the short-term manipulation resulted in severe visible frost damage of in average 30% on shoots of understory dwarf shrubs and delayed their spring phenology by over a week, compared to the control. Surprisingly, this was not present in the long-term manipulation, which could indicate either an adaptation of the vegetation to the long lasting severe frost, or a survival of only those individuals that can cope with these extreme conditions. Data on root growth, mortality and turnover, measured with minirhizotrons, are currently being analyzed and will be incorporated in the final presentation. In general, we show that snow removal significantly affects boreal vegetation both aboveand belowground with implications for soil carbon and nutrient cycling and that some of the more significant and far-reaching ecosystem effects were only seen in the long-term experiment. 46 Rhizosphere and Climate Change Monday 22 June – Poster session 76 Phosphate depletion in Olea europea L. rhizosphere related to mycorrhizal colonization in different climates Malika Boudiaf Nait Kaci*1, Louisa Bouhired2, Sophia Mouas Bourbia3, Noria Smail Saadoun3, Arezki Derridj4 1 Laboratoire Ressources Naturelles Université Mouloud Mammeri, Algeria, 2Université Houari Boumediene, Algeria, 3Laboratoire Ressources Naturelles, Département des Sciences Agronomiques, Faculté des Sciences Biologiques et Agronomiques Université M. Mammeri, Algeria, 4Laboratoire d’Ecologie, Département des Sciences Agronomiques, Faculté des Sciences Biologiques et Agronomiques UMMTO, Algeria In Algeria, olive groves are widespread due to their socioeconomic importance and their rusticity. However, since several decades these groves have been progressively neglected in profit of more yielding cultures. Such policies led to a deterioration of agroecosystems for which currently attempting to restore function. Olea europea L., a perennial plant is able to form arbuscular mycorrhizae. The aim of this work was to study the root induced chemical changes occurring in the rhizosphere of olive that can influence the dynamic of phosphorus. Bulk, rhizospheric soils, roots and leaves were collected in three orchards in summer, from sub-humid, semi-arid to arid regions of Algeria. P-available was obtained by the method of Olsen. The rhizosphere soil showed a significantly higher concentration of organic-carbon and total nitrogen. The concentrations of all phosphorus fractions in the rhizospheric soil were significantly lower than those in bulk soil. A P-available deficiency was measured in bulk and rhizospheric soil and confirmed by P-foliar. The uptake of phosphate by root induced a depletion of all P fractions in the rhizosphere. Philips and Hayman’s technic applied to the root samples showed in the cells of roots characteristic structures of arbuscular mycorrhizae like hyphal coils, arbuscules and vesicles. Arbuscular mycorrhiza is a widespread symbiotic association between plants and fungal microsymbionts that supports plant development under nutrient-limiting and various stress conditions. However their frequency is higher in the root sample of arid orchard. Olive trees surveyed in the three climates showed heavy AM colonization, indicating a high mycorrhizal dependency of olive groves in this environment. 77 Effects of plant-growth-promoting rhizobacteria on barley plants under different CO2 and water regimes Olga Calvo*, Ndubuisi Chimelue Nwabufo, Andreas Fangmeier University of Hohenheim. Institute of Landscape and Plant Ecology, Germany Rising CO2 concentrations in combination with drought stress are likely to influence not only aboveground growth, but also belowground plant processes. Plant-growth-promoting rhizobacteria (PGPR) colonize the rhizosphere of many plant species and confer beneficial effects under environmental stresses. Furthermore, root exudates play a role in interactions between plant roots and other organisms present in the rhizosphere. Only few reports have been published on PGPR as elicitors of tolerance to abiotic stresses, such as drought. Furthermore, little is known about the influence of environmental factors on root exudation patterns. Therefore, this study was conducted in order to investigate the effect of two commercially available PGPRs on the growth and root exudation of barley (Hordeum vulgare L.) under different CO2 and water treatments. 47 Rhizosphere and Climate Change Monday 22 June – Poster session In a growth chamber experiment climatic conditions of a field site close to Stuttgart were simulated. Barley plants were grown in pots filled with sand and exposed to ambient (380 ppm) or elevated (550 ppm) CO2. Plants received the normal daily amount of rainfall in the region of Stuttgart or 33% less. Plants were harvested at the stem elongation growth stage and when the inflorescences emerged. At both dates, data were collected on above and belowground variables. In addition, analyses of water use efficiency, stomatal conductance, SPAD values and composition of root exudates were performed. Preliminary results showed significant effects of the factors and their interactions on some of the measured variables. In the context of food security and agricultural sustainability, further studies with other crop plants are needed to demonstrate whether PGPR cause a range of crops to be tolerant to environmental stresses improving crop production. 78 Drought effects on rhizodeposition and ecological implications Alberto Canarini*, Andrew Merchant, Feike Dijkstra The University of Sydney, Australia Drought intensity is predicted to increase in the next decades across many areas of the world. This phenomenon will affect plant production and consequently rhizodeposition with potentially large ecological impacts on soil carbon (C) and nutrient cycling. However, the direction and magnitude of these impacts are still unclear. Here we investigate the effect of drought on rhizodeposition and the connection with C stabilization and nitrogen (N) cycling. We combine stable isotope techniques with chromatography and mass spectrometry analyses in order to understand how drought affects the allocation of C into different soil pools and the nature of compounds invested belowground. We investigate wheat, soybean and sunflower as species of major agroeconomical value due to their wide spread use and potential to cause large ecological impacts. Results illustrate that plants are able to allocate substantial amounts of C belowground through rhizodeposition, which have the potential to increase the amount of mineral associated C (a relatively stable form of soil C), most likely through microbial interactions (indicated by correlation between plant derived C found in microbial biomass and the stable C pool, R2 = 0.54, P < 0.001). Organic compounds obtained from phloem sap, root tissues and exudates collection will then be discussed with reference to the effect of drought on the abundance of these compounds. Samples are being processed using a recently developed method for liquid chromatography – mass spectrometry targeting major metabolites in plants (sugars, amino- and organic-acids), important for different soil processes. These results provide an improved understanding of biochemical exchange in the plant-soil system in a future of reduced water availability for agro-ecosystems. 48 Rhizosphere and Climate Change Monday 22 June – Poster session 79 Characterizing root traits for edaphic stress adaptation Yinglong Chen*1, Yan Fang1, Suiqi Zhang1, Kadambot Siddique2, Zed Rengel3 1 Institute of Soil and Water Conservation, Northwest A&F University, and Chinese Academy of Sciences and Ministry of Water Resources, China, 2The UWA Institute of Agriculture, The University of Western Australia, Australia, 3The UWA Institute of Agriculture, and School of Earth and Environment, The University of Western Australia, Australia Sustainable crop production is challenged by the climate changes with likely increased production limitation and uncertainty in the future. Edaphic stresses, such as drought and low-fertility soils, are the main factors restricting crop production in many counties. Selecting and breeding cultivars with root architecture traits efficient in water and nutrient use becomes an important breeding strategy aiming for increased crop adaptation to edaphic stress. A serial of phenotyping experiments involving novel semi-hydroponic platform, soilfilled columns and rhizotrons were carried out to study phenotypic variability in root traits and root responses to water and phosphorus deficits in narrow-leafed lupin. Large variability among the tested genotypes was observed with 21 (out of 38) root traits had greater coefficient of variation values than 0.5. Multivariate root traits were constructed with Principal Components Analysis resulted in 9 components with eigenvalues greater than one. Observations confirmed the inability of this species in forming functional mycorrhizas nor cluster roots under any phosphorus (P) status. Significant variations were observed among genotypes in root architecture and distribution in response to drought and low-P stresses. Placing P fertiliser deeper enhanced root growth and P-use efficiency. Plants produced greater amounts of carboxylates in the rhizosphere when plants grown in low P environment. Localised P patches had significant impacts on root architecture and exudation compared to uniform P application. Alteration of root distribution and architecture and changes in rhizosphere exudation of carboxylates were important strategies for efficient P acquisition in low P soil. Our study provides detailed description of the phenotypic variability in root architecture and the insight into the mechanistic responses of root traits influencing water and nutrient acquisition in low-fertility drying soils. 80 Microbial invasions - a complex interplay between soil properties and biology Renata Slavikova1, Jan Jansa2, Muhammad Ali1, Emmanuel Frossard1, Hannes Gamper*1 1 ETH Zurich, Agricultural Sciences, Switzerland, 2The Czech Academy of Sciences, Institute of Microbiology, Czech Republic Biological invasions are greatly facilitated by global traveling and trade and are widely recognized as potential threads to local biodiversity and ecosystem functioning. Some introduced species can be devastating. Yet, research has mainly focused on plants and animals. Microbial dispersal, except for crop and human pathogens and parasites, received much less attention so far, despite its potentially important consequences for soil and plant functioning, due to microbe-soil adaptation. Arbuscular mycorrhizal fungi (AMF) mediate elemental fluxes and plant productivity via assisting plants with mineral nutrient acquisition and protecting them from abiotic and biotic stresses. Here we report on the findings of a pot experiment where we simulated, based on a cross-factorial design, gradients of a major biotic 49 Rhizosphere and Climate Change Monday 22 June – Poster session and abiotic perturbation event by mixing two soils from geographically well-separated, but climatically similar sites in Switzerland. We mixed the native living soils in nine different proportions and established control treatments in which one or both soils had their native AMF removed by gamma irradiation. Other native soil microbes smaller than the propagules of AMF were added back. The experiment so far revealed strong impairments to the growth of Plantago lanceolata when the two soils were alive, as compared to their sterilized counterparts. These impairments were to a great extent rectified by addition of just only 5% of foreign sterilized soil. The mechanistic basis behind this phenomenon will be elaborated based on combined elemental flux and microbial community analyses, using stable (13C, 15N) and radioactive (33P) tracers and molecular genetic fingerprinting. 81 Community assembly processes of N2O-reducing prokaryotes in the rhizosphere - effect of edaphic factors and plant species Daniel R. H. Graf*, Christopher M. Jones, Ming Zhao, Sara Hallin Swedish University of Agricultural Sciences, Department of Microbiology, Sweden Nitrous oxide (N2O) is the dominant ozone depleting substance and a potent greenhouse gas. Nearly 70% of global N2O-emissions can be attributed to terrestrial ecosystems, of which 45% originate from agricultural land. The rhizosphere is known as a hotspot for prokaryotes producing and reducing N2O. The only enzyme known to reduce N2O to nitrogen gas (N2) is the N2O-reductase (N2OR), which is encoded by two variants of the nosZ gene, clade I and clade II, the latter of which has been shown to be correlated with a higher N2O sink capacity. To determine the effect of plant species and soil type on the diversity, structure, and functioning of N2O reducing communities in the rhizosphere and on the root surface, we conducted a pot experiment growing sunflower (Helianthus anuus) and barley (Hordeum vulgare) in a clay and a silt soil. We observed higher potential N2O production rates in rhizosphere in the clay soil compared to the silt soil, and rates were higher when planted with barley than sunflower. In agreement, potential N2O production measurements on washed roots revealed a significantly higher production rate by attached microbial communities or endosymbionts of barley roots as opposed to sunflower roots. Using quantitative PCR, we demonstrate that low N2O production correlates with low nosZ clade I/clade II abundance ratios in the rhizosphere soil. The ratios overall were more dissimilar between the soils than between the plants, even on the roots, indicating a stronger effect of soil type than plant on the community assembly of N2O-reducing prokaryotes in the rhizosphere. Amplicon sequencing of both nosZ clades from rhizosphere soil and root surfaces was performed and additional analyses will be presented on the relative influence of soil and plant species on diversity and composition of N2O reducing communities in the different compartments. 50 Rhizosphere and Climate Change Monday 22 June – Poster session 82 The Bryosphere as a regulator of peatland carbon balance in response to climate warming Vincent Jassey*1, Constant Signarbieux2, Alexandre Buttler3, Bjorn Robroek2 1 Research Institute WSL, Switzerland, 2School of Architecture, Civil and Environmental Engineering (ENAC), Ecole Polytechnique Fédérale de Lausanne EPFL, Ecological Systems Laboratory (ECOS), Switzerland, 3Swiss Federal Institute for Forest, Snow and Landscape Research (WSL), Switzerland In analogy to the rhizosphere, the bryosphere describes the interactions between bryophytes and their associated organisms and forms an important transitional zone mediating aboveground-belowground linkages in northern ecosystems such as peatlands. In particular, Sphagnum mosses are important players for ecological processes in peatlands, especially for carbon (C) cycle. For example, Sphagnum contributes substantially to aboveground leaf area and biomass, and therefore plays an important role in C assimilation. Furthermore, this moss is tightly linked to a wide range of microorganisms and microfauna, which interact and form a complex food web responsible of C cycling. Hence, Sphagnum-bryosphere can influence both primary production and decomposition processes in peatlands. Using a five-year field warming experiment in a Sphagnum-dominated peatland, we studied the response of the Sphagnum-bryosphere to temperature increase. We showed that the response of Sphagnum-bryosphere affects a series of peatland functions with implications for C recycling. We found that a moderate warming (ca. +1°C) exacerbated the effect of drought on Sphagnum-photosynthesis. While rate of Sphagnum-photosynthesis declined by 70% compared to wet conditions with drought alone, it became negative under the combined effect of drought and warming, indicating a carbon loss. In parallel, we found that warming strongly decreased the biomass of microbial predators (-50%) after 5 years of warming, especially mixotrophic predators (i.e. organisms combining autotrophy and heterotrophy). Such decrease of mixotrophs, in turn, modified the structure of the microbial food web and shifted the microbial food web towards a bottom-up control. This implies potentially a faster C turnover, and enhanced organic matter decomposition. Interestingly, our findings also highlighted an important role for mixotrophs declining to overall Sphagnum-photosynthesis, which could explain the differences we found between controls and warmed plots. Overall, our findings suggest that warming-effects on Sphagnum-bryosphere will alter the carbon cycle in peatlands. 83 Use of light-use efficiency functions to describe CO2 uptake at a semi-arid site, role of leaf-area index and leaf density Georgia Koerber*1, Wayne Meyer2 1 University of Adelaide, Australia, 2Adelaide University, Australia The Calperum-Chowilla OzFlux site (34°00.163S 140°35.261E) was established in July 2010 located about 20 km north-west of Renmark in South Australia within an extensive semi-arid Mallee woodland. Eddy covariance flux of carbon dioxide and water from a 20 m tower were recorded during the height of the millennium drought, its subsequent breaking during 2011 and then a wild fire in January 2014. 51 Rhizosphere and Climate Change Monday 22 June – Poster session During 2011 the perennial vegetation gradually increased in leaf area and hence sequestered carbon With future projections of increased fluctuations in temperature and rainfall events in Autumn, we want to assess the role of perennials and the soil in carbon sequestering that is likely to have a long residence time. Monthly leaf area index (LAI) using digital cover photography, leaf mass per area, thickness, soil respiration and above ground biomass additions and carbon budgets have been constructed splitting 24 hours into night and day using a shortwave radiation threshold of 10 W m-2. Nighttime flux filtered by insufficient turbulence and corrected to match daytime temperatures was subtracted from daytime flux to estimate photosynthesis. Light use efficiency curves were solved for the light compensation point and then respiration in the light and dark as described by the “Kok effect”. With improved estimates of night and day respiration and photosynthesis we regressed ecosystem respiration against monthly LAI. We hypothesize the y-intercept is plant independent respiration or heterotrophic soil respiration. With this component we will be able to derive a more accurate estimate of this ecosystems accumulation or loss of carbon. We want to track this with prevailing environmental conditions such as water and temperature/seasonality and compare this with other ecosystems. We also want to see if changes in leaf area or leaf characteristics play a part in continual adjustment and adaptation to the variable environmental controllers, especially water. 84 Deciphering transcriptomic profiles and Verticillium wilt development in olive cultivars under different scenarios of climate change Blanca B Landa*1, Miguel Montes-Borrego1, Carmen Beuzón2, Juan A. Navas-Cortés1 1 Institute for Sustainable Agriculture-CSIC, Spain, 2University of Málaga, Spain Plant disease epidemics result from interactions of a susceptible host, a virulent pathogen and conducive environment. Shifts in any component can change disease expression to still unknown directions. Verticillium wilt (VW) of olive caused by the soilborne fungus Verticillium dahliae (Vd) is of major concern for olive industry in the Mediterranean basin. We carried out experiments using olive cvs. Picual and Arbequina, which grew in soil infested by the defoliating (D) or non-defoliating (ND) Vd pathotypes at 20, 24 and 28ºC and CO2 levels of 386, 550 and 750 ppm, representative of current and future SRES-IPCC A2 and B2 scenarios for southern Spain. Surface response models quantified the combined effects of temperature and CO2 on VW intensity and plant growth, demonstrating a differential effect of biotic and abiotic factors. Optimum VW development occurred at 20-24°C and current CO2, being faster and severe in 'Picual'/D. Raising CO2 delayed VW and Vd infection. The transcriptomic profile of olive cultivars in response to Vd infection and the three climate SRES scenarios was assessed using a 12-plex microarray of 37,449 olive unigenes (OLEAGEN project). Main differences in the expression profiles were due to olive genotype, followed by the climatic conditions at the three SRES scenarios and in a lesser extent by Vd infection. Thus, plants growing at current climate showed higher number of differentially expressed genes compared to plants growing at SRES-IPCC A2 and B2 scenarios with a general trend to 52 Rhizosphere and Climate Change Monday 22 June – Poster session decrease the number of significantly transcribed genes as temperature and CO2 increased. Furthermore, a higher number of genes (up to 10x) were expressed differentially in 'Arbequina' than in 'Picual' in response to environmental changes as well as to infection by the D pathotype with a low number of genes being common for both cultivars. 85 Root exudates can control soil N dynamics Adrian Langarica Fuentes*1, Susan Mitchell1, Marta Manrubia Freixa2, Tim Daniell1 1 The James Hutton Institute, United Kingdom, 2Netherlands Institute of Ecology (NIOOKNAW), Netherlands Agriculture represents the dominant source of the potent greenhouse gas nitrous oxide (N2O). This is largely due to the conversion of added nitrogen-based fertiliser through the action of the microbial nitrogen cycle in soil. Denitrification represents the main source of nitrous oxide emission and is a carbon driven process, as it maintains respiration under low oxygen conditions with nitrogen oxides acting as alternative electron acceptors. Initial experiments using barley plants (Hordeum vulgare) indicated that different cultivars support significant variation in N2O emission from denitrification from associated soil and that these effects are connected to root exudation difference rather than direct interaction or litter effects. In this study, a controlled microcosm experiment was designed to explore the interaction between root exudation and denitrification in terms of N2O production and community dynamics. Different quantities of artificial root exudates were added daily to soil at three different water regimes. N2O emissions were measured and community dynamics were assessed using T-RFLP and real time PCR. Results suggest that N2O emissions are driven by nitrous oxide reductase activity (consumption) rather than nitrite reductase activity (production). Soil condition, through differential aerobic status and variability, also appeared to have a marked effect both on flux through denitrification and community dynamics. Further work is underway to dissect the role of exudation quality in driving the alteration in nitrogen cycle flux and the end product of denitrification. 86 Impact of chemical fertilization and organic amendment on carbon and nitrogen lability between rhiz. and non-rhiz. ecosystems Ibrahim Ortas*1, Gulistan Köskeroglu2, Refik Islam3 1 Cukurova Universitey, Turkey, 2Ç.Ü, Turkey, 3Ohio University, United States Soil carbon and nitrogen stoichiometry is important for evaluating soil organic matter quality. Our study was conducted to evaluate the impact of chemical fertilization and various organic amendments on biological, chemical and physical C and N pools to assess soil C and N dynamics under wheat (Triticum aestivum L.) - corn (Zea mays L.) rotation in semi-arid Mediterranean climate of south-western Turkey. Treatments included control, chemical fertilization (NPK), compost, animal manure, and compost+mycorrhizal inoculations. Soil samples were randomly collected from 0 to 15 and 15 to 30 cm depth and analyzed for rhizosphere and non-rhizosphere total organic carbon (TOC) and nitrogen (TN), extractable phosphorus (EP), active carbon (AC), particulate organic C (POC) and nitrogen (PON), and soluble carbon (SC) fractions to calculate C and N lability and management indices (CMI). 53 Rhizosphere and Climate Change Monday 22 June – Poster session Results showed that the organic amendments such as compost and manure applications significantly increased TOC, TN, EP, AC, POC, PON and SC as compared with the chemical fertilization and control, respectively. In 0-15 cm depth AC in control treatment was 2.37 mg kg-1, however in mineral fertilizer, manure, compost and compost +mycorrhiza treated soils 252, 272, 302, and 274 mg kg-1 respectively. Similar repose was got in non-rhizosphere soil. The C and lability was higher in the rhizosphere than in the non-rhizosphere ecosystems. Our results suggest that higher C and N lability is associated with organic amendments to improve soil quality for crop production. 87 Plant parasitic nematodes attraction by root exudates of range expanding plant species and native congeners Julio Carlos Pereira da Silva*, Rutger Wilschut, Wim H. van der Putten NIOO-KNAW, Netherlands Global climate changes enable many plant species to shift their ranges to higher latitudes and altitudes. Range expansion leads to novel interactions with a non-co-evolved multitrophic belowground community in the new range. It is possible that the novel traits of rangeexpanding plant species make them unattractive, toxic or even not recognized as a new food source to the enemies in the invasive range, resulting in reduced herbivore pressure. This novelty, however, may go at the expense of indirect control of enemies by higher trophic level organisms, as these will not recognize the cues emitted by the novel plant species. So, it may be an advantage of range-expanding plant species if novel enemies do not recognize and respond to chemical signals of host, but it will be a disadvantage when the enemies recognize the host plants, but their natural enemies not. Our study makes a first step in unravelling bottom-effects on root-feeding nematodes in the rhizosphere of range-expanding plant species and comparing this to congeneric plant species that are native in the invaded range. We present pilot results of two such pairs of congeneric plant species and two types of root-feeding nematodes (ectoparasitic or endoparasitic species). We performed experiments on petri dishes and in a greenhouse in order to study root attraction of ectoparasitic and endoparasitic nematodes. Our hypotheses are that (1) nematodes are more attracted by native plant species than by related range expanders and (2) ectoparasitic nematodes, which have less specific relations with the roots than endoparasitic nematodes, will show a less strong preference for native plants than endoparasites. 88 The role of plants in methane flux of upland soils and their influence on archaeal community composition in the rhizosphere Nadine Praeg*, Paul Illmer University of Innsbruck, Institute of Microbiology, Austria Methane is an important greenhouse gas that is produced and consumed in soils by microorganisms. Until recently, studies of the contribution of plants to the global methane flux have focused on the role of plants as conduits for soil-borne methane emissions from 54 Rhizosphere and Climate Change Monday 22 June – Poster session wetlands, but pretty barely examined to date are the influence of plants on methane flux and the presence of methanogenic and methanotrophic microorganisms in aerobic upland soils. The present study used soils from grassland sites from siliceous and calcareous bedrock located in Northern Tyrol, Austria at ~750 m a.s.l. Besides in situ methane measurements and profound soil microbiology analyses, lab-scale gas measurements with two different sitetypical grassland plants were conducted at different temperatures. Gas samples were withdrawn and subsequently analyzed via gas chromatography. To characterize the archaeal community structure in bulk and rhizosphere soil of the respective plants and soil sites, nextgeneration sequencing was conducted. In our investigations we could prove a distinct influence of plants on net methane emissions out of upland soils. Our data proved not only a clear influence of plants on methane flux in comparison to uncovered soil but between the investigated plants as well. On a lab-scale basis temperature was a crucial factor that influenced the effect of plants on methane fluxes significantly. Community analyses revealed clear differences within the archaeal community of the investigated rhizosphere and bulk soils. Furthermore, the two plant species led to different archaeal community shifts within the soils and the shifts were different between siliceous and calcareous bedrock. The study emphasizes the need to better resolve the influence of plants on the methane cycle and its involved microorganisms. 89 Drought affects spatial distribution of enzyme activities in the rhizosphere Baharsadat Razavidezfuly*1, Muhammad Sanaullah2, Evgenia Blagodatskaya2, Yakov Kuzyakov2 1 George-August-University of Göttingen, Germany, 2University of Göttingen /Agricultural Soil Science, Germany Due to high inputs of easily degradable organic compounds from the roots, the rhizosphere is a very important and dynamic hotspot of microbial activity in soil. Consequently, the enzyme activities in the rhizosphere are a footprint of complex plant-microbial interactions and may reflect functional response to climate changes. We modified the in situ soil zymography for identification and localization of hotspots of βglucosidase activity in the rhizosphere of maize under drought stress (30% of field capacity). Zymographic images showed highlighted spots of β-glucosidase activity along the roots. The β-glucosidase activity was especially high at root tips and was much stronger under drought as compared with optimal moisture (70% of field capacity). The high β-glucosidase activity under drought was confirmed by enzyme assay based on fluorogenically labelled substrates applied to the root exudates. The activity of β-glucosidase of root exudates (produced by roots and root-associated microorganisms) was significantly higher by drought stressed plants as compared with optimal moisture. In contrast, the β-glucosidase activity in destructively sampled rhizosphere soil was lower under drought stress compared with optimal moisture. Without roots, drought did not affect β-glucosidase activity in soil. Consequently, the release of organic compounds (such as mucilage) by roots under drought increased βglucosidase activity in the rhizosphere. Thus, the zymography visualized the distribution of βglucosidase activity and allowed assessment of consequences at the root55 Rhizosphere and Climate Change Monday 22 June – Poster session soil interface under drought. Furthermore, coupling of zymography and enzyme assays in the rhizosphere and non-rhizosphere soil enabled not only the precise mapping of the twodimensional distribution of enzyme activities, but also allowed quantitative assessment of soil depth imaged on the zymograms. 90 Does fungal community matter to drought tolerance of cork oak (Quercus suber L.)? Francisca Reis*1, Paula Baptista2, Rui Tavares1, Teresa Lino-Neto1 1 Biosystems & Integrative Sciences Institute (BioISI), Plant Functional Biology Center (CBFP), University of Minho, Portugal, 2Mountain Research Center (CIMO), Polytechnic Institute of Bragança, Portugal Mediterranean forests are one of the biodiversity “hotspots”. The cork oak (Quercus suber L.) forest (“montado”) is an unique and emblematic resource for Portugal, both social and economic manner, protected by EU (Habitats Directive 92/43/EEC). Portugal is the greatest cork producer with 50% of cork extraction worldwide. Nowadays, cork oak faces a severe global climate change and a reduction in water availability is expected for the near future, which is expected to decrease cork oak growth and productivity. Plant benefits from many symbiotic relations that occur between microorganisms and roots that are able to enhance nutrient and water supply. However, root colonization efficiency and fungal community structures are dependent on environmental conditions, such as water availability. The main goal of this work was to evaluate the relationship between fungal abundance and diversity conferred by different drought scenarios. The effect of drought in cork oak fungal ecosystem was accessed by studying cork oak field trees in 7 different Portuguese forests. Five different locations (Gerês, Macedo de Cavaleiros, Vimeiro, Grândola and Moura) were evaluated according to a gradient of water-availability. Samples from extreme conditions, the driest (Moura) and wettest (Gerês) places were sampled in two different sites. Soil samples (35) were analysed by ITS barcoding on ECM tips and metabarcoding using Illumina platform. The obtained results will help to understand not only the importance of fungi to drought tolerance in cork oak forest, but also which are the main fungal colonizers. 91 Combined effects of climate change induced shifts in the plant community and drought on the structure and functioning of the rhizosphere microbiome Thomas Reitz*1, Mika Tarkka2, Jessica Gutknecht3, Tesfaye Wubet1, Francois Buscot1, Erik Welk4, Alexandra Weigelt5 1 Helmholtz Centre for Environmental Research, Germany, 2HelmholtzCentre for Environmental Research, Germany, 3University of Minnesota, United States, 4MartinLuther-University Halle, Germany, 5University of Leipzig, Germany The rhizosphere microbiome and its functioning are directly affected by plant root exudates. Given the fact that these exudates differ between plant species, climate change induced shifts in the abundance of plant species changes the structural and functional composition of the microbial soil community. Besides this, increasing drought is expected to directly (due to reduced soil moisture) and indirectly (due to stress-induced, altered plant root exudates 56 Rhizosphere and Climate Change Monday 22 June – Poster session production) affect the soil microbiome. In order to study the combined effects of drought and altered plant community composition on the functioning of the rhizosphere community, we performed a common garden experiment. Two plant pools were defined and sown on experimental plots. The first pool included plant species that are predominantly distributed in southwestern Europe and, in the context of climate change, may potentially become more dominant in central Europe. The second pool included species mainly distributed in northeastern Europe, which are predicted to become increasingly displaced in central Europe. To simulate drought half of the plots were roofed for five weeks in early and late summer. After each roofing period we determined different plant root traits as well as structural and functional responses of the rhizosphere community. We observed drought-induced, morphological changes in the root structure whereas stronger effects occurred after the second roofing period. Microbial activity indices (nitrification potential and soil enzymes) as well as selected activities of isolated microorganisms were generally reduced by drought, but the extent of the reduction was related to the plant community, indicating different buffering capacities against drought stress. In terms of phosphate solubilization, contrasting responses to drought of bacteria and fungi suggest functional redundancies in the rhizosphere microbiome. In conclusion, plant root morphology was strongly affected by drought. Microbial traits were also highly affected by drought but these changes were possibly mediated through plant community responses. 92 Influence of climate change and heavy metal on the plant-microbe interactions Sukamal Sarkar*, Krishnendu Ray, Hirak Banerjee Bidhan Chandra Krishi Viswavidyalaya (State Agricultural University), India The various biotic and abiotic stress factors can affect the growth and development of crops. Particularly, the change of climatic condition and also heavy metal stress like lead (Pb), arsenic (As), cadmium (Cd), copper (Cu) etc. influence various plant physiological and metabolic process as well as development and yield of the crops. The change of climate especially the elevated atmospheric CO2 condition enhance the biomass production and also metal ion accumulation in most of the plants and help plants to support greater microbial populations and protect the microorganisms against the impacts of heavy metals. Besides, the indirect effects of climate change on the function as well as structure of plant roots, diversity and activity of rhizosphere-microbes would lead to altered metal bio-availability in soils and thus affect plant growth. However, the effects of rising of global temperature, drought or combined climatic stress on the growth of plant and metal accumulation vary substantially across physical, chemical and also biological properties of the environment (e.g., soil pH, heavy metal type and its bio-available concentrations, microbial diversity, and interactive effects of climatic factors). Overall, direct and indirect effects of climate change on heavy metal mobility in soils may further hinder the ability of plants to adapt and make them more susceptible to stress. In this study we have discussed how the various climatic parameters including atmospheric CO2, temperature and drought influence the plant–metal interaction in polluted soils. The effects of climate change and heavy metals on plant– microbe interaction, heavy metal phytoremediation and safety of food have also been narrated. This review shows how the plant–metal interaction responds to changing climatic 57 Rhizosphere and Climate Change Monday 22 June – Poster session condition and how is it beneficial in selecting crops that would be able to produce more yields without accumulating toxic heavy metals for future food security. 93 Influence of drought on the antioxidative stress defense and root exudation of european grassland species Peter Schröder*1, Christian Huber2 1 Helmholtz Zentrum Muenchen GmbH, Germany, 2Helmholtz Zentrum Muenchen, Germany Extreme weather situations like prolonged periods of drought as well as heavy precipitation within short periods of time as a consequence of climate change have an impact on our terrestrial ecosystems. It´s likely that these conditions will not only affect the vegetation but also plants associated rhizosphere bacteria and fungi. Grassland vegetation fulfills a large number of ecosystem functions that need to be preserved. With its high biodiversity, it forms natural habitats for animals and bird species. It serves as an important source for animal fodder, protects the environment from erosion and leakage of nutrients as well as it maintains our countryside and landscapes. Plants actively influence the chemical properties of the rhizosphere by exudating a variety of compounds via their root system. Many of these compounds, mostly low molecular weight organic acids, alter distinct processes within rhizospheres that might have an impact on the interactions between different plants species as well as the interactions between plants, microbes and fungi. In this study we investigated the influence of drought on the antioxidative stress defense system of typical European grassland species (Plantago lanceolata, Lotus corniculatus). For collection and analysis of root exudates we introduce a rhizotron–microsuction cup–system that allows for a continuous and noninvasive collection of exudate samples while growing the plants in a natural soil system. Analysis of exudates was done by a combination of HPLCUV/VIS, ion chromatography and LC MS/MS techniques. Analysis of different biomarkers (e.g. stress enzyme activities, thiol contents) suggests the formation of reactive oxygen species in water stressed plants. Qualitative analysis of root exudates revealed the occurrence of different phenolic and aliphatic acids with the potential to affect rhizosphere processes and its biota. 94 The complex roles of lignin and suberin in plant micronutrient uptake Tânia S. Serra*1, Kasra Talebi1, Ikram Blilou2, Mark G. M. Aarts1 1 Wageningen University, Laboratory of Genetics, Netherlands, 2Wageningen University, Plant Developmental Biology Laboratory, Netherlands Roots are specialized tissues with a central function in water and mineral nutrient uptake from the soil, to support the progressive plant growth and development. Radial transport across the root to the vascular system occurs through the apoplastic, symplastic or transmembrane pathway. To regulate the non-selective apoplastic transport, a ring-like 58 Rhizosphere and Climate Change Monday 22 June – Poster session structure composed of lignin, named Casparian strip (CS), surrounds the endodermis cells. As the root develops, transmembrane transport is blocked through the deposition of suberin at the endodermal layers. The specific mechanism underlying the generation of this diffusional barriers is not fully known, but a few studies show that defects in the barrier composition and structure can have major effects in transport of water and solutes. Thus, understanding the biosynthesis and function of such barriers will allow the future manipulation of root systems in important crops, to improve water and nutrient uptakes. To assess the role of CS/lignin and suberin in plant adaptation to mineral stress we selected several Arabidopsis mutants affected in the composition and/or structure of these polymers, such as esb1, ralph and horst. The root and shoot phenotype are studied under both deficiency and excess concentrations of zinc, copper, manganese and iron, as well as toxic amounts of cadmium. By integrating the mineral stress-response in the different root properties of each mutant, we will further understand how root structure affects metals metabolism. In addition, analysis of root structure in the natural heavy metal hyperaccumulator Noccaea caerulescens showed additional cell layer adjacent to the endodermal cells displaying lignin characteristics. Previous studies showed that several genes involved in lignin and suberin metabolism are upregulated in N. caerulescens compared to Arabidopsis plants. Thus, modifications of the root structure may account for the extraordinary ability of N. caerulescens to tolerate and accumulate high metal concentrations. 95 Hypoxia induces changes in root maintenance costs of Prunus rootstocks Guillermo Toro*1, Paula Pimentel1, Manuel Pinto2 1 Centre for Advanced Studies in Fruit, Chile, 2Institute of Agricultural Research, Chile Low oxygen in the rhizosphere (hypoxia) induces alterations in root respiration. Under hypoxia, the regulation of the energy requirements by roots seems to be necessary to cope with the energy unbalanced produced by hypoxia. Maintenance requirements can be one of them, however no much studies have been done on this matter particularly in rootstocks. In this work, we investigated whether the hypoxia tolerance in Prunus rootstock is associated to changes in the maintenance costs. Three Prunus rootstocks with contrasting tolerance to hypoxia: Marianna 2624 (tolerant), CAB6P (semi-tolerant) and Mazzard F12/1 (sensitive) were grown in nutrient solution and exposed to normal (normoxia: 7.8 mg O2 L-1) and low (hypoxia: 0.5 mg O2 L-1) oxygen concentration. Hypoxia treatment was achieved by bubbling gaseous nitrogen into the nutrient solution (0.1% agar). The root respiration components were obtained by multiple regression analysis of the root oxygen evolution and the root biomass accumulation. Under hypoxia, tolerant Prunus rootstock showed a significant decrease in root maintenance respiration from 1.8 mmol O2 g-1DW d-1 under normoxia to 0.7 mmol O2 g-1DW d-1 under hypoxia (60% reduction). This reduction in maintenance costs was paralleled by an increase of the ion uptake respiration from 5.6 mmol O2 mmol-1N under normoxia to 9.4 mmol O2 mmol1 N under hypoxia (70% increase). In this case no changes were observed in growth respiration, which in average was 7.8 mmol O2 g-1DW in both oxygen conditions. In the sensitive rootstock the maintenance respiration increased from 2.3 mmol O2 g-1DW d-1 under normoxia to 3.8 mmol O2 g-1DW d-1 under hypoxia (65% increase). But the growth respiration, 59 Rhizosphere and Climate Change Monday 22 June – Poster session decreased to 1.8 mmol O2 g-1DW under hypoxia (65% reduction). In this case, no differences were observed in ion uptake respiration. These results showed that sensitivity to hypoxia in Prunus rootstock is related to the respiratory maintenance costs. 96 Climate warming effects on plant belowground biomass allocation in tundra Peng Wang*1, Monique Heijmans1, Liesje Mommer1, Frank Berendse1, Trofim Maximov2 1 Wageningen University, Netherlands, 2The Institute for Biological Problems of Cryolithozone, Siberian Branch of the Russian Academy of Sciences, Russia Climate warming is known to have large impacts on tundra ecosystems. Increases in aboveground plant biomass in response to climate warming in the Arctic have been widely reported. However, little is known about the belowground. In order to improve our understanding of the climate warming effects on the belowground part of tundra vegetation, we did both a meta-analysis and a field experiment. We collected belowground biomass and annual temperature data from 38 published studies which were performed in tundra vegetation. Aboveground biomass increased with increasing temperature, as earlier observed. However, the belowground temperature response was significantly lower than aboveground. Moreover, belowground temperature response differed between vegetation types, as in shrub dominated vegetation belowground biomass significantly increased with temperature, but in graminoid dominated vegetation it did not change. These findings have important implications for the C and nutrient cycling in tundra ecosystems as we expect that a shift in vegetation types will occur in the future warmer conditions. We also performed a soil warming experiment in northeastern Siberia by using heating cables connected to solar panels. The experiment lasted for 5 summers and the warming treatment significantly increased soil temperature and active layer thickness during the growing season. We will present aboveground and belowground biomass results for deciduous shrubs, evergreen shrubs and graminoids harvested at the end of the growing season of 2014. We expect that the warming treatment changed the biomass allocation of different plant functional types along soil depth and thus affected the competitive relationships. 97 Impacts of atmospheric CO2 on root exudation chemistry and rhizospherecolonizing bacteria Alex Williams*, Pierre Pétriacq, T.E. Anne Cotton, David Beerling, Jurriaan Ton University of Sheffield, United Kingdom Atmospheric CO2 concentrations have fluctuated in historic times to present day and are set to increase due to anthropogenic emissions. Little is known about the effects of altered CO2 60 Rhizosphere and Climate Change Monday 22 June – Poster session levels on interactions with beneficial root-colonizing microbes in relation to the chemical composition of root exudates. This study aims to address the impact of low (200ppm), ambient (400 ppm) and high CO2 (1200 ppm) on root exudation chemistry and root-microbe interactions in the model plant species Arabidopsis thaliana. Our data indicate that CO2 influences root colonisation activity of the rhizobacterial strains Pseudomonas putida KT2440 and Pseudomonas fluorescens WCS417r, and these effects are associated with changes in root exudation chemistry. Additionally, previous data indicates that there is a positive relationship between CO2 concentrations and ontological factors that influence the speed at which plants develop. We have therefore considered the effects of developmental stage of the plant on root colonisation and exudation chemistry. Further investigations aim to reveal the physiological mechanisms and ecological consequences of these responses, as well as the impact of atmospheric CO2 on global rhizosphere communities. 98 Comprehensive analysis of rhizosphere effect in top and deep horizons of podzolic soil under spruce trees Ilya Yevdokimov*1, Alla Larionova1, Tatiana Sokolova2, Inna Tolpeshta2 1 Institute of Physicochemical and Biological Problems in Soil Science RAS, Russian Federation, 2Faculty of Soil Science, Moscow State University, Russian Federation Global warming is known to affect the carbon cycle in forest ecosystems through the intensification of soil organic matter (SOM) decomposition. It is well established that rhizodeposits stimulate CO2 efflux from the topsoil, whereas the role of rhizosphere effect in deep horizons in C and nutrients cycling has not been sufficiently estimated. We hypothesized that the rhizosphere effect is significantly different in top and deep soil horizons. C turnover rate and nutrients stoichiometry were determined in rhizosphere and bulk soil sampled from the experimental plots with podzolic soil (Albeluvisol) under spruce trees (Picea abies L.) in the Central Forest Reserve (Tver region, Russia). The rhizosphere factors (Rf) expressed as a ratio of soil characteristics in rhizosphere to that in bulk soil were determined in the top AEL (3 – 15 cm) and deep EL horizons (15 – 46 cm). We subdivided soil characteristics into 2 groups according to Rf value. The group I with 1.05 < Rf < 1.5 includes SOM, acidity, available N, P and exchangeable K. The group II with Rf > 1.5 includes microbial biomass, basal respiration and SOM turnover rate. Increased SOM turnover rate is connected with better SOM quality in rhizosphere soil as evidenced by the increase in polysaccharides portion in rhizosphere SOM revealed by 13C-MAS-NMR spectroscopy. C:N:P stoichiometry in microbial biomass might also affect the SOM susceptibility to decomposition; C:N ratio in rhizosphere microbial biomass was higher than that in bulk soil, while C:P displayed the opposite tendency. The most drastic differences between top and deep rhizosphere were found in basal respiration and SOM turnover rates. Rf for SOM turnover in top soil was about 1.5, while in the deep soil horizon it was as high as 6. Thus, deep rhizosphere was found to be the more pronounced hot spot of biological activity than top one. 61 Rhizosphere and Climate Change Monday 22 June – Poster session Metabolomics 100 Genomic, metabolomic and functional characterization of beneficial Burkholderia species from natural disease suppressive soils Victor J Carrion*1, Desalegn W. Etalo2, Viviane Cordovez2, Kazuki Fujiwara3, Irene de Bruijn2, Victor de Jager2, Jos M. Raaijmakers2 1 Netherlands Institute of Ecology (NIOO-KNAW), Netherlands, 2Netherlands Institute of Ecology (NIOO-KNAW), Netherlands, 3National Agriculture and food Research Organization, Japan Disease-suppressive soils are ecosystems in which crop plants suffer less from specific diseases than expected owing to the activities of antagonistic rhizosphere microorganisms. For most disease-suppressive soils, however, the beneficial microbes and underlying mechanisms involved in pathogen control are largely unknown. In previous studies, we identified key bacterial taxa involved in suppression of the fungal root pathogen Rhizoctonia solani by PhyloChip-based metagenomics of the rhizosphere microbiome of sugar beet seedlings. Members of the Proteobacteria, Firmicutes, and Actinobacteria were found to be consistently associated with disease suppression. Here we focus on the β-Proteobacteria, specifically on the Burkholderia genus. We isolated approximately 50 Burkholderia strains from the rhizosphere of sugar beet seedlings grown in a soil suppressive to R. solani. Based on MLST sequencing, these isolates were classified as B. caledonica, B. graminis, B. hospita, B. pyrrocinia and B. terricola. These five Burkholderia species showed different activities. B. graminis inhibited R. solani via volatile organic compounds (VOCs), whereas B. caledonica, hospita, pyrrocinia and terricola showed antifungal, antioomycete activity and antibacterial activity. Strains of all five species exhibited in planta activity against R. solani. Comparative VOCs profiling of all five species revealed that sulphur-containing compounds were unique for B. graminis. By comparative genomics of the five fully sequenced species/strains, three unique gene clusters were identified in the B. graminis genome that are most likely involved in the biosynthesis of these sulphur-containing VOCs. Comparative genomics also revealed the presence of four non-ribosomal peptide synthetase gene clusters in B. caledonica, hospita, pyrrocinia and terricola that may contribute to the observed broad-spectrum antimicrobial activities. Mutagenesis, cloning, heterologous expression and chemical analyses are ongoing to resolve the functions of these novel gene clusters identified in the genomes of the five Burkholderia species from disease suppressive soil. 62 Metabolomics Monday 22 June – Poster session 101 Mining rhizobacteria-induced metabolome reprograming in plants Desalegn Etalo*1, Judith van de Mortel2, Je Seung Jeon1, Ric de Vos3, Henk Gude4, Thierry Janssen5, Jos Raaijmakers1 1 Department of Microbial Ecology, Netherlands Institute of Ecology, NIOOKNAW, Netherlands, 2HAS University of Applied Sciences, Netherlands, 3Plant Research International, Wageningen University and Research Centre, Netherlands, 4Flower Bulbs, Applied Plant Research, Wageningen University & Research Centre, Netherlands, 5MicroLifeSoilutions, Netherlands The colonization of plant roots, stem, leaves or fruits by beneficial microorganisms can protect plants from diseases, promote growth and enhance yield/biomass. Application of the beneficial rhizobacterium Pseudomonas fluorescens strain SS101 to roots of Arabidopsis thaliana resulted in a distinct shift in the plant transcriptome as compared to non-treated plants, with approximately 1150 plant genes differentially regulated. Gene set enrichment analysis (GSEA) revealed that genes involved in sulfur metabolism were among the most overrepresented gene clusters that were up-regulated in plants treated with the beneficial rhizobacterium. Furthermore, the upregulated gene category included also genes involved in sugar metabolism and auxin biosynthesis. In line with the transcriptome data, the metabolome analyses showed that sulfur containing metabolites were among the metabolites that accumulated in Arabidopsis plants treated with the rhizobacterium. Based on the observed transcriptome and metabolome changes associated to sulfur metabolism in Arabidopsis, experiments are ongoing to boost the production of specific compounds, including sulforaphane, in other cruciferous plants like broccoli. Furthermore, root, bulb and sprout samples obtained from two daffodil cultivars (Carlton, Geranium) treated with rhizobacterial strain SS101 were subjected to untargeted metabolomics. Our analyses of these bulb crops indicated that specific groups of metabolites like spermidine-conjugates were induced only in the sprout of both cultivars that were treated with the rhizobacteria. Collectively, these results show that specific rhizobacteria can (re)program the plant metabolome. 103 Discriminating disease suppressive soils for cereal pathogen Rhizoctonia solani AG-8 using metabolomics Helen Hayden*, Simone Rochfort, Vilnis Ezernieks, Pauline Mele Department of Environment & Primary Industries, Australia Metabolomic analyses were used to investigate possible functional mechanisms for disease suppression in soils known to be suppressive to the fungus Rhizoctonia solani AG-8, which infects cereal crops. Disease suppression refers to a lack of disease manifestation even in the presence of the pathogen, host plant and favourable environmental conditions. Currently the only way to identify fields with disease suppression of R. solani AG-8 is by doing glasshousebased pot trials. Soil samples were collected from two adjacent fields, one known to have a high disease suppression and the other low disease suppression resulting in infected cereal crops. Samples were collected at different times throughout the cropping cycle over two years and analysed for their metabolite profiles. Differentiation of the high and low suppression fields was carried out using multivariate analyses of liquid chromatography mass spectrometry (LC-MS) data, acquired in both the positive and negative ionisation modes, and 63 Metabolomics Monday 22 June – Poster session nuclear magnetic resonance (NMR) data. Several peaks were significantly more abundant in the high suppression soil for the positive and negative ionisation modes. Potential LC-MS biomarkers for the high suppression soil were identified and corroborated by analysing two years of soil samples. The structure of these LC-MS biomarkers was elucidated using accurate mass data and MS fragmentation spectrum information. Analyses of the NMR loadings plots identified soils with high suppression have a greater abundance of polyols and terpenes. Two dimension NMR identified sugar biomarkers in the high suppression soils. Metabolite biomarkers with high abundance in disease suppressive soils were shown to match standards of macrocarpals, which are phloroglucinol containing compounds. Suppression of Rhizoctonia solani AG-8 may be occurring through an antibiotic mode of action as shown for soils suppressive to take all disease of cereals (Gaeumannomyces graminis var. tritici) or through microbial effects upon the plant-pathogen interaction. 104 Effects of microbial signaling molecules on the growth and secondary metabolism of Arabidopsis thaliana Katharina Sklorz*, Michael Bonkowski University of Cologne, Zoological Institute, Germany All plants are simultaneously colonized by a multitude of microorganisms of very different taxonomic affiliation. It is still uncertain how plants orchestrate the complex interaction with the surrounding microbiome, but it is clear that specific signaling molecules must exist that mediate the communication between plant species and their root microbes. We investigated the role of bacterial autoinducers (N-Acyl-Homoserine Lactones, AHL´s), which are employed in quorum sensing systems in various gram negative bacteria, on root growth and the secondary metabolism of Arabidopsis thaliana. We will discuss the observed patterns in the light of plant defense and interkingdom communication. 106 Presence of a growth-promoting endophyte affects the composition of plant secondary metabolites and root exudates in Arabidopsis thaliana Katja Witzel*1, Nadine Strehmel2, Susanne Baldermann1, Susanne Neugart1, Dierk Scheel2, Monika Schreiner1, Rita Grosch1, Silke Ruppel1 1 Leibniz Institute of Vegetable and Ornamental Crops, Germany, 2Leibniz Institute of Plant Biochemistry, Germany We have isolated the strain Kosakonia radicincitans (DSM 16656) from the phyllosphere of winter wheat under temperate conditions. Growth promotion of root and shoot, along with increased yield, was conferred by inoculation of different crop and model plant species. Endophytic plant growth promoting bacteria are discussed to impact significantly plant physiology, but regulatory pathways and biochemical alterations are still unclear. Therefore, transcriptome and metabolome investigations were conducted under controlled conditions to gain a better understanding. A global transcriptome analysis of Arabidopsis thaliana plants inoculation with Kosakonia radicincitans identified an over-representation of genes involved in secondary plant metabolism. Profiling the glucosinolate, carotenoid and phenylpropanoid 64 Metabolomics Monday 22 June – Poster session content of leaves and roots revealed a specific response to endophytic colonization. As some secondary plant metabolites are known to act as signaling molecules in the rhizosphere, such as flavonoids and coumarins, root exudates of control and inoculated plants were collected and analyzed. More than 50 primary and secondary metabolism compounds were differentially enriched when plants were colonized by K. radicincitans. The results and possible implications of these analyses on plant-endophyte interactions are discussed. 65 Metabolomics Monday 22 June – Poster session Root Endophytes 107 Consortia of ACC deaminase-producing bacteria (both endophytic and rhizospheric) isolated from avocado plants mitigate salt stress of wheat plants Patricio Barra*1, Nitza Inostroza1, Maria de la Luz Mora1, David Crowley2, Milko Jorquera1 1 Universidad de La Frontera, Chile, 2University of California Riverside, USA Plants growing under salinity stress conditions increase ethylene production, which triggers inhibition of root elongation. Bacterial enzyme 1-aminocyclopropane-1-carboxylate deaminase (ACCD) cleave the ethylene precursor, 1-aminocyclopropane-1-carboxylate, thus decreasing ethylene levels and consequently their detrimental effects. Some bacterial strains also produce phytohormone indol acetic acid (IAA), which increases cell elongation. Therefore, we hypothesize that ACCD- and IAA-producing bacteria (both endophytic and rhizospheric bacteria) isolated from avocado plants are able to mitigate salt stress effects of wheat plants; and we also hypothesize than bacterial location (both endophytic and rhizospheric), as well as, the levels of ACCD and IAA production by bacteria have different effects on salt stress response of wheat plants. Twelve bacterial strains were isolated from avocado plants, and four bacterial consortia were formulated, each composed of three strains, as follows: 1) Endophytic bacteria with higher ACCD and IAA production; 2) Endophytic bacteria with lower ACCD and IAA production and 3) Rhizobacteria with higher ACCD and IAA production; 4) Rhizobacteria with lower ACCD and IAA production. Wheat seeds were inoculated with the bacterial consortia, and then seeds were grown under salt stress conditions. Length, dry weight and superoxide dismutase (SOD) activity of wheat shoot and roots were determined. The results showed that at lower levels of bacterial IAA and ACCD production, the endophytic bacteria were more efficient than rhizobacteria consortia mitigating salt stress effects. Between rhizobacteria consortia, only those with higher production were able to promote the growth of stressed plants. Both endophytic bacteria and rhizobacteria consortia with higher production were able to increase SOD activity. Finally, bacterial strains isolated from avocado plants mitigate plant stress and therefore have the potential to be used as commercial inoculum of avocado plants. 108 Stenotrophomonas rhizophila SPA P69: deep insights into an endophytic stress protecting agent Gabriele Berg*, Henry Müller Graz University of Technology, Austria Stenotrophomonas rhizophila is able to promote plant growth of many crops and to protect roots against biotic and a-biotic stresses. We studied mechanisms associated with osmotic stress using transcriptomic and microscopic approaches. In response to salt or root extracts, the transcriptome of S. rhizophila SPA P69 (syn. DSM14405T) changed drastically. We found a notably similar response for several functional gene groups responsible for general stress 66 Root Endophytes Monday 22 June – Poster session protection, energy production, and cell motility. However, unique changes in the transcriptome were also observed: the negative regulation of flagella-coding genes together with the up-regulation of the genes responsible for biofilm formation and alginate biosynthesis were identified as a single mechanism of S. rhizophila against salt shock. However, production and excretion of glucosylglycerol (GG) were found as a remarkable mechanism for the stress protection of this strain. For S. rhizophila treated with root exudates, the shift from the planktonic lifestyle to a sessile one was measured as expressed in the down-regulation of flagellar-driven motility. These findings fit well with the observed positive regulation of host colonization genes and microscopic images that show different colonization patterns of oilseed rape roots. Spermidine, described as a plant growth regulator, was also newly identified as a protector against stress. In addition to both the changes in life style and energy metabolism, phytohormons, and osmoprotectants were also found to play a key role in stress protection. Risk assessment studies reveal no health risks. This is mainly because SPA P69 is unable to growth at the human body temperature, 37°C due to the absence of heat shock genes and a temperature-regulated suicide mechanism. Taken together SPA P69 is a promising endophytic stress protecting agent (SPA) ready for commercial applications. 109 Better efficiency of biofertilizers in combination with N2-fixing endophytes in low quality sandy soil Borbala Biro*, Zita Szalai, Tamás Kocsis, Zsolt Kotroczó Corvinus University of Budapest, Hungary Biofertilizer and/or soilconditioner products are containing very often the various types of the Nitrogen-fixing and P-mobilising microorganisms, as only one or 2-3 types of species components. Question arises if single or combined products are more efficient and what is the main driving force for the efficiency in a low quality sandy soil? Tomato Solanum lycopersicon Mill. ’Mobil’ was used in a field experiment, inoculated with Trichoderma harzianum T-22 and TDM, Hungarian Trichoderma inoculums, including also of Azotobacter and Azospirillum N2-fixers in one product. There were patentkali and calcinit fertilizers applied at 1200 kg/ha doses also to the soil. Inoculation treatment was performed twice during the vegetation, at the sowing and at the time of the plantation of tomato seedlings. Growth of tomato, shoot and root biomass was assessed and general soil characterization, including physical-chemical parameters, the MPN counts of some beneficial physiological groups in tomato rhizosphere, outside and inside the plants. Results were evaluated by statistical probes. The beneficial effect was realised with the combined tomato inoculation in comparison with the only Trichoderma strain application. Effect of Trichoderma TDM inoculation was improved with free-living and associative Nitrogen-fixers and thus provided better nutrition. The relatively high P-content (430 mg/kg) was found in the slightly humous sandy soil, where the N2-fixing microbes might able to improve the N,P, K ratio for the more efficient tomato growth. Plant microbe interaction is largely dependent on the soil nutrient-status and the balance among main nutritive elements. In this process the multifunctional inoculums might providing greater plant-growth promotion beyond the single strain effects. 67 Root Endophytes Monday 22 June – Poster session 110 A survey of bacterial root endophytes associated with vegetation at a bitumen impacted site Natalie Blain*1, Bobbi Helgason2, James Germida1 1 University of Saskatchewan, Canada, 2Agriculture and Agri-Food Canada, Canada Bacterial root endophytes can help alleviate plant stress and promote plant growth, and the potential use of these endophytes to assist land reclamation is receiving increased interest. Bitumen is a heavy oil that influences plant growth due to its hydrocarbon composition. The Bitumount provincial historical site is considered the location of two of the world’s first oil sands (bitumen) extraction plants. Operations at Bitumount began in 1923 and ceased completely in 1958. Through natural reclamation, vegetation has re-colonized the area including hardened bitumen. The Bitumount site offers a unique opportunity to study plantmicrobe associations that have co-adapted to this stressed environment. Sampling locations were established in June 2014, and plant and soil samples collected. A total of 6 different plant species were identified and sampled based on their abundance at each sampling location. They included: Bromus inermis, Equisetum spp., Agropyron trachycaulum, Poa pratensis, Fabaceae spp. and Fragaria virginiana. Soil samples were analyzed for hydrocarbon content, which ranged from 330 to 24,700 mg kg-1 throughout the site. Culture dependent and independent methods were used to characterize bacterial root endophytes. The number of culturable root endophytes varied significantly (p<0.05) throughout the site and between plant species, with values ranging from 4.3 to 6.3 log CFU g-1 of fresh roots. Phylogenetic identification of isolated endophytes and Illumina MiSeq bacterial profiling revealed diverse communities associated with different plant types. The function of these endophytes is currently under investigation. 111 Desert root endophytes enhance the growth of Arabidopsis thaliana under salt conditions Ameerah Bokhari*, Juan S. Ramirez, Axel de Zelicourt, Feras F. Lafi, Maged Saad, Intikhab Alam, Vladimir Bajic, Heribert Hirt Center for Desert Agriculture, King Abdullah University of Science and Technology (KAUST), Saudi Arabia Understanding the communication and regulation between host plants and endophytic plant growth promoting bacteria might lead to improved agricultural outputs. Here, our aim is to study root endophytes from desert pioneer plants that ensures plant growth in different extreme conditions. As part of the DARWIN21 project (http://www.darwin21.net) we isolated endophytic bacteria from desert plants in Saudi Arabia, Jordan and Pakistan. We used cultivation-based techniques followed by molecular characterization via sequencing 16S rRNA genes. Our in-house screening assay was developed to select strains for inducing plant growth promotion and/or abiotic stress tolerance on the model plant Arabidopsis thaliana. To date we have isolated 984 bacteria which belong to four phyla, Proteobacteria, Firmicutes, Actinobacteria and Bacteroidetes. Our screening assay revealed several bacterial strains which promote the growth of Arabidopsis thaliana under salt stress conditions. So far, we have sequenced the genomes of nine of these endophytes and we are currently sequencing several more distinct bacterial strains that display the latter properties. Our comparative genomic, transcriptomic and proteomic approach should help to unravel the mechanisms of 68 Root Endophytes Monday 22 June – Poster session conferring salt stress tolerance to plants. As a future goal we aim to create a catalog of microbial strains that can help to improve crop resistance to different biotic and abiotic stresses. 112 Bacterial diversity of native Acacia sp. nodular endophytes in Tamanrasset Zineb Faiza Boukhatem*1, Chahinez Merabet Merabet1, Abdelkader Bekki Bekki1, Sonia Sekkour Sekkour1, Odile Domergue Domergue2, Antoine Galiana Galiana3 1 Oran 1 Ahmed Ben Bella University, Algeria, 2INRA, UMR LSTM, France, 3CIRAD, UMR LSTM, France Tamanrasset located in the heart of the Sahara desert in southeastern Algeria, is the only region where are localized five indigenous species of Acacia: A. ehrenbergiana Hayne, A. nilotica (L.) Delile, A. seyal Delile, A. tortilis (Forssk.) Hayne, and A. laeta Delile, these rustic leguminous trees are surviving under extremely harsh conditions of drought in dry oueds. Some preliminary observations led us to investigate the diversity of nodular endophytes which is poorly documented especially those associated with Acacia species. Among 79 isolates purified from nodules of the five Acacia sp., 24 representative strains were characterized and identified as belonging to nine bacterial genera, namely: Paenibacillus, Ochrobactrum, Stenotrophomonas, Pseudomonas, Microbacterium, Rhizobium, Agrobacterium, Brevibacillus,and Advenella. The isolates of these nodular endophytes revealed a strong tolerance profile to salinity and high temperatures accordingly to their natural ability to survive to harsh edaphoclimatic conditions. PCA confirmed that no correlation existed between bacterial tolerance to maximum growth temperature and depth of sampling. On the other hand, there was no relationship either between in vitro tolerances of rhizobial strains to NaCl concentration and high temperature and the corresponding edaphoclimatic characteristics of the sampling sites. 113 Bacterial root endophytes associated with agricultural crops in Saskatchewan, Canada Jorge Cordero Elvia*1, Renato de Freitas2, James Germida2 1 University of Saskatchewan, Canada, 2University of Saskatchewan / Department of Soil Science, Canada Endophytes influence growth and productivity of crops in agricultural ecosystems, and are an important component of the plant-microbiome. Because of the potential benefits of endophytes, it is important to investigate their diversity and function in different plant species and how edaphic factors influence this relationship. This study investigated the diversity of bacterial root endophytes associated with wheat (Triticum aestivum), canola (Brassica napus), lentil (Lens culinaris) and field pea (Pisum sativum) grown in three Chernozem (Orthic Brown Chernozem Calcic Kastanozem, Orthic Dark Brown Calcic Chernozem and Orthic Black Calcic Chernozem) soils in Saskatchewan, Canada. These soils differed mainly in organic matter content, pH and texture. Culturable bacterial root endophytes (n=298) were isolated using non-selective culture media and identified by Sanger sequencing. In addition, endophyte community profiles were analysed using culture 69 Root Endophytes Monday 22 June – Poster session independent denaturing gradient gel electrophoresis (DGGE). Culture dependent and independent techniques indicated that some endophyte communities varied depending on crop and soil types. Analysis of culturable bacteria by crops revealed that the predominant genus in canola was Stenotrophomonas, whereas Bacillus and Rhizobium were dominant in pea. The most common genera in lentil were Bacillus and Pantoea, whereas Xanthomonas was predominant in wheat. Culture independent DGGE analyses revealed that band patterns were distinct for each crop, with wheat yielding the largest number of endophyte bands. Similarly, soil type influenced endophytic community structure as demonstrated by higher number of culturable endophytes and DGGE bands in the Orthic Brown Chernozem Calcic Kastanozem, followed by Orthic Dark Brown Calcic Chernozem and Orthic Black Calcic Chernozem. Our results suggest that crops select specific bacterial endophyte species, but soil properties also influence their community structure. 114 Endophytic protists in plant roots Michael Bonkowski, Kenneth Dumack* University of Cologne, Germany Protist root pathogens such as Plasmodiophora brassicae the agent of club root disease in Brassicaceae, or Spongospora subterranea the agent of powdery scab in potatoes are well known to phytopathologists. However, scarce knowledge exists on the identity of protists that colonize roots without causing visible disease symptoms. We conducted a systematic screening for root-colonizing amoeboid protists in soil and isolated a new species of the Viridiraptoridae (Glissomonadida) from an agricultural field in Göttingen, Germany. Viridiraptoridae have been recently described as limnic protist predators of filamentous algae, but environmental sequencing studies suggested that they were also common and highly diverse in terrestrial habitats. To investigate the feeding preferences of our isolate, we offered bacteria, fungi, algae, nematodes, and sterile plant seedlings as a possible food sources. The protists multiplied exclusively, but strongly, in roots of various plants like Arabidopsis thaliana and Lolium perenne, and to a lesser extend in dead nematodes. To overcome the plant defense system endophytic protozoa must possess highly adapted signalling mechanisms. Deciphering these interactions may strongly improve our understanding of interactions of plant roots with protozoan pathogens. 70 Root Endophytes Monday 22 June – Poster session 115 Potato endophytic compartments - a hidden reservoir for promising biocontrol agents against Ralstonia solanacearum Tarek Elsayed*1, Stéphane Compant2, Rita Grosch3, Kornelia Smalla1 1 Julius Kühn-Institut – Federal Research Centre for Cultivated Plants (JKI), Institute for Epidemiology and Pathogen Diagnostics, Germany, 2Austrian Institute of Technology, Austria, 3Leibniz Institute of Vegetable and Ornamental Crops, Department of plant Health, Germany Ralstonia solanacearum (Rs, biovar2, race3) is an epidemic soil-borne phytopathogen. Endophytic antagonists might be a solution to control this pathogen. In this study, we assessed the diversity and abundance of naturally occurring bacterial antagonists of Rs, in nine potato microenvironments across three soil types. A total of 2016 isolates was screened, and 215 out of 480 isolates with in vitro antagonistic activity were further characterized. BOXPCR showed similar genotypes in different microenvironments and soils. Endophytic compartments had the highest proportion of antagonists, and most of these antagonists were affiliated to Pseudomonas. Potential modes of action were in vitro tested and phosphate solubilization and siderophore production were dominant traits. Rhizocompetence of 11 green fluorescent protein (gfp) tagged antagonists, representing all microenvironments, was tested in a greenhouse experiment with tomato. Plate counts revealed good rhizosphere competence. A selection of seven antagonists was tested for biocontrol activity against Rs (B3B) in a new greenhouse experiment. Five different consortia, each consisting of three antagonists were prepared (P. chlororaphis, P. brassicacearum in addition to one variable isolate). Based on the wilting symptoms, P. koreensis and P. helmanticensis showed a good biocontrol activity. Confocal laser scanning microscopy (CLSM) confirmed the colonization of root tips. Plate counts and qPCR showed good rhizosphere competence and a reduction in Rs population on the rhizosphere. Denaturing gradient gel electrophoresis (DGGE) of 16S rRNA gene fragments amplified from TC-DNA showed pronounced shifts in bacterial communities. UPGMA analysis revealed distinct clusters based on the inoculated isolates and the severity of infection. P. koreensis and P. helmanticensis showed a good colonization and biological control activity and, in addition, plant growth promotion, suggesting the future use of these isolates as biocontrol agents against bacterial wilt in regions where Rs is endemic. 116 Effect of Azospirillum brasilense strains on Lotus japonicus root growth and development Estrella Elvira-Ramirez*1, Simona Radutoiu2, Dorthe B. Jensen3 1 Aarhus university, Denmark, 2Aarhus Univeristy, Denmark, 3Aarhus University, Denmark Azospirillum brasilense, a nitrogen-fixing bacteria, is broadly present in soils around the world. Strains of this bacterium can live isolated in the soil or in the rizosphere of a wide plant families. The beneficial effects of Azospirillum on some crops, such as maize or wheat are well documented and have been mainly associated with the increase of root system and therefore, the enlarged area available for mineral uptake. The beneficial effect of Azospirillum on crops proved to be, however from time to time inconsistent, and environment dependent. On the other hand, the molecular mechanisms behind the beneficial or less favourable interactions with the different hosts are largely unknown. 71 Root Endophytes Monday 22 June – Poster session We set to better understand the molecular basis of the interaction between Azospirillum and the model legume host, Lotus japonicus. Specific growth conditions in which Azospirillum has a detrimental effect on the legume host have been identified. We are currently using such defined conditions to better understand the molecular mechanism involved in the different responses mounted by the two interacting partner leading to contrasting responses in the host. We will present here our progress on this study based on Lotus mutants analyses and host gene expression investigations. 117 Characterization of free-living and endophytic diazotrophs from rice plants grown on different type of Uruguayan soils Lucia Ferrando*1, Andrea Martinez2, Ana Fernández Scavino3 1 Universidad de la República- Uruguay/ School of Chemistry, Uruguay, 2School of Chemistry- Universidad de la Republilca, Uruguay, 3School of Chemistry- Universidad de la República, Uruguay The soils used for agriculture suffer important changes over the course of the successive cycles of crops, affecting fertility and soil biodiversity. The great contribution of endophytic bacteria to plant growth promotion is well known, in particular, for diazotrophs. Endophytes mainly come from the soil, so soils with different characteristics could determine or affect microbial communities established in plant tissues. The abundance, diversity, and composition of diazotrophic communities from seven Uruguayan soils and endophytic communities established in roots from rice plants grown in them under controlled conditions were addressed using the nifH as marker gene. The abundance of nifH gene in leaves was similar among those from different type of soil and a hundred-fold lower than in rice roots or soils. Rice leaves seem to support a numerically stable and independent population poorly affected by soil type. On the other hand, the nifH copy number present in soils was similar to the retrieved from roots. The most fertile soil presented significant higher nifH abundance. Diazotrophic communities present in the soils were significantly more diverse than those established inside rice roots, according to the Diversity Indexes obtained. However, Clustering analysis from data obtained by T-RFLP of nifH gene revealed that diazotrophic communities from different soils presented a greater similarity among them (45%) than root endophytic communities (20%). Pyrosequencing of nifH gene is being performed for a better understanding of these communities. Diazotrophic populations inhabiting rice roots would be similarly abundant than the ones from soils. However, diazotrophic communities from soil were more diverse and homogeneous among the different soils than the communities established in rice roots. These results suggest that rice plant could be playing an important role in the selection of a restricted group of diazotrophic species which effectively colonize the root achieving to establish themselves as endophytes. 72 Root Endophytes Monday 22 June – Poster session 118 Microbispora spp. are the dominant actinobacterial endophytes of Australian rice plants Christopher Franco*1, Fitri Widiantini2 1 Flinders University, Australia, 2Flinders University, Australia Microbispora spp. were detected as the predominant endophytic actinobacterial population making up 85% of the two hundred and sixty four endophytic actinobacteria isolates obtained from rice plants grown in Yanco, over two consecutives rice growing seasons, whereas they constituted less than 2% of the 287 actinobacteria isolated from the source rice paddy soils. The results were confirmed by culture-independent methods. The Microbispora isolates were categorised into 16 different morphological groups that were morphologically dissimilar to the nearest neighbour type strains based on 16S rRNA gene and gyrB gene sequence similarity. The Microbispora isolates differentiated further based on BOX-PCR fingerprinting. At least 5 strains demonstrated a distinct relationship between each other as well as with the type strains. Further polyphasic studies were able to characterise their novelty. Further confirmation of these unexpected findings was achieved by growing the original and 3 other types of rice cultivars (Langi, Doongara, Amaroo and Nipponbare) in 4 different soils which included the original rice paddy soils. Approximately 94% of the total isolates were Microbispora-like isolates. Challenge experiments were carried out with the rice endophytes Streptomyces sp. R32 and Saccharothrix sp. OS6 added as seeding coatings on rice seeds prior to sowing, or adding these cultures as seed drenches onto rice seed sown in rice paddy soils. In all these experiments Microbispora spp. constituted more than 60% of all endophytes isolated from these plants at 6 weeks after sowing. These results revealed that the rice plants growing in Australia preferentially harbour a high degree of diverse Microbispora. 119 Endophytic bacterial communities in different tissues of mountain sorrel (Oxyria digyna) Cindy Given*, Riitta Nissinen University of Jyväskylä, Finland Endophytic bacteria live inside the plants without harming them, and can provide benefits for their hosts. Endophytes have been studied especially in the crop plants, but very little is known about endophytes from extreme environments like the Arctic. Mountain sorrel (Oxyria digyna) is used in this study as a model plant. It is a non-mycorrhizal, arcto-alpine species, that can grow in very low nutrient soils and is a typical pioneer plant species. In this study, we want to understand the acquisition and functioning of endophytic community in different tissues of O. digyna. 73 Root Endophytes Monday 22 June – Poster session In 2013, 80 endophyte-free micropropagated O. digyna plantlets were planted in the field site on fell Jehkas, in Kilpisjärvi (69°’N; 20°50’E), northwestern Finland. Half of the plants were harvested after 30 days (summer) and the rest were left to over-winter, and harvested in spring 2014. Endophytic bacterial communities in leaves and roots were analyzed by community amplicon sequencing. Additionally, the culturable endophytic bacteria were isolated from the summer samples. The isolates were identified by 16s rRNA gene sequencing and cross-compared with the molecular data. Most of our isolates were closely related to bacteria isolated from other cold environments. Several bacterial species showed a tight association with the host, as they had been previously isolated from seeds and vegetative tissues of O. digyna. The preliminary results showed that Pseudomonas sp. and Microbacterium sp. were the dominant genera in both leaves and roots. Some genera, however, were tissue-specific: Sphingomonas spp. were found only from leaves and Flavobacterium spp. were isolated only from roots. Strains representing the dominant endophyte groups in different tissues were phenotypically profiled in order to detect putative tissue specific and plant beneficial traits. 120 Is the root colonizing endophyte Acremonium strictum an ericoid mycorrhizal fungus? Gisela Grunewaldt-Stöcker*, Henning von Alten Leibniz University of Hannover/ Institute of Horticultural Production Systems/ Section Phytomedicine, Germany Acremonium strictum was associated with roots of several ericaceous plants and thus regarded as a potential mycorrhizal fungus. However, its mycorrhizal traits were not proven yet. A. strictum is an endophyte without visible effects on the host plant, non-systemic and restricted to the root system, with multifunctional traits, and thus can be classified as type II. Entomologists revealed its antagonistic feature to affect herbivorous insects in feeding behaviour and development by altering phytosterols in the host plants. In our previous investigations on flax and tomato it could induce disease resistance towards Fusarium-wilts. Moreover, in our microscopic studies on host-fungus interactions with wild-type and gfptransformants we found hyphal structures of A. strictum in the rhizodermal cells of greenhouse cultured flax plants and later in hair roots of inoculated Rhododendron plantlets in sterile liquid culture with striking similarity to hyphal coils and colonizing mycelia of ericoid mycorrhizal fungi. Therefore the hypothesis of the mycorrhizal nature of A. strictum was revived. Structures bearing resemblance to ericoid mycorrhiza at a low colonization level were shown by confocal laser scanning microscopy of fuchsine acid-stained hair roots of inoculated Rhododendron spec.. Real progress to verify or reject our hypothesis was achieved by using the fluorochrome dye FUN-1® in unfixed tissue to observe the vitality status of the host cells in colonized hair roots. In inoculation trials with in vitro raised mycorrhiza-free Rhododendron plants - tested in sterile liquid and in greenhouse pot culture - A. strictum was never observed in living epidermal or cortex cells. As compared to the mycorrhizal fungi 74 Root Endophytes Monday 22 June – Poster session Oidiodendron maius and Rhizoscyphus ericae that invaded metabolic active host cells, A. strictum was found with elongated or coiled hyphae only in disordered or visibly dead cells and in the apoplast. This corroborates its non-mycorrhizal endophytic attribute. 121 Density and diversity of endophytic bacteria associated with Salicornia europaea L. and Aster tripolium L. in saline soils Katarzyna Hrynkiewicz*1, Silke Ruppel2 1 Nicolaus Copernicus University, Poland, 2Leibniz-Institute of Vegetable- and Ornamental Crops Grossbeeren/Erfurt e.V., Germany Worldwide salt affected soils cover 800 million hectares (6% of the land in total) with still increasing areas, expanding the scale of this phenomenon. Plants evolved a wide range of adaptation mechanisms, which allow them to grow and develop in environments of high soil salinity. We expect a halophyte specific selection of the plant microbiome in respect to the biological atmospheric nitrogen fixing community. To elucidate the question if halophytic plants retrain specific diazotrophic bacterial communities we evaluated S. europaea L. (herbaceae) and Aster tripolium L. growing at two saline sites in central Poland: a salty meadow in the vicinity of a soda factory (anthropogenic salinity) and an area affected with natural brine (landscape park). Leaves, stems and roots were investigated separately. Total bacterial 16S rDNA- and nifHgene copy numbers as well as phylogenetic bacterial strain comparison showed both a plant species specificity and soil salinity impact on the plant microbiome composition. Higher bacterial density and diversity was detectable at test sites with higher salinity and generally numbers of diazotrophic bacteria increased in the following order: leaves > shoots ³ roots. The ratio of nifH-gene to 16S rDNA-gene was higher at naturally saline test site (0.6) compare to the test site with anthropogenic salinity (0.4). Most of isolated diazotrophs were identified to the phylum Proteobacteria and Actinobacteria. 122 Reaction of perennial ryegrass (Lolium perenne L.)/Glomus spp. symbiotic association on infection by Fusarium poae and Rhizoctonia solani Malgorzata Jeske*, Dariusz Panka, Marcin Juda, Karol Lisiecki, Katarzyna Koczwara, Monika Młynska UTP University of Science and Technology, Poland Perennial ryegrass is one of the most important grasses in Poland, used for pasture and for turf. It is often attacked by numerous pathogens, which can decrease yield and its quality. Use of fungicides for plants protection against diseases is often limited. The other way of protection of perennial ryegrass can be use of arbuscular mycorrhizal fungi (AMF) of the Glomus genus. These fungi belong to the most commonly occurring soil microorganisms of the world and are associated with at last 80% of plants of the Earth. Arbuscular mycorrhizal fungi increase the root absorptive area and hence the plant nutrition. Additionally, AMF increase the tolerance of plants to heavy metals, water stresses, as well as pathogenic 75 Root Endophytes Monday 22 June – Poster session fungi. The exact mechanism of higher resistance of AMF infected plant is yet not fully understood, but it can be assumed that PR (Pathogenesis-related) proteins have a significant role. Detailed experiments in controlled conditions were conducted to determine the impact of AMF on induction of specific defense mechanism, including production of PR proteins: chitinases and β-1,3-glucanases in perennial ryegrass. Severity of plant’s infection was estimated 2, 4, 6, and 8 days after inoculation with F. poae and R. solani. The level of chitinases and glucanases in plants was determined 0, 2, 4, 6 and 8 days after infection with use of Abeles et al. (1970) method with modifications. The specific activity of chitinases and glucanases in the extracts was expressed as moles of reducing sugars released in one minute of incubation per one milligram of total protein in the extract. Occurrence of Glomus spp. affected the amount of chitinases and β-1,3-glucanases in perennial ryegrass. The amount of enzymes was also dependent on the time after infection. Results indicate the role of PR proteins in resistance of plants exposed to infection. 123 Genetic analysis of root endophytic Pseudomonas putida bp25 and chemoprofiling of its antimicrobial volatile organic compounds Aundy Kumar*1, Neelam Sheoran1, Agisha Valiya Nadakkakath2, Vibhuti Munjal1, Aditi Kundu3, Kesavan Subaharan4, Vibina Venugopal4, Suseelabhai Rajamma2, Santhosh Eapen2 1 Division of Plant Pathology, ICAR-Indian Agricultural Research Institute, India, 2Division of Crop Protection, ICAR-Indian Institute of Spices Research, India, 3Division of Agricultural Chemicals, ICAR-Indian Agricultural Research Institute, India, 4Division of Crop Protection, ICAR-Central Plantation Crops Research Institute, India Black pepper (Piper nigrum) associated bacterium BP25 was isolated from root endosphere of apparently healthy cultivar, Panniyur-5 that protected black pepper plants against Phytophthora capsici and Radopholus similis -the major production constraints. The bacterium was genetically characterized and mechanisms of its antagonistic action against major plant pathogens elucidated. The polyphasic phenotypic and genotypic analysis revealed its identity as a strain of Pseudomonas putida (PpBP25). Multi Locus Sequence Typing revealed that the bacterium shared gene sequences with several other isolates representing diverse habitats. Tissue localization assays exploiting green fluorescence protein expression clearly indicated that PpBP25::gfp endophytically colonized not only its host -black pepper, but also other plants such as edible ginger (Zingiber officinale) and a model plant, Arabidopsis thaliana. Coupled with PpBP25 colonies enumerated from internal plant tissues four weeks post inoculation indicated its stable establishment and persistence in the plant system. Strikingly, the bacterium inhibited broad range of economically significant plant pathogens representing diverse taxonomic groups such as Phytophthora capsici; Pythium myriotylum; Giberella moniliformis; Rhizoctonia solani; Athelia rolfsii; Colletotrichum gloeosporioides; Magnaporthe oryzae; Ralstonia solanacearum; Xanthomonas axonopodis pv. punicae; Xanthomonas oryzae pv. oryzae; and plant parasitic nematode, Radopholus similis by its volatile organic compounds (VOCs). Gas Chromatography/Mass Spectrometry (GC/MS) based chemical profiling revealed presence of Heneicosane; Tetratetracontane; Pyrrolo [1, 2-a] pyrazine-1, 4-dione, hexahydro-3-(2-methylpropyl); Tetracosyl heptafluorobutyrate; 1,-3Eicosene, (E)-; 1-Heneicosanol; Octadecyl trifluoroacetate and 1-Pentadecene in PpBP25 76 Root Endophytes Monday 22 June – Poster session metabolites. Dynamic head space GC/MS analysis of airborne volatiles indicated the presence of many aromatic compounds such as Dimethyl trisulfide; Dimethyl disulphide; Pyrazine, 2,5dimethyl-; Pyrazine, methyl-; Pyrazine, 2-ethyl-5-methyl-; Pyrazine, 2-Ethyl-3,6-dimethyl; Heptamethyl-2-nonene; β-Naphthol; Octadecyl vinyl ether; Tetradecane, 2,6,10-trimethyl; Cyclobutene, 2-propenylidene-; Heptamethyl-1-nonene; 1,8-Nonadien-3-ol; Octadecanal,2bromo; Isoamyl alcohol; and 1-Undecene. The work paved way for profiling and consequent identification of several broad spectrum anti-oomycetes, antifungal, antibacterial and nematicidal VOCs in black pepper root endophytic bacterium for next generation plant disease management. 124 Evaluation of ACC deaminase-producing endophytic bacteria isolated from organic products by mung bean assay Hiroaki Matsuoka*1, Yoshinari Ohwaki1, Junko Terakado-Tonooka2, Fukuyo Tanaka Tanaka1 1 National Agriculture and Food Research Organization Agricultural Research Center, Japan, 2Saga University, Japan Bacterial endophytes have been associated with the growth promotion of various crops. In particular, 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase-producing bacteria have received widespread attention because of its ability to improve the stress tolerance of plants by lowering ethylene level under environmental stress. We supposed that organic crops which are exposed to frequent attacks of pathogen and pest may establish the beneficial interaction with bacteria with such property. In this study, therefore, we isolate and evaluate ACC deaminase-producing bacteria from organic crops in order to find new candidates of biocontrol agents in practical use for cultivated crops. Bacterial strains were isolated from the fruit or root of organic crops including carrot, aroid, turnip, sweet pepper, and apple by using Dworkin–Foster minimal-salt medium with ACC as a sole nitrogen source. The isolated strains were further selected from amplifying the ACC deaminase synthase (acdS) gene by colony PCR. Moreover, the strains possessing acdS genes were examined for enzyme activity by measuring the production of α-ketobutyrate from ACC ,and bioassay with growth-response of mung bean in the presence of ACC under gnotobiotic condition. Twenty-four strains were found to have acdS genes in all the 80 strains isolated on the medium containing ACC. Of those, 10 strains were observed ACC deaminase activity by enzyme assay. Among them, 4 strains were shown to promote the shoot elongation of mung bean in the bioassay. Interestingly, the ACC deaminase activity in the isolated strains showed different trends with growth response of mung bean. Our data indicate that selected bacteria in organic crops produce ACC deaminase and have potential for plant growth promotion. We will also report the results of inoculation of selected 3 strains into cultivated crop. 77 Root Endophytes Monday 22 June – Poster session 125 Influence of plant host species on the root associated microbial community and functional redundancy of the root microbiome under phosphate starvation Tatiana Mucyn*1, N. W. Breakfield2, G. Castrillo1, S. H. Paredes1, S. M. Yourstone1, C. Hunter1, J. L. Dangle1 1 University of North Carolina, Chapel Hill, NC, USA, 2NewLeaf Symbiotics, St Louis, MO, USA Plants harbor a specific and complex microbiota at the surface of the root (rhizosphere) and within the root (endophytic compartment) which influences plant health and productivity. Various factors such as soil type, abiotic/biotic stress, host developmental stage, and host species shape the root microbiome. The extent to which the host selects its root microbial community remains poorly understood. To determine the influence of plant host species on the assembling of both the rhizospheric and endophytic microbiome, we are comparing the well-studied bacterial root bacterial microbiome of Arabidopsis thaliana Col-0, with those of two monocot species Brachypodium distachyon Bd21 and Setaria viridis A10-1 from plants grown in the same wild soil, using high-throughput bacterial 16S rDNA, as well as fungal internal transcribed spacer 2 profiling. We are characterizing both core and species-specific root microbiomes and are exploring the hypothesis that plants more closely related to each other share more similar microbial communities. The Dangl laboratory has isolated ~600 bacterial strains from the rhizoplane and endophytic compartments of Arabidopsis thaliana and is evaluating the effect of these isolates on plant health under various nutrient starvation stresses. This bacterial collection is currently being tested on Setaria viridis focusing on the identification of isolates that rescue phosphate starvation stress to establish to whether the core microbiome may also present functional conservation across species. We are in parallel establishing a collection of bacterial strains isolated from the root of Setaria viridis to refine our characterization of both the core and potential species-specific microbiome. 126 ACC deaminase-expressing endophyte increases plant tolerance to FD phytoplasma infection Elisa Gamalero1, Elisa Bona1, Giorgia Novello*1, Cristina Marzachì2, Luciana Galetto2, Flavio Veratti2, Nadia Massa1, Bernard Glick3, Simone Cantamessa1, Giovanni D'Agostino4, Graziella Berta1 1 Università del Piemonte Orientale, DiSIT, Italy, 2CNR, Italy, 3University of Waterloo, Canada, 4Mybasol srl, Italy Flavescence dorée (FD) is an epidemic yellows disease of grapevine caused by a phytoplasma (FDP), living in the phloem for which there is currently no cure. We have previously shown that treatment of plants with rhizobacteria leads to mitigation of phytoplasma-induced disease. The alleviation of a variety of disease symptoms by rhizobacteria may involve the reduction of the stress-related plant ethylene via the 1-aminocyclopropane-1-carboxylate (ACC) deaminase. We tested whether the endophytic bacteria Pseudomonas migulae 8R6, able to synthesize ACC deaminase, can limit the damages induced by FDP. Since FD-infected grapevines show symptoms the year after infection or later, the FDP host plant periwinkle was used as model. The use of a mutant of strain 8R6, lacking the ability to produce ACC deaminase, may 78 Root Endophytes Monday 22 June – Poster session demonstrate the involvement of ethylene in the development of FDP symptoms in the plant. The strain 8R6 colonized the internal tissue of periwinkle, thus confirming its endophytic aptitude and significantly reduced the number of symptomatic plants (53% vs. 93%). A lower number of symptomatic plants was also detected in plants inoculated with the 8R6 mutant compared to controls (73% vs 93%) thus suggesting that ACC deaminase is not the only mechanisms involved in the increased tolerance to FD infection. Quantification of phytoplasma inside the leaves was not affected by both bacterial strains. However, FD phytoplasma title was under the quantification threshold in 38% of plants inoculated with the strain 8R6. This value was more than what recorded for FD infected plants (14%). ACC deaminase activity seems to be involved, as FD phytoplasma population was under the quantification threshold in 8% of the plants inoculated with the mutant. In conclusion, ACC deaminase activity of P. migulae 8R6 might help the plant to regulate the level of the stressrelated hormone ethylene, potentially leading to improved tolerance to phytoplasma infection. 127 The roles of root colonizing bacterial endophytes on suppression of Fusarium oxysporum f. sp. cucumerinum and growth promotion of cucumber plants Hatice Ozaktan*, Ayşe Gul, Bi̇ rsen Çakir, Lalehan Yolageldi, Mustafa Akbaba, Sahika Akat Ege University, Turkey The aim of this study was to find that endophytic bacterial (EB) isolates obtained from healthy cucumber plant tissues could promote the plant growth and marketable fruit yield and suppression of soilborne plant pathogen of cucumber plants caused by Fusarium oxysporum f. sp. cucumerinum (FOC). Cucumber plants with treated EB as seed coating and soil drenching (108–109 CFU ml-1). were transplanted to peat inoculated with FOC spore suspension (105 spore ml-1 ). According to growth chamber test results, 38% of tested EB strains exhibited the disease reduction between 30 to 60 %, comparing to only FOC inoculated plants. In the next step of this research, EB strains applied as seed coating and soil drenching inhibited the Fusarial wilt development at the rate of 49 to 52% compared to only FOC inoculated cucumber plants for two months growing period in soilless growing system. Moreover, EB treatments, without being any disease pressure, increased the total marketable fruit yield at the rate of 5 to 28% compared to non-treated cucumber plants for 2 months growing period in soilless cultivation. The population density of some EB strains in the root tissues was determined as approximately 103-105 CFU plant g-1 75 days after seed bacterization. It was determined that colonization rate of EB strains in the root tissues was higher than shoot tissues. So, the application of EB as seed coating and soil drenching was recorded as effective and practical techniques in terms of biological control, growth promotion of cucumber plants and root colonization of bacteria inside the plant tissues. 79 Root Endophytes Monday 22 June – Poster session 128 Siderophore production and iron phosphate solubilization by root endophytic bacteria isolated from maize Vitoria Palhares*1, Ivanildo Marriel2, Eliane Gomes2, Ubiraci Lana2, Crisia Abreu3, Cássia Almeida1, Christiane Oliveira2 1 UniFEMM, Brazil, 2Embrapa Mayze and Sorghum, Brazil, 3Federal University of Minas Gerais, Brazil Tropical soils generally exhibit acidic condition with predominant phosphate immobilized in insoluble forms of iron (P-Fe). Many soil microorganisms solubilize P from P-Fe by siderophores and organic acids production. The microorganisms that live in the rhizosphere and in the interior of plants (facultative endophytic) with potential for phosphate solubilization are important to enhance the production of the maize crop in order to reduce the use of soluble fertilizers. The objective of this work was to isolate efficient facultative endophytic bacteria solubilizing iron phosphates and evaluate their production of siderophores. Maize plants were harvested at flowering stage and 113 bacteria were isolated from roots (54.9%), leaves (20.4%) and sap (24.8%). Fifty-eighty selected strains, most root endophytic, were evaluated in liquid culture medium containing iron phosphate, after 9 days of incubation. Soluble phosphorus was determined using modified ammonium molybdate method. The production of siderophores was through inoculation of microorganisms in solid medium containing the indicator cromoazurol. Strains were identified based in the 16S rDNA gene. Maize plants had a high diversity of endophytic bacteria solubilizing iron phosphate and siderophore producing. One strain of Pantoea and two of Bacillus showed the highest P solubilization, releasing 68.7; 64.1 and 64.08 mgP.L-1, respectively. The siderophore produced by 65% of the evaluated microorganisms was the type carboxylate, and the most strains producing siderophores are efficient in solubilizing P associated with iron. The use these facultative endophytic solubilizing microorganisms as inoculant may be considered a promising strategy in environmental and economic terms for maize production. 129 Effect of GFP-labeled Paenibacillus polymyxa on growth of agricultural crops Kiran Preet Padda*, Akshit Puri, Christopher Chanway University of British Columbia, Canada Paenibacillus polymyxa strain P2b-2R, previously isolated from internal stem tissue of a naturally regenerating pine seedling, fixes nitrogen in association with lodgepole pine seedlings and promotes their growth. This strain has been shown to colonize lodgepole pine seedling tissues endophytically using a green fluorescent protein (GFP)-labelled derivative of the wild type. We wanted to see if the GFP labelled derivative of P2b-2R would fix N and promote growth of agricultural crops such as corn and canola in ways similar to the wild type strain. We inoculated corn and canola seeds with wild type P2b-2R or the GFP labelled derivative and grew seedlings for 40 days in an N-limited soil mix. Seedlings were harvested 20, 30 and 40 days after inoculation and evaluated for biological nitrogen fixation and growth promotion. 80 Root Endophytes Monday 22 June – Poster session Seedlings inoculated with the GFP labelled P2b-2R strain derived small amounts of N from the atmosphere (up to 17%) but were 40% taller and accumulated 70% more biomass than those treated with wild type P2b-2R. We conclude that GFP modification of strain P2b-2R resulted in a significant enhancement of the wild type’s growth promotion efficacy on corn and canola and facilitated some in situ biological nitrogen fixation with these two crop species. 130 Next generation sequencing analysis of soil and plant associated fungal assemblages in sub-Arctic sand dune ecosystem Anbu Poosakkannu*, Riitta Nissinen, Minna-Maarit Kytöviita University of Jyväskylä, Finland Our research focusses on plant associated microbial communities in the sub-arctic inland primary successional site, located in an Aeolian sand dune area in subarctic Northern Fennoscandia (68° 29' N). One of the very few species capable of colonizing these Arctic sand dunes and enabling ecosystem restoration is the grass Deschampsia flexuosa. In the study site, the early successional stage is characterized by D. flexuosa growing as monoculture in the blow-out areas. Late successional stage is mountain birch forest vegetation with continuous ground cover vegetation composed of abundant D. flexuosa together with other plants and under the cover of mountain birch trees. This goal of this study was to assess the community composition of fungal assemblages in soil (bulk and rhizosphere) and in the internal tissues (endophytes) of D. flexuosa growing in the two different successional stages. Endophytic fungal studies in cold environment especially in arctic are very limited, although they can improve host nutrient acquisition and protect the plants from different stresses. Deschampsia flexuosa (leaf and root), rhizosphere and bulk soil samples were collected from four different blow-out areas between 150 and 2250 meters apart. We collected eight biological replicates (two samples per blow out area and successional stage). Soil and plant associated fungal community structure was studied using Next generation Ion torrent sequencing of partial internal transcribed region (ITS) amplicons. Mothur based bioinformatics analysis of ITS sequences revealed 99217 quality-filtered sequences which are abundant (OTUs with less than 10 sequences were removed) that were separated into 2781 species-level OTUs. Of fungal taxa, Ascomycota (67.1%), Basidiomycota (18.9 %), encompassed the largest proportion of OTUs. The phylum Ascomycota was the predominant phylum identified and present in all samples followed by Basidomycota, Zygomycota and Glomeromycota. Endophytic samples are dominated by phylum Ascomycota. The compositional variation was mainly accounted for by successional stages. 81 Root Endophytes Monday 22 June – Poster session 131 Can an endophyte isolated from lodgepole pine trees reside inside agricultural crops and fix N? Akshit Puri*, Kiran Preet Padda, Christopher Chanway University of British Columbia, Canada Several Paenibacillus strains that were able to fix nitrogen were isolated from extracts of surface-sterilized lodgepole pine seedling and tree tissues. One strain, Paenibacillus Polymyxa P2b-2R, was found to fix high amounts of nitrogen when reintroduced to the gymnosperms, lodgepole pine and western red cedar. We wanted to determine if Paenibacillus polymyxa P2b-2R could colonize, fix N and promote the growth of important agricultural crops such as corn and canola. We inoculated corn and canola seeds with P. polymyxa strain P2b-2R and grew seedlings for 40-60 days. Corn seedlings were harvested 10, 20 and 30 days after inoculation and canola seedlings were harvested 20, 40 and 60 days after inoculation for evaluation of endophytic and rhizospheric colonization. Seedlings were also evaluated for biological nitrogen fixation and growth promotion at these harvest intervals. P2b-2R colonized rhizosphere as well as inside tissues of root (endophytically). Corn and canola seedling growth was promoted significantly by inoculation with P2b-2R with an increase of up to 35% in height and 30% in biomass from the controls. P2b-2R also provided more than 20% of foliar nitrogen. These results suggest that Paenibacillus polymyxa P2b-2R has a broad range of plant hosts and is able to fix N and promote the growth of at least certain agricultural crops. 132 Maize roots endophytic bacteria and their potential as plant growth promoting and biological control agents João A. C. Vieira1, Natalia L. S Alves1, Christiane A. O. Paiva2, Vera Lúcia Santos*1 1 Laboratory of Applied Microbiology, Microbiology Department, Institute of Biological Science, Federal University of Minas Gerais, Brazil, 2Embrapa Milho e Sorgo Empresa Brasileira de Pesquisa Agropecuária (Embrapa), Brazil Studies have shown that many endophytic microorganisms, which lives asymptomatically within plant causing no signs of harm to the host, may work as plant-growth promoters and/or biocontrol agents which has made them valuable for agriculture for improving crop performance. In this work, endophytic bacteria were isolated from roots of Pioneer 30F35 Herculex hybrid corn crops, cultivated with and without phosphorus fertilization. After identification by partial sequencing of the 16S rDNA gene, a total of 80 bacteria was evaluated regarding solubilization of inorganic phosphate (CaPO4), antagonizing bacterial (Bacillus subtilis and Pantoea ananatis) and phytopathogenic fungal (Fusarium verticillioides and Coletotrichum graminicola) growth, mineralization of phytate and production of IAA (indole-3-acetic acid). There was an equal distribution of the isolates from Actinobacteria, Firmicutes and Proteobacteria clades, and only one isolate (Flavobacterium acidificum RT3B41) of Bacteroidetes phylum was found. The strains were grouped into 26 genera, which the 82 Root Endophytes Monday 22 June – Poster session most frequent were Bacillus, Leuconostoc, Pseudomonas, Serratia and Enterobacter. From the total, 32 isolates were able to solubilize inorganic phosphate (dosages between 10 and 527 mg/l) and 45 showed production of IAA (4.5 to 111 μg/ml). In the plate antagonism test, 10 isolates inhibited the growth of B. subtilis, 6 of the gram negative pathogen P. ananatis, 11 of the fungus C. graminicola and 4 of F. verticillioides. In plate qualitative tests, 52 isolates also had the ability to mineralize phytate. This work demonstrates the enormous potential application of these isolates, which must still be confirmed by in vivo and field tests. 133 Development of a bacterial cell enrichment method for the analysis of the endophytic microbiota in sugarcane stems Stefan Schwab*1, Carlos dos Santos2, Daniel de Souza2, Dayana Rosa2, José Baldani1 1 Embrapa Agrobiologia, Brazil, 2Universidade Federal Rural do Rio de Janeiro, Brazil Sugarcane is an important culture in Brazil, with high economic support and social relevance. Sugarcane plants are rich in endophytic bacteria, which can promote and modulate plant growth through diverse mechanisms, such as phytohormone production, antagonistic activity against phytopathogens, and improvement of nutrient utilization by the plant; however, most bacteria are yet uncultivable. Therefore, cultivation-independent strategies are essential to better understand their genetic diversity and functional characteristics. Metagenomic surveys of endophytic microbiota currently represent a challenge, due to the low number of bacterial cells in relation to the host plant. In this work a bacterial cell enrichment procedure was established, cultivation-independently, from the inner tissues of the base of sugarcane stems. Results showed that the enriched material contained bacterial cells that are colony-forming, and can be visualized by bright-field microscopy or Gram stain test. DNA content analysis of the enriched material revealed efficient elimination of plant DNA, and results of PCR and ARDRA showed that bacterial DNA is predominant. Preliminary results of 16S rDNA sequencing revealed the presence of alpha-, beta- and gamma-Proteobacteria. In order to obtain a deeper insight of the bacterial taxa, 16S rDNA sequencing of the enriched material on Illumina platform is being conducted, and results will be presented. The developed method may allow, in the future, accessing the sugarcane endophytic microbiome, and reveal bacterial genetic resources with agrobiotechnological applications. 134 Indirect stimulation of the vegetative growth of ‘Elkat’ strawberry plants by the bacterium Pantoea sp. (N52AD) Pawel Trzciński*, Lidia Sas Paszt, Edyta Derkowska, Michał Przybył The Research Institute of Horticulture, Poland The most important problems of agriculture are: increasing the abundance of macro- and microelements in the soil, maintaining adequate moisture levels, and protection from diseases and pests. Excessive use of fertilizers (mineral and manure) and pesticides may reduce soil fertility and biodiversity, and contaminate surface and ground waters. One way of reducing the use of fertilizers and chemical plant protection products is to employ beneficial microorganisms which can decrease the number of pathogens in the soil, increase the availability of macro- and microelements to plants, and improve the resistance of plants to 83 Root Endophytes Monday 22 June – Poster session stress caused by, for example, drought. Among the most common and universal microorganisms used are arbuscular mycorrhizal fungi. By colonizing the roots of the host plant, these fungi increase the uptake of water, macro- and microelements by the plant and protect it from soil-borne pathogens. The degree of the symbiosis is affected by some microorganisms, for example, mycorrhiza helper bacteria. Application of such bacteria can improve the colonization of plant roots by autochthonous mycorrhizal fungi or those present in commercial products, thus allowing plants to make a more efficient use of nutrients and consequently contributing to a reduction in the amounts of the fertilizers and pesticides used. The aim of this study was to evaluate the impact of the bacteria from the genus Pantoea (strain N52AD) on the degree of colonization of strawberry roots by mycorrhizal fungi and on the vegetative growth of strawberry plants. Frigo plantlets of the cultivar Elkat were planted in pots filled with the soil collected from an experimental field in Dąbrowice. In the experiments conducted in 2012-2013 in greenhouse conditions, the plants inoculated with the N52AD strain were colonized by arbuscular mycorrhizal fungi to a greater extent and produced a greater mass of leaves than the control plants (non-inoculated with N52AD strain). 135 Improvement of soil fertility and plant production in the Democratic Republic of Congo by implementation of arbuscular mycorrhizal fungi and Sebacinales Jolien Venneman*, Danny Vereecke, Geert Haesaert Department of Applied Biosciences, Faculty of Bioscience Engineering, University Ghent, Belgium A growing world population is associated with an increased demand for food. This trend is most pronounced in developing countries where the availability of fertile arable land is becoming rather limited. An important reason is the typical physical and chemical properties of tropical soils, e.g. P-fixation and low cation exchange capacity (CEC), in addition to soil degradation caused by unsustainable agricultural practices. Since the access to mineral fertilizers is very restricted in tropical regions, the mentioned problems should be addressed through the application of an integrated soil fertility management. This includes the use of sustainable cultivation techniques in combination with adapted plant genetic material, micro-doses of chemical fertilizers, and maximal amounts of organic matter. In our research, we particularly focus on the implementation of growth promoting micro-organisms, which are isolated from soil and plant root samples collected in the Democratic Republic of Congo, region of Kisangani. Arbuscular mycorrhizal fungi and basidiomycetes belonging to the order Sebacinales are two groups of root endophytes that attracted our attention. To reveal the mechanisms behind the plant-fungus interactions we combine molecular diversity analysis with functional tests. In addition, we plan to set up onfarm based systems for mass production of inoculum in Kisangani in order to provide local farmers with the knowledge on how to maintain the beneficial indigenous micro-organisms. 84 Root Endophytes Monday 22 June – Poster session 136 Monitoring and quantifying Bacillus mycoides in the potato rhizosphere Yanglei Yi*, Jan Spoelder, Marielle van den Esker, Oscar Kuipers University of Groningen, Netherlands Bacillus mycoides is a rod-shaped soil bacterium belonging to the B. cereus species-group. On agar plates, a fungus-like shape forms resulting from cells linked end to end. B. mycoides is reported to have plant growth-promoting effects on sugar beet, cucurbits and tobacco. B. mycoides is believed to be an important bacterium for potato growth as well, due to its abundance in potato rhizosphere and endosphere without causing visible signs of infection. Unraveling mechanisms mediating plant host-endophytes recognition, establishment and colonization dynamics requires a reliable method to monitor these processes. Visualization of cells in the rhizo-/endo-sphere using fluorescent protein (FP) as a marker has been adopted by many researchers to study plant-microbe interactions. However, many laboratories struggle with incorporating foreign DNA into Gram-positive bacteria including some wild isolates of Bacillus. Here, we describe a method to transform the microbe by electroporation with a plasmid encoding a constitutive promoter FP, resulting in fluorescent cells. Combination of quantitative and qualitative data achieved by both molecular and microscopy methods are probably the best choice to monitor endo- and rhizo-bacteria, with the advantages of each technique complementing the drawbacks of the other. 85 Root Endophytes Monday 22 June – Poster session Root Development 137 Architectural analysis of date palm root systems (Phoenix dactylifera L.) Mimoun Asma*1, Stokes Alexia2, Rey Hervé3, Lecoustre René3, Jourdan Christophe4, Bennaceur Malika1 1 Oran 1 University, Algeria, 2INRA, Montpellier, France, 3AMAP/CIRAD BIOS, France, 4CIRAD-Eco&Sols, France Architectural traits were used to describe the structure and development of the date palm (Phoenix dactylifera L.), root system. To characterize root system architecture, two parallel experiments were conducted: one in a rhizotron whereby root growth was measured over time and one in a nursery bags from which roots were sampled regularly. These roots were sectioned and examined under a light microscope in order to determine how the anatomical structure of roots changes as they age. Double staining was performed on roots at different ages and from diverse root zones. The topology (arrangement of axes relative to each other) and typology (classification of different root axes based morphological and functional criteria) of root systems was measured in the first rhizotrons. Root diameter and root elongation were measured regularly on seedlings grown from seed (0 to 6 months). RhizoDigit software (©CIRAD) was used to digitize root observations and to compile data for statistical analyses. Results showed that during this developmental period, the root system architectural unit comprises seven root types having distinct characteristics and spread over three different topological orders (primary, secondary and tertiary). The establishment of different types of root axes evolves over time and depends on the root category. Root growth was variable depending on the root type and branching order of the diameter according to its class. There was an absence of both radical mortality and the arrest in growth of short secondary roots during this period. The anatomies of the radical and adventitious roots were typical of monocotyledonous roots. Radical anatomy also had a characteristic spatial and temporal development during the juvenile stage. We will use these results to develop three-dimensional models for use in agricultural management. 139 Root architecture phenotyping of different quinoa (Chenopodium quinoa Willd) accessions José Correa*1, Phil Pstrong2, Francisco Pinto2, Kurt Ruf1, Iván Matus1, Kerstin Nagel2, Fabio Fiorani2, Manuel Pinto1 1 Instituto de Investigaciones Agropecuarias, Chile, Chile, 2Forschungszentrum Jülich, Germany Quinoa has received an increasing attention because of its nutritional value, adaptability to different environmental constraints, and its vast biodiversity. All these crop aspects are relevant for global food security. Root system architecture plays an important role in determining the ability of a plant to explore the soil and foraging for water and nutrients. These aspects linked to root architecture are not yet well characterized in quinoa accessions and genetically diverse panels. According to that, two pilot experiments were carried out to evaluate the applicability of a rhizotron system for root assessment in quinoa. Plants were grown under greenhouse conditions in rhizotrons at IBG2: Plant Sciences at 86 Root Development Monday 22 June – Poster session Forschungszentrum Jülich GmbH, Germany. The first experiment focused to establish whether the visible root traits at the interface of the rhizoboxes correlate with root and shoot traits measured destructively and the second one to analyze the effect of water stress on root architectural parameters. To represent the diversity of quinoa, three Chilean wild ecotypes and three commercial varieties graciously supplied by Wageningen breeding program, were used. During the second experiment the water stress treatment was performed on the more vigorous ecotypes. The following traits were measured: length of primary, lateral and tertiary roots; root system depth, width and area; and root dry weight. Correlations and differences among ecotypes for all traits were found. In addition, the effect of water stress on root architecture was mainly manifested by a decrease of root system length, width, laterals, and area. In conclusion, there are relationships between traits measured in rhizotrons and traits of the plant/root system allowing analyzing the variation observed in the plant/root system and providing details about the effect of water stress on root development. These findings indicate that protocols for screening germplasm in pre-breeding scenarios could be developed specifically for quinoa. 141 First root allometric model for shifting cultivation systems in PDR Lao Iain McNicol1, Nicholas J Berry1, Thilde B Bruun2, Andreas de Neergaard*2, Casey Ryan1 1 University of Edinburgh, Scotland, 2University of Copenhagen, Denmark Shifting cultivation remains an important land use across of Southeast Asia and other parts of the tropics. This practice creates complex mosaic landscapes with regrowing fallows of various age interspersed with active fields and patches of mature forest. Quantifying root carbon stocks in these secondary forests is limited by the availability of reliable allometric models for these systems, inhibiting the development of policies aimed at reducing the intensification of shifting cultivation systems (REDD+). We developed new allometric models for prediction of both tree stem and root biomass in shifting cultivation systems based on a destructive harvest of 150 trees from Luang Prabang Province, Laos PDR. This study is the first to develop allometric models of root biomass for shifting cultivation landscapes, which we hypothesised would be a major carbon pool given that resprouting, and associated high root biomass, is a common physiological trait. Tree height was less important for estimating root biomass with “diameter only” models performing best. Resprouting trees exhibited significantly greater root biomass compared to trees growing from seed. Our best-fit allometric models were subsequently applied to 12 nearby plots which span a chronosequence of fallows to calculate the impact of accounting for resprouting allometry on forest biomass. We found that root biomass stocks (Mg/ha) were around 58% (22 - 85%) higher after accounting for resprouting, resulting in an average 9.5% (4 - 13%) increase in total biomass stocks, thus demonstrating the need to correctly account for re-sprouting trees in shifting cultivation fallows. Our analysis suggests that using our models will substantially improve the accuracy of tropical estimates of tree biomass and its distribution among different pools in shifting cultivation fallows. We also find that models fit using non-linear regression provide equally good fits to the data compared to the traditional approach of logtransforming biomass data. 87 Root Development Monday 22 June – Poster session 145 Root dynamics of two grapevine cultivars differing in their hydraulic behavior Jhonathan Ephrath*1, Brian Hoefgen2, Uri Hochberg2, Shimon Rachmilevitch2 1 Ben Gurion University of the Negev, Jacob Blaustien Inst. for Desert Research, Israel, 2Blaustein Institutes for Desert Research, French Associates Institute for Biotechnology and Agriculture of Drylands, Ben Gurion University of the Negev, Israel The interaction between scion and rootstock in grafted grapevines is extensively investigated. Nonetheless, the effect of canopy on root morphology has not been widely tested in field conditions. We tested root morphology in response to two irrigation regimes for two hydraulically different cultivars Shiraz (SH) and Cabernet Sauvignon (CS) grafted onto the same rootstock, 140 Ruggeri. These two grapevine cultivars have different aboveground hydraulic behavior (SH near-anisohydric and CS near-isohydric), and it is assumed that these differences may also cause belowground differences. This research was conducted in a four year old experimental vineyard at Ramat Negev, Israel. The grapevines underwent different irrigation regimes (50% and 25% of crop evapotranspiration). Aboveground and belowground physiological parameters were measured for a period of 16 months. Transparent tubes were installed to a depth of 180 cm at distances of 25 and 75 cm from the trunk. Root photographs were taken during different developmental stages of the year using a minirhizotron. Our results indicated that scion hydraulic behavior affected root dynamics. Larger differences were seen in SH than compared to CS in response to the irrigation treatment. It seems that SH invested more growth directly below the trunk, while CS roots were more evenly distributed throughout the soil profile. 146 Modelling of root dynamics in split-root rhizoslides reveals strong selective root placement of maize in response to nitrogen Dina in 't Zandt*1, Chantal Le Marié2, Norbert Kirchgessner2, Eric J.W. Visser1, Andreas Hund2 1 Radboud University Nijmegen, Netherlands, 2Swiss Federal Institute of Technology Zurich, Switzerland The plant’s root system is highly plastic, and can respond to environmental stimuli such as high nitrogen in patches. A root generally responds to a nitrogen patch by selective placement of new lateral roots within the patch to increase nitrogen uptake efficiency. This is a desirable trait in breeding programs, since it may decrease NO3 - leaching and N2O emission. Roots of maize (Zea mays L.) were grown without nitrogen in split-root rhizoslides, a system that enables non-destructive root measurements via direct imaging of the root system. Half of the root system was subjected to high nitrogen after 15 days, and root growth was traced for a subsequent 15 days and modelled. The elongation rates of crown axile roots on the nitrogen-treated side followed a logistic increase to a 5.3 cm d-1 maximum, 95% of which was reached within 4 days. On the untreated side, axile root elongation dropped linearly to 1.2 cm d-1 within 6.4 days and stayed constant thereafter. Twice as many lateral roots were formed on the crown axis on the nitrogen side compared to the untreated side. Furthermore, the elongation rates of laterals exposed to high nitrogen increased linearly with 88 Root Development Monday 22 June – Poster session most of the roots reaching an asymptote approximately 8 days after start of the nitrogen treatment. The laterals without nitrogen did not elongate. Furthermore, it was shown that the crown root system had a greater influence on shoot performance than the seminal root system 15 days after nitrogen application. 147 Effect of phenanthrene exposure on apoplastic barriers formation in maize Pierre Leglize*, Joan Dupuy, Quentin Vincent, Ivan Zelko, Christian Mustin, Stéphanie Ouvrard, Thibault Sterckeman Université de Lorraine, France Management of Polycyclic Aromatic Hydrocarbons (PAH) contaminated soils through phytoremediation process is promising in controlled conditions. However, in situ experiments were not so successful due to plant growth limitation induced by PAH toxicity. Improving revegetation of contaminated site appears therefore crucial in phytoremediation application but requires a better understanding of contaminant’s impact on plant functioning. PAH exposure affects plant physiology such as mineral nutrition. Because the nutrient uptake is linked with the maturation of exodermis and endodermis, we focused on (i) the effect of PAH on the suberin lamellae formation and (ii) the localization of phenanthrene in root. Maize plants were grown in contaminated sand (50 and 150 mg PHE kg-1 dry sand) for 10 or 20 days. Epi-fluorescence microscopic observation of root sections was used to assess PHE localization within the root and maturation of exodermis and endodermis by Fluorol yellow 088 detection of suberin. For 10 days of cultivation, suberization of exodermis and endodermis of maize exposed to PHE was more extensive and PHE were only observed within suberized exodermis. This could be related to an inducer effect of PHE on the deposition of hydrophobic compounds within exodermis, which may act as barrier against PHE penetration. However after 20 days of exposure, exodermis and endodermis of non-exposed roots were totally suberized whereas PHE-exposed roots where less suberized. After 20 days, PHE seemed to inhibit root maturation, which could be caused by its toxicity as it is related with the plant biomass reduction. Furthermore, PHE patches were located only within suberized exodermis and endodermis, which may therefore act as retention zone where the hydrophobic PHE accumulates during its radial transport. 148 Responses of root development and zinc uptake to heterogeneous soil water distribution in lupin and cucumber Huifang Ma*1, Michael Wassilios Evangelou1, Peter Vontobel2, Rainer Schulin1 1 ETH Zurich, Institute of Terrestrial Ecosystems, Switzerland, 2Paul Scherrer Institute, Switzerland The distribution of moisture is typically heterogeneous in soil even at the scale of a root system, and plant root systems are known to adapt to such heterogeneity. In this study we investigated how root growth allocation of two common crop plants, white lupin (Lupinus albus) and cucumber (Cucumis sativus L.), responded to lateral heterogeneity in soil moisture 89 Root Development Monday 22 June – Poster session over time with neutron radiography and how this affected zinc (Zn) uptake in a low-Zn sandy soil with patchy Zn distribution. Single plants were grown in Al-containers (inner size: 27 cm length × 27 cm height × 1.2 cm thickness). To one lateral section of each container 20 mg Zn/kg were added. Soil water heterogeneity was achieved by mixing coarse sand into the soil in one lateral section of the respective containers, either on the same (cis) side as or on the side opposite (trans) to the Zn-enrichment, while no sand was added to control containers. The same amount of water was added to all containers. As a result the soil water content decreased in the order: sections without sand in heterogeneous containers > control containers > sections with sand in heterogeneous containers. In lupin, root growth (primary root length, cluster root number and length, cluster root length/total root length, and root dry weight) was reduced in the heterogeneity treatments, with fewer roots being produced in sand than in no-sand sections, while Zn uptake was not affected. In cucumber, the heterogeneity treatment increased root growth (root length density and total dry weight) and shoot Zn uptake. Lupin roots responded already after 6 days to the heterogeneity treatment, cucumber roots only after 35 days. 149 Effect of drought on surface and deep root dynamics of teak (Tectona grandis) and rubber (Hevea brasiliensis) trees in mainland South East Asia Jean-Luc Maeght*1, Corentin Clément2, Santimaitree Gonkhanmdee3, Oloth Sengtaheuanghoung4, Alexia Stokes5, Alain Pierret6 1 UMR Iess Paris ; IRD, France, 2Iess Paris IRD Nafri, Lao People's Democratic Republic, 3Faculty of Agriculture, Thailand, 4NAFRI, Lao People's Democratic Republic, 5INRA UMR-AMAP, France, 6UMR Iess Paris ; IRD - NAFRI, Lao People's Democratic Republic We present the main results of two experiments conducted in a teak (Tectona grandis) tree plantation in Northern Laos and a rubber (Hevea brasiliensis) tree plantation in North East Thailand. We aimed at documenting the response of fine roots to seasonal and induced drought and quantify the corresponding carbon stocks. Fine root growth of 20-year old teak trees and 18-year old rubber trees were monitored at monthly intervals over 2 and 3 years, respectively. Observations were made using root windows installed at 50 cm depth increments, to a depth of 4.5 m. A rainfall exclusion experiment was performed in the teak plantation. Destructive samples were used to quantify fine root-related carbon stocks. In the case of teak, fine roots were found to a depth of 12 m. In both species, root dynamics is linked to transient changes in water availability. This work shows that deep fine roots are organs of foremost functional importance and we demonstrate their instrumental role in deep-water extraction during critical dry periods. The presence of significant amounts of deep roots in the soil profile further indicates that they should be taken into account when quantifying long-term carbon storage. 90 Root Development Monday 22 June – Poster session 150 The characteristics of root cell components of plants adapted to acidic soil Eriko Maejima*1, Toshihiro Watanabe1, Tadao Wagatsuma2, Mitsuru Osaki1 1 Research faculty of Agriculture, Hokkaido University, Japan, 2Faculty of Agriculture, Yamagata University, Japan Melastoma malabathricum and Melaleuca cajuputi naturally grow in highly acidic soil in tropical and subtropical zones. They are both reported to be highly aluminum tolerant, but their tolerance mechanisms still remain to be explained. Melastoma malabathricum accumulates high concentration of aluminum, whereas Melaleuca cajuputi accumulates low concentration of aluminum. Plasma membrane and cell wall compositions, organic acids released from root tips, and/or phenolics in root cells have often been reported to affect aluminum tolerance and aluminum accumulation in roots. In this study, we focused on woody species in Melastomataceae and Myrtaceae and investigated their root cell components possibly responsible for aluminum tolerance and/or accumulation. Melastoma malabathricum (Melastomataceae), Tibouchina urvilleana (Melastomataceae), and Melaleuca cajuputi (Myrtaceae) were grown in hydroponic culture and transferred to an aluminum treatment solution (phosphorus-free nutrient solution containing 0 or 500 μM AlCl3) for 1 week. Aluminum concentration in roots and leaves, lipid and cell wall composition, and organic acid and phenolics concentrations in roots were determined. We also investigated rice (Oryza sativa L.), an aluminum-tolerant crop, for comparison. Aluminum concentration in Melastoma malabathricum and Tibouchina urvilleana was higher than that in Melaleuca cajuputi. Melastoma malabathricum and Melaleuca cajuputi contained much lower proportion of phospholipid in root cells than that in rice, suggesting that the lipid composition in the plasma membrane in root cells is involved in aluminum tolerance mechanisms in such highly tolerant plants. Moreover, all woody plants investigated contained the highest concentration of phenolics compared with that in rice. Although Melaleuca cajuputi contained highest concentration of phenolics in roots, the extracted phenolics did not have the capacity to chelate aluminum. In contrast, phenolics extracted from Melastoma malabathricum formed a stable chelate with aluminum. These results suggest that phenolics in roots are likely to be involved in the detoxification of aluminum in roots of Melastoma malabathricum. 151 Root system of crops under different soil management systems in sugarcane field reform Gustavo Mateus*1, Felipe Giglio Bernardoni2, Denizart Bolonhezi2, Rafael Müller3, Carlos Alexandre Costa Crusciol3, Humberto Sampaio Araújo2 1 São Paulo Agency of Agribusiness Technology, Brazil, 2São Paulo Agency of Agribusiness Technology - APTA, Brazil, 3Department of Crop Science, College of Agricultural Sciences - FCA, Sao Paulo State University - UNESP, Brazil Sustainable agricultural production systems that improve the quality of soil and water and reduce the emission of greenhouse gases, are essential in modern agriculture. This work was developed in the São Paulo Agency of Agribusiness Tecnhology, Andradina, São Paulo, Brazil. The aim of this study was to evaluate the effect soil management systems and different 91 Root Development Monday 22 June – Poster session rotation system in sugarcane field reform on the root dry matter and root length of rotated crops. The experimental design was randomized blocks in split plots with four replications. The plots treatments consisted of three soil management systems, (conventional tillage, minimum tillage and no-tillage). The split plot consisted of four comercial crops, corn, peanut, sorghum and soybeans, a choice of green manure (Crotalaria juncea + jack beans), and fallow system. Roots were sampled in the flowering plants. No significant interactions were found among the soil management systems and different crops rotation in sugarcane field reform, for all variables. This can be explained by the dry spell and high temperatures occurring during crop development, which allowed the species presented similar behavior for the root dry matter and root length. Thus the volume of soil explored by the roots was the similar to the species regardless of the tillage system. The yield of root dry matter was observed values of 2.053, 2.496 and 1.195 g m-3 in the layers 0-10, 10-20 and 20-40 cm, respectively. Regardless of soil management systems and rotated crops was found that 80.7% of the root system concentrated at between the layers 0-20 cm. 152 Rooting of jujube (Ziziphus jujube mill) Li variety cuttings using some root promoting micro-organism and plant growth regulators Nabil Omar* Agricultural Research Centre, ARC, Egypt This study was conducted at the experimental nursery of the Horticulture Research Institute, Giza, Egypt during 2008 and 2009 seasons. Semi-hard wood cuttings were taken from mature 15 years old trees of jujube (Ziziphus Jujube Mill) Li. Roots treatments inoculated with some plant growth promoting rhizobacteria strains (PGPR) 1- Bacillus polymyxa, 2- Bacillus circulans, 3- Bacillus megaterium, 4- Bacillus pasteurii, 5- Pseudomonas fluorescens, 6- Yeast strain (Saccharomyces cerevisiae), and mixed inoculants from previous PGPR strains. In addition, indole butyric acid (1000 and 2000 ppm), naphthalene acetic acid (1000 and 2000 ppm) were also tested as compared with untreated (control). The obtained results showed that, the effect of Bacillus megaterium as PGPR resulted in the highest significant rooting percentage (60% and 50%). On the contrary, the lowest significant effect of treatments was found as a result of naphthalene acetic acid at 1000 ppm and control during the two seasons of the study. Histological studies revealed that the callus originated from the cambial and phlom parenchyma cells below the cork cells from these protrusions the adventitious roots were developed. In conclusion the use of PGPR offers an attractive way to replace chemical fertilizer and supplements most of the isolates result in a significant increase in plant height, root length, and dry matter production of shoot and root of plants. 92 Root Development Monday 22 June – Poster session 153 Effect of Streptomyces sp. isolated from acidic cultures of minerals in root development in environments polluted with mercury Sara Liz Pacheco Huerta*, Jasmin Elena Hurtado Custodio, Patricia Sheen Cortavarria Universidad Peruana Cayetano Heredia, Peru Two strains of Streptomyces sp. (E1 and K2) and one strain of Streptomyces variabilis (K1A) were isolated from acidic cultures of mineral ores from Peruvian highlands. Molecular and phenotypic identification of the isolate strains was performed. The isolates were also evaluated to determine their resistance to metals, growth at different concentrations of sodium chloride, growth to different pH and exposed to different temperatures. The Streptomyces sp. strains (E1 and K2) were able to grow in 100 ppm of mercury and Streptomyces variabilis (K1A) was able to grow in 50 ppm of mercury. The three strains were evaluated to determine their ability to contribute to the development of roots in Lactuca sativa and Medicago sativa when exposed to mercury. L. sativa showed regular root development in 10 ppm mercury. However, when treated with Streptomyces sp (E1 and K2) and Streptomyces variabilis (K1A), L.sativa showed a remarkable root development in length in up to 50 ppm of mercury. Moreover, M.sativa has the ability to tolerate concentrations of mercury up to 100 ppm allowing a significant root development. In presence of Streptomyces sp (E1 and K2) the root development was in up to 200 ppm of mercury. The results obtained in this study allowed us to identify the ability of these strains to contribute to the development of plants in contaminated areas and therefore allow the restoration of polluted environment. 154 Chili pepper growth and drought tolerance are regulated by Bacillus vallismortis strain EXTN-1 producing volatile organic compounds Kyungseok Park*, Sarnalee Dutta National Academy of Agricultural Science, South Korea The plant growth-promoting rhizobacteria (PGPR), especially Pseudomonas spp. and Bacillus spp., colonize roots of monocots and dicots, and directly or indirectly promote plant growth and elicit induced systemic resistance, among which Bacillus vallismortis strain EXTN-1 plays a pivotal role in enhancing induced systemic resistance (ISR) against multiple pathogens in a variety of crops. Despite of accumulating evidence the role of EXTN-1, the precise underlying mechanism of volatile organic compounds (VOCs) emitted by EXTN-1 has not been conclusively studied. In this study, in order to elucidate the potential physiological role of VOCs from EXTN-1 we investigated the plant growth and development in response to VOCs. Any phenotypic changes were not observed in aerial parts of VOCs-exposed chili pepper plants. However, root architecture was significantly altered in which increased root biomass was obtained in VOCs-exposed plants. Interestingly, a combination treatment between EXTN1 and VOCs from EXTN-1 had a negative effect on pepper growth. In addition, root colonization rate in rhizosphere was higher in VOCs-exposed plants, relative to control plants. These results suggest that increased population density of Bacillus may positively modulate the VOCs-mediated plant growth promotion. Intriguingly, improved drought tolerance was exhibited in plants exposed to VOCs derived from EXTN-1. Our data clearly propose that 93 Root Development Monday 22 June – Poster session VOCs from EXTN-1 contribute to augmentation of plant growth and that VOCs are normally required for abiotic stress tolerance. 155 Response of plant roots and mycorrhizal fungi to soil hypoxia Yasmine Piñuela Samaniego*1, Peter Železnik2, Hojka Kraigher2, Irena Maček1 1 University of Ljubljana, Biotechnical Faculty, Slovenia, 2Slovenian Forestry Institute, Slovenia Root growth and colonization with plant symbiotic arbuscular mycorrhizal (AM) fungi was measured in barnyard grass seedlings exposed to geological CO2 and hypoxia. Barnyard grass (Echinochloa crus-galli (L.) Beauv.) is an annual C4 grass and a cosmopolitan weed. It has been shown to be tolerant to soil hypoxia and flooding. Our experiment was conducted in the area of natural CO2 springs (mofettes) in Slovenia, and was designed to follow the growth of Echinochloa in three different CO2 soil exposures: control (<1 % CO2), medium CO2 and high CO2 exposed plot with peak CO2 concentrations 7.7 ± 2.0 % CO2 and 34.5 ± 6.7 % CO2, respectively. For each location, holes in the ground were dug and were filled with homogenized soil from a control site. In June 2014, 288 root ingrowth soil cores where installed. Individual cores were sampled in August, September and November 2014. Samples were soaked in water and sieved through sieves of different sizes (mesh size 2 mm and 2 µm). Roots where scanned in water on an optical scanner and root parameters were measured with WinRHIZO® (Regent Instruments Inc., CA) software. Subsequently, the roots were dried in an oven and weighed. Additionally, root colonization with AM fungi was assessed. Preliminary analyses of root biomass show a distinct decreasing trend in root biomass from August to November. At the control site, root biomass was significantly higher compared to the medium and high CO2 exposed site in the August sampling. In samples collected later in the year significant differences were found only between root biomass at control and high CO2 exposed plot. We found colonization with AM fungi in all the treatments. Interestingly, despite reduced plant growth and lower root biomass in CO2 exposed sites, in the most extreme conditions high levels of root fungal colonization were found. 156 Local and systemic effects of metals on root growth: towards revealing molecular mechanisms and optimizing growth Tony Remans*, Stefanie De Smet, Ann Cuypers, Jaco Vangronsveld Hasselt University, Centre for Environmental Sciences, Belgium Diffuse contamination by excess metals affects large areas worldwide, and using these areas to produce biomass may bring them back into sustainable use. However, plants unavoidably take up excess metals and are subjected to their toxic effects. Molecular mechanisms of metal uptake, sequestration and detoxification have been intensively investigated. Root placement in the soil is another important factor determining metal uptake, given the heterogeneous distribution of metal contamination in the soil, but little is known on the molecular mechanisms of interference of metals with intrinsic root developmental pathways. 94 Root Development Monday 22 June – Poster session We use Arabidopsis thaliana as a model and investigate local and systemic effects of metal exposure on root architecture when plants are heterogeneously exposed in split-root vertical agar plates. As local effects, cadmium (Cd) and copper (Cu) stimulated the outgrowth of lateral roots, but the elongation of these lateral roots was much stronger inhibited by Cd than by Cu. Excess zinc (Zn) inhibited the emergence of primordia and meristem activation. Systemic effects of metals on root development may either inhibit or stimulate the colonization of less-contaminated soil patches. A systemic inhibitory effect on primary and lateral root growth was observed when part of the root system was exposed to excess Zn, but not when exposed to Cd or Cu. These observations indicate the existence of metal-specific local and systemic interference with intrinsic root developmental pathways. Hormone production and signaling mutants are being studied to start identifying underlying molecular mechanisms. Knowledge on molecular mechanisms of growth-activating and –inhibiting effects of metals can support strategies for improving plant growth to the purposes of phytoremediation or safe biomass production, for which root development in contaminated zones is desired or should be avoided, respectively. 157 The evaluation of root system architecture in rice plant using the data of root distribution Ryosuke Tajima*, Toyoaki Ito, Masanori Saito Tohoku University, Japan Root system architecture has been shown to be important in agricultural systems. It is extremely difficult to identify the parameters of root system architecture, such as root elongation rate, from field experiments. Here, we tested to determine the parameters using the data of vertical root distribution in the field with root modeling. We evaluated five rice lines, Hitomebore (a modern lowland cultivar in Japan), IR8 (a semi-dwarf lowland rice), IRAT109 (an upland rice), NERICA3(an interspecific hybrid rice) and IR65564-44-51 (a lowland rice, New Plant Type), cultivated in flooded condition. At harvesting, one soil monolith (10 cm diameter) was taken from each plot to a depth of 10 cm. Then, the monolith was divided to five layers (0-2 cm, 2-4 cm, 4-6 cm, 6-8 cm and 8-10 cm depth) and each root length density was measured. In addition, we measured the panicle number, the nodal root number and the minimum value of initial angles in nodal roots of each plant, which can be easily measured. In these results, the panicle and nodal root numbers of Hitomebore and IR8 were much larger than those of the other lines. The minimum values of initial angles in nodal roots of IRAT109, NERICA3 and IR65564-44-51 were larger than those of Hitomebore and IR8. In the vertical root distribution, IR8 had a shallowest root system in the five lines, while the deep-root ratio (the percentage of the deep part in the total) of IRAT 109 was highest. Using these datasets, we determined an average root elongation rate and gravitropism of each plant with root modeling and nonlinear optimization. Both of the root elongation rate and the gravitropism of Hitomebore were predicted to be higher than that of IR65564-44-51, although the prediction was changed depending on initial parameters. 95 Root Development Monday 22 June – Poster session 158 Effect of soil type on the beech fine root distribution Marie-Pierre Turpault*1, Christophe Calvaruso1, Stéphane Uroz1, Paul-Olivier Redon2, Gil Kirchen1 1 INRA, France, 2Andra, France Different parameters as climate, tree age, stand management and soil properties can affect the distribution of tree roots in soil. In the experimental site of Montiers (north-east of France) managed by INRA and Andra, all these parameters are constant including a dominance of beech (Fagus sylvatica L.), except for the soil properties. Indeed soils significantly differ from alocrisol-brunisol, bruni-calcisol to rendisol. The alocrisol-brunisol is deep (> 2 m) and acidic while the rendisol is shallow (<30 cm) and alkaline. Two methods were used to quantify the fine roots (< 2 mm diameter) at different depths in these three soils: the root mass and the root impact methods. The root mass method consists in the collect of a same volume of soil at different soil depths. The roots are manually extracted from the total volume of soil and weighed. For each soil depth, the density of roots in the soil is estimated. The root impact method corresponds to the use of a calibrated grid placed in the soil pit and was used to count the number of root impacts in each square, to obtain ten measures of root density by soil thickness of 10 cm. Our analyses revealed that despite its shallowness, the rendisol presents a significantly higher quantity of fine roots compared to that in the alocrisol-brunisol. Furthermore, roots colonize the calcareous rock beyond two-meter deep, potentially by dissolving the limestone (root imprints observed). In the rendisol, which is comparatively to the other studied soils the lessdeep and the most sensitive to water deficits and to P and K deficiencies, trees seem to strongly invest in the development of fine roots in the soil horizons (higher density), but also surprisingly in the calcareous rock (for the same depth, higher density in the rock compared to that in the alocrisol-brunisol). 96 Root Development Tuesday 23 June – Poster session Nutrient Acquisition 2 Iron deficiency-induced root exudation of coumarins in Prunus rootstocks grown at high pH Ana Alvarez-Fernandez*, Yolanda Gogorcena, Javier Abadia, Anunciacion Abadia Aula Dei Experimental Station-CSIC, Spain Plant Fe deficiency is a problem in calcareous soils, due to the low solubility of Fe(III) hydroxide. Peach trees are among the fruit crop species most sensitive to Fe-deficiency. Prunus rootstocks differ widely in their susceptibility to Fe deficiency. Phenolics root secretion is associated to Fe deficiency tolerance in different plant species. Recent studies have shown that the fluorescent coumarin scopoletin along with other coumarins are exudated by the roots of the model species Arabidopsis and tomato in response to Fe deficiency. However, no information on Fe deficiency-induced root phenolics secretion has been reported for any fruit tree species so far. In the present work, two Prunus rootstocks (GF677 and Cadaman) with different tolerance to lime-induced chlorosis were grown for two weeks in half-strength Hoagland solution buffered at 7.5 pH, including either 0 (Fe-deficient plants) or 90 μM Fe(III)– EDDHA (Fe-sufficient plants). Iron deficiency induced the root secretion of fluorescent compounds in both genotypes, and leaf chlorosis was less intense in the Fe-efficient rootstock (GF677) than in the Fe-inefficient one (Cadaman). Root extracts and exudates were analyzed by liquid chromatography coupled to mass spectrometry. Several glycosides of different coumarins accumulated in GF677 roots in response to Fe deficiency. Also, several coumarins and coumarin derivatives were secreted in response to Fe deficiency by GF677 roots. The concentration of these compounds in root extracts and exudates of Fe-deficient Cadaman plants were lower than those from GF 677 roots. Phenolics removal from growth medium of Fe-deficient plants, by circulating the nutrient solution through a C18 resin column, led to a more advanced leaf chlorosis in GF677 but not in Cadaman. This supports that the root secretion of phenolics by Fe-efficient Prunus rootstocks could play an important role in Fe deficiency tolerance at high pH. 3 Rhizosphere P is affected by N sources applied to corn Elialdo Alves de Souza*1, Camila da Silva Grassmann2, Janaina Matias Pereira da Silva2, Sara Isa Vieira Kuchta de Almeida2, Ciro Antônio Rosolem2 1 São Paulo State Universtiy - Department of Crop Science, Brazil, 2São Paulo State University - Department of Crop Science, Brazil Nitrogen (N) source can affect rhizosphere activity and phosphorus (P) uptake in neutral/alkaline soils. The aim of this work was to investigate the impact of ammonium and nitrate-N on P availability in corn rhizosphere in an acidic soil. Corn plants were grown in 12 L pots with soil or in plant growth containers to build a root mat. Nitrogen was applied at 240 mg kg-1 (first experiment) or 120 mg kg-1 (second experiment) as calcium nitrate and ammonium sulfate (plus dicianodiamide-DCD), combined with 0, 40, 80, 120 and 160 mg kg-1 of P doses. Corn was grown and clipped 45 days after emergence. In the first experiment adhering soil (rhizosphere soil) was separated from roots by gently shaking the soil off the roots, and in the second experiment the soil was sliced at 0-0.5; 0.5-1.0; 1.0-2.0; 2.0-3.0; 3.097 Nutrient Acquisition Tuesday 23 June – Poster session 4.0; 4.0-5.0; 5.0-10.0 mm from the root mat. Then Labile-P was determined using anion exchange resin (AER Pi) and 0.5 M sodium bicarbonate (BIC Pi and BIC Po). In presence of ammonium-N corn responded to higher P rates, with higher dry matter yields with 120 and 160 mg kg-1 of P. However, root dry matter was higher when nitrate-N was applied, resulting in higher root/shoot ratio. Ammonium-N resulted in lower labile-P in rhizosphere soil up to 80 mg kg-1 of P. Lower values of Labile-P were found from 0 to 0.5 mm from the root mat, but from 0.5 to 10.0 cm labil P was generally higher with nitrate-N. Ammonium-N increases P availability, corn growth and response to high P fertilization rates in acidic soils. 4 Phosphatase activity and phosphorus lability in the rhizospheric region of sugarcane genotypes Bruna Arruda*, Marcos Rodrigues, Gabriel Novoletti, Paulo Sergio Pavinato Esalq/USP, Brazil Phosphorus (P) is easily adsorbed on the soil solid phase, reducing its availability. When inorganic P (Pi) availability is low in the soil, plants and microorganisms exude organic compounds that can mineralize the organic P phase (Po) by phosphatases activity, supplying in this way plant demands. This study aimed to evaluate the dynamic of P in the rhizosphere of four distinct sugarcane genotypes under low and high P levels. The experiment was conducted in greenhouse at Department of Soil Science - ESALQ – USP, Piracicaba - Brazil, in a sandy soil following a randomized block design with four replications. It was compared the genotypes most used in Brazil: i) RB92-579; ii) RB85-5156; iii) RB86-7515; iv) RB96-6928 and also no plants (witness), with and without P fertilizer application (STP). At harvest, the soil adhered to the root system (rhizospheric soil) was sampled for analysis. When without plants, the bulk soil was sampled to compare to the rhizospheric one. It was determined the phosphatase enzyme activity in the soil freshly sampled and the fractions of labile P by Hedley´s procedure. The data were statistically analysed and means compared by LSD test at 5%. In the witness there was no difference between P fertilization in phosphatase activity, but in the rhizospheric soil of the four sugarcane genotypes when fertilized it was observed higher phosphatase activity than without fertilizer. No differences between genotypes or witness were observed without P fertilization. However, when P fertilized without plants it has presented a lower enzyme activity, 218 mg kg-1 h-1, much lower compared to the rhizospheric soil of the genotypes cultivated. Between genotypes, RB96-6928 had the lowest activity, 245 mg kg-1 h-1 compared to the other ones, whose mean was 280 mg kg-1 h-1 and do not show any difference between them. 98 Nutrient Acquisition Tuesday 23 June – Poster session 5 Effect of zinc solubilizing bacteria on growth promotion and zinc nutrition of wheat Azadeh Bapiri*1, Ahmad Asgharzadeh2, Hesam Mojallali3, Kazem Khavazi2, Ebrahim Pazira3 1 Department of Soil Science, Savadkooh Branch, Islamic Azad University, Savadkooh, Iran, 2Department of Soil Microbiology, Soil and Water Research Institute, Tehran, Iran, 3Department of Soil Science, Science and Research Branch, Islamic Azad University, Tehran, Iran Plant growth promoting rhizobacteria (PGPR) are the soil bacteria inhabiting around/on the root surface and are directly or indirectly involved in promoting plant growth and development via production and secretion of various regulatory chemicals in the vicinity of rhizosphere. One of the most important direct mechanisms is the increase of nutritional elements availability specially zinc in rhizosphere. Zinc deficiency is generally universalized in agricultural fields especially in calcareous soils. In this research, the solubilizing ability of one low-soluble zinc compound (ZnCO3) was studied by 11 Fluorescent Pseudomonads strains in a greenhouse experiment. The plant growth promotory properties such as Zn solubilization of the isolates were checked in a previous study. These isolates were showed effective in significantly increasing the root dry weight (97%), shoot dry weight (8.70%), number of tillers (9%), length panicle (4.25%) and the total Zn uptake (44.75%) as compared with the uninoculated control. The PGPR colonized wheat plants were more efficient in acquiring Zn from either added or indigenous source, than non-colonized plants. One of the hopefully methods to zinc deficiency remedy is to apply bacteria which are ability to soluble low- soluble and insoluble compounds of this element. 6 Role of native microorganisms in shoot biomass and p uptake of maize in low phosphorus soil Pratapbhanu Singh Bhadoria*, Ishita Paul Paul Indian Institute of Technology, Kharagpur, India To study the phosphate solubilizing microorganisms from maize rhizosphere in low P soil was carried out. Among several strains grown on Pikovskaya media modified with ferric phosphate, a pigmented, Gram negative strain of bacteria was isolated. A second pot culture was run in which factorial combinations of inocula from screened phosphate solubilizing bacteria (PSB) and vesicular arbuscular mycorrhiza (VAM) were applied in presence and absence of 100 mg phosphorus per kg soil. Samples of 21 days after sowing showed that PSB treatment led to 12% increase in shoot dry weight over control when aided by phosphorus application. On the other hand, interaction between VAM treatment and phosphorus application enhanced shoot phosphorus content 33% more than only phosphorus application. Without P fertilizer, PSB and VAM individually enhanced shoot weight by 94% and 49% respectively. When compared at 39 DAS, individual treatments with VAM and PSB gave 186% and 74% increase in shoot dry weight unaided by applied phosphorus while interactions between VAM and PSB in joint application with phosphorus gave 32% increase in shoot weight and 26% increase in shoot phosphorus concentration over only phosphorus application. Results showed that shoot weight and shoot phosphorus concentration were significantly enhanced at each additional interaction level. Determination of soil fractions of 99 Nutrient Acquisition Tuesday 23 June – Poster session aluminium, iron and calcium bound phosphorus led to the inference that the bacterial strain mobilized unavailable phosphorus and locked up excess soluble phosphorus in rhizosphere and slowly released it in soluble form as depletion zones formed round the roots; while the VAM fungi rapidly dissolved any mineral phosphorus to maintain steady uptake of phosphorus. The results suggest that microbial inhabitants of the soil favorably affect crop growth in the long term. This study recommends supplementation of phosphorus fertilizers applied to acid soils with dual inoculation of VAM and PSB. 7 Effect of different nitrogen sources on nitrogen dynamics, mineral elements and microbial communities in rhizosphere soil of different plant species Qingnan Chu*1, Toshihiro Watanabe2, Zhimin Sha3, Mitsuru Osaki2 1 Faculty of Agriculture, Hokkaido University, Japan, 2Hokkaido University, Japan, 3Shanghai Jiaotong University, China Little is known with respect to the variation of microbial metabolism, minerals and metabolites in rhizosphere of different plant species in response to different nitrogen sources. Soybean is able to perform nitrogen fixation and sorghum can utilize organic nitrogen efficiently. We therefore conducted the rhizobox experiment to investigate the effect of different nitrogen sources (ammonium sulfate, manure, without nitrogen fertilizer) on rhizosphere environment in soybean and sorghum. Plant roots and rhizosphere soil were separated from bulk soil with a nylon cloth (300 mesh, wire diameter 30μm) and different soil fractions moving laterally away from root compartment were analyzed for mineral elements, metabolites and microbial community. For plant growth, ammonium-source caused significant decrease of biomass and nitrogen accumulation in soybean nodules but no significant difference was detected for biomass or nitrogen concentration in other soybean organs. Manure application facilitated nitrogen accumulation in sorghum stems. Fertilization without nitrogen resulted in the significant biomass reduction in sorghum roots and leaves, nitrogen concentration in leaves. The Biolog EcoplateTM was analyzed for characterizing microbial communities in soil. By principle component analysis using the Biolog data, 55.6% of total variance was revealed by the first principle component and 22.4% by the second principle component. Two principle components both differentiate the effect on microbial communities in rhizosphere of soybean but did not between different nitrogen sources in sorghum rhizosphere. More efficient carbon source consumption activity excluding amino acid and malic acid was detected in soybean rhizosphere receiving manure application. The distribution of mineral elements, metabolites, microbial carbon source utilization, and their interactions in rhizosphere environment of soybean and sorghum under the influence of different nitrogen sources will be discussed. 100 Nutrient Acquisition Tuesday 23 June – Poster session 8 Phosphorus efflux rates from roots of Trifolium repens, T. uniflorum and T. repens × T. uniflorum interspecific hybrid clovers Jim Crush*, Shirley Nichols AgResearch, New Zealand Phosphorus efflux rates are six times higher in white clover (Trifolium repens) than in perennial ryegrass, and it has been suggested that this may contribute to the poor competitive ability for phosphorus of clover grown with ryegrass. Recent research has shown that some T. repens × Trifolium uniflorum interspecific hybrids backcrossed to T. repens can grow better at low to intermediate levels of soil phosphorus, than both of the parental species. T. uniflorum, a close, wild relative of white clover demonstrates adaptation to low soil phosphorus and intermittent phosphorus enrichment. No information exists on phosphorus efflux rates from T. uniflorum roots. The ecology of the species suggests these may be low and, if so, this may contribute to the observed improved growth of T. repens × T. uniflorum interspecific hybrids when soil phosphorus is strongly limiting for white clover. Phosphorus efflux rates from roots were measured on hydroponically grown plants of two white clover parental cultivars, two T. uniflorum seedlines, and six T. repens × T. uniflorum hybrids that had been back-crossed to the two white clover cultivars. Efflux of phosphorus was measurable for every clover type. One T. uniflorum line had a significantly (P<0.05) lower phosphorus efflux rate than the white clover cultivars (3.87 µg phosphorus g-1 root dry weight h-1 compared with 7.73 or 6.55 µg phosphorus) but the second T. uniflorum line did not differ from the white clovers. Efflux rates in the hybrid lines varied widely and included the highest, and lowest, values recorded. On average less than 0.4% of root phosphorus was lost over the four hour assay, suggesting that variation in phosphorus efflux rates should be a low priority target in breeding for improved phosphorus use efficiency in white clover. 9 Solubilization of inorganic phosphorus from biochar by phosphate-solubilizing microorganisms Aikaterini Efthymiou*, Mette Grønlund, Nelly Sophie Raymond, Dorette Sophie MüllerStöver, Iver Jakobsen University of Copenhagen, Denmark Global reserves of rock phosphate for fertilizer production are declining and this coincides with the generation of phosphorus rich organic wastes. There is a need for strategies to efficiently recycle the phosphorus in waste products. Organic wastes such as sewage sludge are rich in phosphorus and can be pyrolized, which ensures energy recovery, improved hygiene, volume reduction and a biochar product that is a potentially valuable phosphorus fertilizer. The use of phosphate-solubilizing microorganisms is a promising biotechnological strategy for the management of soil phosphorus resources, however, there is limited knowledge regarding their ability to solubilize phosphorus from recycling products. In this study, 14 phosphate solubilizing rhizosphere bacteria and fungi are screened for their ability to solubilize inorganic phosphate from sludge biochar, rock phosphate and Ca3(PO4)2 in vitro and the mechanisms of solubilization are being evaluated. Selected Penicillium spp. and Pseudomonas spp. strains result in higher solubilisation of phosphorus in the biochar. 101 Nutrient Acquisition Tuesday 23 June – Poster session The production of organic acids appears to be the main mechanism for the solubilization of inorganic phosphates and we are currently investigating how the production pattern varies with the combination of microorganism, carbon source and phosphate source. Furthermore, the selected strains are being evaluated for their ability to improve the plant availability of the biochar-phosphorus in pot experiments. 10 Effect of Thiobacillus on nutrient uptake and quality and quantity characteristics of two varieties of turnips forage (Brassica rapa L.) Seyed MohammadReza Ehteshami*1, Neda Moazeni2 1 University of Guilan, Iran, 2Islamic Azad University, Iran In order to investigate the effect of Thiobacillus bacteria on nutrient uptake and quality and quantity characteristics of the forage turnips (Brassica rapa L.), an experiment conducted as factorial arrangement with randomized complete block design with three replications at the research farm of Rice Research Institute Experimental. Investigated Factors include sulfur powder (use or non-use), sulfur-oxidizing bacteria (Thiobacillus use or non-use), Phosphorous fertilizer (use or non-use) and two varieties of forage turnips (white purple and white globe). Results showed that combined application of bio-fertilizer, phosphorus and sulfur powder increased LAI and morphological indices during the growing season. The highest of Chlorophyll a that is the most important role in the biomass production, abserred in the integrated treatment of sulfur powder Thiobacillus + cultivar. The highest of quality parameters, including dry matter digestibility, total ash and water-soluble carbohydrates in shoots and tubers of forage turnip obtained in sulfur powder + bacteria Thiobacillus + cultivar.The highest forage yield (tons / ha) obserred in integrated application of powdered sulfur + Thiobacillus + cultivar. Thiobacillus bacteria increases the oxidation of sulfur and sulfate reducing soil acidity and soil result in increased production, thus increasing the absorption of some nutrients, particulary phosphorus, is seeking. totally, taking into account the results of all evaluated traits, seems to Using a combination of powdered sulfur + Thiobacillus bacteria + phosphorus fertilizer and the chemical Fertilizers way for reducing consumption, Improve soil and improve plant nutrition and moving toward sustainable agriculture. 11 Potentials of rumen digesta as a source of organic manure in Owerri Imo Southeast Nigeria Ibiam Ikwuo Ekpe*1, S.C Asagwara Asagwara1, Emmanuel Nwannebuife Ogbodo2, Bethel Uzoho1, Samuel Okere1, Nnaemeka Anyanwu1, Maureen Nwaigwe3, Agatha Elem Ekpe4 1 Federal University of Technology, Nigeria, 2Ebonyi State University,, Nigeria, 3Alvan Ikoku Federal College of Education, Nigeria, 4Ministry of Lands and Housing,Ebonyi State, Nigeria An assessment of the potentials of rumen digesta as a source of organic manure in Owerri zone in Imo state was studied. the identification of all the Abattoirs in the city, the species of animals slaughtered, the number of each of the ruminant animal species slaughtered per 102 Nutrient Acquisition Tuesday 23 June – Poster session day/month and per year, total rumen digesta yield and possible economic returns from sale of bagged digesta per annum. The results revealed that a total of 65,160 cow, 28080 goat and 13320 sheep were slaughtered annually with a corresponding total rumen digesta yield of 2,952,720kg. When this quantity is processed and bagged into 50kg sizes and sold at ₦600.00/bag, a total financial return of ₦35,432,640.00K was possible. Rumen digesta after decomposition has the ability to replenish soil nutrients, provide cementing agents that bind soil particles together, increase soil moisture content, increase infiltration rate and general increase in crop yield. The quantity of rumen digesta produced in Owerri zone in Imo State is high. One can run a rumen digester enterprise by bagging and selling rumen digesta. The environment will be better, cleaner and healthier while at the same time increasing crop yield and providing employment opportunities to many youths. 12 Actinobacteria with phosphate solubilizing activity in tropical soils Marcela Franco-Correa*1, Luis Daniel Prada-Salcedo2, Claudia Liliana Cuervo Patiño3, Jose Salvador Montaña Lara3 1 Pontificia Universidad Javeriana, Colombia, 2UNIDIA, Colombia, 3Pontificia Universidad Javeriana/UNIDIA, Colombia The evaluation of biofertilizer products are expected to have the capacity to solubilizing nutrients, for this reason the aim of this work was to isolate actinobacteria able to release soluble phosphate in vitro and identify organic acids associated with this activity. We isolated 57 strains of actinobacteria, to screening the isolates of actinobacteria we used two qualitative assays to determine the efficiency of solubilization by measuring the halo of hydrolysis in a Pikovskaya’s agar plate. A second assay was performed on broth with NBRIP medium containing BPB following the protocol of Mehta and Nautiyal (2001). Finally to measure the release soluble phosphate by actinobacteria was quantified using insoluble Ca3(PO4)2 or AlPO4 as sole sources of P. Ten of these strains demonstrated by qualitative and quantitative assessments have a high and significant phosphorus solubilizing activity. Molecular identification was realized for the best isolates, revealed that belong to the genus Streptomyces, (T3A, T1C, T1H, T1J, T3C, P3E, F1A, F2A, V1E and V2B). Addition of new records for the country of the genera Kitasatospora (L3A) and Streptacidiphilus (M2A) was reported. The strain T3A have a very good activity with both sources of phosphate Ca3(PO4)2 or AlPO4, with activities of 500 mgP/l and 14 mgP/l respectively. The phosphorus solubilizing activity of this strain is associated to the release of organic acids, this was evidenced by a pH decrease, used of the sugar source in the medium and the presence of organic acids like gluconic acid, citric acid and oxalic acid identified by HPLC. 13 Transcriptomic data reveal new insights in Si effects on iron acquisition of rice Martin Hinrichs*1, Manfred K. Schenk2 1 Leibniz Universität Hannover, Germany, 2Institute of plant nutrition, Germany Silicon is one of the most abundant elements in the earth crust. Many plants are able to accumulate Si with several beneficial effects. One of these effects is the earlier formation of Casparian Bands (CB), as shown in rice, maize and onion roots. In rice it was demonstrated 103 Nutrient Acquisition Tuesday 23 June – Poster session that this anatomical change resulted in a decreased radial oxygen loss. Investigations of the molecular background of the enhanced CB formation by means of a custom made microarray with 265 genes putatively involved in CB formation resulted in the identification of twelve differentially regulated genes. To get a full insight in Si- induced genes a 45k gene chip approach was used. The experimental design consisted of four biological repeats and two technical repeats of rice plants grown in nutrient solution in a growth chamber. Sections of 26 cm behind the root tip of adventitious rice roots grown with Si (30 mg/l) and without Si (>3 mg/l) were harvested for RNA extraction. The transcriptomic data revealed regulation of genes related to the phenylpropanoidpathway. Surprisingly, also an up regulation of iron acquisition strategy II related genes like OsIRO2, OsNAS2, TOM1 or OsHRZ1 was observed. The Fe-concentration in shoot dry matter was reduced by half in Si fed plants. This shortage of Fe caused by Si supply was reflected in the transcriptomic data. Reason for reduced iron uptake could be a lower Fe flux into the apoplasm, because of enhanced CB development. 14 Performance and yield of maize in response to grinded-sawdust application Chris Ifeanyi Igbozuruike*1, Oliver Opara_nadi2 1 Abia State University, Uturu, Nigeria, 2Abia State University, Nigeria The effect of grinded sawdust on the performance of maize was carried out in 2013 at Federal University of Technology (FUTO) Owerri, Imo state Nigeria. Grinded sawdust was either applied single and in composite with other organic amendments. The treatments were T1 = control, T2= sawdust not grinded, T3=grinded sawdust, T4 =grinded sawdust + cow dung, T5= grinded sawdust + poultry droppings, T6 =grinded sawdust + pig waste, T7=grinded sawdust + NPK 10: 10: 10, T8=Cow dung alone, T9 =poultry manure alone, T10= pig waste alone, T11= NPK 10: 10: 10: alone. Both in single and composite applications, treatments were applied 10tons/ha except where NPK was involved which was applied npk 100kg/ha and sawdust 5tons/ha. These were laid out in randomized complete block design (RCBD). Growth and yield Parameters were taken. Grinded sawdust increased the growth and yield of maize relative to the control (T1) with highest effect occurring in T7 followed by T11, T5, T6, T4, T9, T8,T10, T3,T2. There were yield advantages of about 50 % where grinded sawdust were involved over 40 % of noninvolvement. Also about 35 % yield advantages occured where grinded sawdust were involved over no-grinded sawdust (05 %). Comparative assessment of the seasons saw residual growth and yield highest where grinded sawdust were involved, and also had yield advantages of 70 % (residual) to against 50 % (first season). The results were discussed in light of the nutritive ability of the various components in mixture. 104 Nutrient Acquisition Tuesday 23 June – Poster session 15 Investigation of the molecular mechanisms of phosphate starvation-induced growth of root hairs in Brassica carinata Thomas W. Kirchner*, Kim L. Rössig, Timo Lauterbach, Manfred K. Schenk Gottfried Wilhelm Leibniz Universität Hannover, Institute of Plant Nutrition, Germany The deficiency of inorganic P induces long root hairs in Brassica carinata cv. Bale, but not in cv. Bacho. The aim of the present study was the identification and characterization of genes, which are responsible for this different reaction. Therefore, root tips with a length of two centimeter were harvested from Bale and Bacho plants, grown in nutrient solutions under sufficient (1 mM) and deficient (0 mM) P conditions, respectively. Besides root hair length, genome-wide expression profiles of Bale and Bacho were compared under both conditions by Massive Analysis of cDNA Ends (MACE). Genes, which were differentially expressed in Bale, but not in Bacho, were defined as candidate genes. After validation of the expression of these genes by qRT-PCR, the genes were further characterized. To investigate, if these genes are involved in rather local or systemic signaling pathways, their expression was determined in samples obtained from a split-root experiment. Furthermore, nutrient specificity of gene expression was explored in potassium and nitrogen deficient plants. 16 Nutrient depletion from rhizosphere solution by maize grown in soil with longterm compost amendment Leandro Bortolon1, John Kovar*2, Michael Thompson3, Clesio Gianello4 1 EMBRAPA, Brazil, 2USDA Agricultural Research Service, USA, 3Iowa State University, USA, 4Federal University of Rio Grande do Sul, Brazil Improved understanding of rhizosphere chemistry will enhance our ability to model nutrient dynamics and on a broader scale, to develop effective management strategies for applied plant nutrients. With a controlled-climate study, we evaluated in situ changes in macronutrient concentrations in the rhizosphere of juvenile maize (Zea mays L.) grown in soil (fineloamy, mixed, superactive, mesic, Typic Hapludoll) amended with cattle (Bos taurus L.) manure compost. Soil was collected from two depths within a 10-yr. no-till study, so that nutrient stratification could be investigated. Maize seedlings at the two-leaf stage were transplanted into mini-rhizotrons, and grown for five days. Moist soil (550 g dry weight) collected from the 0-5 cm layer was placed in one compartment of a vertically divided mini- rhizotron, and soil from the 5-10 cm layer was placed in the other compartment. Micro- capillaries (15 per chamber) were used to collect rhizosphere and bulk soil solution at 24-h intervals. Samples were analyzed for pH and phosphorus (P), potassium (K), calcium (Ca), and magnesium (Mg) concentrations. Shoot dry matter production was similar in the two soil materials (0.65 g/plant in the 0-5 cm layer and 0.60 g/plant in the 5-10 cm layer), suggesting that nutrient availability was not limiting. Phosphorus concentrations in solution were similar in the two soil layers, but were lower at the root surface. Potassium concentrations were higher in the surface soil material, but did not change with distance from the root surface. Solution pH and Ca and Mg concentrations did not differ among the soil layers and distances from the root surface. From a practical perspective, results indicate that significant differences in rhizosphere solution chemistry can develop quickly. From a crop production point of view, the use of compost can be a means to reduce the inputs of mineral P and K fertilizer. 105 Nutrient Acquisition Tuesday 23 June – Poster session 17 Characterizing the variation of Brassica napus root system architecture: an in vitro screening approach László Kupcsik*, Hugues de Gernier, Christian Hermans Université libre de Bruxelles / LPGMP, Belgium Agricultural productivity relies quantitatively on nitrogen fertilization, which in excess is detrimental to the environment. Hence, ameliorating the nitrogen use efficiency of crops is an important approach for breeding future in order to reduce nitrate leaching and nitrous oxide emission via denitrification. We study Brassica napus (oilseed rape) which has small recovery of applied N in the seed. Our goal is to ameliorate its nitrogen use efficiency by redesigning a more branched root system that explores a larger soil volume for better N capture. As a first step, we examined the existing root trait variation between cultivars. We screened 57 cultivars from the Belgian Catalogue of Agricultural Plant Varieties upon in vitro culture at two N supplies (0.01 vs. 10 mM nitrate). After one week treatment, organ biomass and multiple root traits were measured. These outcome variables were interrelated, so principal component analysis was used to reduce data complexity. We showed that the first two principal components were responsible for 55% of the total variation. The primary source of variation (45%) was root system ramification, which consisted of length and number of lateral roots, and root biomass. The second most important component (10%) reflected the primary root morphology: its length and the position of the lateral root zone. These two principal components clearly separated certain groups of cultivars, and were also influenced by the nitrate availability in the culture medium. These results demonstrate that the most important difference between the surveyed cultivars lies indeed in the lateral expansion of their root system. The method described here also enables us to select contrasting genotypes, and thereby to investigate the genetic regulation of root system architecture in Brassica napus. 18 Maize nutrient uptake, rhizosphere pH and root architecture affected by soil compaction and application of biogas-digestate Daniela Piaz Barbosa Leal*1, Stephan Blossfeld2, Nicolai Jablonowski2 1 Forschungszentrum Jülich (IBG-2: Plant Sciences), Germany, 2Forschungszentrum Jülich, Germany Application of biogas-residues as fertilizers may promote enhanced cycling of nutrient resources. Plants obtain water and mineral nutrients from the soil due their capacity to develop extensive root systems. However, soil compaction may restrict deep root growth. Thus, we evaluated the effects of the biogas-digestate applied as a fertilizer on root architecture, rhizosphere pH, nutrient uptake and biomass development of maize growth when subjected to soil compaction, in comparison to NPK fertilizer application and noncompacted soil condition. Experimental factors were: fertilizers - biogas-digestate (40 Mg ha1 ), NPK (equivalent amount) and biogas-digestate (20 Mg ha-1) + NPK (equivalent amount); and soil compaction or non-compaction. The upper compacted layer (25 cm) received an equivalent pressure of 1.2 kg cm2. The biogas-digestate (elemental composition: 41.1% C, 106 Nutrient Acquisition Tuesday 23 June – Poster session 3.2% N, 1.5% P and 3.8% K) was composed of maize silage as the major feedstock, and minor amounts of chicken manure. Fertilizers were mixed into the soil (50%v. arable field soil + 50%v. peat substrate). Plants were grown for 28 days under greenhouse conditions (16h per day of light period, day/night temperature of 22°C/17°C and 60% of humidity). At harvest, the measurements included: shoot and root dry mass and nutrients content (C, N, P and K); leaf area; root architecture; and soil elemental analyses. Rhizosphere pH was measured with planar optodes along the experimental period. Preliminary results showed variation of the rhizosphere pH along the experimental period for all treatments. The lowest rhizosphere pH (4.2) was observed for the biogas-digestate + NPK/soil compaction treatment and the highest rhizosphere pH (5.3) was observed for the NPK/soil compaction treatment. Similar shoot and root biomass and nutrient uptake were observed between the treatments. These findings can enhance our understanding of rhizosphere pH dynamics and root architecture subjected to soil compaction aiming to improve soil fertilization practices. 19 Dynamics of competitive N and P uptake between intercropped wheat and faba bean in a calcareous soil Chunjie Li*, Haigang Li, Fusuo Zhang Center for Resources, Environment and Food Security (CREFS), China Agricultural University, China Cereal/legume intercropping is well known to be complementary and facilitative on N and P uptake because of N2 fixation and P mobilized by intercropped legumes. However, the dynamics of N and P uptake by intercropped plants during the co-growth period were rarely documented in a N- and P-deficient soil. The aim of present study is to reveal these dynamics. Wheat and faba bean were grown isolated or together in pots contained 2.0 kg N- and Pdeficient soil. Biomass, N and P content, carboxylates and phosphatases activity were measured in 13 destructive samplings. Data of biomass and N, P content were fitted to the logistic model to derive the cumulative biomass and N, P uptake curves and relative interaction index (RII). There was gradually increased competition for N and P uptake by intercropped wheat and faba bean along with growing time. In terms of dynamic competitive N uptake, the RII of wheat (from -0.06 to –0.26) was always higher than that of faba bean (from -0.10 to -0.20) during the whole growth stage, which may resulted from soil N depletion for wheat but faba bean has the ability to fix N2 from atmosphere (39 mg N/plant). Although competition for P uptake was observed between intercropped wheat and faba bean, the RII of faba bean (from -0.13 to -0.22) was hardly changed while the RII of wheat increased from -0.12 to -0.33 and decreased to -0.10 in the final sampling. From the analysis of rhizosphere processes, faba bean has higher ability to exudate carboxylates and acid phosphatase activity, which is helpful for it to adapt to the depletion of soil P and facilitate P uptake of wheat. This study provided experimental evidence about the dynamic N and P uptake by intercropped plants and the relationship with rhizosphere process. 107 Nutrient Acquisition Tuesday 23 June – Poster session 20 Physiological and biochemical responses of olive (Olea europea L.) to aluminum Adamo Domenico Rombolà1, Enrico Maria Valentino Liquori*2, Michele Di Foggia2, Gianfranco Filippini2, Annamaria Pisi2, Sergio Bonora2 1 University of Bologna - Department of Agricoltural Sciences, Italy, 2University of Bologna, Italy Crops native of alkaline-calcareous soils (e.g. barley) are more often cultivated on acid soils with high levels of soluble aluminum, which can cause heavy adverse effects on root growth and mineral nutrition. The aim of this work was to investigate the response mechanisms of olive to the presence of aluminum (Al) in the growing medium. Micro-propagated plants (cv Leccino) were grown in hydroponics with a 1/4 Hoagland solution adjusted to pH5. After a week of acclimation, plants were subjected, at the same pH, to 5 different Al concentrations: 0, 50, 250, 500 and 1000 µM. pH of the nutrient solution, stem and root length and number were daily measured. Nutrient solutions were renewed every 4 days. During the experiment, root exudates were collected and analyzed for organic acids and phenols (HPLC) and their complexes with Al (IR and RAMAN). After 1 month (end of the experiment), root apexes were excised and analyzed by SEM. Noteworthy, root and shoot growth occurred even at the highest Al concentration. Plants were able to increase the pH of nutrient solution. At SEM, the root epidermal layer of plants exposed to the highest Al concentration looked more compact, less defined and the morphology of the cells in cross section was altered. In contrast, root epidermis of low aluminum-treated plants was more preserved and very similar to that of controls. The main organic acids detected in root exudates were oxalic and formic. Their concentration resulted higher in 500 and 1000 µM Al treatments. Spectroscopic measures conducted on exhausted nutrient solutions showed the formation of Al-complexes, mainly under the form of Al-oxalate. 21 Morphological variation is the main adaptive response in maize roots to low phosphorus availability Haitao Liu*1, Chunjian Li2 1 China Agricultural University, China, 2Department of Plant Nutrition, China Agricultural University, China Different plants show different adaptive responses in roots to phosphorus (P) deficiency, include morphological and physiological traits. To clarify the adaptive mechanisms of maize root to low P availability, hydroponic experiments under P-sufficient (HP, 250 µM) and Pdeficient (LP, 1 µM) conditions were conducted, with two legumes (Vicia faba L. and Lupinus albus L.) as references. The results showed that LP treatment increased root biomass, root to shoot dry weight ratio (R/S), and total root length (TRL) of maize and faba bean with more dramatic change in maize than in faba bean in R/S and TRL. However, the same treatment for 12 d did not cause any variation in shoot and root growth in white lupin. The measured root exudation of carboxylates including malate and citrate and acid phosphatase (APase) activity on maize root surface showed no significant difference, while dramatically increased in both legumes when they grew under LP, compared with that under HP. Rhizospheric alkalization (agar technique) or net H+ influx (ion-selective electrode technique) on maize root surface was observed regardless of P levels. By contrary, increased rhizospheric acidification or net H+ efflux on root surface of both legumes were determined under LP compared with that under 108 Nutrient Acquisition Tuesday 23 June – Poster session HP. P inflow rate (mg P m-1 root length d-1) and P uptake efficiency (mg P m-1 root length) in legumes were remarkable high than maize under LP. However, P use efficiency (mg dry weight mg-1 P) of maize compared with both legumes was much high. The results suggest that differing from the physiological-type and morpho-physiological-type response in white and faba bean respectively, morphological variation is the main adaptive response to low phosphorus availability. 22 Relationship between phosphorous deficiency tolerance and rhizosphere management in Vicia genus Wissal M'sehli*, Haythem Mhadbi Centre de Biotechnologie de Borj Cedria CBBC, Tunisia Plants resort many adaptive strategies in response to abiotic environmental stresses such as soil phosphorous deficiency. These adaptive mechanisms include changes in the rhizosphere processes. Objective of the present study was to assess inter and intra-specific variations in Vicia genus to P deficiency response and to elucidate the implication of rhizospheric processes on phosphorus (P) deficiency tolerance. Experiments in controlled condition for Vicia sativa and two Tunisia Vicia faba varieties (Locale and Saber 2) were conducted. Seedlings were growing hydroponically under three treatments: + P (360 mM); - P (5 mM); + P +Bicarbonate (360 mM + 10 mM NaHCO3). It was revealed that P deficiency (-P and + bicarbonate treatments) restricted the whole plant RGR (the relative growth rate) in both species of vicia, particularly in – P treatment. This depressive effect was more pronounced in Vicia sativa (- 66.6%) than in Vicia Faba (var. Locale: -23.8%; var. Saber 2: - 44.8%). In our experiments, an increase in the root/shoot P content, the P acquisition and use efficiency was found in the two varieties of Vicia faba compared to Vicia sativa. The highest increase was found in the tolerant cultivar Locale both at low P levels and in the presence of bicarbonate. The rhizosphere management underling the differential responses of the vicia species and cultivars to P deficiency were explored. Exposure to P-deficient medium led to an increase in root biomass and the intracellular root phosphatase activity. This enhancement was more pronounced in the tolerant cultivar Locale. In our study, we proposed that root-induced acidification of the rhizosphere by proton release and the exudation of phosphatases acids as a mechanism of facilitation P acquisition in deficient medium. Our results showed that these two mechanisms were more pronounced in the tolerant cultivar Locale. 109 Nutrient Acquisition Tuesday 23 June – Poster session 23 The ability of beech seedlings (Fagus sylvatica) to shape their rhizosphere in order to cope with low phosphorus availability Sonia Meller*1, Beat Frey1, Emmanuel Frossard2, Marie Spohn3, Jörg Luster1 1 Swiss Federal Institute for Forest, Snow and Landscape Research WSL, Switzerland, 2Swiss Federal Institute of Technology ETH, Switzerland, 3Bayreuth University, Germany The German priority research program SPP 1685 aims to better understand the adaptation of forest ecosystems to low availability of nutrients, in particular P. The leading hypothesis is that an ecosystem facing depletion of mineral P sources appears to decrease P losses by a more effective P recycling. Within this program we address the question to what extent beech (Fagus sylvatica) is able to enhance the mobilisation of specific P sources in its rhizosphere. We hypothesize that direct adaptation mechanisms such as root exudation of carboxylates are governed by the internal demand of P status – and thus the overall availability of soil P - rather than by the actual relative availability of organic and inorganic P. Thus, P uptake efficiency could be enhanced, by investing carbon to stimulate the growth of P mobilising soil microbes that are better adapted to local nutrient availability. Beech seedlings were grown in different acid forest soils using rhizoboxes. Specific visually identified microzones, e.g. the rhizosphere around a root tip, were assessed for potential P mobilisation. We analysed P in soil solution („available“ soil P fractions), P mobilising substances / mechanisms (phosphatase activity, filter collection of carboxylates, pH changes using optodes) and composition of microbial communities (amplicon sequencing using Illumina MiSeq). In this presentation we will show the results from a first experiment with one-year old beech seedlings originating from a soil poor in mineral P growing in a soil relatively rich in both mineral and organic P. Preliminary data show lower phosphatase activity and pH around root tips and elongation zones of non-mycorrhizal long roots than in bulk soil. This may indicate that beech seedlings are not suffering from P limitation. This interpretation will be evaluated using data on rhizosphere modifications by roots and on phosphatase activity in plant cells as indicator of P status. 24 Different root exudation patterns affect the mobilization of Fe from soil minerals Rebeka Fijan1, Roberto Terzano2, Concetta Eliana Gattullo2, Youry Pii3, Luca Medici4, Stefano Cesco1, Tanja Mimmo*1 1 Free University of Bolzano, Italy, 2University of Bari, Italy, 3Free University of Verona, Italy, 4CNR, Italy Plants can cope with Fe deficiency by either acidifying the rhizosphere and enhancing the ferric chelate reductase activity (strategy I) or by releasing high affinity complexing compounds known as phytosiderophores (strategy II). In this research, tomato, barley and cucumber plants were grown hydroponically in an Fe free (–Fe) nutrient solution using the RHIZOtest. Fe-deficient (–Fe) plants were grown on a calcareous agricultural soil and root. Root exudates were determined quali-quantitatively using HPLC and colorimetric analyses. The accumulation of nutrients in plant tissues was measured by ICP-OES. Soil mineral modifications were assessed by XRD and SEM-EDX. 110 Nutrient Acquisition Tuesday 23 June – Poster session Root exudates obtained by the hydroponic system were higher than those obtained from the soil extracts; plant uptake and adsorption by soil particles could be, at least in part, responsible of this result. After a 6-day soil contact the plants show a visible recovery from Fe deficiency symptoms at leaf level suggesting the efficacy, of root exudate release in the mobilization of Fe into soluble soil forms and its uptake. Moreover, significant soil mineral modifications were observed. At last, organic ligands released in Fe-deficient soil conditions show very complex exudation patterns both in monocotyledonous and dicotyledonous plants. For instance, barley plants besides the dominant release of phytosiderophores also significant amounts of amino acids and traces of organic acids were found. The results of this research will enable to better understand the soil influence on the release of root exudates, the dynamics of mineral weathering in the rhizosphere and the capability of plant species to mobilize and take up poorly soluble Fe forms. 25 Potential of rhizospheric and endophytic microorganism to enhance P availability from organic and inorganic phosphate and phytohormones production Christiane Oliveira*1, Eliane Gomes1, Ivanildo Marriel1, Maycon Oliveira1, Ubiraci Lana1, Vitoria Palhares1, Astrid Beinhauer2, Ralf Greiner2 1 Embrapa Mayze and Sorghum, Brazil, 2Max-Rubner-Research Institute, Germany Rhizosphere microorganisms contribute significantly to solubilization of inorganic and organic forms of phosphorus fixed in soil, but only a few endophytic microorganisms are known in this respect. Since the amount of organic phosphorus in soil with no-tillage farming is increased and use of organic fonts, microorganisms efficient in the mineralization of organic P are more promising as plants inoculants. Apart from the P solubilizing abilities, the phosphate solubilizing microorganisms have the ability to produce plant growth hormones and enzymes for mineralization of organic phosphorus. The aim of this work was to investigate the potential of phosphate solubilization and mineralization of rhizosphere and endophytic microorganisms in order to select bacteria as plants inoculants. The production of phytase and phytohormone auxin indole-acetic acid (IAA) were also evaluated. Nine rhizobacteria from Embrapa collection and 113 endophytic bacteria isolated from maize roots, leaves and sap were evaluated for calcium and phytate phosphate bio-solubilization. The soluble P produced was quantified by a modified ammonium molybdate method after 3,6 and 9 days of bacterial growth. The most efficient phosphate solubilizing bacteria were identified based in the 16S rDNA sequence and evaluated for phytase and IAA production. IAA production was evaluated in a liquid culture medium supplemented with tryptophan. The rhizobacteria Bacillus subtillis strain B70 showed the highest calcium-P (167.8 mg P.mL-1) and phytate-P releasing (93,79 mg P.ml-1). The best intracellular phytase producer were identified as Pantoea sp. 1931 (85 mU.ml-1) and the above mentioned B. subtillis B70 (64mU.mL-1). The highest IAA production (150 µg.mL-1) was observed with the above mentioned Pantoea sp. 1931 and a non-identified endophytic bacteria (isolate 1913). It was concluded that bacterium from corn rhizosphere and endophytic microbiome have promising features of bio-solubilization and plant promotion of growth. Furthermore, Bacillus isolates were identified as promising candidates for future inoculation studies in plants. 111 Nutrient Acquisition Tuesday 23 June – Poster session 26 Response of maize mycorrhizas to green manure David Ortiz*, John Larsen Universidad Nacional Autónoma de México, Mexico Green manure crops are employed in most plant production systems, but still limited information is available on the effects of green manure on important plant growth promoting microorganisms such as arbuscular mycorrhizal fungi. In the present study the response of maize mycorrhizas to different types of green manure crops was examined in a greenhouse pot experiment. The experiment had a fully factorial design with two main factors: 1) Green manure with four levels (without, rape, oat, vetch) and 2) Mineral fertilization with two levels (without and with). All eight treatments had four replicates giving a total of thirty two experimental units. Green manure crops were grown in 3 liter pots with an agricultural soil low in P (4 ppm) mixed with sand (1:1, w/w) for six weeks and the green manure mixed in to the soil and left for eight weeks to decompose. Maize hybrid seeds were then sown in the soil with the different types of green manure and mineral fertilizer treatments and the maize plants were harvested after another ten-week period. Variables measured included plant growth performance (shoot and root dry weight), shoot nutrient content (N and P) and arbuscular mycorrhiza fungus root colonization. The green manures oat and rape reduced maize plant growth performance as well as total N and P shoot content, except for vetch, which on the contrary increased maize plant growth and total N and P shoot content in treatments without mineral fertilization. Oat and rape green manures also reduced the arbuscular mycorrhizal fungus root colonization with 20% and 49% respectively, whereas vetch had no effect on mycorrhiza formation. In conclusion, green manure crops not only affect growth and nutrition of maize, but also maize mycorrhiza formation, which seems to be important to consider when planning crop rotation programs. 27 Rhizospheric labile phosphorus fractions affected by phosphate source and filter cake Paulo Pavinato*, Bruna Arruda, Aline Santos, Marcos Rodrigues University of São Paulo/College of Agriculture Luiz de Queiroz, Brazil In tropical soils the adsorption of phosphorus (P) is higher onto Fe/Al oxides, reducing plant availability. In this way, the soil mass exploitation by the root system becomes much more important in rhizospheric region for adequate plant nutrition. This study aimed to evaluate the dynamic of labile and moderately P in the soil sugarcane rhizosphere under sources of phosphate and filter cake. The experiment was conducted in a sandy soil under greenhouse conditions at Department of Soil Science, ESALQ – USP, Piracicaba-SP-Brazil, in randomized blocks with four replications. The treatments followed a factorial design of 2x3: a) with and without filter cake (10 t DM ha-1) and b) no P fertilizer; soluble phosphate and rock phosphate (both at the rate of 180 mg soluble P2O5 kg-1 soil). Soil samples were obtained from the rhizospheric soil, 0-2 mm from the rhizoplane (root/soil separation system), using PVC columns with a horizontal mesh for root exclusion, in a tube with 0.15 m diameter and 0.30 m tall. These samples were collected 45 days after seedlings planting. It was determined in laboratory the labile P fractions by anion exchange resin (PAER) and bicarbonate (PBIC) extractors, and moderately labile fractions by NaOH and HCl. The data was analysed and means were compared by Tukey test at 5%. The average of PAER for all treatments was 10.0 112 Nutrient Acquisition Tuesday 23 June – Poster session mg kg-1 and PiBIC was 27.9 mg kg-1. There was a significant interaction between P source and filter cake use for both PAER and PBIC. Both fractions under soluble phosphate presented the highest values of labile P, regardless the absence or presence of filter cake. Without P fertilization and under rock phosphate, the presence of filter cake increased both labile P fractions. Other fractions will be presented and discussed at the conference. 28 Urea and nitrate: a reciprocal interaction affecting nitrogen acquisition by maize plants Laura Zanin1, Anita Zamboni2, Nicola Tomasi1, Zeno Varanini2, Stefano Cesco3, Roberto Pinton*1 1 University of Udine - Dip. Scienze Agrarie e Ambientali, Italy, 2University of Verona Dip. Biotecnologie, Italy, 3University of Bolzano - Faculty of Science and Technology, Italy Although urea (U) and nitrate (Ni) are the two main nitrogen forms applied as fertilizers in agriculture and occur concomitantly in soils, the reciprocal influence of these two nitrogen sources on the mechanisms of their acquisition is poorly understood. In the present work, the molecular and physiological aspects of U and Ni uptake were investigated in maize, a crop plant consuming large amounts of nitrogen. Morphological investigation allowed comparing the root architecture of plants fed for 1 week with U, Ni or U+Ni. Under U and U+Ni treatments, the presence of U in the external solution promoted the development of the root system showing an extensive proliferation and elongation of the roots, more than under Ni alone. This stimulatory effect might allow plants to increase the volume of the rhizospheric soil explored by the roots promoting nutrient acquisition. The capacity of maize plants to acquire urea and nitrate was evaluated by net-uptake assay at time intervals during 24 hours of root exposure to the two sources. Like Ni, also U is taken up by maize roots by an inducible high-affinity transport system. However, the induction of Ni and U high-affinity transport mechanisms were reciprocally limited when both sources were present in the root-external solution. Root transcriptomic analyses revealed that the simultaneous presence of U and Ni promoted the activation of both the plastidial GS2/GOGAT cycle and a cytosolic nitrogen-assimilation pathway (cytosolic glutamine synthetase and asparagine synthetase). Furthermore, the induction of the aromatic amino-acid synthesis, that would sustain the production of secondary metabolites, was observed. Based on these results, the transcriptional modulation induced by the presence of both nitrogen sources would determine an increase in nitrogen metabolism promoting a more efficient assimilation of taken-up nitrogen. 113 Nutrient Acquisition Tuesday 23 June – Poster session 29 Do nitrogen-fixing plants show higher root phosphatase activity on phosphoruspoor soils Guochen Kenny Png*1, Patrick Hayes1, Hans Lambers1, Benjamin Turner2, Etienne Laliberté3 1 The University of Western Australia, Australia, 2Smithsonian Tropical Research Institute, Panama, 3Institut de recherche biologie végétale, Canada Symbiotic nitrogen fixation in nitrogen-fixing plants may enhance plant performance on nitrogen-poor soils, but may not be favoured on phosphorus-poor soils, due to its high phosphorus costs. Yet surprisingly, nitrogen-fixing species are abundant in ecosystems with nitrogen-rich soils such as lowland tropical rainforests, where phosphorus is likely to limit plant growth. A prominent hypothesis seeking to explain this paradox is that nitrogen-fixing plants have a greater ability to acquire organic phosphorus through higher root phosphatase activity. However, evidence to support this hypothesis remains limited. We measured extracellular root phosphomonoesterase activity from 18 species of nitrogen-fixing (including legumes and two non-legume Allocasuarina species) and non-nitrogen-fixing species along a soil age gradient in Western Australia that shows a ~40-fold decline in total soil phosphorus from the youngest to the oldest soils, leading to some of the most phosphorus-impoverished soils found in any terrestrial ecosystem. In support of the hypothesis, we found that nitrogenfixing legumes had higher phosphomonoesterase activity than co-occurring non-legumes on all sites, and that the difference in phosphomonoesterase activity between legumes and nonlegumes increased with declining soil phosphorus concentration. However, phosphomonoesterase activities of both nitrogen-fixing Allocasuarina species (which form associations with Frankia) were consistently low across all soils which do not support the hypothesis. We conclude that the high root phosphatase activity of legumes on phosphoruspoor soils is likely a phylogenetically conserved trait that is not necessarily linked to their nitrogen-fixing ability. 30 Mobilization of trace elements by phytosiderophores in contaminated soils Markus Puschenreiter*1, Barbara Gruber1, Eva Oburger1, Walter W. Wenzel1, Yvonne Schindlegger1, Stephan Hann1, Walter D.C. Schenkeveld2, Stephan M. Kraemer2 1 University of Natural Resources and Life Sciences Vienna, Austria, 2University of Vienna, Austria Phytosiderophores (PS) are a group of root exudates released by grass species (Poaceae) for the mobilization of iron. PS have high complex stabilities with iron, but also with other trace elements, such as Zn, Cu and Ni. In trace element-contaminated soils, the PS-induced mobilization rate of Fe might be reduced by the presence of competing elements. A rhizotest experiment has been conducted with wheat exposed to four different contaminated soils. Three soils were enriched by Zn, Cd, and Pb due to atmospheric deposition in the vicinity of metal smelters, whereas the fourth soil was a serpentine soil naturally enriched by nickel. Prior to the soil contact period, the wheat plants were grown in Fe-deficient or normal conditions. In addition, a trace element mobilization experiment was carried out by adding different PS concentrations to an extraction solution. An increase of root exudation and of PS release was found for Fe-deficient wheat. Due to the enhanced PS concentrations in the rhizosphere, also the mobilization of other elements than Fe was increased, leading to higher 114 Nutrient Acquisition Tuesday 23 June – Poster session Zn and Cu concentrations in wheat shoots. Ni concentrations were higher in Fe-deficient wheat exposed to the serpentine soil. No changes were found for Cd and Pb. Likewise, in the extraction experiment Cd and Pb were hardly mobilized, whereas a significant solubilisation was found for Fe, Zn, Cu and Ni, in particular for PS concentrations of 100 and 1000 µMol/L in the extraction solution. Our results show that grass species grown on trace element-enriched soils might accumulate pollutants under conditions of Fe deficiency. 31 IPNM vis-à-vis soil-plant-water environment on diversified wetland crops for yield stability and quality - sustainable for rural livelihoods in India Anandamoy Puste*1, Kalyan Jana2, Krishnendu Ray2, Dorashila Nagarjuna2 1 Bidhan Chandra Krishi Viswavidyalaya (State Agricultural University), India, 2Bidhan Chandra Krishi Viswavidyalaya, India The science ‘IPNM (integrated plant nutrient management)’ combines & supply of all key, macro and micronutrients in proper doses, resulted good growth, production potentiality and quality of produces, more effective in developing country like India. With this significance importance, case studies on IPNM at farmers’ field during 2009 - 2012 on diversified valuable wetland crops [water chestnut, makhana, Cyrtosperma chamissomis, water-lily] under wetland ecosystem were undertaken in the different agro-zones. Major and other essential plant nutrients were combined and balanced through organic and inorganic sources including spray [F1 - Control, (farmer’s practice), F2 - F4 consisting of FYM/vermicompost/neem oilcake + NPK (60:30:30 kg ha–1), F5 - F7 composed of F2 - F4 along with Zn (chelamin) spray at 20, 40, 60 DAT (days after transplanting) + NPK spray at 60 DAT] and FYM @ 3.0 t and vermicompost and neem oilcake was used @ 0.2 t ha–1 and Zn + NPK @ 0.5% each], respectively. Root rhizosphere (growth, proliferations and establishment) as well as food quality were analyzed following standard methods. Results shows that yields of water chestnut, seeds of makhana, lati or stolons of Cyrtosperma and flower-stalks of water-lily (11.06, 2.98, 37.43 and 12.07 t ha–1, respectively) were remarkably influenced and highest by IPNM (F7) with optimum and balance form of plant nutrients that favours soil-plant-water interrelations including its quality (carbohydrates, starch, sugar, protein, minerals). Among various organic sources, neem oilcake performed better applied with proper dose and in time. From the experimental results pertaining to the field performance, it may be concluded that this vast unused wetlands, particularly in north-eastern part of the country may be utilized for aquatic crops, more precisely with balanced and economic sources of plant nutrients for crop productivity, making congenial soil-plant environment and sustainability to the farming communities in the regions. 115 Nutrient Acquisition Tuesday 23 June – Poster session 32 Yield improvement, indigenous nutrient supply, and nutrient use efficiency of hybrid maize cultivars in an inceptisol of West Bengal, India Krishnendu Ray*1, Hirak Banerjee2, Anandamoy Puste2, Sukamal Sarkar2 1 Bidhan Chandra Krishi Viswavidyalaya (State Agricultural University), India, 2Bidhan Chandra Krishi Viswavidyalaya, India Indian soil has greater diversity from poor to humus-rich, somewhere more scarce due to acute nutrient loss, even from inherent soil sources due to prevalence of sub-tropical to tropical environment. In this context, site-specific nutrient management (SSNM) has a pivotal role for supplying balanced and in-time nutrients for plant growth and development. Plant based approach for nutrient requirement determination requires omission plot technique (OPT), which facilitates development of decision support tool (DST) for implementation of SSNM at field levels and formulates fertilizer guidelines with a clear estimation of indigenous nutrient supply (INS). Present study was conducted during consecutive winter seasons of 2012-2014 to develop SSNM package for hybrid maize at the new alluvial soils of West Bengal, India. Target yield was fixed using the Nutrient Expert (NE) software. The experiment was laid out in strip-plot design with three hybrid maize cultivars (V1: P-3522, V2: P-3396 and V3: Rajkumar) in the horizontal strips and nine different fertilizer doses [F1: 50% RDF; F2: 75% RDF; F3: 100% RDF (Recommended dose of fertilizer 200:60:60 kg N: P2O5: K2O / ha is the State recommendation for hybrid maize); F4: 125% RDF; F5: 150% RDF; F6: 100% of P2O5 and K2O (N omission); F7: 100% of N and K2O (P omission); F8: 100% of N and P2O5 (K omission); F9: Absolute control (NPK omission)] in vertical strips replicated thrice. Nitrogen proved to be most limiting nutrient followed by K for P-3522 and P-3396, and P for Rajkumar. Results showed that the indigenous supply of N and P was highest for P-3396; however, P-3522 witnessed highest indigenous K supply. Application of 100% NPK (RDF) showed significantly higher growth, yield attributes and yields of the cultivars over omission plots, thus suggesting higher use efficiency of nutrients owing to better root plastic responses in the nutrient-rich plots. 33 Modelling nutrient uptake by roots of crops in fixed and variable volume of soil Juan Carlos Reginato*1, Jorge Luis Blengino1, Domingo Alberto Tarzia2 1 Departamento de Física, Facultad de Ciencias Exactas Físico-Químicas y Naturales, Universidad Nacional de Río Cuarto, Argentina, 2Departamento de Matemática– CONICET, Facultad de Ciencias Empresariales, Universidad Austral, Argentina Nutrient uptake by roots of crops in fixed (pots) and variable soil volume (field conditions) is modelled through a moving boundary problem. Thus, we evaluate the nutrient uptake for different situations and compare results with those obtained by 1D and 3D fixed boundary models. Nutrient uptake model is formulated through a one-dimensional moving boundary problem, both in the case of fixed soil volume fixed as the variable soil volume. To compute influxes on root surface, the problem is solved by immobilisation of the domain and application of the adaptive finite element method (FEM). To estimate the cumulative nutrient uptake a generalised and verified formula is used. The results obtained show that fixed and moving boundary models produce similar results for ions without limitations of availability, low variation of root density and low Peclet numbers. For low concentrations, large variations of root density and low numbers of Peclet the moving boundary produces better predictions 116 Nutrient Acquisition Tuesday 23 June – Poster session particularly for K. For P the moving boundary produces better predictions only at low concentrations being these predictions comparable to the obtained by 3D-dimensional architectural models. Obtained improvements are mainly due to three factors: the use of a generalised formula for the cumulative nutrient uptake, the use of a same dynamics to obtain influxes and the cumulative uptake in the moving boundary model, and the use the finite element method. Moreover, only our model satisfies the mass balance. Finally, in the light of these findings, conclusions drawn by previous papers could be reinterpreted. 34 Modelling nutrient uptake by roots of crops in fixed and variable volume of soil Juan Carlos Reginato*1, Jorge Luis Blengino1, Domingo Alberto Tarzia2 1 Universidad Nacional de Rio Cuarto, Argentina, 2Universidad Austral, Argentina Nutrient uptake by roots of crops in fixed (pots) and variable soil volume (field conditions) is modelled through a moving boundary problem. Thus, we evaluate the nutrient uptake for different situations and compare results with those obtained by 1D and 3D fixed boundary models. Nutrient uptake model is formulated through a one-dimensional moving boundary problem, both in the case of fixed soil volume fixed as the variable soil volume. To compute influxes on root surface, the problem is solved by immobilisation of the domain and application of the adaptive finite element method (FEM). To estimate the cumulative nutrient uptake a generalised and verified formula is used. The results obtained show that fixed and moving boundary models produce similar results for ions without limitations of availability, low variation of root density and low Peclet numbers. For low concentrations, large variations of root density and low numbers of Peclet the moving boundary produces better predictions particularly for K. For P the moving boundary produces better predictions only at low concentrations being these predictions comparable to the obtained by 3D-dimensional architectural models. Obtained improvements are mainly due to three factors: the use of a same dynamics to obtain influxes and the cumulative uptake in the moving boundary model, the use of a generalised formula for the cumulative nutrient uptake and the use the finite element method. Moreover, only our model satisfies the mass balance. Finally, in the light of these findings, conclusions drawn by previous papers could be reinterpreted. 35 Can Fe-chelating bacteria increase bioavailable Fe-content in lentil (Lens culinaris L.)? Mortuza Reza*, Fran Walley, Renato de Freitas University of Saskatchewan, Canada Iron (Fe) deficiency is the most common human nutrient deficiency worldwide, particularly in developing countries. Legumes including lentil (Lens culinaris L.) are an important and relatively inexpensive source of protein and are consumed worldwide. Iron bio-fortification of legumes can be an effective approach to increase Fe-bioavailability in areas affected by human Fe deficiencies. The objective of this study is to obtain natural Fe-chelating bacteria associated with lentil roots and assess their potential to improve crop Fe uptake. Lentil (cv. CDC Milestone) was grown in a growth chamber using soils collected from eight locations in Saskatchewan, Canada. Iron chelating bacteria (n=553) were isolated from the rhizosphere 117 Nutrient Acquisition Tuesday 23 June – Poster session (n=271) and endorhizosphere (n=282) using CAS selective medium and in vitro siderophore production was subsequently assessed in a Fe-free M9 minimal medium. Additionally, siderophore concentration was measured in cell free supernatants using a SideroTecTM Assay kit. Preliminary results indicate that selected isolates of both rhizosphere and endorhizosphere bacteria produced siderophores in concentrations ranging from 0.04 to 119.54 µg∙mL-1, although this population represented less than 1% of the total culturable heterotrophs. Of these, 14 isolates exhibited relatively high siderophore production i.e., >50 µg∙mL-1. These putative Fe-chelating bacteria isolates will be further identified using Sanger sequencing. Our results demonstrate that Fe-chelating bacteria colonize both the rhizosphere and endorhizosphere of lentil and likely play a role in the iron nutrition of the host plant. Studies designed to assess the effect of Fe-chelating bacteria on iron acquisition of lentil are currently in progress. 36 Potassium dynamics in ruzigrass rhizosphere as affected by K rates Ciro Rosolem*, Amanda Gomes São Paulo State University, Brazil Ruzigrass (Brachiaria ruzisiensis) has been used in Brazil as a cover crop because it has a vigorous root system and is efficient in cycling nutrients in the system. It has been shown to take up soil K considered as non-exchangeable. This study assessed exchangeable and nonexchangeable K dynamics in the rhizosphere of ruzigrass as affected by long term K fertilization. Samples were taken from an 11-year field experiment where plots have been fertilized with a total of zero to 1980 kg ha-1 of K2O as KCl. The soil is a Rhodic Hapludox with 210 g Kg-1 of clay, 78 % of kaolinite and less than 3 % of gibbsite. Ruzigrass was grown for 48 days in pots divided in half by a nylon screen. The top half was filled with sand and the bottom half with soil from each field plot. The soil from the bottom was sliced and analyzed for exchangeable (resin) and non-exchangeable (hot HNO3) K. Potassic fertilization had no effect on ruzigrass dry matter yields, but K uptake responded linearly to K rates. Rhizosphere exchangeable K was not affected in K absence. With 60 Kg ha-1 per year, K was depleted in the rhizosphere, which was increased with 120 Kg ha-1 of K per year or more. Nonexchangeable K was higher than in bulk soil where K fertilizer was applied. The K considered as non-exchangeable is affected by K fertilization and plays a role in ruzigrass nutrition. The use of high rates of K in a sandy loam rich in kaolinite results in a higher proportion of K transported in soil by mass flow, avoiding K depletion in the rhizosphere of ruzigrass. If K rate is high enough there is accumulation of the nutrient in the rhizosphere both in exchangeable and non-exchangeable forms. 118 Nutrient Acquisition Tuesday 23 June – Poster session 37 Rock phosphate solubilization by rhizobacteria and endophytic associated with maize (Zea mays L.) Ubiana Silva1, Luiz Freitas1, Julliane Medeiros2, Christiane Oliveira3, Sara Cuadros2, Eliane Gomes3, Vera Santos*1 1 Laboratory of Applied Microbiology, Microbiology Department, Institute of Biological Science, Federal University of Minas Gerais, Brazil, 2René Rachou Research Center, Brazil, 3National Research Center Mayze and Sorghum, Brazil Phosphate fertilizers are largely used for improving soil fertility. However, the use of less reactive phosphates requires the activity of phosphate solubilizing microorganisms. Some bacteria possess the ability to solubilize phosphate and increase the bioavailability of this nutrient in the soil-plant system. The aim of this work was to isolate and to characterize the phosphate solubilization potential of rhizobacteria and endophytic bacteria from Zea mays. Rhizobacteria were isolated from Zea mays rhizosphere soil cutivated without phosphate fertilizer and with Araxá rock phosphate. Thirty five bacteria obtained were evaluated for phosphate solubilization of rock Araxá jointly 10 endophytic bacteria obtained from collection of National Research Center mayze and Sorghum. Solubilized phosphate was quantified by molybdenum blue method at 880 nm after 72 h. The pH medium was also measured. Strains were identified based in the 16S rDNA gene. Furthermore, was evaluated the organic acid production for strains more efficient. The results were subjected to variance analysis and compared by Scott-Knott tests (P<0.05). Pantoea ananatis and Pantoea sp. showed major phosphate solubilization capacity, releasing 75 mg L -1. Other strains of the Enterobacter and Klebsiella genera, released 54 until 69 mg L -1, but some strains these genera showed also low potential for solubilization. Lower values of pH and acid production occurred in the samples with more phosphate released. It was found citric, acetic and latic acids. Citric acid was the most produced by efficient strains, aproximated 120 mg L -1. Thus, the phosphate solubilization capacity was influenced by strain evaluated. Moreover, the acid production certainly contributed to the dissolution of phosphate from rock Araxá, as has been reported in several experiments. 38 A window for iron uptake in Strategy II iron acquisition Walter Schenkeveld*1, Martin Walter1, Eva Oburger2, Yvonne Schindlegger2, Stephan Hann2, Markus Puschenreiter2, Stephan Kraemer1 1 University of Vienna, Austria, 2Boku, Austria Graminaceous plants (grasses including staple crops like wheat and barley) exude so-called phytosiderophores (PS) into the rhizosphere in a diurnal pulse release for acquiring the essential micronutrient iron (Fe). Phytosiderophores are chelating agents that are able to bind and solubilize soil-Fe and make it available for plant uptake. When PS are released, they participate in rhizosphere processes including adsorption, degradation and complexation of Fe and competing metals. In a series of batch interaction experiments with a calcareous clay soil from Santomera (Spain) to which the PS 2’-deoxymugineic acid (DMA) was added at various concentrations, the influence of rhizosphere processes on Fe mobilization was explored. It was found that these processes draw up a time and concentration window, during which DMA increases the 119 Nutrient Acquisition Tuesday 23 June – Poster session Fe concentration above background levels: the window for Fe uptake. This size of the window proved to strongly depend on the DMA concentration. The window was further constrained in time by biodegradation and metal competition (particularly with copper (Cu)). Our work shows that the extent to which PS are able to provide plants with Fe depends on environmental and rhizosphere soil conditions, which affect thermodynamically and kinetically controlled processes. The influence of these conditions on the success or failure of Strategy II Fe acquisition can be interpreted by means of the window concept. In this context, it will be discussed how soil parameters including temperature, rhizosphere pH, electrolyte type and concentration, and abundance of soil reactive surfaces and competing metals affect the effectiveness of Strategy II Fe acquisition. 39 Biostimulant and PGPR treatment efficacy for P-solubilisation and winter wheat growth in large pot trial Shekhar Sharma*1, Chris Selby1, David Nelson1, Graham McCollum2, Eugene Carmichael2, Fiona Clarke2, J R Rao2 1 Agri-Food Biosciences Institute, United Kingdom, 2AFBI, United Kingdom The efficacy of phosphorus (P)-solubilising bacterial soil treatments comprising commercial formulations ABiTEP Bacillus simplex + FZB42 Bacillus amyloliquefaciens, foliar biostimulant Alga Vyt, Agreges and P solubiliser bacteria were evaluated. The treatments were assessed for their effects on growth and grain yield of winter wheat grown on either a regularly cultivated soil or a nutritionally poorer soil that has not received fertiliser for several years. Seventy litre pots were filled with each soil type, and winter wheat seeds were sown to a depth of 1.5 cm in open air. The soil P solubilising bacterial preparations were applied as a 20 ml spray to the soil surface of each pot at the two leaf stage and later on fully grown plants. The AFBI-P solubiliser was an equal mixture comprising two Rahnella aquatalis and one Burkholderia spp. selected from low input soils. The foliar biostimulant Alga Vyt (1.5 g/l) was applied weekly, 4 times. Soil samples were collected for analysis. The addition of bacteria and treatment with foliar biostimulant did not exhibit apparent growth/development enhancement of wheat plants. A total count of P-solubilisers at the start and end of trial remained unchanged. However, closer scrutiny of the results indicated that wheat matured marginally faster on the standard input soil compared to the low input soil without increasing ultimate grain yield. Such subtle interactions between soil type and the biostimulants on plant growth and development that were often not recognised by standard plant analyses prompted us to study P-solubiliser activities in the soil/root rhizosphere. A protocol using an anion exchange membrane technique demonstrated that membrane matrix yielded better soil P-recovery than traditional direct soil chemical extraction methods and may impact the final analyses for validation of results. 120 Nutrient Acquisition Tuesday 23 June – Poster session 40 Rhizosphere interactions – Competition and environment regulate plant nitrogen acquisition Judy Simon* University of Konstanz, Germany Plants have developed different mechanisms to optimise the utilisation of limited N resources, for example by N uptake via mycorrhiza symbionts and / or acquisition of organic N sources. However, the complex interactions between different vegetation components, mycorrhizal fungi, and free-living soil microorganisms with regard to the competition for N in the rhizosphere are currently only little understood. Thus, the research presented here provides new insights into the understanding of the processes involved in the regulation of belowground competition for N between the different players in temperate forest ecosystems (i.e. seedlings vs. adult trees, different woody species, tree vs. soil microorganisms). For example, competition for N is regulated by N source (organic vs. inorganic), signals in the rhizosphere (i.e. nitric oxide and/or carbon dioxide as well as rhizodeposition) and shifts depending on environmental factors (i.e. soil N availability). Furthermore, plants acquire different N forms (i.e. organic, inorganic) when growing in competition. In addition, plants do not have to rely on the support of soil microorganisms, such as mycorrhizal fungi, to gain access to organic nitrogen but could have a competitive advantage over other plant species via exudation of proteases to obtain organic nitrogen directly from the humus layer, thus forgoing the microbial loop of nitrogen turnover processes. 41 Simultaneous use of gypsum and legume crops reduces soil pH and alters soil carbon on rhizosphere of calcareous alkaline soils Ehsan Tavakkoli*1, Pichu Rengasamy2, Glenn McDonald2 1 NSW Department of Primary Industries, Wagga Wagga Agricultural Institute, Australia, 2Waite Research Institute, School of Agriculture Food and Wine, The University of Adelaide, Australia Large areas of the world’s arable land have alkaline soil and in the southern cropping region of Australia alkaline sodic soils, often with high concentrations of carbonate salts, are widespread. High pH presents many nutritional challenges to crop production that influence both direct and indirectly, the accumulation of soil organic C. The use of legumes in conjunction with the supplying additional Ca to soils has the potential to lower pH on highly alkaline soils. To examine the potential to lower pH, two short-term rotation experiments at South Australia and Victoria were established on alkaline calcareous soils. In addition an experiment was conducted to investigate plant and gypsum effects on the chemistry of the rhizosphere in a growth chamber experiment The experiments examined the effects of legume biomass and gypsum application on soil pH, soil C and productivity of wheat grown in the following year. Applying 2.5 t/ha of gypsum reduced soil pH by between 0.2 and 0.4 units to a depth of 30 cm after 12 months. The changes in pH were associated with a decline in dissolved organic C at both sites and with higher organic C. Biomass production by legumes did not affect these soil properties. We also used C (1s) near-edge x-ray absorption fine structure (NEXAFS) and synchrotron-based Fourier transform infrared (FTIR) spectroscopy to investigate the C and N speciation in the rhizosphere of alkaline soils. Together, this 121 Nutrient Acquisition Tuesday 23 June – Poster session information is not only useful in terms of understanding the mechanisms responsible for SOC retention/dissolution but also in terms of nutrient bioavailability, as this is strongly related to SOC speciation. In this presentation, analyses of SOM chemistry in relation to its speciation and stability and implications for sequestration and nutrient cycling will be discussed. 42 Zinc biosorption mediated by exopolysaccharide in plant growth promoting Pseudomonas fluorescens Psd Anamika Upadhyay*1, Mandira Kochar2, Manchikatla Venkat Rajam3, Sheela Srivastava3 1 Department of genetics, University of Delhi South Campus, India, 2TERI Deakin Nanobiotechnology Centre, The Energy and Resources Institute (TERI),, India, 3Department of Genetics, University of Delhi South Campus, India Zinc is an essential trace element required for the growth and development of all organisms including bacteria, but may exert toxic effects at higher concentrations. Extracellular biosorption is the mechanism of zinc resistance in the plant growth promoting bacterium, Pseudomonas fluorescens strain Psd. Our study aims to identify the key players involved in Zn2+ biosorption and their effect on the plant-growth promoting potential of the strain. The increased Zn2+ accumulation by the strain was accompanied by an increase in the various plant-growth promoting parameters like siderophore, phenazine production and phosphatesolubilization. IAA production, however, was found to decrease with increasing Zn2+ concentrations. Higher Zn2+ accumulation also led to increase in total exopolysaccharide content. The compositions of the exopolysaccharides were measured using different biochemical assays and FT-IR spectroscopy. Expression analysis was carried out for genes responsible for biosynthesis of two of the important exopolysaccharides, alginate (alg8) and psl (psl). Quantitative RT-PCR analysis revealed an increase in expression of alg8 with increased Zn2+ accumulation by the strain, whereas pslA expression levels remained unchanged. Biofilm formation was also found to be increased with enhanced exopolysaccharide biosynthesis. Our study explores the property of plant-growth promoting rhizobacteria to secrete exopolysaccharides under metal-rich/contaminated environments, which aids in bacterial survival as well as its attachment to the root surface ensuring better plant-rhizobacterial association. 43 Exudation mechanisms in strawberry plants as affected by iron and phosphorus deficiency Fabio Valentinuzzi*, Youry Pii, Stefano Cesco, Tanja Mimmo Free University of Bozen-Bolzano, Italy Strawberries are among berries a very popular fruit, especially for their beneficial effects for human health. However, their bioactive compound content is strictly related to the nutritional status of the plant and might be affected by nutritional disorders. To overcome the nutrient shortages plants evolved different mechanisms, which often involve the release of low molecular weight compounds known as root exudates. The biochemical and molecular 122 Nutrient Acquisition Tuesday 23 June – Poster session mechanisms underlying root exudation and its regulation are yet still poorly known, in particular in woody plants. The aim of this work was therefore to characterize the pattern of root exudation of strawberry plants grown either in phosphorous (P) or iron (Fe) deficiency. In addition, phylogenetic analyses were carried out to identify the sequences of putative genes involved in the release of organic acids and protons. This is the first study that shows the release of citric acid from strawberry roots. In particular, this release significantly increased with time being +250% and +300% higher in Fe and P deficient plants, respectively, compared to the control. Furthermore, concomitantly, a significant acidification of the growth medium was observed in both treatments. Phylogenetic analyses allowed the identification of five proteins which clustered in the MATE sub-family characterized for the transport of citrate in response to Fe and aluminum stresses. Moreover, also putative strawberry PM H+-ATPases were identified. In conclusion the response differs in function of the nutritional disorder (Fe vs P) and the extent of organic acids release together with the extrusion of protons depends on plant growth stage and treatment. These results will allow managing agronomical practices towards a more sustainable agriculture. 44 Arbuscular mycorrhizal fungi increase plant phosphate uptake from phytate Xinxin Wang*1, Ellis Hoffland1, Gu Feng2, Thomas Kuyper1 1 Wageningen University, Netherlands, 2China Agricultural University, China Phytate constitutes the largest pool of soil organic phosphorus (P). Increased P nutrition of arbuscular mycorrhizal plants after phytate addition has been repeatedly reported; however, earlier studies assessed acid phosphatase rather than phytase as an indication of mycorrhizal fungi-mediated phytate use. Earlier studies also disregarded the potential effect of phytate addition on mobilization of mineral P, due to competitive adsorption of phytate and concomitant desorption of phosphate ions on metal (hydr-)oxides. We investigated the effect of mycorrhizal hyphae-mediated phytase activity on phytate mineralization and subsequent P transfer to the host plant. Two maize (Zea mays L.) genotypes, non-inoculated or inoculated with the AM fungi Funneliformis mosseae or Claroideoglomus etunicatum, were grown in twocompartment rhizoboxes. The soil in the hyphal compartment was supplemented with 20, 100 and 200 mg P kg-1 soil as calcium phytate. We measured activity of phytase and acid phosphatase in the hyphal compartment, hyphal length density, P uptake and plant biomass. Our results showed: (1) phytate addition increased phytase and acid phosphatase activity, and resulted in larger P uptake and plant biomass; (2) increases in P uptake and biomass were correlated with phytase but not with acid phosphatase; (3) lower phytate addition rate increased, but higher rates decreased hyphal length density. We calculated that at the higher rate, competitive desorption of phosphate is substantial, providing an explanation for the decrease in hyphal length density at high phytate addition P. We conclude that P from phytate can be a major source for arbuscular mycorrhizal plants. 123 Nutrient Acquisition Tuesday 23 June – Poster session 45 Growth condition and P mobilizing properties of cluster roots of Helicia cochinchinensis (Proteaceae) at Miyajima Island, Japan Jun Wasaki*, Taiki Yamauchi, Jin Takahashi, Hayato Maruyama, Shinji Uchida, Seiji Mukai, Hiromi Tsubota Hiroshima University, Japan Some specific plants form bottle brush-like root structures, so-called cluster roots, during P deficiency. The family Proteaceae are well known group for cluster root-forming plants. Helicia cochinchinensis Lour. is only one species present in Japan among the Proteaceae, and is distributed in the range from southwest Japan to the Indochina Peninsula. It is still unknown what properties of soils distributing H. cochinchinensis and whether this species forms cluster roots. The aim of this study is to investigate the growth conditions, morphological properties of roots, and phosphate mobilizing abilities of H. cochinchinensis growing in Miyajima Island, Hiroshima, Japan. Available P in soils growing H. cochinchinensis grew was very poor; ranged 0.46 – 3.7 mg-P/kg-soil (Olsen-P). We found cluster roots from P deficit H. cochinchinensis in natural and hydroponic conditions. It was shown that acid phosphatase activity was increased in the rhizosphere soil. Activity staining revealed a higher activity of acid phosphatase in half-part from root tip of each rootlet. Strong decrease of pH in the rhizosphere of matured cluster roots was also shown, suggesting that the cluster root enhanced organic acid exudation like other Proteaceae plants. P concentration of leaves of H. cochinchinensis was low level; those of mature and senesced leaves ranged 0.34 - 0.69 mgP/gDW and 0.15 - 0.29 mg-P/gDW, respectively. This range was some extent high than that of other Proteaceae plants growing under very low P soils in Western Australia, but it was very low level among general plant species. It was concluded that the P mobilizing capacity of cluster roots and the tolerance to low P content supported the growth of H. cochinchinensis under low P soil at Miyajima Island. 46 Ionomic variation in plant species growing in various soil environments Toshihiro Watanabe*1, Yoshinobu Kusumoto2, Sayaka Morita2, Tomoyo Koyanagi2, Mitsuru Osaki1, Syuntaro Hiradate2 1 Laboratory of Plant Nutrition, Research Faculty of Agriculture, Hokkaido University, Japan, 2National Institute for Agro-Environmental Sciences (NIAES), Japan Ionomics is the study of all metal, metalloid, and nonmetal accumulation in living organisms. Plant ionomics has been applied to various types of study in the last decade. For example, many ionomics studies have been conducted to investigate the relationship between the ionome and genome in model plants, such as Arabidopsis thaliana or Lotus japonicus. In our previous report of a broad survey of various plant species, we investigated higher-level phylogenetic effects influencing the leaf composition of a wide range of elements. However, the effect of changes in the rhizosphere environment on the plant ionome in different species has hardly been investigated. Therefore, in the present study, we analyzed concentrations of 25 elements in leaves of various plant species growing under different soil environments to evaluate the ionomic variation among different soil environments and to compare the variation among different plant species. Twenty-three herbaceous species were sampled at 80 sites in Shiozuka highland, Japan. Concentrations of 25 elements in leaves were determined by inductively coupled plasma mass spectrometry. Chemical properties of soils, 124 Nutrient Acquisition Tuesday 23 June – Poster session including the ammonium acetate- or water-extractable concentrations of the same 25 elements, at 80 sites were also analyzed. The coefficients of variation in concentration of each element among different sampling sites varied among plant species, particularly in nonessential elements, which showed higher coefficients of variation than those in essential ones. No significant correlations were observed between concentration in leaf and extractable concentrations in soil in most elements. However, in some elements, a positive correlation was observed under some phylogenetic influence. With respect to the correlation between the vegetation cover rate and extractable concentration of each element in soil, Miscanthus sinensis showed significant positive correlations for aluminum, sodium, and several heavy metal elements and a negative correlation for phosphorus, possibly associated with its higher adaptation capacity to acidic soil. 47 Tree roots: plastic in architectural traits, but not in morphological and biotic traits Monique Weemstra*1, Frank Sterck1, Eric Visser2, Thomas Kuyper1, Frits Mohren1, Liesje Mommer1 1 Wageningen University, Netherlands, 2Radboud University Nijmegen, Netherlands Functional traits are important drivers of resource acquisition and growth of trees. Yet, knowledge on plastic responses in belowground traits is still limited, impeding our understanding of relationships between root traits, nutrient acquisition, and tree growth. This study is among the first to examine plasticity in root functional traits of large forest trees. We compared root traits of beech and Norway spruce trees on a nutrient-rich clay soil and on a nutrient-poor sandy soil in the Netherlands. We hypothesized that trees adjust their roots to nutrient-poor conditions relative to rich soils, by 1) increasing the size of their root system, 2) changing their root morphology, and 3) investing more in mycorrhizal symbioses. Trees of both species had a higher root length density and higher root growth rates at the poor than at the rich soils. Root morphology (specific root length and root tissue density) however, did not differ between soils. Mycorrhizal mycelium abundance was also larger at the sandy soil, but this difference was not significant when correcting for the higher root length present. On the root tips, however, we observed a larger number of mycorrhizal exploration types that form mycelia on the sandy soil, while we only found mycorrhizal exploration types that do not make mycelia on the clay soil. Our study demonstrates that on poor soils, trees adapt their roots by enhancing the length of their root system, and not by changing their root morphology. The larger number of mycorrhizal mycelia on the poor soils seemed partly driven by larger root length, but the larger abundance of mycorrhizal types that form mycelia may point at increased tree investments in mycorrhizal fungi that are carbon-costly but more efficient in soil exploration and resource uptake. 125 Nutrient Acquisition Tuesday 23 June – Poster session 48 Root morphological traits and rhizosphere processes of maize with increasing shoot P concentration on a calcareous soil Zhihui Wen*, Jianbo Shen, Haigang Li China Agricultural University, Department of Plant Nutrition, China The modifications of root growth and rhizosphere processes in maize (Zea mays L.) induced by phosphorus (P) deficiency were well documented in previous studies. However, the continuous changes of these modifications with increasing shoot P concentration have not been investigated. We hypothesized that the changes in modifications of root growth differ from rhizosphere processes with increasing shoot P concentration. A broad-gradient shoot P concentration of maize (0.96-3.96 mg g-1) was founded in a pot experiment by application of a series of P fertilizer rates (0, 2.5, 5, 10, 25, 50, 75, 150, 300, 600 and 1200 mg P kg-1 soil) on a calcareous soil. Root morphology and rhizosphere processes were determined after 28 day growth. Maize reached to maximum shoot biomass when shoot P concentration was 2-3 mg g-1, above which biomass did not increase any more, and below which biomass reduced because of P deficiency. Total root-length (TRL), specific root-length (SRL) and the proportion of fine root (diameter < 0.2 mm) to total root-length (PFR) showed the opposite patterns. TRL, SRL and PFR of maize maintained a low level at P sufficient status (>2-3 mg g-1), and these root parameters showed an increasing trend with decreasing P supply until shoot P reduced to 1.1-1.3 mg g-1, below which these traits exhibited a rapid decrease. In contrast, rhizosphere acidification, acid-phosphatase activity and carboxylates accumulation in rhizosphere were enhanced with increasing shoot P concentration, and had no reduction even if shoot P concentration reached up to 4.96 mg g-1 that was seldom found in field. In conclusions, we observed the modification trajectory of root morphological traits differs from rhizosphere processes of maize with increasing shoot P concentration, indicating that maize had a strong root morphological alteration in response to P deficiency compared to root exudation in the rhizosphere. 49 Improving phosphorus acquisition efficiency in rice: The role of mycorrhizal versus plant-induced uptake mechanisms Matthias Wissuwa*1, Lidia Campos Soriano2, Blanca San Segundo2, Stephanie WattsWilliams3, Asako Mori1, Erina Shimamura1, Vincent Pujol1, Adam Price4 1 JIRCAS, Japan, 2CRAG, Spain, 3Monash University, Australia, 4University of Aberdeen, United Kingdom Phosphorus (P) is a limited resource and price hikes have made P fertilizers increasingly costly. Breeding P-efficient rice varieties has therefore become an objective in many national rice breeding programs. We previously identified the major P uptake QTL Pup1, and showed that the underlying gene OsPSTOL1 improves P uptake via higher root growth. Here we report on the identification of novel P uptake donors and QTLs for enhanced P acquisition efficiency (PAE), the capacity for improved P uptake per given unit root size. Field experiments on a P fixing soil identified sets of rice genotypes contrasting for total P uptake and PAE. High PAE genotypes took up as much as 40 µg P m-2 root surface area compared to an average of 18 µg P m-2. To examine whether arbuscular mycorrhizae (AM) are associated 126 Nutrient Acquisition Tuesday 23 June – Poster session with PAE we used three AM detection methods: a common staining method, quantification of AM-specific DNA within rice roots, and detection of OsPT11 gene expression (OsPT11 being a P transporter specifically expressed upon infection of rice roots by AM). Root samples taken from the field did show OsPT11 expression in all genotypes, possibly indicating P transfer from fungus to plant occurring very commonly. The staining and DNA methods also detected the presence of AM in all roots but colonization rates varied from 10-70 % between genotypes. However, this variation was not associated with genotypic differences in P uptake. Additional pot experiments in soil-sand mixtures with rock-phosphate or phytate as main P sources confirmed genotypic differences in P uptake. Together these results suggest plant induced changes in the rhizosphere rather than associations with AM to be underlying causes of superior PAE. Whether this rhizosphere effect is related to root exudation and direct P solubilization or to indirect effects via beneficial changes in soil microbial populations is discussed. 50 Effect of different levels of organic manure and chemical fertilizer inputs on crop yield and soil nutrient content in winter wheat-summer maize rotation system Zhang Xiaoning*, Liu Ruili, Zhang Hongyan China Agricultural University, China Since the autumn of 2007, a field experiment was carried out in Quzhou Experimental Station of China Agricultural University to compare the different influences of organic manure (OM) and chemical fertilizer (CF) fertilization on crop yield and soil nutrient content in winter wheat (WW) - summer maize(SM) rotation system. There were one CK treatment, four OM treatments and four CF treatments in the experiment. The OM treatments were fertilized by cattle manure of 3, 6, 9 and 12t DM/crop/year/ha, respectively. By contrast, the CF treatments were designed with the same NPK input as that in the four OM treatments, respectively. The preliminary results showed that the 7 year averaged WW grain yields in all the OM treatments were significantly lower than that in the CF treatments at the same NPK input level. At the same NPK input levels, the 7 year averaged SM grain yield in the OM treatments of 3 and 6 t DM/ha/growing season were significantly lower than that of CF treatments, although the 7 year average grain yield in the OM of 9 and 12 t DM OM/ha/growing season were the same as that in CF treatments.The yield differences between OM and CF treatments at the same NPK input level reduced gradually with planting years. After 7 year rotation, soil organic matter content increased with the increase of OM inputs but did not changed with the increase of CF input. Olsen-P, available K and Nmin content in soil increased with the increasing of both OM and CF inputs. Generally, the Olsen-P content in OM treatments were higher than that in CF treatments while Nmin content in CF treatment were higher than that in OM treatments with the same NPK input level. 127 Nutrient Acquisition Tuesday 23 June – Poster session 51 Effect of phytase adsorbed on montmorillonite on the root activity of Malus hupehensis and soil enzymes and microorganism in root-zone Hongqiang Yang*, Pingping Yang Shandong Agricultural University, China Phytate is rich in phosphorus, but it cannot easily be absorbed by the root directly. Phytase can catalytic the release of phosphorus in phytate, but it is easily inactivated directly applied into the soil. Montmorillonite has good adsorption properties of enzyme and other organic matter. In this experiment, the phytase adsorbed on montmorillonite (PAM) was prepared by the liquid phytase sprayed onto montmorillonite and added it to the soil of potted Malus hupehensis seedlings. The result show that the montmorillonite can effectively adsorb the phytase and delay the decline of phytase activity in soil, the activity of soil phytase by PAM treated was 18.97% higher than that by liquid phytase treated on the 50th day after two kinds of phytase added. The activity of soil phosphatase, urease and dehydrogenase, the quantity of soil microorganisms, the content of available phosphorus in root-zone soil, and the root activity of Malus hupehensis seedlings were all increased significantly after applying PAM and liquid phytase to the soil, and all of which increasing with the increase of the applying dosage. With the time prolonged, the activity of soil phytase became stable after dropping rapidly, the activity of soil phosphatase rose gradually in the mass, and the content of available phosphorus declined slowly after rising fast. In addition, PAM suggested more effective and longer validity than that of liquid phytase. 52 Iron deficiency in Zea mays: transcriptomic changes and acquisition of different Fe-sources Laura Zanin*1, Silvia Venuti1, Nicola Tomasi1, Anita Zamboni2, Stefano Cesco3, Zeno Varanini2, Roberto Pinton1 1 University of Udine - Dip. Scienze Agrarie e Ambientali, Italy, 2University of Verona Dip. Biotecnologie, Italy, 3University of Bolzano - Faculty of Science and Technology, Italy Plants react to iron (Fe) deficiency using different adaptive strategies. Under limited Fe availability maize, a model species for Strategy II plants, improves Fe acquisition through the release of phytosiderophores (PS) into the rhizosphere and the subsequent uptake of the FePS complex into root cells. Aim of the present work was to study, at physiological and transcriptional level, the Fe-deficient response in a commercial maize hybrid widely grown in Europe. Eleven-days-old maize plants (Zea mays L., PR33T56) were fed for 6 days with a nutrient solution containing 0 (Fe-deficient) or 100 μM Fe (Fe-sufficient). At the end of the growing period, visible symptoms of Fe deficiency were observed in the interveinal yellowing of young leaves and proliferation of lateral roots and root hairs, with increase in the diameter of the sub-apical root zone. Transcriptomic analysis on root tissues (NimbleGen microarray) revealed that the Fe-deficiency modulated the expression levels of 376 genes (289 up- and 87 down-regulated, respectively). As expected, roots of Fe-deficient maize plants 128 Nutrient Acquisition Tuesday 23 June – Poster session overexpressed genes involved in the synthesis and release of 2’-deoxymugineic acid (the main PS released by maize roots). Moreover, a strong modulation of those genes involved in the regulatory aspects, Fe translocation, root morphological modification, in primary metabolic pathways and in hormonal metabolism were induced by the nutritional stress. The capacity of maize plants to respond to Fe-deficiency was further evaluated exposing roots to soluble or poorly soluble Fe-sources for up to 24 hours. Real time RT-PCR analyses and 59Fe uptake experiments showed that the mechanisms involved in Fe acquisition were induced by the nutritional stress; while the pathways involved in the translocation and distribution of the micronutrient within the plant were not yet activated in Fe-deficient plants. 53 The influence of phosphorus sources on the growth and rhizosphere soil characteristics of two wheat (Triticum aestivum L.) genotypes Shuxiang Zhang*, Xiaoyng zhan Institute of Agricultural Resources and Regional Planning, Chinese Academy of Agricultural Sciences, China A rhizobox experiment was conducted to investigate the effects of phosphorus (P) sources on the rhizosphere soil characteristics of two wheat genotypes, namely Xiaoyan54 (P-efficient) and Jing411 (P-inefficient), which were colonized with arbuscular mycorrhizal fungus and grown on Cumulic Haplustoll. The four P sources included a control (no P), OP (organic P: Naphytate), IP (KH2PO4) and OPIP (Na-phytate plus KH2PO4). The results showed that when no P was added, the shoot biomass for Xiaoyan54 was significantly higher than Jing 411 by 28%. The rhizosphere soil acid phosphatase activity was significantly depressed when inorganic P was added; the acid phosphatase activity for Xiaoyan54 was higher than that of Jing411 in most of the layers for all the P sources. Inorganic and organic P fertilizer applications did not significantly impact the soil pH relative to the control, and the pH did not lead to significant variations between the two genotypes. In conclusion, significantly greater soil acid phosphatase activity but not acidification promoted greater shoot biomass in Xiaoyan54 than Jing411 when no P was added. The rhizosphere soil pH was not affected whether choosing different wheat genotypes or various P sources, but the greater acid phosphatase activity was found in the control and OP treatment compared with IP and OPIP, meanwhile the acid phosphatase activity of Xiaoyan54 was higher compared with Jing411 for all the P sources. The response mechanism of rhizosphere soil acidification is different from that of acid phosphatase exudation to different P sources application under arbuscular mycorrhizal fungus inoculation. 129 Nutrient Acquisition Tuesday 23 June – Poster session 54 Positive interactions between a P-mobilizing (Embothrium coccineum) and a Nfixing species (Sophora microphylla) on volcanic soils in Southern South America Alejandra Zúñiga-Feest*1, Susana Valle2, Angela Bustos-Salazar2, Mabel Delgado2, Frida Piper3 1 Instituto de Ciencias Ambientales y Evolutivas, Universidad Austral de Chile, Chile, 2Universidad Austral de Chile, Chile, 3Centro de Investigación en Ecosistemas de la Patagonia, Chile Young volcanic soils are frequently found in Southern South America. These soils have limited availability of phosphorus (P) and nitrogen (N), however some species have developed root physiological strategies for P and N acquisition; Sophora microphylla (Fabaceae) is a tree with symbiotic N-fixing bacteria, and Embothrium coccineum (Proteaceae) is a tree with cluster roots (CR) which exude high rates of carboxylates (malate and citrate) improving their P acquisition. It have been proposed that P-mobilizing species improve nutrient acquisition to neighbors; however, scarce evidence exists for Southern South America environments. We studied the interaction between E. coccineum and S. microphylla to test if both species growing together have higher growth and better nutritional status than growing separately. Seedlings of both species were grown on volcanic or alluvial soils (with or without P and N limitations, respectively), for one growing season, in pots and watering at field capacity, at full sun light conditions in Valdivia (40°S), Chile. We evaluated photosynthesis rate, growth, biomass distribution, and N and P content in plants and in rhizospheric and non-rhizospheric soils. Both species grew less in volcanic than alluvial soil, without differences between “growing together” or “separately”. E. coccineum showed higher photosynthesis rate in alluvial soil, higher size of individual cluster roots in volcanic soil and higher cluster roots biomass distribution in alluvial soil, when it grew together. Higher P concentration was observed in roots of S. microphylla growing together. E. coccineum showed higher N and P content growing together than separately in alluvial soil. The rhizospheric soil P availability increased in pots of E. coccineum, in both separate and combined treatments. These results imply that P-movilizing and N-fixing species grown in a combined system could lead to facilitation. 130 Nutrient Acquisition Tuesday 23 June – Poster session Human Pathogens in the Rhizosphere 55 First pictures and key features support a role of Pseudomonas fluorescens injectisomes in interactions with rhizosphere or clinical hosts Dorian Bergeau*1, Xavier Latour2, Sylvie Mazurier3, Marie-Laure Follet-Gueye4, Victorien Decoin1, Annabelle Merieau1, Nicole Orange5, Marc Feuilloley5, Philippe Lemanceau3, Maïté Vicré-Gibouin4, Xavier Latour5 1 Normandie Université, Laboratoire de Microbiologie Signaux et Microenvironnement EA 4312 & IRIB, France, 2Normandy University (University of Rouen), France, 3INRA Dijon, UMR 1347 Agroécologie, France, 4Normandie Université, Laboratoire de Glycobiologie et Matrice Extracellulaire Végétale - EA 4358, France, 5Normandie Université, Laboratoire de Microbiologie Signaux et Microenvironnement - EA 4312, France Many plant and animal pathogenic Gram negative bacteria use a type III secretion system (T3SS) as a molecular syringe to inject effector proteins directly into the host cell. Two different classes of T3SSs are well described in Pseudomonas, a bacterial genus in which members are frequently encountered in the rhizosphere: (i) a short needle involved in translocation of exotoxins by the human pathogen Pseudomonas aeruginosa and, (ii) a long and flexible pilus able to pierce the thick plant cell wall carried by phytopathogenic P. syringae. T3SSs were detected more recently in the species P. fluorescens, including in (i) strains considered as saprotrophic or as plant growth-promoting rhizobacteria, known as devoid of pathogenic potential, and (ii) strains associated with clinical cases (blood bacteremia, cystic fibrosis). The role of these secretion systems in P. fluorescens strains is still unclear. In order to elucidate the nature of the interactions mediated by P. fluorescens T3SSs, the structure of the secretion system was apprehended in representative strains by transmission electron microscopy in comparison with P. aeruginosa and P. syringae T3SSs. For these strains, ability to interact with animal and plant cells, and expression of T3SS genes by qPCR were also studied. The results together with ecological data reveal potential interactions mediated by T3SS with rhizospheric partners, noteworthy at the level of myccorhizosphere, and with animal cells. These results support the hypothesis that P. fluorescens T3SSs would be implied in interactions with human host, thereby promoting opportunistic infections, but also with fungi leading to beneficial effects on the plant. To the best of our knowledge, we unveil here the first electron micrographs of T3SS apparatus observed within the P. fluorescens species. 131 Human Pathogens in the Rhizosphere Tuesday 23 June – Poster session 56 Factors influencing the fate of human pathogens in the plant environment Eva Fornefeld*, Kornelia Smalla Julius Kühn-Institut, Federal Research Centre for Cultivated Plants (JKI), Institute for Epidemiology and Pathogen Diagnostics, Germany Diseases caused by human pathogens (HP) on vegetables are increasingly reported. However, so far knowledge is scarce about factors influencing the persistence of HP in the plant environment. In this study, we analysed the influence of sludge, preadaptation of HP and presence of nematodes on the survival of HP and their internalisation into plants. Therefore, two greenhouse experiments with lettuce plants (Lactuca sativa ‘Tizian’) and Salmonella enterica serovar Typhimurium LT2 were performed to analyse the effects of the presence of sludge, preadaptation of HP and coinoculation of plant-parasitic nematodes. Preadaptation of HP was simulated by cultivation under non-optimal conditions in a self-made culture medium. The first experiment included six different treatments: soil amended with sludge or not and each inoculated with Salmonella, preadapted Salmonella or no inoculum. Soil was sampled regularly, and numbers of Salmonella were monitored using culture-dependent and -independent methods. Salmonella counts in soil decreased overall from about 106 to 103 per g dry soil within five weeks. Direct plating showed significantly higher numbers of Salmonella in the treatment with preadapted Salmonella without sludge compared to the other treatments from 10 days post inoculation (dpi). Significant differences were confirmed using qPCR for 14 and 21 dpi. Salmonella was detected in soils up to 175 dpi using PCR-Southern blot hybridisation and enrichment culture and subsequent plating. In a second greenhouse experiment, the presence of the plant-parasitic nematodes Meloidogyne hapla and Pratylenchus penetrans did not lead to internalisation of Salmonella through lettuce roots. Despite a rapid decline of Salmonella in soil our data showed a long-term survival at low abundance. Preadaptation promoted the survival in soil while the presence of sludge reduced the survival. The presence of nematodes did not seem to promote internalisation of Salmonella into lettuce roots. 57 Transcriptomic analysis of enterohaemorrhagic Escherichia coli O157:H7 in response to plant extracts Robert Jackson*1, Louise Birse2, Ian Toth3, Carol Wagstaff1, Simon Andrews1, Yannick Rossez3, Nicola Holden3 1 University of Reading, United Kingdom, 2James Hutton Institute and University of Reading, United Kingdom, 3James Hutton Institute, United Kingdom Enterohaemorrhagic Escherichia coli (EHEC) are a group of food and contact-borne pathogens responsible for haemorrhagic colitis. The bacteria can be transmitted by contaminated meat, but importantly, also by plants and have been involved in many largescale produce associated outbreaks. The bacteria can use plants as a secondary host, where they associate with both the leaves and the roots. Colonisation in the roots & rhizosphere of plants is thought to be the main habitat for colonisation. Global gene expression changes of EHEC O157:H7 strain Sakai were measured in response to plant extracts such as leaf lysates, root exudates and leaf cell wall polysaccharides from spinach and lettuce. A significant change in expression of 17% of genes on exposure to leaf lysates of spinach was determined by microarray. A more selected response was seen to spinach leaf cell wall polysaccharides 132 Human Pathogens in the Rhizosphere Tuesday 23 June – Poster session with only a 1.5% change. In contrast, when exposed to lettuce leaf cell wall polysaccharides a higher change of 4.8% was seen, indicating plant species-specific responses from the bacteria. The different metabolic pathways induced upon exposure to polysaccharides of the different plant species involved the utilization of plant-specific polysaccharides. As well as this, several pathways were induced that are involved in different aspects of adaptation of the bacteria to the plant environment in many of conditions tested. Expression of selected differentially regulated genes was validated in planta by qPCR, and targeted for functional analysis by mutagenesis. These results provide insight to the colonisation of different plants by human pathogenic EHEC and provide the foundation for further work towards developing strategies to mitigate pathogen contamination of food produce. 58 T3SS and virulence markers highlight similarities and differences between human and plant-associated Pseudomonas fluorescens related isolates Sylvie Mazurier1, Annabelle Merieau2, Dorian Bergeau2, Victorien Decoin2, Daniel Sperandio2, Alexandre Crépin2, Corinne Barbey2, Katy Jeannot3, Maïté Vicré-Gibouin2, Patrick Plésiat3, Xavier Latour2, Philippe Lemanceau*1 1 INRA, France, 2Normandie Université, France, 3Université de Franche-Comté, France Opportunistic bacterial pathogens are a major concern for human health since they are able to cause or aggravate serious clinical conditions. Among these bacteria, isolates related to the species Pseudomonas fluorescens remain little known although they appear to be regularly found associated with bacteremia or in the respiratory tract of patients suffering from cystic fibrosis. Meanwhile, knowledge on environmental P. fluorescens related isolates is increasing, especially that on their ability to interact with eukaryotic hosts via Type III Secretion Systems (T3SS). Plant-associated and clinical isolates were compared for a series of genotypic and phenotypic traits and two bacterial groups were delineated. Isolates of the first group only included clinical isolates from blood infections and were showed on the basis of 16S rRNA phylogeny to belong to the P. putida complex and not to P. fluorescens as initially identified in hospitals. T3SS sequences of these isolates were highly conserved and belonged to the Ysc-T3SS family known to include P. aeruginosa. The presence Ysc-T3SS genes in clinical isolates belonging to the P. putida complex, described here for the first time, is proposed to be used for the identification of these bacteria which are potential human pathogens as supported by virulence assay using an amoeba. The second group included plant-associated and clinical isolates from patients suffering from cystic fibrosis. They all belonged to P. fluorescens and harbored T3SS genes of Hrp1-T3SS family which is commonly found in plant-associated P. fluorescens. Clinical and plant-associated isolates of this group could be differentiated neither on their 16S rRNA and T3SS gene phylogenies, nor on their virulence, some plant-associated isolates expressing a positive response in virulence assay. Therefore, no identification test could be proposed to discriminate clinical and environmental isolates in this second group commonly found in the rhizosphere. 133 Human Pathogens in the Rhizosphere Tuesday 23 June – Poster session 59 Investigation of the colonization of the root system of Festuca arundinacea by Listeria monocytogenes Pascal Piveteau*1, Gilles Boussemart1, Laurent Gal2, Dominique Garmyn1 1 Université de Bourgogne, France, 2AgroSup, France Listeria monocytogenes is a ubiquitous opportunistic human pathogen responsible for listeriosis, a life-threatening food-borne disease. L. monocytogenes is detected in many habitats spanning from the farm environment (soil, vegetation) to the food industry (foodstuff, drains, working surfaces) and the gastrointestinal tract of animals and humans. Contamination of raw farm products such as vegetables and crops is a major problem as a source of contamination of raw and processed foods. A better understanding of the adaptation of L. monocytogenes to soil and plants is clearly necessary. In the present study, we investigated the rhizosphere as a potential habitat of L. monocytogenes. Soil mesocosm experiments clearly demonstrated that the presence of Festuca arundinacea improved survival of L. monocytogenes. This prompted us to investigate colonisation of the root system of this monocotyledon by L. monocytogenes. Plants were grown aseptically on Hoagland plates and incubated in plant culture cabinets. The root system was inoculated with L. monocytogenes after 7 days of growth of the seedlings. Growth was observed and L. monocytogenes reached around 2x106 CFU/root after 4 days of incubation. The population then stabilised until the end of the experiment, 7 days after inoculation. Organisation of the listerial cells was followed in situ by confocal microscopy during colonisation. Plantassociated biofilms were observed during colonisation of the root system. We further investigated intrinsic factors that could facilitate root colonization. Interestingly, InlA and InlB, two surface proteins of the internalin family were involved during root-associated growth of L. monocytogenes. These two surface proteins are known virulence factors required for adhesion to enterocytes during infection of the mammalian host. Further evidence points out to a global reshaping of the physiology of L. monocytogenes during plant-associated growth. 134 Human Pathogens in the Rhizosphere Tuesday 23 June – Poster session Below and Above Ground Interactions 61 The effect of grazing on root morphology and mycorrhiza formation in simulated ryegrass (Lolium rigidum) pasture Lynette Abbott*1, Jing-Wei Fan2, Zakaria Solaiman1, Bede Mickan1, Yan-Lei Du2, FengMin Li2 1 University of Western Australia, Australia, 2Institute of Arid Agroecology, Lanzhou University, China Defoliation of shoot tissue by grazing can alter the structure and function of grassland ecosystems. Grazing modifies morphology of plant root systems. There are contradictory reports of impacts of defoliation on colonization of roots by arbuscular mycorrhizal (AM) fungi. Defoliation has been shown to both decrease and increase mycorrhiza formation when assessed as % root length colonized by AM fungi. We investigated mycorrhiza formation in simulated pasture swards of ryegrass (Lolium rigidum cv. Wimmera) grown for 6 months in a glasshouse in response to three defoliation regimes. Plants were sampled 5 times during this period. Our objective was to determine effects of moderate and severe defoliation. The correspondence of several root traits (root length, root mass and specific root length) to nutrient uptake, rhizosphere soluble carbon and mycorrhiza formation was evaluated in response to defoliation. We hypothesized that (i) shoot growth would be reduced in the severe defoliation treatment and less so with moderate defoliation; (ii) root biomass and root length would be reduced by both defoliation treatments, (iii) defoliation would increase rhizosphere soluble carbon, and (iv) defoliation intensity would alter the relative abundance of AM fungi in roots. Both shoot and root biomass decreased with defoliation intensity and remained stable from 87 days after sowing, but both continually increased in non-defoliated plants. Defoliation had a major impact on root traits. The proportion of roots colonized by AM fungi (measured as % root length colonized) increased with defoliation severity. In contrast, the length of root that was colonized by AM fungi was reduced by defoliation. Soluble carbon concentration and microbial biomass carbon in rhizosphere soil were reduced by defoliation intensity. Root length colonized by AM fungi, rather than % root length colonized by AM fungi, was more closely related to soluble carbon and soil nutrient status of rhizosphere soil. 62 Biotic plant-soil feedback effects on germination and seedling growth of agricultural and semi-natural plant species Janna Barel*, Jingjue Wang, Gerlinde De Deyn Wageningen University, Netherlands Plant-soil feedbacks co-determine plant productivity and vegetation dynamics and are increasingly being studied. However, studies on germination and seedling growth are underrepresented. Moreover, comparisons between agro-ecosystem and natural grassland plant-soil feedbacks are rare although they could offer valuable mechanistic insights. We aimed to understand biotic plant-soil feedback effects on germination rate and success, relative growth rate and biomass production. We studied four agricultural plants and four 135 Below and Above Ground Interactions Tuesday 23 June – Poster session grassland congeners (respectively Lolium perenne, Vicia sativa, Raphanus sativus, Cichorium endivia and Festuca rubra, Vicia cracca, Raphanus raphanistrum, Cichorium intybus). In a climate cabinet we tested soil legacy effects on germination and whether crops germinate better than their semi-natural counterparts. In a greenhouse, we verified if crops grow faster, and whether soil legacy left by conspecific plants would suppress plant productivity more than soil legacy of heterospecific plants. Overall, crops were expected to respond more negatively to conspecific soil conditioning than semi-natural species, and more positive to heterospecific conditioning. As hypothesized, crops germinated faster with a higher success rate than semi-natural plants. However, soil legacy didn’t affect germination. Relative growth rate differed between plant species but not between agricultural and semi-natural species. Soil legacy effects on biomass production gave contrasting results: agricultural species responded similarly to soil conditioning, with soil legacies of C. endivia and L. perenne promoting plant growth in comparison to unconditioned soil. While, semi-natural plant productivity was driven by the identity of the response plant. Overall our results demonstrate that soil legacy effects on subsequent plant performance can depend on both the identity of the preceding species generating the legacy and on the species that responds to the legacy. In agro-ecosystems preceding species play a main role, while the responding species are important determinants in the semi-natural system. This indicates that the two systems don’t seem to mirror each other’s functioning. 63 Can plant growth-promoting rhizobacteria mitigate P-starvation stress in Brachypodium distachyon? Caroline Baudson*, Benjamin M. Delory, Patrick du Jardin, Pierre Delaplace University of Liège, Gembloux Agro-Bio Tech, Belgium Phosphorus (P) is a macronutrient essential to plants but mainly present in unavailable forms in soils. Although it is abundant, P is often a limiting factor for crop productivity due to its poor mobility. While the fossil P reserves are diminishing, new strategies allowing a better exploitation of the soil P resources have to be developed. In this regard, biotic interactions occurring between plant roots and soil microorganisms are increasingly considered. Among those microorganisms, beneficial rhizospheric bacteria (called plant growth-promoting rhizobacteria, PGPR), are able to mitigate P deficiency stress in plants by several mechanisms (e.g. solubilization and mineralization of unavailable P forms, modification of root morphogenesis). In this context, the current research project aims at studying the effects of three PGPR strains (Bacillus subtilis AP305-GB03, Pseudomonas fluorescens Pf29Arp and Azotobacter vinelandii A60 - F08 19) on Brachypodium distachyon (L.) Beauv. Bd21 response to P deficiency, in an ex-vitro co-cultivation context. The first step of the project consisted in investigating Bd21 response to P deficiency and allowed the identification of contrasted P conditions, from strong stress to optimal growth conditions. Based on these results, P limiting conditions are used to reveal potential mitigation effects of the selected PGPR strains on P deficiency stress in Bd21. Plants and bacteria are co-cultivated in sand with different P supplies. Biomass production and allocation, root system architecture parameters and P 136 Below and Above Ground Interactions Tuesday 23 June – Poster session content will be measured in Bd21. This study will help us to find out if there is a beneficial bacterial activity improving P availability, P uptake capacity and plant development in our experimental conditions. The presented results will be discussed from fundamental and applied viewpoint. 64 A new methodology for assessing root amino acid exudation in soil and interactions with the rhizosphere Hélène Bobille*1, Anis L. Limami2, Richard Robins3, Gaétan le Floch4, Joëlle Fustec1 1 Ecole Supérieure d'Agriculture (LEVA), France, 2University of Angers, IRHS, France, 3University of Nantes, CEISAM, France, 4University of Brest, LUBEM, France Legumes release substantial amounts of nitrogen into soil via rhizodeposition and constitute a sustainable source of nitrogen incorporation in cropping systems. In addition, the exudation of small molecular weight compounds such as amino acids may significantly contribute to defining the composition of rhizospheric microbial communities. Our aim is to unravel the adaptive response of legumes to changes in the rhizosphere in terms of amino acids exudates, since root exudate composition is likely to be influenced by biotic and abiotic factors. However, studying exudation from plants grown in unsterilized soil is challenging due to rapid uptake of exudated molecules by the microorganisms or their sorption on soil mineral particles. In the present work, we have developed a new methodology that allows us to establish relationships between exudated free amino acids in the rhizosphere, the physiological status of the plant, the substrate type and the microflora. Medicago truncatula plants were grown in controlled conditions, either in unsterilized soil, or in sterilized sand and amino acids were extracted in formic acid (2 mM). Biomass and diversity of the microbial communities were also characterized. Results on the legume rhizosphere showed that the presence of a plant substantially modifies the total amino acid content of the rhizospheric soil. The highest content of amino acids was observed at the end of the vegetative stage. There was a great diversity in the amino acid profile in rhizospheric sterilized sand: the major amino acids were glycine, alanine, glutamate and serine. In rhizospheric unsterilized soil there was less diversity in amino acids profile with asparagine representing 38% of the total. These results highlight that it is crucial to work with unsterilized soil in order to obtain reliable information on the dynamics of amino acids in the rhizosphere in relation to plant physiology and diversity of the soil microbial communities. 65 Sugarcane root system affected by insecticide and plant growth regulator Denizart Bolonhezi*1, Adriano Mastro2, Caio Santili2, Tais Lima da Silva3, José Custódio de Souza4, Eglairto Veloso de Carvalho4, Edmar Veloso de Carvalho4, Roberto Botelho Ferraz Branco4 1 APTA -IAC, Brazil, 2Syngenta Company, Brazil, 3Moura Lacerda University, Brazil, 4APTA, Brazil In Brazil the sugarcane crop is cultivated in almost 9.2 million hectares, which 60% of plantations are concentrated in Sao Paulo state. Nowadays, the intense mechanization increases the soil compaction; consequently the sugarcane root system is affected. Some 137 Below and Above Ground Interactions Tuesday 23 June – Poster session insecticides and plant growth regulator normally used in commercial field could help to improve the root characteristics in sugarcane crop. The objective of this research was to quantify the effect of Moddus® (trinexapac) and Actara® (thiametoxam) on the sugarcane root characteristics in three different field conditions. The trials were installed in three different soils, located at Colorado Mill (Miguelopolis city) and Maringa Mill (Araraquara city), both in Oxisoil and Noble Mill (Catanduva city) under Ultisol. Roots were sampled with probe, in two dates defined by the layers down to a depth of 1,0 m. The evaluations were done 30 (effect of trinexapac) and 180 (thiamethoxan) days after harvesting of the third ratoon, respectively December 2012 and April 2013. After washing, sieving and cleaning 2800 samples collected, the images of roots were analysed by Safira® software in order to determine the diameter, area and volume. After that samples were dried to know the dry biomass. Results showed that the association of Moddus® and Actara® increased the dry biomass of roots, the area and root length, respectively in 2,3 Mg ha-1, 60 m2 and 8 m per trenches. It could be concluded that, the application of trinexapac associated with thiamethoxam improve the sugarcane root system, consequently the influence of water stress could be reduced as well as the nutrient uptake. 66 Soil bacterial and fungal community composition along a plant species richness gradient Sigrid Dassen*1, Henk Martens1,2, George Kowalchuk1,3, Wim van der Putten1,2, Gerlinde De Deyn1,2 1 Netherlands Institute of Ecology, Netherlands, 2Wageningen University and Research Centre, Netherlands, 3Utrecht University, Netherlands Abundance and diversity of microbes in soil is extremely large, yet their distribution is not totally random. Several studies have shown how particular microbial groups respond to increasing plant species richness but it remains unclear how the soil microbial community, in terms of microbial richness, identity and abundance changes in response to plant species richness and composition. We hypothesised that 1) the fungal community is more responsive to variation in plant communities than the bacterial community, 2) α-diversity (richness within communities) of soil microbes increases and 3) β-diversity (turnover of species between communities) decreases with plant species richness. To test these hypotheses we sampled soil from the long-term Jena biodiversity experiment comprising a plant species richness gradient of 1, 2, 4, 8, 16, and 60 grassland species, replicated in 4 blocks. We used 454pyrosequencing of 16S and 18S to identify the soil microbial community composition. Most bacterial species occurred in few plots and at low relative abundance, while a minority of species were highly abundant and occurred in nearly all plots. In contrast several fungal species occurred in only few plots and at high relative abundance, indicating that fungal species respond stronger to the local plant community than bacteria, confirming the first hypothesis. α-diversity of bacteria and fungi was highly variable across the plant communities, but variation could not be explained by plant species richness, rejecting the second hypothesis. β-diversity of bacteria, but not fungi, decreased with increasing plant species richness, partly accepting the third hypothesis. Additionally, the variation in α- and βdiversity of bacteria and fungi could partly be explained by plant functional group identity (grasses, legumes, small and tall herbs). Lowest α-diversity observations are mainly from 138 Below and Above Ground Interactions Tuesday 23 June – Poster session grass dominated plots, and regarding β-diversity it was found that legume monocultures cause homogeneity in both the bacterial and fungal community composition. 67 Effect of plant cover diversity on soil organisms: a preliminary study Baptiste Drut*1, Nathalie Cassagne1, Mario Cannavacciuolo1, Gaëtan Lefloch2, Joëlle Fustec1 1 LUNAM Université Angers - Groupe ESA - LEVA, France, 2LUBEM, Université de Bretagne Occidentale, France In low input agrosystems, positive interactions between diversified plant cover and soil organism communities could be powerful tools to increase yields. The aim of this study was to assess how a genetic and/or specific plant cover diversification affects the performance of Triticum aestivum L. and soil organisms. In a greenhouse experiment, we investigated the effects of plant diversity on the rhizospheric fungal biomass and the nematofauna. We also tested whether genetic or specific diversity has an effect on wheat biomass and nitrogen status. Plants were sown in mesocosms filled with soil added with five endogeic earthworms. We tested three levels of plant diversity: single wheat genotype, mixture of three wheat genotypes and the three genotypes mixed with Trifolium hybridum L. Nitrogen concentration in roots and shoots were measured and the percentage of legume nitrogen derived from biological fixation was determined using the 15N natural abundance method. Within wheat-clover association, the performance of wheat (biomass, nitrogen content) was improved by higher N availability thanks to complementarity effects. Indeed, the nitrogen content of clover shoots was mainly due to atmospheric nitrogen fixation. The fungal biomass tended to be higher in wheat mixture than in single wheat. The gradient of plant diversity positively increased the abundance of bacterial-feeding nematodes especially the family of Rhabditidae. The abundance of phytophagous nematodes (Meloidogynidae) in each plant cover modality increased in time. Overall, these results suggest that plant cover diversity influences soil organisms and wheat performance. Root systems of several wheat genotypes seem to favor fungal biomass. Plant cover diversity increases especially opportunistic bacterial-feeding nematodes abundance indicating a higher bacterial biomass. Abundance of phytophagous is related with higher density of plant cover during the experiment. Finally, plant diversity seems to influence soil nematode community structure however more information is needed. 139 Below and Above Ground Interactions Tuesday 23 June – Poster session 68 Monitoring the impact of Bt maize events on the rhizosphere microbial communities cultivated in Brazil Ubiraci Lana, Eliane Gomes*, Fernando Santos, Lucas Silva, Christiane Oliveira, Fernando Valicente Embrapa Maize and Sorghum, Brazil Genetically modified (GM) maize expressing the insecticidal protein from Bacillus thuringiensis (Bt) has been commercially cultivated in Brazil since 2008. The monitoring of transgenic plants after commercial release should be done in order to assess and evaluate possible environmental effects on non-target organisms. Microorganisms in soil can be into contact with Bt proteins when they are released from root exudates of Bt maize or from decomposing plant tissue, and the impact of these proteins on the composition of the rootassociated microbiota is still poorly understood. The objective of this study is to evaluate the impact of commercial events of transgenic maize expressing Bt proteins on the rhizosphere soil bacterial community. To this end, the transgenic maize MON 810, Bt 11, Herculex, MON 89034, VTPro II and Viptera, their respective isogenics treated and not treated with chemical insecticides were assessed using Biolog EcoPlates™ and DGGE (Denaturing Gradient Gel Electrophoresis). The rhizosphere soil was collected at 30 and 60 days after germination in two environments (Sete Lagoas and Janaúba - MG), for three consecutive years (2010/2011, 2011/2012 and 2012/2013), under field conditions. The activity and metabolic diversity evaluated using the Biolog EcoPlates™, after 72 h of incubation, showed no differences between transgenic and non-transgenic maize. The values of Shannon diversity index demonstrated a high level of diversity in the rhizosphere bacterial community for all the treatments. However, the statistical analysis indicated no significant differences among transgenic, isogenic counterparts and samples sprayed with chemical insecticide. The DGGE analysis also did not show difference between transgenic and non-transgenic treatments, but the samples were grouped by environment, showing the importance of soil modulating the microbial community. In conclusion, these results demonstrated that the events of maize Bt cultivated in Brazil showed no significant impact on the soil bacterial community structures based on the parameters studied. 69 Using a whole plant crop model to assess phosphate fertiliser use in barley James Heppell*1, Sevil Payvandi1, Peter Talboys2, Konstantinos Zygalakis1, David Langton3, Roger Sylvester-Bradley4, Robin Walker5, Davey Jones2, Tiina Roose1 1 University of Southampton, United Kingdom, 2Bangor University, United Kingdom, 3Agrii, United Kingdom, 4ADAS, United Kingdom, 5Scotland's Rural College, United Kingdom The role of phosphorus (P) is essential to the agricultural world providing a valuable nutrient for crop production. Its future scarcity, along with escalating concerns over climate change, could greatly harm global food security. In order to improve P efficiency in farming, we have developed a whole plant crop model that allows us to assess a range of P fertiliser and soil cultivation strategies. The model estimates plant P uptake and leaf mass throughout the crop life cycle, given uncertain climate conditions. Two barley field trials are used to validate the model and enable a forecast for other possible fertiliser and soil cultivation strategies. 140 Below and Above Ground Interactions Tuesday 23 June – Poster session The whole plant crop model combines an above ground leaf model (to estimate leaf mass) with our existing below ground root and soil model (to estimate plant P uptake). The leaf model includes the role of photosynthesis to estimate carbon mass stored by the plant. Carbon mass in conjunction with temperature is used to set the root growth rate. With the addition of the leaf model we achieve a better estimate of two sets of barley field trial data for leaf mass and plant P uptake, compared with just the root and soil model alone. There is a positive feedback effect from the two models i.e., an increase in plant P uptake from the root system increases carbon production via photosynthesis in the leaves, and vice versa. The whole plant crop model is sensitive to the initial state of P and its distribution within the soil profile; experimental parameters which are rarely measured in the field. The combination of modelling and experimental data provides useful agricultural predictions for site specific locations. 70 Long-term management effects on root biomass and carbon rhizodeposition of field grown maize Juliane Hirte*, Jens Leifeld, Hans-Rudolf Oberholzer, Jochen Mayer Agroscope ISS, Switzerland Below ground carbon (C) inputs, i.e. dead roots and C rhizodeposition, by agricultural plants into the soil are an important variable in soil C modelling and are mostly estimated from above ground biomass. Agricultural management practices affect above ground biomass considerably; however, their effects on below ground C inputs are only poorly understood. Our aims are therefore to determine management effects on below/above ground C ratios, total root biomass, C rhizodeposition, and vertical root distribution of field grown maize. We hypothesise that, with increasing management intensity, (i) total root biomass is not affected but (ii) below/above ground C ratios, C rhizodeposition, and deep root biomass decrease. In 2013, we conducted field experiments with maize on two Swiss long-term trials (“DOK”, Basel and “ZOFE”, Zurich) covering seven different management treatments. Four individual plants per treatment were grown in microplots and pulse-labelled with 13C-CO2 in weekly intervals throughout the growing season. After harvest, the microplot soil was sampled in three layers to 0.75m depth, coarse- and fine root biomasses were determined by picking and wet sieving, and all samples were analysed for their δ13C values. Preliminary results reveal a decrease of below/above ground C ratios with increasing management intensity on both sites and range from, on average, 0.38 to 0.22 on the “DOK” site and 0.32 to 0.18 on the “ZOFE” site (trends only). Total root biomasses and C rhizodeposition do not differ between treatments on both sites. While the proportion of subsoil (0.25-0.75m) root biomass of the total (0-0.75m) root biomass decreases with increasing management intensity on the “DOK” site, this trend is not visible on the “ZOFE” site. These findings show that below ground C inputs cannot easily be estimated from above ground biomass and input data for soil C models should be differentiated according to the management system. 141 Below and Above Ground Interactions Tuesday 23 June – Poster session 71 Small but important fraction of DOM: exudation of plant dominants in swamp forest Eva Kastovska*1, Tomas Picek1, Keith Edwards1, Jakub Borovec2, Hana Santruckova1 1 University of South Bohemia, Faculty of Science, Czech Republic, 2iology Centre of the Academy of Sciences of the Czech Republic, Czech Republic Swamp forest is a common type of peatland, its herbal floor typically co-dominated by Sphagnum mosses, Eriophorum vaginatum and Vaccinium myrtillus distributed in patches tracking the differences in microtopography and hydrology of the terrain. As with other peatlands, swamp forests are characterized by formation of high amounts of dissolved organic matter (DOM). In the DOM, mostly derived from accumulated soil organic matter of low biodegradability, regular daily input of fresh plant-derived compounds (rhizodeposition) can represent the key C and energy source for soil microorganisms and form the most reactive part of DOM. Therefore, we aimed to quantify and compare the “exudation rate” from roots of Eriophorum and Vaccinium and stalks of Sphagnum girgensohnii (sampling of exudates from living roots of plants and fresh moss in intact peat monoliths), quality of the released compounds (C and N content, size exclusion chromatography and HPLC) and contribution of the released compounds to DOM and their biodegradability (13CO2 pulse-labeling of plants placed in fresh natural DOM). We found that plant-derived compounds formed only 2-3% of the DOM within the rooting zone, with the exudation rate largest for Sphagnum and lowest for Vaccinium (per m2 of the particular vegetation cover). Sphagnum “exudates” were of very different quality than in the other two species, poorer for simple compounds but relatively rich in oligosaccharides, had larger C/N ratio and lower biodegradability. The difference in the quality of the compounds released from Sphagnum and the other two herbal dominants suggests that the patchy distribution of vegetation in the understory of the swamp forest ecosystem could significantly contribute to the high spatial diversity of the soil microbial community and of the rates of C and N processing in its upper soil layer. 72 Carbon input into soil originating from fine root and foliage litter in two Norway spruce forests Jaana Leppälammi-Kujansuu*1, Lasse Aro2, Maija Salemaa2, Karna Hansson3, Dan Kleja Berggren4, Heljä-Sisko Helmisaari1 1 University of Helsinki, Finland, 2Natural Resources Institute Finland, Finland, 3INRA, Centre de Nancy, Biogéochimie des Écosystèmes Forestiers, France, 4Swedish University of Agricultural Sciences, Sweden Fine root litter forms a significant carbon input to soil. Knowledge of the quantity of fine root litter is scarce – but highly valued in C budget calculations. We quantified fine root litter production in two Finnish Norway spruce stands and compared it to foliage litter. The stands differed in their geographical location and environmental conditions. Finally, we added data from four previously published spruce stands in Sweden for determining the above- and belowground litter ratios along latitudinal and soil fertility gradients. 142 Below and Above Ground Interactions Tuesday 23 June – Poster session The amount of belowground litter was estimated from fine root turnover with the minirhizotron method and biomass from soil cores. Foliage litter was collected with litter traps and ground vegetation was sampled. In the less fertile, northern stand, the fine root biomass per stand basal area was almost double and the tree fine root lifespan longer than in the southern stand. The annual production of tree foliage litter was less than in the southern stand, but the total amount of litter (including trees and understorey, above- and belowground) was similar at both sites, as was the ratio between the above- and belowground litter production. The increased contribution of understorey vegetation towards low fertility sites was clearly seen when regressing the aboveground:belowground litter production -ratio of the Finnish and Swedish sites against the C:N-ratio of the organic layer in the sites. In boreal forest soil, the input of carbon via fine roots at least equals to foliage litterfall and the share of understory vegetation is also substantial. This shift in the litter production pattern from above to belowground in the least fertile sites may have an impact on litter C quality and soil C storage and should be studied further. 73 Rhizosphere adaptive strategies of three plant species of high-mountain environment under periglacial conditions (Majella Massif, central Italy) Luisa Massaccesi*1, Gian Maria Niccolò Benucci1, Giovanni Gigliotti2, Mauro De Feudis1, Stefania Cocco3, Giuseppe Corti3, Alberto Agnelli1 1 Department of Agricultural, Food and Environmental Sciences, University of Perugia, Italy, Italy, 2Department of Civil and Environmental Engineering, University of Perugia, Italy, Italy, 3Department of Agricultural, Food and Environmental Sciences, Polytechnic University of Marche, Italy, Italy The rhizosphere is a highly dynamic interface for chemical, physical and biological interactions between plants and soil. As rhizosphere processes have been widely investigated, little is known about the rhizosphere adaptative strategies of pioneer plants in soils of periglacial environment. The rhizosphere processes in these severe and nutrient-poor environments may provide a key in the prediction of plant growth and health, soil quality and nutrients availability. We selected three plants (Helianthemum nummularium (L.) Mill. subsp. grandiflorum (Scop.), Dryas octopetala (L.), and Silene acaulis (L.) Jacq. subsp. cenisia (Vierh.) P. Foum) that sparsely occupy deglaciated areas of the Majella Massif (central Apennines, Italy), with the aim to assess how they overcome harsh pedoclimatic and nutritional conditions, we investigated physico-chemical properties together with microbial community structure, abundance and activity for both rhizosphere and bulk soil. The three plant considered in this study have shown different adaptative strategies addressed to force changes in soil and nutrient uptake. Helianthemum induces a strong rhizosphere impact through a synergistic effect between the roots activity and a well-adapted rhizosphere microbial community. For Dryas, which does not foster a microbial community structure specifically designed for its rhizosphere, an intense consumption of the energetic resources supplied by the plant is needed to make nutrients available. The scarce rhizosphere effect produced by Silene suggested that also the 143 Below and Above Ground Interactions Tuesday 23 June – Poster session root morphology influences the intensity of the soil changes induced by roots and associated microorganisms. In the case of Silene, the ability to colonize harsh environments may be mostly linked to the shape and functions of its canopy rather than to a functional rhizosphere effect. This study showed that the activity of the roots and associated microbial community are decisive in modifying the soil properties, so to create a suitable environment where plants are able to grow even under a hostile climate. 74 Linking below ground and above ground phenology of Hybrid Walnut in temperate agroforestry systems Awaz Mohamed*1, Yogan Monnier1, Sylvie-Annabel Sabatier2, Jean-Luc Maeght3, Christophe Jourdan4, Alexia Stokes1, Merlin Ramel1 1 INRA, UMR AMAP Montpellier, France, 2CIRAD, UMR AMAP Montpellier, France, 3UMR Iees Paris, Institut de Recherche pour le Développement (IRD) c/o UMR AMAP, CIRAD, Montpellier, France, 4CIRAD UMR Eco&Sols Montpellier, France Climate models predict that an increase in atmospheric CO2 concentration, precipitation and temperature could affect many biological phenomena and increase the frequency and magnitude of extreme weather events. Root and shoot phenology could be strongly influenced by the variations in soil water content, soil and air temperatures. Changes in plant phenology are considered to be a very sensitive and observable indicator of plant responses to climate change. In contrast, very little is known about the relationship between shoot and root phenology especially in the natural soil environment. Knowledge about this relationship is needed to understand the response of vegetation to climate changes which will give us a better understanding of ecosystem structure and function. The objective of this study is to quantify the dynamics of root and shoot growth of Hybrid Walnut (Juglans regia x nigra) in three temperate agroforests, along a climatic gradient of precipitation and temperature. We also regularly measured soil carbon, nitrogen and phosphorus concentrations. Fine root growth dynamics were measured every 3 weeks in rhizotrons and minirhizotrons. The results of this project will provide data allowing a better understanding and assessment of spatial and temporal impacts of climate on vegetation. 75 Bacterial volatile organic compound that hampers plant growth Jun Murata*, Takehiro Watanabe, Hajime Komura Suntory Foundation for Life Sciences, Japan Selected soil microbes have been shown to trigger alteration of plant growth by releasing a blend of volatile chemicals called volatile organic compounds. Examples of such bacterial strains include a plant growth promoting rhizobacteria Bacillus subtilis GB03 which is known to release 2,3-butanediol that enhances growth of Arabidopsis seedlings. However, it is largely unknown whether such bioactivity of volatile organic compounds can be found only in specialized bacterial strains. Moreover, molecular mechanism of how such volatile organic compounds are recognized by plants still remains elusive. Here we show that the growth of 144 Below and Above Ground Interactions Tuesday 23 June – Poster session Arabidopsis seedlings can be interfered by volatile organic compounds that were released from an experimental strain of Bacillus spp. in a distance-dependent manner. Similar effects upon plant growth were observed when either E. coli or Agrobacterium instead of Bacillus was used as a source of volatile organic compounds, suggesting that the active compound is conserved among various bacterial species. Notably, while Arabidopsis seedlings grown in a position relatively closer to Bacillus showed severe growth retardation, the seedlings grown in a position distal to Bacillus showed rather enhanced growth in their above-ground parts compared to control seedlings without Bacillus co-culture. Through activity-guided purification approach, we identified isovaleric acid as a key volatile that exhibits plant growth retardation activity. Authentic isovaleric acid was capable of inhibiting the growth of Arabidopsis seedlings as in the case of Bacillus volatiles. Bioassay experiments using 14Clabeled isovaleric acid showed that isovaleric acid was incorporated into leaf and root tissues of Arabidopsis seedlings in a manner dependent on the distance from the source of the volatile. These data provide novel insights into the molecular basis of how soil microbes interfere with the growth of plants. 76 Resilience of ectomycorrhizal communities under drought conditions Karin Pritsch*1, Uwe Geppert1, René Kerner1, Franz Buegger1, Martina Peter2, Joerg Luster2, Arthur Gessler2, Frank Hagedorn2, Marcus Schaub2, Rolf Siegwolf3, Matthias Arend2 1 Helmholtz Zentrum München, Germany, 2WSL, Switzerland, 3Paul Scherrer Institut, Switzerland Prolonged drought periods are predicted in most future climate scenarios. Especially for long-lived tree species, genetic adaptation of the plant may be hindered by long generation times. Instead, mycorrhizal symbionts may mitigate drought stress by improving the nutrient and water status of plants. Different mycorrhizal fungi are ecologically beneficial to the plant to very varying extents. We present results from the drought experiment ‘BuKlim’ performed over 3 years in the model ecosystems of WSL (CH). The aim was to study mechanisms of resilience of the whole plants including their ectomycorrhizosphere after strong drought events and upon rewatering. For this purpose, young beech trees (Fagus sylvatica) were 13CO2 labelled at the end of the drought period and after rewatering. We then followed the time course of allocation of 13C through the plant to the ectomycorrhizosphere and into the soil. Aboveground reactions of trees such as reduced carbon uptake during drought and high compensation of photosynthesis upon rewatering were reflected by an enhanced 13C allocation into below-ground parts. A wide range of 13C accumulation was detected in different ectomycorrhizal exploration types indicating which species were resistant and resilient under severe drought conditions. Overall, our results show plastic reactions at the whole root-system level, a strong link between above and below-ground reactions, and different mechanisms that may contribute to the observed resilience of ectomycorrhizal communities under extended drought periods. 145 Below and Above Ground Interactions Tuesday 23 June – Poster session 77 Pseudomonas sp. P482 shows broad spectrum of antagonistic activity towards plant pathogenic bacteria Sylwia Jafra, Dorota Krzyzanowska, Adam Ossowicki, Magdalena Rajewska*, Magdalena Jablonska University of Gdansk, Poland Plant-associated Pseudomonas produce large numbers of biologically-active secondary metabolites. Many of them were demonstrated to have antimicrobial properties allowing the bacteria to survive in a competitive environment. Pseudomonas sp. P482, a tomato rhizosphere isolate, displays antagonistic activity towards a broad spectrum of plant pathogenic bacteria, including pectinolytic plant pathogens – from Pectobacterium and Dickeya genera - which cause important economic losses in potato production worldwide. Spreading of these pathogens calls for development of an efficient and environmentallyfriendly control strategy based on natural compounds and microorganisms. The aims of this study were to verify the potential of the Pseudomonas sp. P482 strain to antagonize bacterial plant pathogens and to analyze the genetic background of this strain’s antimicrobial activity by genome data mining approach. To investigate whether the Pseudomonas sp. P482 could be employed as a biological control agent against soft rot, in planta tests on potato tubers and chicory leaves, involving coinoculation of plant tissue with bacterial mixtures, were applied. Additionally, the ability of P482 to colonize the roots of soil-grown potato plants was tested. Genome sequencing of P482 was performed, followed by bioinformatics analysis with the antiSMASH and BAGEL3 platforms in order to determine the ‘candidate’ genes potentially involved in the antimicrobial activity of P482. In planta analyses revealed that although P482 inhibits growth of bacterial pathogens in a plate assay, the biocontrol effect is pathogen-specific. The P482 is a moderate potato root colonizer and thus can be tested for its biocontrol potential on this host in a greenhouse or field setup. A draft genome sequence of P482 has been obtained and annotated. The BAGEL3 analysis yielded no putative bacteriocins. The antiSMASH analysis revealed five biosynthetic clusters encoding genes potentially involved in the synthesis of antimicrobial factors. Selected genes from these clusters were further investigated. 78 Aboveground insect infestation attenuates belowground Agrobacterium Choong-Min Ryu*1, Geun Cheol Song1, Soohyun Lee1, Jaehwa Hong1, Hye Kyung Choi1, Gun Hyong Hong1, Dong-Won Bae2, Kirankumar S. Mysore3, Yong-Soon Park1 1 KRIBB, South Korea, 2Gyeongsang National University, South Korea, 3The Samuel Roberts Noble Foundation, United States Agrobacterium tumefaciens causes crown gall disease. Although Agrobacterium can be popularly used for genetic engineering, the influence of aboveground insect infestation on Agrobacterium induced gall formation has not been investigated. Nicotiana benthamiana leaves were exposed to a sucking insect whitefly infestation and benzothiadiazole (BTH) for 7 days, and these exposed 146 Below and Above Ground Interactions Tuesday 23 June – Poster session plants were inoculated with a tumorigenic Agrobacterium strain. We evaluated both in planta and in vitro, how whitefly infestation affects crown gall disease. Whitefly infested plants exhibited at least a 2-fold reduction in gall formation on both stem and crown root. Silencing of isochorismate synthase 1 (ICS1), required for salicylic acid (SA) synthesis, compromised gall formation indicating an involvement of SA in whitefly-derived plant defence against Agrobacterium. Endogenous SA content was augmented in whitefly infested plants upon Agrobacterium inoculation. In addition, SA level was three times higher in root exudates from whitefly infested plants. As a consequence, Agrobacterium-mediated transformation of roots of whitefly infested plants was clearly inhibited when compared to control plants. These results suggest that aboveground whitefly infestation elicits systemic defence responses throughout the plant. Our findings provide new insights into insect-mediated leaf-root intra communication and a framework to understand interactions between three organisms, whitefly, N. benthamiana and Agrobacterium. 79 Influence of bacterial N-acyl-homoserine-lactones on growth and defense in barley and yam bean Peter Schröder*1, Christine Götz-Rösch2, Tina Sieper3, Anton Hartmann2, Agnes Fekete4 1 Helmholtz Zentrum Muenchen GmbH, Germany, 2Helmholtz Zentrum Muenchen Gmbh, Germany, 3Helmholtz Zentrum Muenchen, Germany, 4Universität Würzburg, Germany Bacteria communicate with each other and sense their environment in a population density dependent mechanism known as quorum sensing (QS). N-acyl-homoserine lactones (AHLs) are the QS signalling compounds of Gram-negative bacteria which are frequent colonizers of rhizospheres. While cross-kingdom signalling and AHL-dependent gene expression in plants has been confirmed, the responses of enzyme activities in the eukaryotic host upon AHLs are mainly unknown. We investigated the influence of three different AHLs, namely N-hexanoyl(C6-HSL), Noctanoyl-(C8-HSL) and N-decanoyl- homoserine lactone (C10-HSL) on two agricultural crop plants. The AHL-effects on Hordeum vulgare (L.) as an example of a monocotyledonous crop and on the less known tropical leguminous crop plant Pachyrhizus erosus (L) were compared. While plant growth and pigment contents in both plants were less influenced, AHL treatment triggered tissue- and compound-specific changes in the activity of important detoxification enzymes. The activity of dehydroascorbate reductase (DHAR) in barley shoots after C10-HSL treatment for instance increased up to 384% of control plant levels, whereas superoxide dismutase (SOD) activity in barley roots was decreased down to 23% of control levels upon C6-HSL treatment. Other detoxification enzymes reacted similarly within this range, with interesting clusters of positive or negative answers towards the stress. In general, changes on the enzyme level were more severe in barley than in yam bean. This might be due to the different abilities of the plants to degrade AHLs to metabolites such as the hydroxy- or keto-form of the parent signalling compound. 147 Below and Above Ground Interactions Tuesday 23 June – Poster session 80 Effects of silver nanoparticles on soil microorganisms and maize biomass are linked in the rhizosphere Wouter Sillen*1, Sofie Thijs1, Nele Weyens1, Jason White2, Jaco Vangronsveld1 1 Hasselt University, Belgium, 2Connecticut Agricultural Experiment Station, United States Silver nanoparticles form part of an industrial revolution that has two faces. On the one hand, nanotechnology has developed into an industry that is capable of making use in the most efficient and targeted way of the well-known anti-microbial properties of silver. On the other hand, it are the increasing quantities in which silver nanoparticles are produced and applied, as well as the special characteristics caused by their nanodimensions, that have led to the development of the study of nanotoxicology because earlier toxicology studies of bulk silver are not applicable. Silver nanoparticles therefore are highly promising and useful, mainly for combatting microorganisms, but are also a significant reason for concern as they exert their toxicity in natural environments to non-target microbes and plants. Agricultural crops like maize (Zea mays sp.) in particular are likely subjects of silver nanoparticle exposure, as these particles may end up in the agro-ecosystem during manufacturing, incorporation into products, and use of nanoparticle-containing products. Maize growing in hydroponics responds to silver nanoparticle exposure by producing less biomass, while exposure to realistic concentrations in natural soil often leads to an increase in maize biomass. Because of silver’s well-known antimicrobial properties, microbial communities associated with maize are expected to be important mediators in this outcome, although very few studies have investigated the effects of silver nanoparticles in combined plant-microbial systems. Our results confirm the existence of a strong link between the responses of maize and the microbial communities in its rhizosphere to silver nanoparticles. The specific conditions that shape the rhizosphere also alter the effects of silver nanoparticles on microbial communities, compared to the bulk soil. The link between plant and rhizobiome response highlights the role of rhizosphere microorganisms in the welfare of their host plant and its response to contaminants, especially those that strongly target microorganisms. 81 Root exudate cocktails: the link between plant diversity and soil microorganisms? Katja Steinauer*1, Julia Friese2, Nico Eisenhauer1 1 German Centre for Integrative Biodiversity Research (iDiv) Halle-Jena-Leipzig, University of Leipzig, Germany, 2Friedrich Schiller University Jena, Institute of Ecology, Germany Higher plant diversity is often associated with higher soil microbial biomass, which is assumed to be – at least partly – due to higher root exudate diversity. However, little is known about the quantity and diversity of root exudates shaping their specific soil microbial community. In this experiment we tested whether higher root exudate diversity enhances soil microbial biomass in a plant diversity gradient. We set up a plant diversity gradient ranging from monocultures to 2- and 3-species mixtures in microcosms using functionally dissimilar plant species (grass: Anthoxanthum odoratum, small herb: Plantago lanceolata, tall herb: Centaurea jacea) in soil of the Jena Experiment, Germany. After one month of plant growth, exudate cocktails were added to the microcosms combining the most common sugars (e.g., 148 Below and Above Ground Interactions Tuesday 23 June – Poster session glucose), organic acids (e.g., citric acid), and amino acids (e.g., alanine). In total, we used four different exudate cocktails, two exudate diversity levels (low and high), two nutrient availability levels (carbon and nitrogen), and a control with only water addition. We hypothesized the addition of a diverse exudate cocktail to offset significant plant diversity effects on soil microbial biomass. After two months, we found a significant increase of soil microbial biomass with increasing plant diversity in the control treatment. Microbial biomass in the low exudate diversity treatment increased, though not significantly, with increasing plant diversity. As hypothesized, microbial biomass was at similar levels across all plant diversity levels in the high exudate diversity treatment, because microbial biomass increased particularly at low plant diversity. Furthermore, microbial biomass was significantly higher in the carbon treatment than in the nitrogen-rich exudate treatments. This corresponds to previous findings of carbon-limited soil microbial communities at the field site of the Jena Experiment. Our findings provide the first experimental evidence that root exudate diversity represents a crucial link between plant diversity and soil microorganisms. 82 Changes in the root metabolism of Arabidopsis thaliana during mutualistic association with a beneficial rhizobacterium Nadine Strehmel*1, Sylvia Krueger1, Paul Schulze-Lefert2, Dierk Scheel1 1 Leibniz Institute of Plant Biochemistry, Germany, 2Max Planck Institute for Plant Breeding Research, Germany The narrow zone between the root and its surrounding soil is defined as the rhizosphere. Here, hundreds of thousands of bacteria can be found, exhibiting a beneficial, a deleterious or no effect on the performance of the plant. In general, bacteria colonizing the plant root and promoting plant growth are referred to as plant growth-promoting rhizobacteria (PGPR). Arabidopsis thaliana was cocultivated with Rhizobium leguminosarum under sterile conditions in a growth cabinet, which resulted in an increased production of root and shoot dry and fresh weight. In addition, changes in root morphology were observed, as roots of inoculated plants were longer and exhibited more lateral roots than those of control plants. To understand the underlying metabolic processes during this mutualism in the rhizosphere, Arabidopsis thaliana was cocultivated for a defined time period with a plant growth promoting strain of Rhizobium leguminosarum on agar plates. Finally, roots were analyzed for changes in semipolar secondary metabolism following a non-targeted metabolite profiling approach. Comprehensive analysis revealed a decline of major substance classes such as glucosinolates and their degradation products, flavonoids and coumarins. In contrast, signaling metabolites and defense compounds, phenylpropanoids as well as oligolignols increased during cocultivation. To examine whether these compounds are essential for this interaction, cocultivation studies were conducted in a hydroponic system and samples were analyzed for changes in secondary metabolism. This revealed that phenylpropanoids and oligolignols showed a contrasting behavior in root tissue and the corresponding root exudates. These studies are essential to understand microbial community dynamics in the rhizosphere and thus contribute to the understanding of rhizosphere biology. 149 Below and Above Ground Interactions Tuesday 23 June – Poster session 83 Screening for ISR-inducing Trichoderma strains Christine Vos*1, Katrijn Raymaekers2, Kaat De Cremer2, Barbara De Coninck2, Kemal Kazan3, Bruno Cammue2 1 KU Leuven and CSIRO Australia, Australia, 2KU Leuven, Belgium, 3CSIRO Agriculture Flagship, Australia The genus Trichoderma constitutes a promising collection of potential biocontrol organisms (BCOs), reducing plant disease either via direct interaction with plant pathogens and/or indirectly through induced systemic resistance (ISR). The ISR capacity is mostly investigated by performing classical disease assays in which physical contact between the BCO and pathogen is avoided, however, such tests are very labour, time‐ and space-consuming. The discovery of general ISR markers could therefore greatly facilitate the search for novel or more efficient BCOs. We reported that application of Trichoderma hamatum T382 to Arabidopsis thaliana roots resulted in ISR against leaf infection by the necrotrophic pathogen Botrytis cinerea and performed a genome-wide analysis of ISR-related leaf gene expression, both before and after B. cinerea infection. In addition, we recently completed a similar microarray analysis with tomato replacing A. thaliana in the tripartite interaction. Based on the comparison of the transcriptomic analyses in both plants, a series of orthologous genes up-regulated in both tripartite systems was selected for their potential as general markers for Trichoderma-induced ISR. We developed a screening system based on pMarker-GUS lines, allowing fast and effective visual detection of ISR-inducing Trichoderma spp. The potential of the pMarker-GUS lines as a screening tool is demonstrated by the clear correlation between the percentage of disease reduction and the degree of staining of the pMarker-GUS lines. In this presentation we will describe our screening assay and cover the most recent advances that we have made with this system. Hereby we will focus on our ongoing characterization of Trichoderma isolates with so-far unknown ISR capacity, as well as on the extrapolation of our discoveries to tomato. In addition, we will address the question whether the screening system can also be used for BCOs that do not belong to the genus Trichoderma. 84 The critical level of soil available P for the highest grain yield and the lowest AMF colonization is identical in field-grown maize Chao Wang*, Chunjian Li China Agricultural University, China The concentration of soil available phosphorus (P) is important for both plant growth and colonization of arbusular-mycorrhizal fungi (AMF). A critical level of soil P for getting the highest maize grain yield is reported in many studies, whilst that for AM colonization in the field condition is not clear. Here a two-year field experiment (2013 and 2014) using maize at six P application rates from 0 to 300 kg ha-1 on a long-term experiment site was conducted at the Shangzhuang Experimental Station (40o8′20″N, 116o10′47″E), China Agricultural University. The soil is a typical Ustochrept with a silty loam texture, mesic soil temperature regime, and mixed mineralogy. Shoot, root and soil samples in 0-20 (topsoil) and 20-40 (subsoil) cm soil layers were collected at five growth stages over the whole growth period. The results demonstrated that soil Olsen P in the topsoil layer, rather than in the subsoil layer, increased with the increased application rate of P-fertilizers. The shoot biomass, grain yield, shoot P content and concentration increased and reached the highest values when the 150 Below and Above Ground Interactions Tuesday 23 June – Poster session amount of P fertilizers increased from 0 to 75 kg ha-1, and then kept constant, in spite of further increases in P-fertilizer input. By contrast, the AMF colonization rate onto topsoil-layer maize roots decreased with the increased amount of P-fertilizers, and reached the lowest value at the P application rate of 75 kg ha-1, and kept unchanged regardless of the further input of P-fertilizers. The measured Olsen-P concentration corresponding to the P fertilizer rate of 75 kg ha-1 was 6-10 mg kg-1 in both years. The results suggest that the shoot P concentration decided the AMF colonization rate on maize roots in the field. However, the AM colonization rate on subsoil-layer maize roots distributed kept relatively constant at different P application rates. 85 Inoculation of arbuscular mycorrhizal fungi and Gaeumannomyces graminisvar causes asymmetric competition between wheat and faba bean in intercropping system Guangzhou Wang*, Haigang Li, Fusuo Zhang, Junling Zhang China Agricultural University, China The mechanistic understanding of the effect of soil microorganisms to overyielding in intercropping systems can be instructive in optimizing the productivity and ecosystem service. To date the influence of soil microbes on interspecific and intraspecific interactions, and consequently the system productivity is not well understood. In this study, we inoculated arbuscular mycorrhizal fungi (AMF) of Funneliformis mosseae or a mix of six species (Glomus etunicatum, Funneliformis mosseae, Rhizophagus intraradices, Glomus versiforme, Glomus etumicatum and Glomus aggregatum) 10 days prior to the inoculation of wheat take-all fungus of Gaeumannomyces graminisvar. tritici (Ggt) to wheat and faba bean in mono and poly-culture to explore their respective and combined effects on plant growth, nutrient uptake and competitive relationships between the two plant species. No overyielding was observed. The dry biomass, N and P uptake by wheat in monoculture decreased significantly by 18.7%, 20.7% and 26.5% respectively when inoculated with Ggt. When intercropped with faba bean, the competitive disadvantage was aggravated and wheat biomass, N and P uptake decreases by 31.8%, 39.6% and 40.2% respectively. By contrast, inoculation of Ggt improved the dry biomass, N and P uptake of intercropped faba bean compared to those of mono plants. Neither the growth of both plant species nor the disease severity of wheat was significantly affected by the inoculation of one or mix AMF. However, inoculation of Ggt reduced colonization percentage in wheat roots but not faba bean roots in both mono and poly-culture treatments. Whereas inoculation of AMF enhanced P uptake of wheat and faba bean whether inoculating with Ggt or not. Our results show that soil pathogen leads to asymmetric competition between wheat and faba bean in intercropping system, and the beneficial effect of AMF on plant P uptake does not offset the detrimental effects of soil pathogen on the host plant. 151 Below and Above Ground Interactions Tuesday 23 June – Poster session 86 Above and belowground productivity of experimental grassland communities in the field during initial assembly Emanuela W. A. Weidlich*1, Philipp von Gillhaussen2, Vicky M. Temperton1 1 Forschungszentrum Jülich, Germany, 2University of Bayreuth - Department of Disturbance Ecology, Germany The order of arrival of different species can change the trajectory of a community as it assembles. This is known as a priority effect, where the plants that first arrive affect the further development of the system to such an extent that their impact on the system is still detectable many years after their arrival. The more we understand community assembly, the more information we will have on how to return a degraded area to a functioning ecosystem. A large scale field experiment, called Priority Effect was established in 2012 to test whether we could create priority effects by sowing high and low diversity mixtures and changing the sequence in which plant functional groups arrived in the system. We measured above and below ground productivity, soil chemistry and root productivity using the ingrowth core method. In the first year plots with legumes sown first were more productive in aboveground biomass than grasses or forbs sown first, but not more than the control sown at the same time. Belowground we found opposite patterns of productivity: legume-first plots were less productive belowground versus grasses-first as most productive. These results indicate different biomass allocation patterns to above and belowground plant parts depending on which functional group arrived first. We repeated the in-growth core method to assess root biomass and turnover in 2014 and will see whether the belowground priority effect remains or have disappeared with time. Following the development of the species and productivity in this experiment will help us to understand the mechanisms of priority effects in grassland communities in order to use this knowledge in improving grassland restoration as well ensuring a certain level of productivity. 87 Induced systemic resistance in Arabidopsis against Pseudomonas syringae pv. tomato by disease suppressive soils David Weller*1, Johan A. van Pelt2, Corné M. J. Pieterse2, Peter A. H. M. Bakker2 1 USDA-ARS, United States, 2Utrecht University, Netherlands Two-week-old Arabidopsis thaliana ecotype Col-0 seedlings were transferred into an autoclaved sand-soil mixture amended with 10 or 20% (weight/weight) soil that is suppressive to either take-all or Rhizoctonia root rot of wheat from fields in Washington State USA. These soils contain population sizes greater than 105 colony forming units per gram of wheat root of 2,4-diacetylphloroglucinol or phenazine-1-carboxylic acid-producing pseudomonads, respectively. Plants were challenge-inoculated with Pseudomonas syringae pv. tomato three weeks later. Both suppressive soils induced resistance in Arabidopsis against P. syringae similar to the well-studied Pseudomonas strains WCS417r, Q2-87 (produces 2,4diacetylphloroglucinol) and 2-79 (produces phenazine-1-carboxylic acid). Pasteurization of the suppressive soils before adding them into the sand-soil mixture eliminated 2,4diacetylphloroglucinol and phenazine-1-carboxylic acid-producing pseudomonads from the Arabidopsis rhizosphere and significantly reduced induced systemic resistance activity. However, population sizes of total aerobic culturable bacteria were similar in the rhizosphere of plants grown in the mixes with pasteurized and raw suppressive soils. This is the first report 152 Below and Above Ground Interactions Tuesday 23 June – Poster session of induced systemic resistance activity by take-all and Rhizoctonia suppressive soils and the ability of phenazine-1-carboxylic acid to induce resistance. 88 Insights into bacterial communities associated with apple replant disease soil revealed by 16S rRNA gene amplicon sequencing Bunlong Yim*1, Traud Winkelmann2, Guo-Chun Ding3, Kornelia Smalla4 1 Leibniz University of Hannover, Germany, 2Leibniz Universität Hannover – Institute of Horticultural Production Systems, Section of Woody Plant and Propagation Physiology, Germany, 3College of Resources and Environmental Sciences, China Agricultural University, China, 4Julius Kühn-Institut – Federal Research Centre for Cultivated Plants (JKI), Institute for Epidemiology and Pathogen Diagnostics, Germany Apple replant disease (ARD) severely affects apple production worldwide. The possible causes are parasitic nematodes, accumulation of soil borne pathogenic microorganisms or absence of antagonists and beneficial microorganisms, and toxic effects from decomposition of crop residues. The aim of this study was to reveal bacterial communities associated with ARD soils and to analyze the effects of soil treatments with heat and gamma irradiation on plant growth and bacterial community composition and diversity in two different soil types. The results of a biotest with micropropagated apple rootstock M26 plants showed that the plant growth in replant disease soils from two nurseries (Kle and Alv) was reduced in ARD soils (control) compared to soils either treated at 50 oC (H50) or with gamma irradiation (Gamma). Soil attached to roots of the plants after eight weeks of the biotest experiment was collected and submitted to soil total community DNA (TC-DNA) extraction. The TC-DNA was subjected to 454-pyrosequencing of amplified 16S RNA gene fragments. The differences in plant growth were associated with differences in the bacterial communities. The two soil types with different culture practices and management showed different bacterial community composition and diversity. In both soils, the treatments led to an increased abundance of Proteobacteria (Gamma), Firmicutes (H50), and Bacterioidetes (Gamma), whereas members of Acidobacteria (H50, Gamma) and Actinobacteria (Gamma) decreased. Despite of the soil type specific responses a few genera were recorded as responders to the treatments in both soils. Significantly increased relative abundance was recorded for Streptomyces, Bacillus, Paenibacillus and Sphingomonas in the H50 treatments while Mucilaginibacter, Devosia and Rhodanobacter had an increased relative abundance in the Gamma treatments of both soils. The increased abundance of potential degraders of phenolic compounds might contribute the improved plant growth in the H50 and Gamma treatments. 89 Linking shoot and root characteristics with soil microbial communities to better understand the influence of agroforests on soil structure Monnier Yogan*1, Amandine Erktan1, Luis Merino-Martin1, Catherine Roumet2, Yves Le Bissonnais3, Alexia Stokes1 1 INRA, UMR AMAP, France, 2CNRS, UMR CEFE,, France, 3INRA, UMR LISAH, France In European agricultural landscapes, conventional farming is often described as a factor incrementing soil erosion. Recent policies have been encouraging agroforestry practices that 153 Below and Above Ground Interactions Tuesday 23 June – Poster session are believed to provide a number of ecosystem services and improve biodiversity. However research is still required to determine the best plantation and management practices, depending on land use, climate, topography and characteristics of species used. Our project investigates above and belowground interactions in crop/forest systems and changes that occur in the rhizosphere of different cropping systems. Eight field-sites representative of different agroforests with distinct ages and pedoclimatic characteristics were selected throughout France. On each site, measurements were performed to assess the influence of the cultivated tree row, the distance to trees, and the perennial plant cover on shoot morphology, litter cover, soil aggregate stability, root traits, soil microbial activity and microbial metabolic diversity. Preliminary results show that in a number of agroforests, soil aggregate stability is greater along the tree row because root density is higher. However, the magnitude of these effects seems to be related to the pedological characteristics of the agroforest. Results will allow to determine how above and belowground characteristics of plant communities contribute to soil aggregate stability. This research will provide stakeholders and agroforesters with data to determine the best soil management strategies to protect against erosion with regard to above and below traits of cultivated trees. 90 A new conceptual framework show that plant genotype and soil N availability modify both plant nutritional strategies and the associated rhizosphere microbiome Anouk Zancarini*, Christophe Mougel, Anne-Sophie Voisin, Marion Prudent, Christophe Salon, Nathalie Munier-Jolain INRA, France In the context of sustainable agriculture, it is desirable to lower the input of mineral fertilizers. Therefore, we need to select ‘new’ genotypes that are both adapted to ‘low fertilizer inputs’ and more efficient in nutrient use. A better understanding of plant-microbe interactions under low input of fertilizers is now needed. Nevertheless, in microbial ecology, conceptual frameworks or models are used to analyse plant-microbe interactions but plant phenotype is currently viewed as a “black box”. We developed a new conceptual framework to study plantmicrobe interactions using a multidisciplinary approach combining Microbial Ecology and Plant Ecophysiology. The links among plant genotype, soil nitrogen availability and the plant associated rhizosphere microbiome were assessed at both structural and functional level. On the one hand, the influence of both Medicago truncatula genotype and soil nitrogen availability on the genetic structure of the soil microbiome was determined by DNA fingerprint and 454 pyrosequencing. On the other hand, the different nutritional strategies of the plant-microbe interactions were evaluated using an ecophysiological framework. We observed that nitrogen availability affected rhizosphere bacterial communities only in presence of the plant. Furthermore, we demonstrated the existence of a strong Medicago truncatula genotype x nitrogen availability effect on the rhizosphere bacterial communities. Finally, the nutritional strategies of the plant varied greatly in response to a modification of nitrogen availability. Three contrasted structural and functional adaptive responses of plantmicrobe interactions to nitrogen availability were thus identified. 154 Below and Above Ground Interactions Tuesday 23 June – Poster session With recent developments in statistics and high throughput genotyping, this new conceptual framework is now used in a genome-wide association studies (GWAS) to identify plant genetic determinants that may be linked to the selection of more beneficial microbiome for plant under low input of fertilizers. 155 Below and Above Ground Interactions Tuesday 23 June – Poster session Water Relations 92 Comparisons of two root water uptake models based on field observations Gaochao Cai*1, Jan Vanderborght1, Valentin Couvreur2, Harry Vereecken1 1 Agrosphere, Institute of Bio- and Geosciences (IBG-3), Forschungszentrum Jülich, Germany, 2Department of Land, Air andWater Resources, University of California, Davis, California, USA Accurate estimation of water requirements and efficient water management of crops requires appropriate mathematic descriptions of water flow. Modeling root water uptake is an important approach to evaluate water movement in the soil-root system. However, dynamic root distribution and a physically based concept to describe water uptake from soil profiles with vertical variations in soil water availability are often not taken into consideration. Therefore, we incorporated observations of soil moisture and root development within two different modeling frameworks and compared the simulated root water uptake. To do so, we carried out field experiments where soil water content, soil water potential, and root growth were monitored non-invasively for three water treatments: sheltered, rain-fed, and irrigated. Root growth of winter wheat was measured by using 7-m long horizontally installed minirhizotubes at six depths with three replicates per treatment. The observed data were interpreted by Feddes model and an upscaled model based on hydraulic architecture approach that considers compensatory root water uptake and hydraulic conductance in the coupled soil root system. Results showed that root water uptake simulated by Feddes model was affected by root distributions, whereas it was influenced by the conductance of the root system when using upscaled model with the same soil hydraulic properties and boundary conditions. Moreover, root water uptake simulated by the two models tended to deviate in dry and wet conditions due to different water stress functions. The upscaled model showed a promising optional approach for simulations of root water uptake by using 1-D spatial discretization to represent soil water dynamics in soil root system. To validate simulated root water uptake by these models, additional measurements, such as sap flow, might be helpful for the predictions of water requirement of different crops. 93 Mathematical modelling for water movement in ridged plant systems Simon Duncan*1, Tiina Roose1, Paul Sweeney2 1 University of Southampton, United Kingdom, 2Syngenta Ltd., United Kingdom In arable farming a widely used practice of growing crops is the use of row production. The potato is traditionally grown in a ridge and furrow system in which the ground’s surface is adapted to form a periodic series of ridges and furrows to allow water flow through the field and provide water to the plants. The model used compares the saturation levels in two different ground geometries for the growth of a potato crop, these geometries are the ridge and furrow system and a flat ground structure. 156 Water Relations Tuesday 23 June – Poster session The water movement model is based on an adapted Richards equation. The model includes a function of water uptake via root activity in each of the geometries that is responsible for reducing the saturation in the root region. The boundary conditions for the model consist of a flux boundary condition on the ground’s surface to allow the inflow of water, a pressure boundary condition at the base of the soil geometry of pressure value pF2 and finally two zero flux boundaries at the sides of the geometries. We determined that the saturation levels in each of the geometry systems remains similar over long periods of time. However we determine there is a difference in the amount of water uptake by the plants in the two structures. We found that the ridge and furrow system has a greater uptake of water when compared to the flat geometry. We observed the formation of dry regions in the area of root activity for each of the two geometries and determined that there is a larger amount of water uptake by the potato plants in the ridge and furrow system. 94 Seeking water: Rooting depth and drought tolerance of Zea mays genotypes driven by nutrient patches and subsoil fertilization Christian Fritz*1, Eric Visser2, Philippe Hinsinger3, Claude Doussan4, Hans de Kroon2, Dina in 't Zand2, Annette Bérard4, Florian Wichern1 1 Rhine-Waal University of Applied Sciences, Germany, 2Radboud University, Netherlands, 3INRA Montpellier, France, 4INRA Avignon, France Water deficiency upon drought is a major constrain on yields. Increasing rooting depth and the soil volume explored by lateral roots seem effective mechanism to prevent water stress in cereals. This research reports the effects of vertical fertilizer placement on root distribution, water and nutrient acquisition of a selection of genotypes (6) in Zea mays. Experiments were conducted at the Phytotron facility in Nijmegen (NL) using 70 cm deep containers equipped with mini-rhizotrons and drip irrigators over 2 growing seasons (full factorial design experiments). We mixed nutrients into the topsoil (0-30 cm) or subsoil (35-70 cm) at field rates. Additionally, nutrients were added as patches (15 vs. 40 cm deep). Drought was simulated by mid-season drying out of the topsoil without affecting subsoil water supply. Trials were complemented by pot experiments manipulating nitrate patches only. Maize genotypes were selected from the EURoot maize panel. We found that rooting depth of maize was strongly dependent on vertical nutrient availability. Fertilization of subsoil increased averaged rooting depth by 3 to 5 times with highest root densities found nutrient patches (280 m.l-1). Improved rooting depth was mainly due to increased root biomass allocation at relatively stable specific root length (~190 g.m-1). Genotypes, however, could differ strongly in root allocation response to nutrients. Plants receiving subsoil fertilization under drought showed higher rates (18%) of photosynthesis and transpiration. Subsoil fertilized plants acquired significantly more water from deep soil layers especially at the height of drought. Interestingly, water use efficiency was little affected by vertical fertilizer placement or concentrations. We observed a trade-off 157 Water Relations Tuesday 23 June – Poster session between early-stage vigour and late-season drought tolerance in treatments receiving subsoil fertilizers. Nutrient placement plays a dominant role in root system architecture and consequently water uptake. Drought tolerance may be further improved by a better understanding of genotype specific root-soil-nutrient interaction. 95 Imaging of root water uptake by MRI in combination with tracer motion Sabina Haber-Pohlmeier*1, Andreas Pohlmeier2, Jan Vanderborght2 1 RWTH Aachen University, ITMC, Germany, 2Research Center Jülich, Germany Flow processes in natural porous media are often too slow to be monitored by direct flow imaging, therefore the visualization of such fluxes is best performed by tracer tracking. While T1 reducing contrast agents are well known in medical diagnostics, their usefulness in natural porous media is not yet well explored. As pointed out in a preceding study, GdDTPA is most convenient since it is very stable and does not adsorb at the soil matrix. Furthermore, it´s specific relaxivity in the liquid phase is sufficiently high to yield good contrast when used in a strongly T1 weighted pulse sequence. Here, we use a simple procedure for the quantification of tracer concentration in saturated and unsaturated natural porous media, where the reduction of relaxation times by desaturation is compensated for by a reference measurement. The procedure is applied to examples from natural porous media, which are decisive bottlenecks in the water flow from soil to the atmosphere: Root water uptake and evaporation from top soil. While plant roots take up water from the surrounding soil, GdDTPA is first enriched in the neighborhood of some roots, indicating their activity. But the tracer is also enriched in the immediate, some mm thick, layer around the root, the so-called rhizosphere, which appears dark in conventional MRI sequences. Although this layer has been frequently interpreted as water depletion zone, the enrichment of GdDTPA there proves the high permeability for water and solutes. 96 Linking mesoscopic to macroscopic scale modeling of water and osmotic stresses on root water uptake Helena Jorda Guerra*1, Adi Perelman2, Naftali Lazarovitch2, Jan Vanderborght3 1 KU Leuven, Belgium, 2Ben-Gurion University of the Negev, Israel, 3Forschungszentrum Jülich GmbH, Germany Current soil-hydrological models predict the effect of water and salt stresses on macroscopic root water uptake by using so-called transpiration reduction functions. However, the use of these functions is restricted to the environmental conditions at which they were empirically developed. This research intends to develop a macroscopic reduction function based on biophysical knowledge. Simulation experiments are conducted for a range of atmospheric conditions, soil 158 Water Relations Tuesday 23 June – Poster session and plant properties, irrigation water quality and scheduling using a 3-D physically-based model that resolves flow and transport to individual root segments and that couples flow in the soil and root system. The effect of salt concentrations on root water uptake is accounted for by including osmotic water potential gradients between the solution at the soil root interface and the root xylem sap in the hydraulic gradient between the soil and root. Simulation experiments are performed in a soil volume around a single root segment. We discuss how the simulation setup can be defined so as to represent: (i) certain root system characteristics, (ii) plant transpiration rate, (iii) leaching fraction of the irrigation, (iii) salinity of the irrigation water, and (iv) water potential threshold for stress onset. The output of these simulation experiments gives a first insight in the effect of salinity on transpiration and on the relation between the bulk salinity, which is used in macroscopic salt stress functions of models that do not resolve processes at the root segment scale, and the salinity at the soilroot interface, which determines the actual root water uptake. In a next step, simulations considering the whole root architecture will be conducted to evaluate how the outcome of the single root simulation experiments can be upscaled to the whole root system scale. 97 Variability of some important Tunisian Chickpea genotypes (Cicer arietinum L.) response to drought stress Abdelmajid Krouma*1, Chedly Abdelly1, Tatsuhito Fujimura2 1 Centre of Biotechnology, Tunisia, 2School of Life and Environmental sciences, Japan Scarcity of water is a severe environmental constraint to plant productivity. Drought-induced loss in crop yield probably exceeds losses from all other causes, since both the severity and duration of the stress are critical. In the current study, a glasshouse experiment was conducted to assess the effects of a progressive drought stress on plant growth, photosynthetic activity and water relations parameters in three Tunisian chickpea genotypes (Cicer arietinum L.). These genotypes were largely cultivated in Tunisia for their diverse uses, Amdoun with big size seeds shown tolerant to salt stress, Chetoui with small size seeds shown sensitive to salt stress and Kesseb with medium size seeds. A progressive drought stress was applied during 16 days of water starvation. A close relationship between plant growth, photosynthesis, leaf water status and osmotic adjustment was found. In comparison to Chetoui and Kesseb, Amdoun showed the highest plant growth and photosynthetic activity, and an important osmotic adjustment under drought stress. 98 Influence of soil nitrate concentration on plant root water uptake Sophie Maloteau*1, Jan Diels2, Sarah Garré3 1 Université de Liège, Gembloux Agro-Bio Tech, Belgium, 2KULeuven, Department of Earth and Environmental Sciences, Belgium, Belgium, 3Université de Liège, Gembloux Agro-Bio Tech, Belgium, Belgium Many real world and modelling experiments, seeking for a quantification of root water uptake pathways, were conducted under simplified conditions. However, we know that plants are able to regulate their hydraulic conductivity coarsely by increasing root growth in certain areas (long term regulation) and finely by changing the conductivity of specific parts of the 159 Water Relations Tuesday 23 June – Poster session root system along the water flow pathway (short term regulation). This regulation is rarely taken into account in models, and its impact at larger scales has never been studied. Novel experimental and modelling techniques must be developed in order to simultaneously monitor root system characteristics, soil solution concentrations and flow rates along the soilplant continuum in a real soil medium. In this context, we will study the influence of nitrate on the hydraulic conductivity of plants root systems. First, we will adapt an existing root water uptake model (R-SWMS), by adding a nitrate-dependent hydraulic root architecture. Next, we will conduct a destructive experiment to quantify how much water and nitrate is taken up by the plant at different development stages, depending on the nitrate concentration in the root zone. As the measures are realized at different times of the experiment on similar plants, short term regulation of plants water uptake can be distinguished from long term regulation. Finally, a second experiment will be prepared to monitor temporal changes in local root water uptake and transpiration upon locally increased nitrate concentration. Plants will be grown in two-dimensional rhizotrons so that they can be used for neutron radiography imaging of the soil-root and root-root water fluxes upon locally increased nitrate concentrations. The combined information on root system architecture, transpiration rate and local root water uptake will then be used in an inverse modelling to obtain root hydraulic conductivities and describe how nitrate can affect it. 99 Minimize the tillage operation improve the root water and nutrient uptakes Rajesh Nallaiah* Tamil Nadu Agricultural University, India As farm tractors and field equipment become larger and heavier, there is a growing concern about soil compaction. Soil compaction can be associated with a majority of field operations that are often performed when soils are wet and more susceptible to compaction. Heavy equipment and tillage implements can cause damage to the soil structure. Soil structure is important because it determines the ability of a soil to hold and conduct water, nutrients, and air necessary for plant root activity. Now a day all the field tillage operation utilized heavy loaded machines only. Compare the traditional and modern tillage operation soil compact more in modern tillage operation. The plant response to subsoil compaction, as with surface compaction, depends on the crop, soil conditions, and the climatic conditions in a particular year. If plants are already stressed for water, subsoil compaction may add to the stress by limiting the growth of plant roots to additional water. If plants are growing in soils that have aeration problems due to high water content, subsoil compaction will slow drainage and could result in an anaerobic root environment that limits nutrient uptake. it does observe the available water and nutrient uptakes with soil and plant relationship. Recently aware of the minimum tillage operation it’s minimized the soil compact condition compare the maximum tillage operation condition. The short duration crops or shallow rooted crops mostly affected the soil compact. Shallow root systems crop very sensitive the water and nutrient uptake its will affect soil compact condition. If they are given the needful tillage operation based on crop condition its will get good root growth easy to uptake the available moisture and nutrient. 160 Water Relations Tuesday 23 June – Poster session 100 A novel 3-D imaging of water on its passage through the rhizosphere in plantsoil systems Sascha Oswald*1, Christian Tötzke2, Nicole Rudolph-Mohr2, Sabina Haber-Pohlmeier3, Andreas Pohlmeier4, Eberhard Lehmann5 1 University of Potsdam, Germany, 2University of Potsdam, Institute of Earth and Environmental Science, Germany, 3RWTH Aachen, Institute for Technical and Macromolecular Chemistry, Germany, 4Forschungszentrum Jülich IBG-3, Germany, 5Paul Scherrer Institute, Germany We use a combination of neutron tomography (NT), fluorescence imaging and magnetic resonance imaging (MRI) to capture the dynamic behaviour in the root – soil interface in situ. This complementary approach provides detailed information about water distribution as well as oxygen or pH patterns within the rhizosphere. The big advantage is that all these measurements are non-invasive and allow a spatially highly-resolved mapping of the rhizosphere throughout the root systems of young plants, in 2-D and 3-D, and dynamic changes during hours and days. Recently, we started to combine NT with MRI to study the dynamic behaviour at the root–soil interface. The high spatial resolution of NT and its sensitivity for water can be exploited for 3D analysis of root morphology and detailed mapping of water content at the root-soil interface and surrounding soil, while MRI can yield complementary information, that is what fraction of water placed in smaller or larger pores or bound in mucilage. The hypothesis is that beyond this it is possible to use special tracers to investigate the fluxes of water through soil, rhizosphere and roots, working with both imaging methods on the same samples. With NT we explored if these tracers pass the rhizosphere and to which degree its concentrations and fate can be detected. The samples were especially constructed to allow MRI in parallel and some of them were also imaged via MRI. The results suggest that by NT the root system can be identified well, its morphology retrieved and related to the concentration changes of the tracer during water uptake, while MRI can work with the same samples at least to image the root system. A detailed analysis of spatial differences in water and tracer passage is on its way and will be complemented by additional MRI measurements and information from fluorescence imaging. 101 Simulating root trait evolution in a functional structural plant model: effect of spatial and temporal water availability Pieter Poot*, Michael Renton University of Western Australia, Australia Functional-structural plant models (FSPMs) represent the interaction between a plants’ structure, the processes occurring within that structure and the surrounding environment. They usually model the dynamic development of the plant structure with explicit topology and geometry, as well as the dynamic processes occurring within and between the structure and its environment. We developed a FSPM that captures the key processes involved in the growth and development of root structures and that can be used to evaluate optimal structural rooting strategies. The model is flexible enough to allow a wide range of strategies 161 Water Relations Tuesday 23 June – Poster session to be represented and explored but simple enough to be linked with an evolutionary optimization algorithm without becoming computationally infeasible. In recently published work we applied this novel approach to a case study habitat with a spatially and temporally heterogeneous distribution of water: perennial plants occurring on shallow soils in seasonally dry environments that depend on cracks in the underlying rock to access more permanent water. The model successfully simulated the evolutionary dynamics of root structures leading to improved fitness under two different fitness criteria and predicted the evolution of several root traits in a direction that has been observed for plants confined to these habitats. Here we will further refine our approach by incorporating more realistic assumptions in the FSPM, the evolutionary algorithm and the fitness criteria. We will show how the structure of optimal root systems and specific root traits change as dependent on the spatial accessibility of soil water (i.e. the frequency of cracks in rocks or pores in compacted soils), and the frequency and amount of rainfall during the wet season. Results will increase our understanding of the evolution of root structures and root traits and will be highly relevant in the context of our changing climate. 102 Modelling root exploration of biopores in soil Katrin Huber1, Glyn Bengough2, Jan Vanderborght1, Mathieu Javaux1, Magdalena Landl1, Harry Vereecken1, Andrea Schnepf*1 1 Agrosphere (IBG 3), Forschungszentrum Jülich GmbH, Germany, 2The James Hutton Institute, United Kingdom Biopores are soil macropores formed by biological activity that enable plant roots to reach water reservoirs at depth. Here we present an extension of the R-SWMS model for water and solute transport in the soil-plant system in which roots are able to grow in pre-existing biopores in the soil. Root growth is based on vector addition and influenced by the local soil parameters, e.g. penetrometer resistance or nutrient availability, around a growing root tip. Water uptake by roots growing inside air-filled macro pores occurs only from the soil matrix and depends on the contact area between roots and biopore walls. Root architecture and growth velocity are shown to depend on plant growth parameters, in particular tropisms, as well as soil parameters, especially the penetrometer resistance and the inclination of macro pores. This has implications on root water uptake patterns that can be beneficial for plants in drought periods. As biopores are often coated with nutrient rich material, this modelling approach can also be used to investigate the benefits of biopores for plant nutrient uptake. 162 Water Relations Tuesday 23 June – Poster session 103 Effect of mucilage on rhizosphere hydraulic properties: a new experimental approach Nico Schultze*1, Eva Kröner2, Andrea Carminati2, Doris Vetterlein1 1 Helmholtz Centre for Environmental Research GmbH - UFZ, Germany, 2Georg-August University of Goettingen, Germany Roots are hypothesized to alter rhizosphere hydraulic properties by release of mucilage. This mechanism is expected to have strong implications for root water uptake under drought conditions. Direct measurement of rhizosphere hydraulic properties is hindered by the dynamic nature of the components involved; root hydraulics change with ontology; mucilage production, composition and diffusion are not constant; soil water content changes. An experimental approach was developed which enables to simultaneously measure hydraulic conductivity and water retention curve around artificial roots covered with mucilage or modified model substances mimicking individual properties of mucilage. The system accounts for the radial geometry of root water uptake. The set-up consists of a soil-filled cylinder connected via a ceramic plate to a water supply for establishing initial soil water content. An artificial root with known and constant hydraulic properties is placed in the center of the soil cylinder and connected to a pump. A microtensiometer is inserted half way between the cylinder wall and the artificial root for soil matric potential measurement. The installation is placed on a weighing cell for determination of changes in soil water content. The experiment is modelled solving a modified version of the Richards’ equation that includes mucilage dynamics. The functions to be fitted are soil matric potential and water outflow from the artificial roots in analogy to classical multistep outflow experiments. Fitting parameters are the van Genuchten-Mualem parameters of the soil and how mucilage modifies them. First results will be presented for homogeneous distribution of gel throughout the soil cylinder versus gel localized just around the artificial root. 104 Numerical study on the impact of rhizosphere processes on root water uptake Nimrod Schwartz*1, Eva Kroener2, Andrea Carminati2, Mathieu Javaux1 1 Catholic University of Louvain, Belgium, 2Gottingen University, Germany For many years, the rhizosphere is known as a unique soil environment with different physical, biological and chemical properties than those of the bulk soil. Indeed, recent studies showed that root exudates, in particular mucilage, alter the hydraulic properties of the soil and that induce non-equilibrium water dynamics in the rhizosphere during drying and wetting cycles. While there are experimental evidences and simplified 1D model for those concepts, an integrated model that couples rhizosphere processes with a detailed decsription water flow in soil and roots is absent. Therefore, the objective of this work was to develop a 3D physical model of water flow in the soil-plant continuum that takes in consideration root architecture and rhizosphere specific properties. To achieve this objective, we coupled a detailed model of water flow in soil and root system (R-SWMS) with a model of the rhizosphere hydraulic properties. 163 Water Relations Tuesday 23 June – Poster session We simulate wetting and drying cycles and examine the water content distribution around roots and the onset of plant stress. We also performed a scenario analysis in which the impact of different rhizosphere processes on water flow and root water uptake was examined. For the wetting process, the model predicted that after infiltration the water content in the rhizosphere remained lower than in the bulk soil (non-equilibrium), but over time water infiltrated into the rhizosphere and eventually the water content in the rhizosphere became higher than in the bulk soil. During drying, the high water holding capacity of the rhizosphere and the non-equilibrium between water content and water potential delayed the onset of stress. These results are in qualitative agreement with the available experimental data on water dynamics in the rhizosphere and suggest an important role of the rhizosphere in modulating plant water stress during drying and wetting cycles. 164 Water Relations Tuesday 23 June – Poster session Root-Root Interactions 105 Unraveling the positive biodiversity-productivity relationship: linking plant community productivity to root trait diversity Lisette Bakker*, Liesje Mommer, Frank Berendse, Jasper van Ruijven Wageningen University, Netherlands The positive relationship between plant biodiversity and community productivity is well established. However, our knowledge about the mechanisms underlying this positive relationship is still limited. The consensus is that the positive effects on productivity are driven by the functional diversity of the community, i.e. the diversity in functional characteristics ('traits') among species, rather than species richness per se. Communities with a higher diversity in traits are expected to better explore the available resources in space and time, a concept called resource partitioning. However, experimental evidence for specific trait combinations underlying resource partitioning is scarce. This study aims to elucidate the link between trait diversity and the productivity of plant communities. Our aims are 1) to establish the relationship between trait diversity and community productivity, and 2) to identify those traits that best predict productivity. In a common garden experiment in Wageningen, the Netherlands, 16 grassland species are grown in monocultures, 4-species mixtures differing in trait diversity, and 16-species mixtures. Here, we focus on the results from the first growing season. Complementarity effects, indicating resource partitioning, did occur but differed greatly between communities (ranging from + 101% to – 48%). Trait diversity of the communities is calculated based on measurements in the greenhouse and existing trait databases, and linked to community productivity. In contrast to most previous studies, we explicitly incorporate belowground traits in our approach, as the resource partitioning is expected to occur predominantly in the soil. However, diversity in belowground traits such as rooting depth and specific root length could not explain complementarity effects. Instead, positive complementarity effects were associated with the presence of specific species. The implications of these findings for biodiversity-ecosystem functioning research will be discussed. 106 Spatial heterogeneity of plant-soil feedback affects root interactions and interspecific competition Hans de Kroon*1, Marloes Hendriks1, Janneke Ravenek1, Wim H. van der Putten2, Liesje Mommer3 1 Radboud University, Netherlands, 2Netherlands Institute of Ecology, Netherlands, 3Wageningen University, Netherlands Soils are inherently heterogeneous. At a small scales, availability of water and nutrients may differ by an order of magnitude. Spatial variation in soil microbial communities is likely to be large as well, as plant species have been shown to accumulate species-specific soil communities in their vicinity. While responses of roots to nutrient heterogeneity have been studied for decades we hardly know how plants respond to soil biotic heterogeneity. Here we report on an experiment in which we combined a plant-soil feedback approach with a competition experiment, involving two grass and two forb species from temperate 165 Root-Root Interactions Tuesday 23 June – Poster session grasslands. We subjected pairs of plants of the same or different species to patches of soil that were previous conditioned by the same or different species. This was repeated in all possible combinations, resulting in 100 different treatments. This design allowed us to examine root interactions as a function of soil type and soil heterogeneity. All species suffered negative soil feedback, but the strength was species-specific, reflected by decreased root growth in own soil compared to foreign soil. Interestingly, the competitively superior species also suffered the strongest negative plant-soil feedback. Root competition was affected by soil conditioning and interacted with soil biotic heterogeneity. Specifically, in heterogeneous soils, substantial reduction in root growth by the superior competitor in patches with own soil provided opportunities for inferior competitors belowground. Our results suggest that plants do not need to perish due to their own soil enemies, but can escape on small spatial scales to better resorts in terms of plant-soil feedback. Negative plant-soil feedback effects are considered an important mechanism of species coexistence and plant community productivity. Small-scale responses of root systems to soil biotic heterogeneity may be an important factor contributing to these effects that has hitherto been overlooked. 107 Start from scratch: how a cereal and a legume create spatial heterogeneities and hotspots of soil microbial activities in the rhizosphere Philippe Hinsinger1, Sarah Placella1, Josiane Abadie1, Camille Cros1, Gabrielle Daudin1, Esther Guillot*2, Agnès Robin3, Claire Marsden4, Naoise Nunan5 1 INRA, France, 2IRD, France, 3CIRAD, France, 4SupAgro, France, 5CNRS, France Rhizosphere processes are known to generate gradients of biological, chemical and physical properties in the soil as a function of the distance to root surface. This has been rather well documented, while micro-heterogeneities of such properties along roots have been little evidenced and quantified. Sampling at the appropriate (millimetric or less) scale is a methodological challenge, and many of the gradients data have been obtained with microcosms designed to simplify the geometry of the rhizosphere. Here we report of the use of rhizoboxes with 2D access to the soil profile, in which we mapped acid phosphatase activity in situ in chickpea and durum wheat, either grown alone or intercropped, as well as in rhizoboxes containing bare soil (control treatment without plant). This enzymatic activity was actually monitored over 5 weeks with a non-destructive technique based on the use of a membrane sorbing the enzymes, that was applied once a week against the soil profile for a collection period of 3.5 hours. The soil had been sampled in the nill treatment of a long-term fertilizer trial that had not received phosphorus application over 40 years, and thus had very low phosphorus availability. It was sieved and packed so that the spatial heterogeneity in each rhizobox was minimal at the start of the experiment. The presence of roots of both plant species was clearly responsible for the increased heterogeneity of acid phosphatase activity observed relative to bare soil. Greater acid phosphatase activity was generally observed near roots, especially chickpea roots and root nodules. The greatest activities were found in the vicinity of the younger parts of the roots (apices and elongation zones). While there was a clear difference between the two plant species, there was no significant effect of intercropping at the fine spatial scale of investigation reported in the present study. 166 Root-Root Interactions Tuesday 23 June – Poster session 108 Root exudate effects on humic acids and the consequences for belowground plant species interactions Diederik Keuskamp*1, Rob Comans1, Liesje Mommer2, Ellis Hoffland1 1 Department of Soil Quality, Wageningen University, Netherlands, 2Nature Conservation and Plant Ecology, Wageningen University, Netherlands Humic acids are supramolecules that are the major organic components of most soils. They have been shown to induce plant species specific (root)growth responses, such as lateral root formation and higher (relative) root biomass. These root responses depend on the molecular size of the HA. Interestingly, organic acids are known to destabilize HA into smaller fractions. Since root exudates contain several organic acids, such as citric acid or phenolic acids, root exudates may be of extreme relevance for the HA induced growth responses. The quantity and quality of these root exudates are known to be plant species specific. This could mean that the root exudates of a plant can affect the growth of neighbouring plants via their interaction with HA, especially when these neighbouring plants are of another species. Mommer and co-workers showed root aggregation for the grass species Anthoxantum odoratum when in interspecific interaction. In our research project we test the hypothesis whether this root aggregation could be (partly) explained by humic acids (HA) and its interaction with root exudates. We investigate this interaction by growing A. odoratum and Leucanthemum vulgare seedlings in a mixed culture and under the sterile conditions of agar plates. These growth conditions enable us to identify changes in root development (independent of different microbial and/or soil effects) and allow us to add HA at different concentrations to test the effects of these HA. We will also make use of HA pre-treated with root exudates or organic acids, trying to mimic the growth responses in monocultures of each of the species. On the poster we will present the preliminary results of our ongoing research. 109 Effects of different wheat varieties intercropped with faba bean on root morphology of wheat Wang Yuyun, Ren Jiabing, Tang Li* Yunnan Agricultural University, China Wheat and faba bean intercropping is one of the important intercropping systems in China. The effectiveness of this intercropping system on grain yield increasing, nutrients acquisition improvement and disease control has been demonstrated. But the genotypes difference of intercropping has been seldom studied. A field experiment was conducted to investigate the effect of intercropping on wheat root morphologyalong the soil depth. Three wheat varieties YM-47, YM-42 and MY-29, with different resistances to wheat powdery mildew were used by monocropping or intercropping with faba bean. Results showed that, wheat and faba bean intercropping significantly increased the wheat root length density and root surface area, but decreased the root average diameter. Compared with monocropping wheat, the root length density of intercropped YM-47 and 167 Root-Root Interactions Tuesday 23 June – Poster session YM-42 were increased by 225.8% and 301.7%, the root surface area were greater by 235.9% and 236.5% respectively at 0-10cm soil layer, but the intercropped MY-29 showed no significant difference with that of monocropping. There was no significant difference of root length density at 10-20 cm layer between intercropping and monocropping. But the root length density and root surface area of wheat were lower at 20-30 cm layer under intercropping. Compared with monocropping wheat, No significant difference was found at 0-10 cm layer, but the root average diameter in intercropping decreased by 12%, 16% and 11% at 10-20 cm and by 32%, 12% and 14% at 20-30 cm soil layers of YM-47, YM-42 and MY-29, respectively. The root length density, root average diameter and root surface area showed significant positive correlation with soil available P content. Wheat and faba bean intercropping significantly increased wheat yield and showed significant intercropping advantage. The LER was 1.20, 1.14 and 1.01 for wheat YM-47, YM-42 and MY-29, respectively. In conclusion, The intercropping advantages were influenced by intercropped wheat varieties in YM-47>YM-42> MY-29. 110 Plant nitrophily drives plant productivity and plant-microbial interactions Barbara Pivato*1, Hugues Busset2, Florence Deau2, Annick Matejicek2, Philippe Lemanceau2, Laurent Philippot2, Delphine Moreau2 1 UMR Agrécologie - INRA Dijon, France, 2INRA, UMR1347 Agroécologie, France The interest for plant interactions, especially between crop and weed species, in agricultural situations has increased with the pressure to reduce the use of chemical inputs. Thus, a better knowledge is required in plant traits mediating the issue of plant competition and how this competition is impacted by the environment. Nitrophily is a plant trait referring to plant habitat, ranging from oligotrophic species mostly found in soils with low N content to nitrophilic species better adapted to soils with high N content. We hypothesize that nitrophily impacts (i) the issue of plant-plant interactions, through competition for N and light and (ii) plant associations-rhizospheric microbial community interactions. We can expect that microbial community from nitrophilic/oligotrophic roots association is (i) different from community associated to the same plants cultivated alone, and (ii) differently impacted according to the association type. To test these hypotheses, three nitrophilic and two oligotrophic graminaceous species (one crop plant and four weeds) were cultivated in mono- or bi-specific associations in a soil supplemented or not with N. The issue of the plant-plant interactions was assessed by checking their productivity through biomass measures. The impact of the different plant combinations on total bacterial communities was characterized by A-RISA (AutomatedRibosomal Intergenic Spacer Analysis) fingerprinting. 168 Root-Root Interactions Tuesday 23 June – Poster session The biomass was significantly higher in nitrophilic than in oligotrophic species when grown in N supplemented soil. Bi-specific associations showed different root biomass compared to monocultures in both N conditions. Genetic structure of rhizospheric bacterial community differed according to the N level. In higher N availability, bacterial communities structures were very similar, whereas in N limiting conditions, the rhizosphere of bi-specific associations harbored different bacterial communities in comparison to monocultures. These results suggest that nitrophilic plant trait is a driver of plant productivity via plant-plant competition, and this affects rhizospheric microbial community in N limiting conditions. 111 From pots to plots: Hierarchical trait-based prediction of population biomass in a mesic grassland Thomas Schröder-Georgi*1, Christian Wirth2, Karin Nadrowski3, Sebastian T. Meyer4, Liesje Mommer5 1 University Leipzig, Biology, Germany, 2University of Leipzig, Institute of Biology / German Centre for Integrative Biodiversity Research (iDiv), Germany, 3University of Leipzig, Institute of Biology, Germany, 4Technische Universität München, Department of Ecology and Ecosystemmanagement, Germany, 5Wageningen University, Nature Conservation and Plant Ecology group, Netherlands Plant functional traits are powerful tools when it comes to the prediction of plant performance and ecosystem functioning. For a long time ecologists focused on aboveground traits but widely ignored root traits because their sampling and measurement is expensive and labourintensive. A number of newer studies have shown that root traits link not only to belowground (e.g. N cycling) but also to aboveground functions (e.g. biomass production). So far it has never been tested, how the importance of root traits for plant performance depends on the hierarchical level (individual vs. population). We hypothesized that root traits are more important on the population level than on the individual level, due to an increase in process complexity. We used univariate and multiple regression analyses to test the importance of 35 root-, leafand stature traits for the prediction of individual and population biomass production of 59 European grassland species. We found that traits of all three clusters (root, leaf and stature) correlate with both individual and monoculture biomass. The most parsimonious multiple regression model significantly improved for both, individual and monoculture biomass, when root traits are included. However, whereas root traits improved the individual biomass model by only 3% they improved the monoculture biomass model by 28%. Root traits are important for both, individual and population performance but the significance of root traits increases with increasing hierarchical level. 169 Root-Root Interactions Tuesday 23 June – Poster session 112 Research on rhizosphere soil phosphorus fractions and availability in a maizesoybean relay intercropping system Chun Song*1, Xia Xiao2, Lu Mao2, Min Xu2, Taiwen Yong3, Wenyu Yang3 1 Sichuan Agricultural University, China, 2Institute of Ecological and Environmental Sciences, College of Resources and Environment, Sichuan Agricultural University, China, China, 3Key Laboratory of Crop Ecophysiology and Farming System in Southwest China, College of Agronomy, Sichuan Agricultural University, China, China Intercropping is a traditional and sustainable cultivation model based on ecology principle of plant-plant interaction and plant-soil feedback between different biological species. However, the knowledge of crop rhizosphere soil phosphorus forms transformation and availability under intercropping system is still limited. This research focused on the effect of root interaction on crop aboveground biomass, rhizosphere soil phosphorus availability in a maize-soybean relay intercropping system by treated maize and soybean root system with root separation by polyvinyl chloride (PVC) clapboards or without root separation in PVC boxes. Two seedlings of maize and four seeds of soybean were planted per PVC box, and the distances between maize and soybean plants were 60cm. Soluble chemical reagents of CO(NH2)2, KH2PO4 and K2SO4 were applied as N, P and K fertilizer. We collected plant and soil samples in maize silking and maturity stages, and soybean branching, early flowering and maturity stages separately, and then determined the crop aboveground biomass and rhizosphere soil phosphorus fractions, to clarify the mechanism of rhizosphere soil phosphorus fractions transformation and bioavailability in maize-soybean relay intercropping system. The plant biomass data showed that soybean aboveground biomass were significantly higher in treatments without root separation. The phosphorus fractionation results showed that treatments without root separation significantly improved the rhizosphere soil NaHCO3-Po and NaHCO3-Pi content in maize silking stage, whereas decreased NaHCO3-Po content in soybean branching stage, and the NaHCO3-Pi content in soybean early flowering stage. These results indicated that maize had stronger competition for phosphorus uptake than soybean during maize and soybean co-growth stage. Residue P content of maize rhizosohere soil in maturity stage considerably higher than soybean maturity stage, which indicated that soybean had higher soil phosphorus use efficiency. In a word, maize-soybean relay intercropping can improve soil phosphorus use efficiency by root interaction, so as to improve crop yield. 170 Root-Root Interactions Tuesday 23 June – Poster session 113 Root priority effects: Are root-root interactions in grasslands significantly affected by who interacts with whom and who arrives first? Vicky Temperton*1, Philipp von Gillhaussen2, Marco Brendel3, Marc Faget4, Stephan Blossfeld3 1 Plant Sciences (IBG-2), Forschungszentrum Jülich & Leuphana University Lüneburg, Germany, 2Bayreuth University, Germany, 3Plant Sciences (IBG-2), Forschungszentrum Jülich, Germany, 4University of Louvain, Germany Whilst we all know that the timing of interactions between plants has important implications for plant performance, we still know relatively little about how community assembly (either aboveground or belowground) is affected by who arrives first. In a large field experiment we tested the possible role of priority effects of by sowing either grasses, legumes or nonlegume forbs five weeks before the other two functional groups, at the same time as testing effects of sowing a low or a high diversity seed mixture. Intriguingly, the species richness of the mixture did not have much of an effect on aboveground biomass, whereas the arrival time of each functional group had a very significant effect on aboveground as well as belowground productivity. When legumes were sown before forbs and grasses, community root productivity was significantly lower during the first growing season than when grasses were sown first. A repeat measurement of root turnover using in-growth cores in 2014 (data pending) will show whether this pattern was stable and created a priority effect underground or not. Which species interact can also affect rhizosphere pH. In a controlled experiment combining maize with bean, we measured rhizosphere pH and found that if these two species were grown together variation in pH was lower than when each species was growing alone. We also tested whether there were species-specific differences in the extent to which grass species experienced nitrogen facilitation when interacting with the legume Trifolium repens and this also in presence of an invasive non-legume forb Senecio inaequidens. The grass Holcus lanatus benefitted the most from growing near the legume, but pots with Festuca pratensis in had highest root biomass and less pronounced facilitation effects. Overall our data show that plant root performance is clearly affected by the identity and functional traits of the neighbouring plants. 114 Characteristics of phenolic acids exuded by roots in wheat and faba bean intercropping Jingxiu Xiao*1, Yi Zheng2, Li Tang1 1 Yunnan Agricultural University, China, 2Southwest Forestry University,Yunnan Agricultural University, China Phenolic acids play important role in rhizosphere process, and few works have been cited on the effects of intercropping on phenlic acids exudation. Wheat and faba bean is an important planting pattern in southwest of China due to its significant yield advantages and diseases resistance. Hydroponic culture was conducted and the exudates form the roots were collected and examined by HPLC to investigate the effects of wheat and faba bean intercropping (W//F) on the phenolic acids exudation. The results showed that the three 171 Root-Root Interactions Tuesday 23 June – Poster session phenolic acids were identified in root exudates: ρ-hydroxybenzoic, vanillic and syringic acid in both mono and intercropping. Intercropping decreased total amounts of phenolic acids in root exudates by 39.0%-70.1% and 37.56 -57.79% respectively, in comparison with that of mono cropping wheat (MW) and faba bean (MF). The maximum amount of phenolic acids exuded by root was found on 35d after transplanting in both mono and intercropping treatments, and exudation rate tended to decrease with crop growth from35d to 85d. During whole crops growth stages, intercropping significant decreased exudation rate of ρhydroxybenzoic and syringic acid by 41.06%-100% and 37.89%-64.35% respectively in comparison with that of MW. In W//F, vanillic and ρ-hydroxybenzoic acids were not detected from root exudates on 35d and 55d after transplanting respectively, which were detected till to85d in mono cropped wheat and faba bean. 172 Root-Root Interactions Tuesday 23 June – Poster session Root Turnover 115 Root decomposition along a grassland plant diversity gradient Hongmei Chen*1, Arthur Gessler2, Hans de Kroon3, Michael Scherer-Lorenzen4, Liesje Mommer5, Christian Wirth1, Alexandra Weigelt1 1 University of Leipzig, Germany, 2Swiss Federal Institute for Forest, Snow and Landscape Research WSL, Switzerland, 3Radboud University, Netherlands, 4University of Freiburg, Germany, 5Wageningen University, Netherlands Decomposition of plant litter is a key process for carbon cycling and nutrient availability in ecosystems. However, we know very little about decomposition of root compared to leaf litter, especially in the light of biodiversity-ecosystem functioning relationships. To elucidate how plant diversity affects root decomposition in grassland and to disentangle the effects of root litter quality from environmental factors (including both abiotic and biotic aspects), three decomposition experiments using a litter-bag approach were conducted at the Jena Experiment, Germany, on 80 plots differing in species richness (1, 2, 4, 8, 16 species): 1) decomposition of plot-specific roots in their origin plots to assess plant diversity effects; 2) decomposition of plot-specific roots in a common-garden plot to assess root litter quality effects; and 3) decomposition of standard roots (from Lolium perenne) in all plots to assess general environmental effects. The litter bags were installed in April 2014 and retrieved 1, 2 and 4 months afterwards to trace the decomposition progress. Mass loss was determined as a measure of decomposition rate. With increasing plant diversity, decomposition of plot-specific roots decreased both in their origin plots and in the common-garden plot, while decomposition of standard roots did not change along the plant diversity gradient. These patterns remained constant over time. Moreover, in the common-garden plot, there was a negative effect of the presence of grasses on root decomposition, which increased along the diversity gradient. Our results suggest: 1) that the overall effect of plant diversity on root decomposition is negative; 2) that this effect is primarily due to changing root litter quality rather than environmental factors; and 3) that grasses have a substantially higher negative effect on root litter quality and thus decomposition compared with other functional groups. 173 Root Turnover Tuesday 23 June – Poster session 116 Fertilization and root presence of permanent grassland have no effect on microbial biomass and mineralization Christoph Knoblauch*1, Conor Watson2, Nicole Wrage-Mönnig3, Rolf Becker4, Clara Berendonk5, Florian Wichern2 1 Rhine-Waal University of Applied Sciences, Faculty of Life Sciences, Germany, 2Faculty of Life Sciences, Rhine-Waal University of Applied Sciences, Germany, 3Faculty of Agricultural and Environmental Sciences, University of Rostock, Germany, 4Faculty of Communication and Environment, Rhine-Waal University of Applied Sciences, KampLintfort, Germany, 5Chamber of Agriculture North Rhine-Westphalia, Kleve, Germany The release of nitrogen (N) from soil organic matter (SOM) and plant residues is an important source of available N in permanent grassland. Added inorganic fertilizer can result in an alteration of root biomass and therefore also influence the input and turnover of SOM. However, there is a lack of knowledge about interactions of inorganic fertilization and mineralization in grassland soils. The objectives of the current study were to (i) measure the carbon (C) and N mineralization of grassland soils, (ii) to quantify microbial biomass (MB) and (iii) to monitor the effect of roots on C and N mineralization. We initiated a short-term incubation experiment with varying organic and inorganic fertilizer rates and the presence or absence of course roots and measured the effects on C and N mineralization and MB. Soil was taken from a fertilizer field trial on permanent grassland (Lolium perenne L.) and visible roots were removed manually. During incubation, we measured soil respiration at least once a week for a period of 35 days. Inorganic N and MB were determined at the beginning and end of the experiment. The presence of roots did not significantly affect the mineralization processes within the contrasting fertilizer rates. However, inorganic N tended to increase with a higher application rate of the two fertilizer types. Neither cumulative soil respiration nor MB was affected by fertilizer type or application rate. Yet, we determined a hysteresis effect of the pre-equalized water content on soil respiration. Especially at the beginning, water content concealed the importance of fertilizer rate in terms of soil respiration. In conclusion, the heterogeneity of the taken samples within each fertilizer treatment predominated over the amount and type of fertilizer or presence and absence of roots. 117 Long live the roots! Characterizing root lifespan in a grassland biodiversity experiment Natalie Oram*, Liesje Mommer, Frank Berendse, Jasper van Ruijven Nature Conservation and Plant Ecology, Wageningen University and Research Centre, Netherlands Since the discovery of the positive plant diversity-productivity relationship in grasslands, the mechanisms underlying this relationship have become central research questions. It is wellknown that greater plant species diversity results in higher standing root biomass. However, whether this is the result of increased root production or decreased root mortality is largely unknown. Yet, understanding the dynamics of root biomass is crucial with respect to carbon and nutrient cycling. Increased root production could lead to enhanced organic matter inputs into the soil, whereas increased root lifespan may actually slow down these inputs, leading to different soil C and N dynamics in the long term. An increasingly positive plant-soil feedback, as well as changes in microclimate and nutrient uptake that parallel high plant diversity may 174 Root Turnover Tuesday 23 June – Poster session lead to a more favourable environment for roots. We hypothesize that root lifespan will be positively correlated with species richness; greater root lifespan in diverse plots will contribute to the observed increase in standing root biomass. We test this hypothesis in a long term experimental grassland biodiversity experiment in Jena. We aim to quantify root lifespan in a novel and more precise way using repeated imaging of roots through minirhizotrons and quantification of root decomposition with a litterbag experiment. Root morphology and standing root biomass will also be measured. Imaging started in March, 2015, one year after the minirhizotrons were installed. Preliminary results will be presented. 118 Effect of nitrogen form, pH and plant species in the mobilization and acquisition of P from a recycled phosphorus fertilizer Ana Alejandra Robles Aguilar*1, Vicky M. Temperton2, Stefan Blossfeld2, Nicolai David Jablonowski2 1 Forschungszentrum Jülich GmbH, Germany, 2Forschungszentrum Jülich GmbH, Institute für Bio- und Geowissenschaften, Germany World phosphorus resources are limited. Therefore recycling of phosphorus from waste materials is important, and struvite (MgNH4PO46H2O) is a common precipitate recovered from waste water treatments or during anaerobic digestion of manure. Our approach is to evaluate how mobilization of phosphorus may differ when two different species (narrow– leaved Lupin and Maize) are grown in an acidic or alkaline sand with phosphorus added as either struvite or as superphosphate. Nitrogen was applied as ammonium or nitrate as an important factor that could affect phosphorus availability by changing soil pH. The parameters to evaluate the mobilization of phosphorus from struvite were phosphorus uptake, phosphorus present in the soil and comparison of plant performance within the different treatments. Lupines are capable of symbiotically fixing atmospheric nitrogen, as well as to release phosphate-mobilizing carboxylates. These two traits make lupines good candidates forstudying nutrient mobilization in the rhizosphere. In order to observe if citrate (mimicking root exudates) was able to make the P from the struvite more available compared with water, our study also included a flushing experiment with citrate in columns filled with two different sands of acidic and alkaline pH mixed with struvite. We predict that lupine growing on alkaline sand will have better access to phosphorus in struvite (than on acidic sand) due to its ability to acidify the rhizosphere via exudation of carboxylates. Thus we expect lupine to have higher biomass when growing on alkaline sand with an ammonium supply. 119 Plant presence reduces root decomposition rate of non-legume species Sirgi Saar*1, Marina Semchenko1, Janna M Barel2, Gerlinde DeDeyn2 1 Tartu University, Estonia, 2Wageningen University, Netherlands Litter nutrient concentrations are important traits for litter decomposition, which generally proceeds faster with increased nutrient concentrations in the litter. However, the presence of living roots can affect decomposition rates due to a priming effect by releasing root exudates 175 Root Turnover Tuesday 23 June – Poster session which can give saprotrophic microbes an energy boost, enabling them to degrade the litter faster. To test whether and how plant presence affects decomposition of roots with different traits, we used dead roots of seven species (3 grasses, 3 legumes, 1 forb) as litter material and measured the litter mass loss after 8 weeks of incubation in soil with or without a white clover (Trifolium repens) plant. We expected that the decomposition rate would increase in the presence of a living plant, especially in the case of nitrogen-rich litter, because root exudates can provide a source of easily available carbon and stimulate microbial activity while subsequent nitrogen release from the litter could further enhance microbial activity and thus decomposition. On the contrary, we found that the decomposition rate of grass and forb roots decreased in the presence of living roots, while the decomposition rate of legume roots was not significantly affected by the presence of a living plant. Our results show that root decomposition rate can be slowed down in the presence of a living plant, but that this effect depends on the properties of the decomposing roots, with reduction being pronounced in root litter poor in N and P but not in the relatively nutrient-rich legume root litters. 120 Consequences of plant kin recognition for microbial soil feedback and root decomposition Marina Semchenko*1, Anu Lepik2, Sirgi Saar2 1 University of Manchester, United Kingdom, 2Department of Botany, University of Tartu, Estonia Recent studies have shown that plants are able to detect the genetic identity of their neighbours. Some species proliferate roots when grown next to unrelated individuals but apparently avoid direct competition with kin by reducing root growth. Neighbour recognition can be mediated by root exudates and involve complex changes in root growth and morphology. The consequences of such behaviour for ecosystem functioning are as yet unknown. In this study, we examined the consequences of kin recognition for microbial soil feedback and root decomposition. In the conditioning stage, plants of Deschampsia caespitosa were grown in groups of either siblings or unrelated individuals from the same population. In the feedback stage, new seedlings were planted into soil conditioned by either siblings or non-siblings. Conditioned soil included dead roots, allowing estimation of root litter decomposition during the feedback stage of the experiment. We found that microbial soil feedback for seedlings at early stages of development tended to be more negative on soil previously occupied by siblings. However, soil feedback did not differ between sibling and non-sibling soils at later stages of development. We also found evidence for significantly slower decomposition of root litter obtained from plants grown in groups of siblings compared with groups of unrelated individuals. Roots of plants that grew with siblings also had a significantly higher C:N ratio. This may explain slower root decomposition and indicate increased investment into pathogen defence at the expense of competitive ability when interacting with kin. 176 Root Turnover Tuesday 23 June – Poster session 121 Phosphorus availability and its interaction with plant belowground carbon, nitrogen and phosphorus input into Oxisol Pierre Stevenel*1, Astrid Oberson1, Samuel Abiven2, Idupulapati M. Rao3, Emmanuel Frossard1 1 ETHZ - Institute of Agricultural Sciences, Switzerland, 2University of Zürich Department of Geography, Switzerland, 3International Center for Tropical Agriculture, Colombia Phosphorus (P) deficiency is an issue for a large part of tropical soils worldwide. Low P availability in Oxisols is mainly due to strong P sorption and often limits plant growth. This feature affects the carbon (C) cycle by influencing its capture by plants, and the input of belowground C contained in roots and rhizodeposition. Moreover, low P availability influences nitrogen (N) input in soil by limiting symbiotic N2 fixation by legumes. Additionally, plants have developed strategies to cope with P deficiency, such as root system extension or exudation enhancement, which also determine belowground organic matter input. However, a quantitative understanding of combined root and rhizodeposition C and N inputs in response to P availability has not yet been studied. In this project, we want to investigate the role of P availability on the regulation of belowground C, N and P inputs, and its impact on the turnover of these inputs in soil nutrient pools over time. Experiments will be performed with two tropical plant species: the legume Stylosanthes guianensis and the grass Brachiaria decumbens. Plants will be grown in an Oxisol along a gradient of plant available P. A promising tri-isotopes (13C, 15N, 33P) labeling method will be used to quantify belowground C, N, P inputs. The isotopic composition and isotope recovery in shoots, roots and root free soil will be determined. Following this experiment and after removing the aboveground biomass, nutrient turnover will be studied in soils incubated during three months. The incorporation of plant belowground input of C, N and P into soil pools over time will be determined with the isotopic composition. This project will provide a better understanding of the influence of P availability on C and N inputs to soil but also strong foundations on the long term storage of those nutrients in soils. 122 Root activity and dormancy in forest communities along an elevational gradient Yan Wang*1, Zhun Mao2, John Kim3, Christophe Jourdan4, Herve Ray4, Alexia Stokes3 1 Université Montpellier 2, France, 2c IRSTEA, UR EMGR, 2 Rue de la Papeterie, BP 76, 38402 Saint-Martin-d’Hères Cedex, France, France, 3INRA, UMR AMAP, Boulevard de la Lironde, 34398 Montpellier Cedex 5, France, France, 4CIRAD, UMR Eco&Sols – Ecologie Fonctionnelle & Biogéochimie des Sols & Agroécosystèmes (Montpellier SupAgroCIRAD-INRA-IRD), 2 Place Viala, 34060 Montpellier, France, France Belowground processes in plant communities drive the sequestration of carbon into soil, but how these processes are modified by climate remains largely unexplained. Elevational gradients are unique tools for studying the responses of communities to climatic variability in situ. 177 Root Turnover Tuesday 23 June – Poster session We installed rhizotrons in contrasting forest communities at 1400 m, 1700 m and 2000 m. We measured the number of growing roots and elongation every month over 4 years. Results showed that mean daily root elongation rate (RER) was driven by soil temperature between 0-8°C, outside which extreme soil temperatures perturbed growth. RER peaked in springtime, and a smaller peak was sometimes observed in autumn. Cumulated root length was not significantly different between altitudes, although the growing season was significantly shorter at 2000 m. However, when winter snow cover was > 6 months, root growth was severely limited the following year. Root longevity was dependent on altitude and the season in which roots were initiated; roots emitted in the autumn lived significantly longer than those initiated in springtime. Root diameter was a significant factor explaining much of the variability. Few differences in root demography between contrasting plant communities within each altitude were found, and were explained by air or soil temperature. We conclude that at high altitudes and when extreme abiotic events are not restrictive to carbon supply, more resources are invested belowground in a shorter period of time, revealing a plastic response to climatic variables within a community. 178 Root Turnover Tuesday 23 June – Poster session BIOFECTOR 123 Single and tripartite biofectors for tomato (var. Mobil) upscaled from pots to plots Borbala Biro*, Zita Szalai, Anita Dudás, Tamás Gáspár, Heléna Wass-Matics, Zsolt Kotroczó Corvinus University of Budapest, Hungary Living and non-living bioeffectors are beneficial treatments of organic and sustainable agricultural practices. It is a question if single or 2-3-types of microbes in one particular product is the most necessary or how to successfully upscaling the beneficial effects from pots to the fields? Tomato Solanum lycopersicon Mill. ’Mobil’ was used in the experiment. Bioeffector products, as BE1: Trichoderma harzianum T-22 and BE-4: Hungarian Trichoderma TDM, including Azotobacter and Azospirillum N2-fixers, BE2: Pseudomonas sp., BE3: Bacillus amyloliquefaciens Rhizovital 42 F1 were applied on the bases of the supplier’s recommendations. The plants were grown in 2.5 kg soil/pot in 4 replicates. TSP (0,62g), Rock-phosphate RP (1,59g), Patentkali (1,67g) and Calcinit (1,46g) were applied at the pots and 1200 kg/ha from both at the field. BE application was performed both at sawing and at the time of plantation. Growth of tomato, shoot and root biomass was assessed and general soil characterization, including MPN counts of bioeffectors in tomato rhizosphere. Results were evaluated by statistical probes. Upscaling was resulting the similar beneficial effect at Bacillus (BE3) bioeffector, with greater variability and less improvement at the field. Shoot biomass production was less variable parameter than the root or fruit biomass, more particularly also among the rather changeable field condition. Combined tripartite microbial inoculations seem to have better beneficial influence, then the only single bioeffector treatments in general. Effect of BE1 Trichoderma inoculation was improved for instance in combination with free-living and associative Nitrogen-fixers, due to the relatively high P-content (430 mg/kg) of the slightly humous sandy soil. Plant microbe interaction is largely dependent on the soil nutrient-status and the balance among nutritive elements. 124 Soil application of seaweed extracts and micronutrients to improve the cold stress tolerance in maize during early growth Klara Bradacova*1, Nino Weber2, Markus Weinmann2, Günter Neumann2 1 University of Hohenheim, Institute of Crop Science (340h), Germany, 2University Hohenheim, Institute of Crop Science (340h), Germany Low soil temperature in spring is a major constraint for cultivation of tropical crops in temperate climates, associated with impaired seedling development, inhibition of root growth and activity. 179 BIOFECTOR Tuesday 23 June – Poster session In this study, we tested the potential of commercial bio-effector products such as seaweed extracts and rhizobacteria (strains of Bacillus and Pseudomonas) with plant growth promoting potential to improve the tolerance of maize to low root zone temperatures during early growth. Maize (v. Colisee) was cultivated in pots containing 2 kg of a fully fertilized silty-loam soil (pH 6.8). In order to control the root zone temperature (RZT), pots were installed in a cooling system. After germination at 22 °C, the cold stress phase (12-14 °C) started at 14 days after sowing to simulate a cold-period in spring. Bioeffectors were supplied close to the plants with a dosage of 109 CFU kg-1 soil for bacteria and 0.017 g kg-1 soil for seaweed extracts at 0, 2 and 4 weeks after sowing. In two independent experiments, positive BE effects on plant growth and particularly on root development at low RZT were exclusively detected for seaweed extracts with high Zn/Mn contents and similar growth promotions were induced by sole application of Zn and Mn in comparable amounts. This finding suggests that also the seaweed extracts were mainly acting via improved Zn and Mn supply to the plants. Assays of different oxidative stress markers showed that the beneficial effect of Zn/Mn treatments was associated with increased superoxide dismutase (SOD) acitivity in the root tissue, playing a key role in antioxidative defense mechanisms. In this context, Zn/Mn supplementations may effectively improve cold stress tolerance of maize by their function as enzymatic co-factors particularly for SOD. Accordingly, formation of necrotic leaves in cold-stressed plants was associated with a low Zn-nutritional status. 125 The combination of humic substances and inoculation with different microorganisms enhances the growth of maize (Zea mays L.) Vincenza Cozzolino*, Hiarhi Monda, Daniele Todisco, Davide Savy, Giovanni Vinci, Alessandro Piccolo University of Naples Federico II, Italy Plant biostimulants, or bioeffectors (BEs), include diverse substances and rhizosphere microorganisms that enhance plant growth, contributing to improve fertilizer use efficiency, enhancing nutrient uptake, and increasing root health through competition with root pathogens. The well-known direct effect of humic substances (HS) on root growth has produced an expanding market for HS extracted from various composted green wastes, that represent a more sustainable source than those obtained from expensive fossil matrices, such as lignites or peats. The aim of this work was to evaluate the ability of these new biotechnological tools to promote the growth of maize plants in soil with low P availability. A pot trial was performed with the following BE treatments: 1. B0 (no inoculation), 2. B2, Pseudomonas sp plant growth promoting bacteria (Proradix), B3, Bacillus amyiloliquefaciens PGP (Rhizovital 42); MYC Funneliformis mosseae and Rhizophagus irregularis, commercial inoculum of arbuscular mycorrhizal fungi (Aegis); HA, humic acid (HA) from composted artichoke wastes. We evaluated the benefits of each BE alone or in combination. The P addition to soil comprised: P0 (no P addition), and CM (composted cow and buffalo manure at 50 mg P/kg soil). The impact of P fertilization, inoculation and HS were evaluated on biomass production and P and 180 BIOFECTOR Tuesday 23 June – Poster session N content. The plants were grown in pots filled with 5 kg of substrate, prepared using an alkaline clay-loam soil and harvested 8 weeks after sowing. The benefits of the combined action of compost, microorganisms and HS were evident. Inoculation with the bioeffectors enhanced plant growth, as well as N and P content in shoots, especially in treatments where each microorganisms were mixed with HA. This new generation of biological products based on HS and selected beneficial microorganisms provides opportunities for an effective biological support of a sustainable crop production. 126 Combinations of rhizosphere-competent fungal and bacterial isolates improve nutrient acquisition and growth in maize and tomato Joerg Geistlinger*1, Markus Weinmann2, Borbala Biro3, Alessandro Piccolo4, Shekhar Sharma5, Rainer Borriss6 1 Anhalt University of Applied Sciences, Germany, 2University of Hohenheim, Germany, 3Corvinus University of Budapest, Hungary, 4University of Naples Federico II, Italy, 5Agri-Food & Biosciences Institute, United Kingdom, 6ABiTep GmbH, Germany BIOFECTOR (7th EU Framework Programme, grant agreement 312117) works on “an improved understanding and utilization of biological processes supporting soil fertility”, essentially on the soil microbiome and bioactive natural compounds interacting with the root system and/or mutual or symbiotic microbes, termed bio-effectors (“BEs”). Here, we report on the results of Work Package 2: BE combinations and synergisms. Ten partners from 5 EU countries agreed on a corporate standardized experimental design for greenhouse and field trials with tomato and maize as model crops. Local soils received basal fertilization (low in P) and were enriched with standard BEs (BE1= Trichoderma harzianum (TRIANUM-P™), BE2= Pseudomonas spec. (PRORADIX™), BE3= Bacillus amyloliquefaciens (RHIZOVITAL™)), combinations thereof and in combination with newly developed fungal and bacterial strains as well as extraction products from seaweed and compost. Main findings after 2 years of research were in maize: (1) combining standard BEs1-3 in a loamy top soil with low P availability has no positive effects, but growth-promotion was achieved by combining BE2 with seaweed extracts. (2) BEs1-3 have no effects on cold stress induced necrotic leaf areas but necrosis could be completely avoided by combining Zn/Mn micronutrients with seaweed extracts. (3) Composted cow manure with the combination of BE3 and humic acid extracted from compost yielded 65% more dry weight biomass. (4) Cultivar/strain-specific effects between different maize cultivars and Trichoderma strains were confirmed by qPCR. In tomato (1) the combination of Trichoderma with associated N-fixing bacteria had positive effects on yield, (2) the combination of Trichoderma and Gram+ bacteria (Bacilli) doubled yield, (3) early application of Piriformospora indica (Sebacinales) caused positive seedling development and (4) co-colonization of tomato roots by different Trichoderma strains was proven by molecular marker application. Currently new combinatorial BE-products are developed on the bases of Trichoderma, seaweed extracts rich in micronutrients, Bacilli and N-fixing bacteria. 181 BIOFECTOR Tuesday 23 June – Poster session 127 Influence of bioeffectors application on maize growth and yield Martin Kulhanek*, Pavel Tlustoš, Jiří Balík, Zlata Holečková, Jindřich Černý Czech university of life sciences in Prague, Czech Republic Phosphorus will become probably the limiting nutrient in near future. The sources of suitable rock phosphates for phosphorus fertilizer production are restricted. Therefore, the improvement of phosphorus acquisition from less available soil forms is needed. One of these ways is the bioeffectors (BEs) application. The aim of this study was to confirm, that the BEs have a positive influence on soil phosphorus mobilisation leading to better P uptake by plants and their higher yields and quality. In this study, the data from pot experiment were evaluated. Three BEs – Trianum (Trichoderma harzianum), RhizoVital (Bacillus amyloliquefaciens) and Proradix (Pseoudomonas sp.) in different combinations with rock phosphate and triple superphosphate were tested on maize plants. The soils from two sites (Humpolec and Lukavec) with low P level and diluted with sand (2:1 soil:sand) were used for the experiment. Plant height, shoot and root dry mass yield, and P plant contents were measured. The average plant height was significantly higher at all treatments fertilized with triple superphosphate in first two months of the experiment. Thereafter, at harvest after 15 weeks of growth, significant differences among plant heights disappeared. The above ground biomass dry weight was significantly higher at the treatments fertilized with triple superphosphate only at Lukavec site. The significant influence of BEs on plant height and above ground dry weight was not confirmed. The root dry weight was slightly, but not significantly higher at the treatments without P fertilization. On the contrary, the lowest root dry mass weight was measured at the triple superphosphate treatments. From the obtained data is possible to conclude, that used bio-effectors did not significantly influenced the plant height and root and shoot dry biomass amount weight as well. 128 Effect of bio-effective microorganisms on early Maize root development and P uptake under low temperature Jonas Duus Stevens Lekfeldt*, Beatriz Gomez Muñoz, Jakob Magid, Lars Stoumann Jensen, Andreas de Neergaard University of Copenhagen, Denmark Maize is a rapidly expanding crop in Northern Europe, but emergence and establishment can be challenged by low soil temperatures in early spring. Microbial inoculants have been demonstrated to stimulate root development, and may help overcoming initial water and nutrient stress, caused by poor root development. However, if soil nutrient levels are low, early investment of plant biomass in root development may not be a successful strategy from a nutrient acquisition point of view. We tested the ability of selected microorganisms with known root stimulation potential (Penicillium sp; Bacillus sp) and humic acids on maize root development in soils of low and medium P availability at 10 and 16 degrees. The soil used was sandy loam from a long-term depletion trial, one with very low P availability, as well as a moderate P treatment, fertilized with animal manure for the past 10 year. 25% sand was mixed into the soils to enable root re-isolation at harvest, and avoid water-logging. Plants were destructively harvested after 5 and 8 weeks, and analysed for shoot and root biomass, plant P in root, shoot and remaining in the seed. Root architecture parameters from 182 BIOFECTOR Tuesday 23 June – Poster session entire isolated roots were assessed using WinRhizo software. Soil extractable P and pH was measured at planting and harvest. Results from the ongoing trial will be presented at the conference. 129 Microbial bio-effectors combined with manure proliferate plant growth of tomato: investigations on the modes of action Zhifang Li*1, Justus Riemann1, Angelika Lüthi1, Nino Weber1, Markus Weinmann1, Gheorghe Poşta2, Günter Neumann1 1 University Hohenheim, Institute of Crop Science (340h), Germany, 2Banat’s University of Agricultural Sciences and Veterinary Medicine, Faculty of Horticulture and Forestry, Romania When tested under practice conditions, commercial bio-effector preparations induced strong growth improvements (80 % increase in plant height) of tomato seedlings grown on a substrate with composted cow-manure (45 %), garden soil (30 %), peat (15 %), and sand (10 %), but without mineral fertilizers. To study the modes of action how these products based on (Bacillus strains FZB42 and R41, Pseudomonas sp. DSMZ 13134 and Penicillium sp. PK 112) isolates could improve plant growth, pot experiments under controlled conditions were conducted. The question whether the plant growth-promoting effect was due to mobilization of sparingly available mineral nutrients from the humified manure was studied in comparative tests where the portion of manure was replaced by an increased share of peat in the substrate while adding mineral nutrients in easily available forms or not. The role of manure as a carbon source supporting the establishment and activity of the introduced microbial strains was tested through substitution by alternative carbon sources. Root morphological characteritics and colonization by arbuscular mycorrhizal (AM) fungi were assessed to detect if an improved spatial acquisition of mineral nutrients could explain the beneficial effects of the bio-effectors. Contrary to the strong effects observed under practice conditions, weak or no growth responses to the bio-effector treatments were found in plants under controlled conditions. However, the Pseudomonas treatment induced a significant increase in root hair length. The biomass production of tomato plants grown on the substrate with manure raised by 9 % in response to the combined application of the Penicillium preparation with an AM inoculum (Glomus intraradices). Plant growth-promotion by microbial bio-effectors, therefore, the combination of diverse modes of action seems to be involved, whose relative importance may vary depending on the environmental conditions. 183 BIOFECTOR Tuesday 23 June – Poster session 130 The effect of biomass ashes and bioeffectors on soil solution composition Filip Mercl*, Václav Tejnecký, Petra Hubová, Pavel Tlustoš, Pavla Ochecová, Jiřina Száková Czech University of Life Sciences in Prague, Czech Republic Biomass ash, the waste material coming from biomass combustion, is generated in large quantities worldwide. Mineral nutrients, bound in ash, are usually poorly plant-available, therefore there is an effort to enhance their availability using Bioeffectors (BEs). Objectives of this study was to i. assess the influence of biomass ash on soil solution composition and ii. investigate BEs ability to enhance availability of nutrients from ash in soil-plant system. In the pot experiment, plants of wheat (Triticum aestivum L.) and maize (Zea mays L.) were grown in BE inoculated mixture of soil and biomass ash under outdoor precipitationcontrolled conditions. Two different types of ash were used (straw and wood ash), and two different BEs were tested (BE3 - Rhizovital® and BE4 – Bacto_Prof). Soil solution was sampled using suction cups (MacroRhizons, Rhizosphere Research Products B.V., Netherlands) from the root zone of plants five times during the vegetation period. In samples of soil solution, concentrations of phosphorus (P) and major low-molecular-mass organic acid (LMMOA) anions (acetate, formate, lactate, oxalate, pyruvate) were determined. The yield of grain and their P content were also determined. Results showed different influence of ashes on soil solution composition. Application of straw ash significantly increased P concentrations in soil solution throughout the vegetation period. Both types of ashes did not differ in the influence on the grain yield, but straw ash significantly increased P content in wheat grains. We observed no significant influence of tested BEs on parameters listed above. Changes in relative and total amount of individual LMMOAs after ash addition were registered. BE4 significantly changed oxalate content in soil solution for one treatment (wheat + straw ash) in early stage of plant development (27 days after planting). 131 Humic, fulvic and water-soluble organic fractions from different compost structure-bioactivity relationship revealed by chemical-biological characterization Hiarhi Monda*, Vincenza Cozzolino, Riccardo Spaccini, Alessandro Piccolo University of Naples Federico II, Italy The influence of humified organic matter on soil fertility has led to a growing interest in the use of humic substances (HS) as soil amendments, revealing their ability to positively affect biochemical processes underlying plant growth and nutrition. An ecologically based approach to farm management consist in the use of compost, recognized as a natural method for recycling organic wastes, and widely used as soil conditioner and fertilizer. In this study we combine the bright side of both technologies by comparing three different organic extracts, humic acids (HA), fulvic acids (FA) and water-soluble organic matter (WOM), from composts made out of different biomass wastes: tomato (C1), artichoke (CYN), and artichokefennel (CYN+F). We conducted bioactivity assay on maize seedling with the humic extracts, which were thoroughly characterized. All the extracts generally increased the whole plant 184 BIOFECTOR Tuesday 23 June – Poster session biomass and positive effects were observed on root length and chlorophyll content. The molecular composition assessed by CPMAS-NMR spectroscopy and pyrolysis-GC-MS of all original composts appeared generally similar, whereas that of the HA and FA extracts varied somewhat as by hydrophobic and hydrophilic components. However, all HA and FA extracts were generally low in aromatic compounds, while they were abundant in biolabile compounds, such as carbohydrates and lipids. The hydrophobic/hydrophilic ratio seemed to play an important role in determining their bioactivity. Moreover, the biological assay pointed out that the larger the content of carbohydrates and sugar-like compounds, as well as nonlignin aromatic components, and the smaller the amount of phenols groups, the greater was the biological/chemical effects exerted by HS. These results confirm the role of humified organic matter in stimulating plant growth, especially when the content of potentially bioavailable hydrophilic components is predominant. 132 Supplementation of silicon in maize (Zea mays) to improve P acquisition and AMF symbiosis efficiency Narges Moradtalab*1, Markus Weinmann2, Roghieh Hajiboland3, Günter Neumann2 1 University of Hohenheim, Institute of Crop Science (340h), Germany, 2University Hohenheim, Institute of Crop Science (340h), Germany, 3University of Tabriz, Iran Phosphorus (P) deficiency is a major limiting factor for plant growth and development. Based on preliminary observations in strawberry (see: Moradtalab et al., this issue), we hypothesized that an optimum level of silicon (Si) supplementation improves arbuscular mycorrhizal (AM) colonization and P acquisition in maize. In this work, the effects of Si supplementation (as K2SiO3) and a commercial silicone fertiliser (Actisil, Yara, ) on AMF colonization, symbiosis efficiency and P availability were studied in maize (Zea mays L. cv. Colisee) plants grown in organic farming soil under greenhouse conditions with additional AM inoculation. Concentrations of 6, 100, 1000 and 10000 mg Si/kg soil (as K2SiO3 by adding to the soil) and 6 mg Si/kg soil of commercial Actisil were applied to the 3 L pots. Application of Actisil was performed either as start application or by weekly irrigation. No P fertilizer was added to the soil within first 4 weeks but after appearance of stronger deficiency symptoms, a moderate P fertilisation was performed with 50 mg P kg-1 soil. Si-supplied plants showed less P deficiency symptoms and growth stimulation especially at application rates of 6 and 100 mg kg-1 as compared with plants without Si and 1000 and 10000 mg/kg Si. Application of 10000 mg kg-1 Si exerted detrimental effects on plant growth and the plants died after 5 weeks application. Start application with Actisil was more effective than weekly fertigation. Our results indicated an optimal level of Si application at 6 mg Si/kg soil. Evaluation of AM colonisation is currently on the way. 185 BIOFECTOR Tuesday 23 June – Poster session 133 Effects of silicon supplementation in drought-stressed strawberry plants in the presence or absence of arbuscular mycorrhizal fungi Narges Moradtalab*1, Roghieh Hajiboland2, Markus Weinmann3, Günter Neumann3 1 University of Hohenheim, Institute of Crop Science (340h), Germany, 2University of Tabriz, Iran, 3University Hohenheim, Institute of Crop Science (340h), Germany Drought stress seriously affects plant growth and development. In this work the effects of silicone (Si) supplementation (3 mM, as Na2SiO3) were studied in strawberry (Fragaria × ananassa var. Parus) plants. Three levels of irrigation (field capacity, 0.75 field capacity, 0.35 field capacity) and two levels of inoculation with arbuscular mycorrhizal fungi (G. Clarum) (‒AMF, +AMF) were applied in a pot experiment and plants were cultivated for 6 weeks under growth chamber conditions. Plant dry matter production and protein content decreased in drought-stressed plants associated with significant reduction of net photosynthesis. In response to drought, soluble carbohydrates and proline were accumulated in the leaves up to 2 and 10 folds, respectively. Application of Si and AMF significantly increased shoot and root dry weight and relative water content not only in drought-stressed but also in well-watered plants. Both Si and AMF treatments significantly increased leaf photosynthesis rate, protein and relative water content and decreased leaf accumulation of osmolytes and malondialdehyde. Combined Si and AMF treatments exerted stronger effects on the improvement of physiological and biochemical parameters of drought-stressed plants than single applications, associated with significantly higher AMF root colonization of +Si treated plants. Our results suggest a beneficial effect of Si on yield, photosynthesis and water relation parameters as well as AMF responsiveness in strawberry plants. 134 Improved P acquisition in crops by inoculation with P-solubilising microorganisms - vision or reality? Guenter Neumann*1, Mira Kuhlmann2, Nicole Probst3, Mehdi Nkebiwe2 1 University of Hohenheim, Germany, 2University of Hohenheim, Institute of Crop Science (340h), Germany, [email protected], Germany Mobilisation of sparingly-soluble phosphates via pH modifications of the growth medium and release of chelating metabolites are widespread mechanisms for P acquisition in many plant growth-promoting microorganisms (PGPMs) cultivated on artificial media and is promoted also as a measure to improve P acquisition in crops. However, within the BIOFECTOR-Project, more than ten experiments in eight countries with three different crops and nine P-solubilising microorganisms failed to show any crop benefit via acquisition of sparingly-soluble P sources. Therefore, this study aimed to characterise critical factors, determining microbial mobilisation of sparingly-soluble P sources in the rhizosphere. Maize was used as test crop with low adaptive potential for root-induced P-solubilisation. A calcareous Loess sub-soil, extremely low in available P (2-3 ppm) and organic matter (0.1%) but rich in acid-soluble Ca-P was supplied (1) with or (2) without rock phosphate fertilisation and (3) with soluble P as positive control. Plants inoculated with 6 different P-solubilising microorganisms (Pseudomonas sp. Poradix®, Pseudomonas sp., Paenibacillus mucilaginosus, 186 BIOFECTOR Tuesday 23 June – Poster session Bacillus subtilis, Streptomyces spp., Penicillium sp.) were cultivated on this substrate, ensuring that plant P-acquisition in treatments (1 and (2) was only possible after microbial Ca-P solubilisation. However, all microbial inoculants failed to stimulate P-acquisition of the test plants and even exerted inhibitory effects on plant growth. Also simultaneous fertigation with a mixture of glucose and glycine had only marginal effects on crop performance, suggesting that limitation of root exudates as C and N source was not a critical factor. However, successful P acquisition and plant growth promotion was observed after diluting a similar growth substrate with 75 % (w/w) washed quartz sand, drastically lowering the pH buffering capacity and the CaCO3 concentration for unspecific binding of P-solubilising chelators. These findings suggest that the soil-buffering capacity may be a major constraint which limits the P-solubilising potential of many PGPMs. 135 Placement of Pseudomonas sp. PRORADIX around NH4+-based fertilizer depots in maize stimulates root exploitation of the fertilizer depot Peteh Mehdi Nkebiwe*1, Markus Weinmann2, Günter Neumann2, Torsten Müller3 1 University of Hohenheim, Plant Nutrition: Fertilisation and Soil Matter Dynamics (340i), Germany, 2University of Hohenheim, Plant Nutrition: Nutritional Crop Physiology (340 h), Germany, 3University of Hohenheim, Plant Nutrition: Fertilisation and Soil Matter Dynamics (340 i), Germany Combining fertilizer placement and inoculation with plant growth-promoting rhizobacteria that stimulate root-growth and solubilize phosphates may be a strategy to improve root exploitation of fertilizer depots. Moreover, root colonization by such rhizobacteria may be enhanced in the depot zone with intense root development as a consequence of increased availability of root exudates induced by localized supply of root-attracting nutrients like ammonium. After confirming that the rhizobacteria strain Proradix®WP (Pseudomonas sp. DSMZ13134, Sourcon Padena, Germany) grows normally at high ammonium concentrations (50 mM) in the presence or absence of the nitrification inhibitor 3,4-Dimethylpyrazole phosphate, we investigated the effect of placing concentrated ammonium sulfate (64 mg N ml-1) with 3,4Dimethylpyrazole phosphate as a fertilizer depot in combination with PRORADIX as inoculant on growth-stimulation of maize roots (Zea mays L.), rhizosphere pH changes, root colonization by PRORADIX, nitrogen (N) and phosphorous (P) uptake in a rhizobox experiment on a calcareous Loess subsoil, pH 7.6 and in a field experiment on silty loam, pH 7.1. In the rhizobox experiment, increased root length densities around the NH 4+-depot were recorded 8 weeks after sowing, associated with intense root surface and rhizosphere acidification by 2 pH units and to higher N and P uptake per plant compared to the control variant with homogenous NO3 - fertilization. This may be attributed to improved spatial nutrient acquisition and to mobilization of acid-soluble calcium-phosphates, which are usually formed in neutral to alkaline carbonate-rich soils. Particularly in the depot zone, a high root colonization density of Pseudomonas was detected by culturing root extracts on a selective nutrient medium. In the field, root length density was doubled in the ammoniumdepot zone inoculated with PRORADIX at 11 weeks after sowing, as a first indication of improved root exploitation of the fertilizer depot, induced by simultaneous rhizobacteria inoculation under field conditions. 187 BIOFECTOR Tuesday 23 June – Poster session 136 No matter whether dead or alive? An investigation of bio-effector application in tomato and maize experiments under phosphorus limited soil conditions Dinah Reinhardt*1, Marie Spohn2, Sven Marhan1, Namis Eltlbany3, Kornelia Smalla3, Ellen Kandeler1 1 University of Hohenheim/Institute of Soil Science and Land Evaluation, Germany, 2University of Bayreuth/Department of Soil Ecology, Germany, 3Julius Kühn-Institute/Institute for Epidemiology and Pathogen Diagnostics, Germany Bio-effectors are viable organisms and active natural compounds that are able to promote plant growth and health. This ability is enabled, for instance, by mobilizing sparingly soluble mineral nutrients like phosphorus. Particularly plant growth promoting rhizobacteria (PGPR), such as strains from genera Pseudomonas, Bacillus and Rhizobium are among the most powerful phosphate mobilizers but the functional mechanisms are still poorly understood. In the frame of the BioFector project we focus on the effects of phosphorus solubilizing rhizobacteria. We hypothesize: (I) Plant growth promoting effects of PGPR are based on an increasing phosphatase activity (especially acid phosphatase) and here we assume a higher activity in rhizosphere than in bulk soil; (II) PGPR colonization will be displayed in phosphatase activity hotspots. Two rhizobox experiments (tomato and maize) under phosphorus limited soil conditions were performed using Pseudomonas jessenii as the bioeffector. Experiments comprised five control treatments, including inoculation with native soil microorganisms and dead Pseudomonas jessenii cells, respectively. During plant cultivation a soil in situ zymography for determination of spatial and temporal phosphatase activity was conducted. Results showed a significant plant growth promoting effect, but contrary to our expectation, in treatments with living as well as dead Pseudomonas jessenii cells. In both treatments we observed yields that nearly corresponds to the optimally phosphorus fertilized plants; 79% in plant height and 83% in biomass, for example. First evaluation of zymography indicates a higher acid phosphatase activity in rhizosphere than in the bulk soil across all treatments. A particularly high activity was revealed in maize, especially on root tip surface. Additional analyses will identify whether the improved plant growth is based on an increasing nutrient availability and uptake. In addition, we will analyse the cell components of lysed P. jessenii to identify potential plant growth promoting compounds. 137 Max-Res TGA and FTIR evaluation of plant growth promoting rhizobacteria David Nelson, Eugene Carmichael, Graham McCollum, JR Rao, Gary Lyons, Shekhar Sharma* Agri-Food Biosciences Institute, United Kingdom Analysis of bacterial biomass provides valuable information on the cell contents and structural differences in cell wall polysaccharides. The objective is to develop rapid methods for evaluating plant growth promoting rhizobacteria (PGPR) by Fourier Transform Infrared spectroscopy (FTIR) and thermogravimetric analysis (TGA) maximum resolution (Max-Res) protocol. Bacterial strains (BM- Bacillus mucilaginosus, FBZ42- B. amyloliquefaciens A, RLH – B. subtilis S, 4.2 – Burkhoweria sp, H2.6/RC2.5 – Rahnella aquatalis) were inoculated in potato dextrose 188 BIOFECTOR Tuesday 23 June – Poster session broth for 4 days and incubated on a rotary shaker at 160 rpm for 48 h at 25oC. After removing the supernatants by centrifugation at 6000g for 10 min, the bacterial pellets were washed three times with distilled water and lyophilized. FTIR spectra of the samples were obtained using a diamond ATR. TGA was carried out at a heating rate of 20oC/min in nitrogen. TGA heating rates for maximum resolution (Max Res) analysis were programmed at 20oC / min when no weight loss was detected and a heating rate of 5oC / min on detection of weight loss. The six isolates can be distinguished from the differences in infrared functional groups and TGA composition data of the isolates. Max-Res analysis of the isolates showed compositional differences between the six isolates as indicated by differences in the weight loss steps. 138 Development of Quality Assurance protocols for biostimulant products Shekhar Sharma*, Eugene Carmichael, Eugene Conlon, Trevor Martin, Chris Selby Agri-Food Biosciences Institute, United Kingdom Plant biostimulants are usually formulated using a range of components including macroalgae, animal or plant proteins, amino acids, minerals and plants extracts containing a wide range of trace compounds. The objective is to develop quality assurance (QA) protocols for evaluating physico-chemical characteristics of biostimulants. Six formulations (Rygex, Algavyt, Ryzoset, Manek, Ecoryg and Algavyt Zn/Mn – Agriges products) containing algal/plant extracts, humic and amino acids, lipids and inorganic components were assessed for particle size distribution, zeta potential and auxin activity. The particle size range of biostimulant products can be reduced by employing a wet mill and micro-fluidiser. The particle sizes of Rygex, Algavyt, Manek and Ecoryg were less than Rizoset and Algavyt Zn/Mn. Among the six products, Algavyt and Manek showed the highest zeta potential (ZP) values. The formulations were assessed for auxin activity using a mung bean rooting bioassay (cytokinin bioassay result not presented). Measurements of total bean roots per cutting and root length demonstrated the presence of significant auxin activities in some formulations (e.g. Rygex and Ecoryg). The gross differences in the particle size, ZP and composition of the products could be used for developing formulations to improve field performance and stability during transportation and storage. 139 Field performance of barley treated with two biostimulant products Trevor Martin, Fiona Clarke, Chris Selby, Eugene Conlon, JR Rao, Shekhar Sharma* Agri-Food Biosciences Institute, United Kingdom Two types of arable fields were selected on the basis of their nutrient status: (a) standard input field – regularly applied with inorganic fertiliser to maintain the soil nutrient status for good plant growth and (b) low input field - not treated with fertiliser during the past ten years, with the exception of nutrient input grazing animal waste. The objective is to evaluate 189 BIOFECTOR Tuesday 23 June – Poster session if certain biological extracts can influence the yield of spring barley in either a standard input soil or a soil of low nutritional value. Two formulations of biostimulants supplied by Bioatlantis, Ireland (A039F249 and D002G138) were assessed for biological activity and evaluated as foliar treatments. Spring barley was sown at both sites (standard nutrient input soil and low input soil) in randomised blocks. Treatment applications at three rates were applied during the growth of the crop. Mung bean bioassays was carried out to monitor growth hormone activities of the products to ascertain their biostimulant potential. Yields and 1000 grain weights were assessed at harvest. The bioassay of the products using mung bean root extension data showed that two products (A039F249 and D002G138) are biologically active. The application of D002G138 increased the number of mung bean roots, while both products at concentrations 1 and 5 gml-1 initiated greater root length. Field trials with spring barley at the two concentrations showed differing levels of chlorophyll between sites but no significant differences in either the yield of grain or the 1000 grain weight. 140 Response of cold and heat stressed Arabidopsis and wheat plants on the expression of salicylic acid and jasmonic acid Colin Fleming, Thomas Fleming, Colin McRoberts, Stewart Floyd, Shekhar Sharma* Agri-Food Biosciences Institute, United Kingdom The impact of cold and heat stresses on the productivity of arable crops is expected to be a major factor worldwide in reducing yields by 10 to 30% . This study was aimed at developing tools to monitor key biotic and abiotic stress related plant hormones such as salicylic acid and jasmonic acid in wheat and Arabidopsis plants. The objective is to monitor salicylic acid (SA) and jasmonic acid (JA) expression in Arabidopsis and wheat plants in response to cold and heat stress. Heat and cold stresses (4oC and 38oC for 18 hrs) were applied to plants in growth cabinets. SA and JA levels were measured directly using Q-TOF-LC/MS. Total mRNA was extracted and cDNA produced before RT-PCR, targeting the PR1 SA pathway gene and MYC2 JA pathway gene. The conclusions are (a) heat and cold stress significantly affected the concentration of SA and JA in wheat and Arabidopsis plants (b) heat and cold stress changed expression levels of wheat genes in the SA and JA pathways and (c) these models can be used to monitor the effects of stress modifying materials on monocotyledons and dicotyledons. 190 BIOFECTOR Tuesday 23 June – Poster session 141 Evaluation of anion exchange membrane for monitoring of phosphate solubilisation in soil types Chris Selby, Eugene Carmichael, Eugene Conlon, Shekhar Sharma*, JR Rao Agri-Food Biosciences Institute, United Kingdom It is relatively easy to identify soil bacteria capable of solubilising phosphorous by plating them on microbiological media containing an insoluble phosphate (P) and observing rings of media clearing around active colonies. The objective is to estimate phosphorous solubilisation in soils using an anion exchange membrane (AEM) protocol. A method was evaluated to confirm if these organisms including AFBI P solubilisers and Porodix are equally active in agricultural soils. The method involved using sections of AEM (1.5 x 4 cm) to act as “artificial roots” by placing them in soils for a given period then recovering them to assay (molybdate reactive P) captured PO4 3- ions. The conclusions are (a) AEMs can be used to assess the availability of P to plants in agricultural soils and give a good correlation with estimates found using conventional chemical analyses of soils; (b) AEMs can be used to distinguish P availabilities between soils and effects of supplementation of soils with various P salts; (c) No significant increased P solubilization occurred with either the AFBI P solubilisers or Porodix regardless of P supplementation or soil type and (d) Future work will focus on why the bacterial inoculae do not appear to be solubilising P in soils. In the first instance this will involve providing additional supplements such as increased organic matter or organic compounds common in root exudates (eg glutamic and aspartic acids and their amines) to provide the bacteria with a more readily available energy source. 142 Evaluation of ryegrass extracts for plant bio-stimulant activities Chris Selby, Eugene Carmichael, Eugene Conlon, Shekhar Sharma* Agri-Food Biosciences Institute, United Kingdom Plant fibres are generally regarded as being the major product of the bio-refining of grasses. However, this process also produces large volumes of aqueous extract that is potentially useful to the agricultural and horticultural industries. Here we compare the plant defence elicitor activity (French bean cell suspension cultures) and auxin-like activity (mung bean hypocotyl rooting test) of extracts produced from high pressure screw-pressing with those prepared by homogenising foliage in water for. The objective is to evaluate if ryegrass extracts have the potential to act as crop bio-stimulants by assessing their biological activity in a series of bioassays. Two growth hormone (auxin and cytokinin) bioassays were used to compare screw-pressed juice with extracts prepared by homogenising foliage with water. In both assays, all six extracts were equalised to the lowest sugar content sample, as assayed using the anthrone reagent, by dilution with water. Extracts of perennial ryegrass contained potent elicitors of plant defence that caused rapid and intense browning of cultured French bean cell suspensions. This is indicative of the 191 BIOFECTOR Tuesday 23 June – Poster session synthesis of phenolic compounds, many of which have phytoalexin activity. Extracts produced by screw-pressing were marginally but significantly more active than those produced by homogenisation of tissues in water. This was illustrated by the significant interaction between extraction method and treatment concentration. Differences between ryegrass varieties were small and insignificant, particularly if attempts were made to equalise sample strengths using a measure such as sugar concentration. Root induction (a) and growth (b) in hypocotyl cuttings of mung bean indicate that the ryegrass extracts are rich in “auxin-like” or auxin cofactor activity. Studies are in progress to characterise the extract components responsible for the biological activities reported here. 143 Quantitative tracing of two Pseudomonas strains in the roots and rhizoplane of maize, as related to their plant growth-promoting effect in contrasting soils Carla Mosimann1, Thomas Oberhänsli1, Dominik Ziegler2, Dinah Reinhardt3, Ellen Kandeler3, Thomas Boller4, Paul Mäder1, Cécile Thonar*1 1 FiBL (Research Institute of Organic Agriculture), Switzerland, 2Mabritec AG, Switzerland, 3University of Hohenheim, Institute of Soil Science and Land Evaluation, Germany, 4Zürich-Basel Plant Science Center, University of Basel, Switzerland Plant growth-promoting rhizobacteria (PGPR) are able to facilitate plant nutrient acquisition and can act as biocontrol agents by suppressing soil-borne diseases. Efficient strains can be formulated as microbial inoculants and their successful use for field application often requires a certain ability of the strains to survive in the soil where they are inoculated. In this respect, there is a need to create tools enabling the tracing of inoculated PGPR which can also serve to monitor their spread in space and time. In this poster we describe the development and application of a molecular method allowing the quantitative detection of two Pseudomonas strains (Pseudomonas fluorescens Pf153 and Pseudomonas sp. DSMZ 13134) contained in commercial formulations. The method is based on a Taqman qPCR assay targeting two polymorphic regions of the bacterial genome in order to ensure the specificity of the detection. Inoculation experiments with maize were conducted in three contrasted soils. Eight weeks after planting and inoculation, Pf153 could still be detected and its persistence in root and rhizoplane was shown to be the best in the organic soil (versus the two other conventional soils). On the other side, DSMZ 13134 could not be detected anymore after 8 weeks but its persistence after 4 weeks in root and rhizoplane samples was equally good in the three tested soils. Combined with the plant responses, our data indicate that the persistence of the two strains cannot clearly explain the measured plant biomass and nutrient acquisition (nitrogen and phosphorus) due to inoculation of the two strains. The discrepancy observed between the level of colonization by the strains and their effects on plant indicate that more research is needed to elucidate the mechanisms and conditions leading to successful application of PGPR. The developed tools will definitively contribute to this objective. 192 BIOFECTOR Tuesday 23 June – Poster session 144 Metabolites Profiling of Zea mays inoculated with the Bacillus amyloliquefaciens as Bioeffector Vincenza Cozzolino, Giovanni Vinci*, Hiarhi Monda, Daniele Todisco, Alessandro Piccolo University of Naples Federico II, Italy Bacillus amyloliquefaciens represents the largely studied plant growth-promoting rhizobacteria (PGPR) that competitively colonize plant roots and can act as either biofertilizers or antagonist biopesticides or both. PGPR enhance growth and yield of cereals through mechanisms that include production of growth stimulating phytohormones, solubilization and mobilization of phosphate and induction of plant systemic resistance to pathogens. For this reasons, PGPR represent biofertilizers well recognized as efficient tool for sustainable and safe agriculture. In this study, we investigated the effects of Bacillus amyiloliquefaciens when inoculated on plants of Zea mays under different P amendments. In order to improve our understanding we determined the metabolic profiling followed by multivariate analysis. Gas chromatography– mass spectrometry (GC-MS) was employed to screen potential differences among metabolic leaf extracts. A number of 56 primary polar metabolites, comprising amino acids, organic acids, sugars, phenolic acids, amino sugars, sugar alcohols, and 7 unknown compounds, were identified. The PCA showed a robust reproducibility among five replicates, revealed a significant separation between control and treatments with Bacillus a. Moreover, the different P amendments (no P addition, triple super phosphate, rock phosphate, composted cow manure, composted horse manure) showed a substantial difference in plants metabolic profiles. A significant variance from control was found for plants undergone to both Bacillus a. inoculation and compost treatment, whereby the difference in metabolic profile was accounted by a large abundance of sugars and organic acids metabolites. These results indicated that the different P amendments and the presence of Bacillus amiloliqefaciens strongly influenced the presence and distribution of primary metabolites. However, more detailed analyses of the metabolome (secondary metabolites) will better elucidate how Bacillus a. affect the metabolic pathways and consequently the performance of maize plant. 145 Bioeffectors promoting Bioeffectors - Seaweed extracts as prebiotics for plant growth-promoting rhizobacteria? Nino Weber*1, Markus Weinmann1, Peteh Mehdi Nkebiwe2, Kristin Dietel3, Sarah Symanczik4, Günter Neumann1, Uwe Ludewig1 1 University Hohenheim, Institute of Crop Science (340h), Germany, 2University Hohenheim, Institute of Crop Science (340i), Germany, 3ABiTEP GmbH, Germany, 4FiBL, Switzerland A major constraint for a successful practical application of plant growth-promoting microorganisms is the limited reproducibility of the desired effects, depending on often unknown external factors. One important determinant for effective application of microbial bioeffectors is their rhizosphere competence and effective root colonization of the host plant, 193 BIOFECTOR Tuesday 23 June – Poster session frequently promoted by repeated inoculations with high inoculum densities. However, under practical conditions, this approach is not always economic. Using the prebiotic potential of various natural compounds such as seaweed extracts to stimulate microbial growth may be an alternative strategy. Our in vitro studies with bacterial suspensions of low CFU showed highly prebiotic potential of selected seaweed extracts on strains of Bacillus amyloliquefaciens and Pseudomonas sp. “Proradix” which could not be attributed to carbon source effects as demonstrated by control treatments with glucose. In a pot experiment with maize, combinations of Proradix® and seaweed extracts led to significantly higher plant biomass compared to the untreated control, even on a well fertilized field soil. Based on these results, promising product combinations were tested in a field experiment with maize on a silty loam (pH 6.8) and fertilization according to farmers practice. For tracing of the bacterial strains in the field selective plating assays and RT-qPCR with specific primers were employed. Plant sampling at four weeks after sowing revealed no effect of the selected seaweed extracts (Superfifty, Algafect) either on root colonization of Proradix or on the total cell number of B. amyloliquefaciens. However, the proportion of Bacillus spores was increased by Algafect. No significant effects on biomass production and final grain yield of maize were observed. The results suggest that seaweed extracts may exert prebiotic effects on plant growth-promoting bacteria but the characterization of conditions determining positive interactions with the host plant still needs further investigation. 146 Strategies for bio-effector application to improve the growth and mineral nutrition of maize under field conditions Markus Weinmann*1, Mehdi Peteh Nkebiwe2, Nino Weber1, Klara Bradacova1, George Fora3, Torsten Müller2, Günter Neumann1 1 University Hohenheim, Institute of Crop Science (340h), Germany, 2University Hohenheim, Institute of Crop Science (340i), Germany, 3Banat’s University of Agricultural Sciences and Veterinary Medicine, Romania For many bio-effectors, such as plant growth-promoting microorganisms, the ability to support the mineral nutrition of plants has been proven under controlled conditions. Mobilization of mineral nutrients, stimulation of root growth and activity, and beneficial interactions with other microorganisms are among the most noticeable modes of action. These properties could be effectively utilized for the development of more environmentally friendly and sustainable plant nutrition strategies that are less dependent on mineral fertilizers. A critical task is to meet the demand of high yielding crops for phosphate and other essential minerals that are sparingly supplied by the soil or from alternative fertilizers. However, the agricultural implementation of such approaches is still limited and this is likely due to a lack of appropriate application techniques ensuring the functioning of bio-effectors under adverse environmental conditions. In the present work, commercial preparations of bacterial (Bacillus strains FZB42 and R41, ABiTEP GmbH, Berlin, Germany; Pseudomonas sp. DSMZ 13134, Sourcon Padena GmbH & Co. KG, Tübingen, Germany) and fungal (Penicillium sp. PK 112, Bayer CropScience Biologics 194 BIOFECTOR Tuesday 23 June – Poster session GmbH, Malchow/Poel, Germany) bio-effectors were tested with different application methods to improve the phosphorus nutrition of field grown maize. No significant improvements in growth or mineral nutritional status of the plants were found when the bio-effectors were applied by seed incrustation, broadcast distribution with soil incorporation, or spray application on top of the plants. These findings suggest that the application of these microorganisms as discrete measures is not decisive for the productivity of maize on fertile soils. Advanced application strategies, such as the combination of bioeffectors with fertilizer placement and/or the use of recycling fertilizers are proposed to provide viable alternatives to the current practice of mineral fertilization. 147 Microbial bio-effectors combined with manure proliferate plant growth of tomato: investigations on the modes of action Zhifang Li1, Justus Riemann1, Angelika Lüthi1, Nino Weber1, Markus Weinmann*1, Gheorghe Posta2, Günter Neumann1 1 University Hohenheim, Institute of Crop Science (340h), Germany, 2Banat’s University of Agricultural Sciences and Veterinary Medicine, Faculty of Horticulture and Forestry, Romania When tested under practice conditions, commercial bio-effector preparations induced strong growth improvements (80 % increase in plant height) during the early development of tomato seedlings grown on a substrate prepared with composted cow manure (45 %), garden soil (30 %), peat (15 %), and sand (10 %), but without mineral fertilizers. To study the modes of action how these products based on bacterial (Bacillus strains FZB42 and R41, Pseudomonas sp. DSMZ 13134) and fungal (Penicillium sp. PK 112) isolates could improve plant growth, pot experiments under controlled conditions were conducted. The question whether the plant growth-promoting effect was due to mobilization of sparingly available mineral nutrients from the humified manure was studied in comparative tests where the portion of manure was replaced by an increased share of peat in the substrate while adding mineral nutrients in easily available forms or not. The role of manure as a carbon source supporting the establishment and activity of the introduced microbial strains was tested through substitution by alternative carbon sources. Root morphological characteritics and colonization by arbuscular mycorrhizal (AM) fungi were assessed to detect if an improved spatial acquisition of mineral nutrients could explain the beneficial effects of the bio-effectors. Contrary to the strong effects observed under practice conditions, weak or no growth responses to the bio-effector treatments were found in plants under controlled conditions. However, the Pseudomonas treatment induced a significant increase in root hair length. The biomass production of tomato plants grown on the substrate with manure raised by 9 % in response to the combined application of the Penicillium preparation with an AM inoculum (Glomus intraradices). In plant growth-promotion by microbial bio-effectors, therefore, the combination of diverse modes of action seems to be involved, whose relative importance may vary depending on the environmental conditions. 195 BIOFECTOR Tuesday 23 June – Poster session Food-web Interactions 148 Carbon flow and enzyme activities during protozoan grazing in rhizosphere and detritusphere Sebastian Löppmann*1, Fionn Clissmann2, Anna Gunina1, Johanna Pausch1, Robert Koller3, Michael Bonkowski2, Yakov Kuzyakov1 1 University of Göttingen, Büsgen-Institute, Germany, 2University of Cologne, Institute of Zoology, Germany, 3Forschungszentrum Jülich, Institute of Bio- and Geosciences, Germany The differences in complexity and accessibility of plant provided carbon (C) sources for the soil food web result in two major decomposition pathways based on 1) root and shoot litter, and 2) rhizodeposits (especially exudates). The amount and quality of substrates entering the soil mainly controls microbial processes in the rhizosphere and detritusphere. Furthermore, soil fauna has important functions in regulating microbial activity and enzymatic substrate utilization. A triple-labeling (13C, 14C and 15N) experiment was conducted focusing on the identification of C resources (rhizodeposited C by 14C and root litter by 13C) that fuel microbial-protozoan interactions in two soil hotspots: rhizosphere and detritusphere. Soil was taken from an arable field planted with maize, autoclaved and re-inoculated with a microbial community extracted from the same soil. The following treatments were established: 1) no addition of plant C, 2) addition of sterilized 13C /15N labeled maize root litter, representing detritusphere 3) 14CO2 pulse labeling of growing maize plants, representing rhizosphere. To determine the effects of faunal grazing on nutrients (by 15N) release each treatment was setup with and without amoeba. Enzyme kinetics was implemented as indicator for microbial activity and 13C flux to the microbial pool was determined by 13C-PLFA analysis. We analyzed14C, 13C and 15N in the microbial biomass, soil, plant, root and CO2. Additionally, microbial biomass was assigned by dsDNA extraction. Between 65-89% of soil released 14CO2 was respired by roots and microorganisms in the first 3 h after the 14CO2 pulse. First results showed higher 14C activity in the microbial biomass in presence of protozoa. The dsDNA-extracted microbial biomass was significantly higher in litter amended and planted soil compared to the unplanted control. Furthermore, the DNA contents were increased in the presence of protozoa. Clear effects of enzymatic affinity to the specific substrate were elucidated during protozoan grazing. 196 Food-web Interactions Tuesday 23 June – Poster session 149 Litter decomposition and home-field advantage of range-expanding plant species Marta Manrubia Freixa*, Ciska Veen, Wim van der Putten NIOO-KNAW, Netherlands The current climate warming enables many native plants to expand ranges to higher altitudes and latitudes. Plants develop in close interaction with soil organisms in a direct (e.g. via pathogenesis) and indirect way (e.g. via the detritus food web). During range shifts, these specific plant-soil interactions might become temporally disrupted since soil organisms have limited dispersal capacity. Consequently, range-expanding plants are expected to benefit from the release of soil-borne pathogens and suffer from losing positive interactions with specialized decomposer organisms. The present research aims to study local specialization by decomposers of range-shifting plant species along latitudinal gradients and determine whether specialization results in ‘home field advantage’. We tested the hypothesis that soil microbial communities from the native range have higher affinity with litter of range-expanding plants than communities in the new range. We collected senesced leaves and soil of two range-expanding plants (Centaurea stoebe and Lactuca serriola) and a native congener (Centaurea jacea) in the native and expansion range. We set up a 97-day microcosm incubation experiment for each plant species. Litter was reciprocally transplanted to all soils within and between ranges and we measured CO2 fluxes over the incubation period. Soil heterotrophic respiration response to litter addition was used as a proxy for decomposition activity. We found that soil and litter origin had the strongest effect on soil heterotrophic respiration for all 3 plant species, whereas no local specialization effect was observed in litter transplants within or between ranges. High within-range variability suggested that litter decomposition controls operate at a highly local scale. 150 A root-feeding insect in the shallow soil zone significantly alters yield, mortality, and size structure of Lolium perenne populations Tomonori Tsunoda*1, Naoki Kachi2, Jun-Ichirou Suzuki2 1 Tokyo Metropolitan University / Yokohama National University, Japan, 2Tokyo Metropolitan University, Japan We evaluated the effects of vertical distributions of a larva of a root-feeding insect, Anomala cuprea, on the yield, mortality, and size structure of Lolium perenne populations. We conducted a growth experiment with a two-way factorial randomized block design with nine replications. Factors included plant density (36 plants per pot, and six plants per pot) and vertical distribution of A. cuprea. Pot soil was divided into three distinct vertical zones (top, middle and bottom) with a wire gauze, and each pot received a larval insect into one zone. There were two additional treatments, one with a larva but without a partition and one without an insect. Yield of L. perenne significantly decreased in the treatments with herbivory, with the largest decrease in the top feeding zone, i.e. the shallowest soil. Yield was also significantly reduced at low plant density, but was more affected by herbivory. Plant mortality occurred only when the herbivore was in the top feeding zone. Plant density did not significantly alter the number of dead plants. At low plant density, the shoot biomass of the 197 Food-web Interactions Tuesday 23 June – Poster session three largest plants significantly decreased with a larval herbivore, but the three smallest plants not. Thus, the standard deviation in shoot biomass significantly decreased under herbivory. At high plant density, shoot biomass was not significantly affected by herbivory, irrespective of plant ranks. The standard deviation in the shoots was smaller at high density than at low density. In contrast with aboveground herbivory, which is known to increase size variance in plant populations, the root-feeding of an insect in shallow soil decreased the size variance in a sparse plant population. 151 Top-down and bottom-up control of generalist root-feeding nematodes in the rhizospheres of range-expanding plant species and native congeners Rutger Wilschut*, Stefan Geisen, Wim van der Putten NIOO-KNAW, Netherlands As a consequence of climate warming many plant species are expanding their range to higher latitudes and altitudes, thereby leaving behind their rhizosphere soil organisms. It is unknown how new rhizosphere communities become assembled in the new range. Generally, rangeexpanding plant species are less negatively affected by soil communities in the new range than congeneric native plant species. This may be due to a lower amount of herbivores and pathogens that are able to attack the range expanding plants in the new range, either because of a lack of co-evolutionary history or by stronger direct or indirect defense mechanisms. We study how specific multitrophic interactions develop in the rhizospheres of rangeexpanding plant species. In a greenhouse experiment we exposed two range-expanding plant species and their congeneric natives to different generalist root-feeding nematodes and inoculated microbial communities that may contain antagonists of the nematodes. Subsequently, we examined if nematode population growth differed between rangeexpanding and native plant species and if top-down control of nematodes by the microbial communities was plant species-dependent. We tested the hypotheses that 1) if top-down control of root-feeding nematodes is plant-mediated, this will be stronger in native species than in range-expanders, as native plants have a shared evolutionary history with the local soil community and 2) range-expanding plants are better defended against generalist rootfeeding nematodes than congeneric natives, because of strong direct defense mechanisms. Our results suggest that these defense mechanisms are species-specific, rather than depending on range shift capacity. 198 Food-web Interactions Tuesday 23 June – Poster session Engineering the Rhizosphere 152 Severe rhizosphere oxygenation optimising the carbon footprint of Paludiculture // (Sustainble biomass production on peat soils) Christian Fritz*1, Eric Visser2, Alfons Smolders3, Leon Lamers2, Florian Wichern1, Bikila Dullo4, Theo Elzenga5, Veronica Pancotto6, Ab Grootjans5 1 Rhine-Waal University of Applied Sciences, Germany, 2Radboud University, Netherlands, 3B-Ware Research Center, Netherlands, 4Addis Abeba University, Ethiopia, 5Rijksuniversiteit, Netherlands, 6CADIC, Argentina Water-logging is a major threat to plant health and causes substantial methane emissions from crops and natural vegetation. Flood adapted plant species influence soil processes by releasing root exudates and degradable litter. This can increase methane production and nutrient availability. In contrast, little is known to what extent root derived oxygen caps methane and nutrient release. Our research tested which growth conditions are necessary to maintain oxygen release at sufficient rates to dominate rhizosphere processes. Root-methane-soil interactions were studied by comparing methane emissions, stock and oxygen availability in depth profiles below stands of cushion plants and paludicrops. We followed rhizosphere nutrient-availability along depth profiles in Ethopia, Argentina and the Netherlands. We investigated nutrient cycling by 15N field experiments and N:P ratios of plant organs. Cushion plants, Eriocaulon schimperi and Astelia pumila, formed extensive root systems up to 150 cm deep. Root biomass (3590 g.DW.m-²) dominated the belowground biomass of cushion plants but resulted in a higher nutrient demand. Roots of paludicrops, Typha spec. and Phragmites spec., were in general shallow (< 70 cm) and root proliferation was sensitive to vertical nutrient availability. In contrast, soils surrounding cushion plant roots were depleted in methane and plant-available nutrients. Main finding is that methane emissions were then cut to zero high root densities promoting soil oxygen (> 1.5 mg.l-1). High soil N:P ratios and low temperatures seemed major controls on root density and rhizosphere oxygenation. Increase in shallow root length was associated with higher 15N recycling Our study shows that roots of cushion plants and biomass crops can dominate biogeochemical processes at the ecosystem scale given certain growth conditions. Oxygen release from the dense root biomass was sufficient to cause a thorough soil oxidation. Soil nutrient ratios seem promising in reducing greenhouse gas emissions from water-logged fields by tuning the root system architecture. 199 Engineering the Rhizosphere Tuesday 23 June – Poster session 153 Biocontrol practices alter rhizosphere community structure in pepper and suppress Phytophthora blight Lori Hoagland*, Dan Egel, Natasha Cerruti, Jyothi Thimapuram, Clayton Colling, Ketaki Bhide Purdue University, United States Phytophthora blight has become one of the most serious threats to the vegetable industry in the Midwest U.S. Controlling Phytophthora capsici, the pathogen responsible for Phytophthora blight is difficult because it has a broad host range, spreads rapidly under ideal environmental conditions, and produces resilient spores that can survive indefinitely in soil. There is also insufficient resistance in crop varieties, and P. capsici is now resistant to many fungicides. Biocontrol practices that are thought to suppress soil-borne pathogens via changes in soil microbial communities could help control Phytophthora blight, though the mechanisms are not well understood limiting the effective deployment of these practices. In this study, we evaluated the potential for various cover crop species grown alone, or in combination with a biochar amendment made from hardwood forest species to suppress Phytophthora blight in two naturally infested field soils. Changes in the rhizosphere community structure of a subsequently planted pepper crop were quantified with semi- selective media and 16S sequencing performed by Illumina MiSeq and analyzed using QIMME. A wheat cover crop and biochar, alone and in combination, increased pepper root and shoot biomass, and increased the abundance of Pseudomonas fluorescens in the pepper rhizosphere in both soils. In the low organic matter soil, P. capsici abundance in the pepper rhizosphere was reduced most by treatments that received the wheat cover crop, whereas in the high organic matter soil P. capsici was reduced most by treatments that received biochar. These results indicate that including a wheat cover crop and applying biochar can improve pepper growth in soil infested with P. capsici, though the suppression mechanisms and most effective biocontrol strategy could vary given soil type. Results of on-going sequencing analyses will provide additional insights to the potential mechanisms regulating P. capsici suppression and enhanced pepper growth observed in this study. 154 Beneficial interaction between tomato plant and Flavobacterium strains isolated from tomato rhizosphere Seon-Woo Lee*1, Soo Yeon Choi1, Eun Joo Jung1, Ju Yeon Song2, Jihyun Kim2 1 Dong-A University, South Korea, 2Yonsei Unversity, South Korea Plant and microbe interaction in rhizosphere may influence various plant functions such as plant growth, plant development and tolerance to stresses. Microbial community analysis of tomato rhizosphere from our study revealed that members of phylum Bacteroidetes were abundant in tomato rhizosphere compared to bulk soil. We hypothesized that those bacteria dominating tomato rhizosphere contribute to the plant function. A pair of specific primer targeting those bacteria in class Flavobacteria was designed and used to select bacterial isolates in a member of the class Flavobacteria by colony PCR of bacterial colonies derived from tomato plant rhizosphere. A number of bacteria were isolated and examined for phenotypes relevant to plant-beneficial interaction, such as biofilm formation, seedling growth promotion, and antimicrobial activities. Five strains were selected for further study because they showed plant-beneficial effects. One of the isolates, F. daejeonensis RCH33, 200 Engineering the Rhizosphere Tuesday 23 June – Poster session showed the delayed occurrence of bacterial wilt in tomato plants, when RCH33 strain was treated in tomato rhizosphere prior to the bacterial wilt pathogen Ralstonia solanacearum inoculation by soil-soaking method. Genomes were analyzed for all five strains in Flavobacterium and revealed that they harbor genes for plant growth promotion, such as auxin biosynthesis and ethylene removal, and for secondary metabolites which may be involved in beneficial effect to plants. However, one of novel species of Flavobacterium TCH32 did not contain any known genes for beneficial interaction with plant, while plant growth promotion effect and the capability of biofilm formation were remarkable. We are under investigation of bacterial gene expression in planta for these Flavobacterium strains. 155 The right set of circumstances – Selecting microbial bio-effectors for applications in alternative fertilisation systems – the BIOFECTOR project Guenter Neumann* University of Hohenheim, Germany Practical application of plant growth-promoting microorganisms (PGPMs) as so-called biofertilisers to improve growth and nutrient acquisition of crops is frequently biased by low reproducibility of the desired effects. Various environmental factors including biotic and abiotic stresses, competition with the indigenous microflora and genotypic differences in host compatibility may limit the rhizosphere competence, the survival and thus the efficiency of the microbial inoculants. BIOFECTOR is a collaborative project, located within the 7th EU Framework Programme with the aim to select suitable microbial partners for applications in various alternative fertilisation systems. It is expected that a strategic combination of microbial strains adapted to the culture conditions characteristic for the respective fertilisation systems will reduce the variability of host plant responses and thus the efficiency of the plant-microbial interaction. First promising results but also drawbacks and open questions are summarised with respect to PGPM use for improved nutrient acquisition from organic recycling fertilisers, in fertiliser placement strategies and mobilisation of sparingly soluble nutrients. 156 The engineered rhizosphere bacterium help plant sense a hazards chemical Choong-Min Ryu*, Hae-Ran Lee, Soohyun Lee KRIBB, South Korea Synthetic biology is a new field of biological engineering that generates new biological modules and synthesizes novel pathways to reprogram organisms. It is valuable to the development of reporter plants to provide a rapid, low-cost, and in situ monitoring of environmental hazards and plant diseases. The reporter plants are also useful for the ecological risk assessment of industrial chemicals. We attempted to develop a reporter plant that senses hazardous aromatic compounds such as toluene. Bacterial two-component signal transduction system such as TodST of a rhizosphere bacteria Pseudomonas putida is useful to construct artificial genetic regulatory module for synthetic biology. For developing toluene sensing reporter plant, we exploited root-rhizosphere bacteria (rhizobacteria) interaction. First, we generated P. putida KT2440 to produce indole-3-acetic acid (IAA) depending on 201 Engineering the Rhizosphere Tuesday 23 June – Poster session toluene concentrations through TodST system. Secondly, we manipulate Arabidopsis and tobacco plant to elicit RNAi of the magnesium chelatase (ChlH) gene induced by concentration dependent manner of IAA produced by P. putida under presence of toluene. Collectively, we provide new two-step chemical sensing system which turn over signals from chemical sensing rhizobacteria to plant indicator. 157 Plant-Sediment Microbial Fuel Cells: electron transfer mechanisms and their energy efficiencies David Strik*, Koen Wetser, Cees Buisman Wageningen University, Netherlands Wetlands can be used to generated electricity by means of Plant-Sediment-Microbial Fuel Cells (PSMFC). In this since 2008 emerging technology, solar energy is converted into electricity. Basically the technology consists of: 1) an anode which is placed in the vicinity of plants roots to take-up electrons; 2) an electric circuit to harvest energy out of released electrons.; and 3) a cathode placed at an oxygen rich environment to deliver electrons for oxygen reduction. Experimental work supports that electrochemical active bacteria are responsible for catalysis of anodic oxidation and cathodic reduction reactions. The PSMFC provides a new solid electron acceptor and donor in the wetland. Plants and sediments both provide a wide pool of (potential) electron donors (e.g. exudates, lysates, sulfide) and electron acceptors (e.g. oxygen, metals, nitrate, sulfate, carbon dioxide). Electron donors can be converted and transferred via different direct and indirect mechanisms. E.g. micro-organisms can oxidise acetate and directly deliver electrons to the anode. Also, microbial or plant-based mediators can be used to transfer electrons from micro-organisms to the solid electrode. Based on thermodynamics and actual conditions, one can calculate the energetically most attractive route for electron transfer at different sites like root tips and root-turn over locations. Aim of this study was to determine (novel) theoretical pathways of electron transfer mechanisms in the Plant-Sediment Microbial Fuel Cells and reveal their energy efficiency. With this work we present the working principles of novel electron transfer pathways and provide in-sights on effective placement of anodes and cathodes. Past experimental results were linked to the outcome of the model. By further experimental elucidation of the proposed model, the actual processes and efficiencies in Plant-Sediment Microbial Fuel Cells can be quantified. 202 Engineering the Rhizosphere Tuesday 23 June – Poster session 158 Rhizogenesis: Exploring the physical development of the emerging root:soil interface Jonathan Helliwell1, Craig Sturrock*1, Sacha Mooney1, Anthony Millar2, Richard Whalley3 1 University of Nottingham, United Kingdom, 2John Innes Centre, United Kingdom, 3Rothamsted Research, United Kingdom The rhizosphere is a distinct zone of soil directly influenced by a plant root, with all below ground resources passing through this dynamic zone prior to capture by plants. Therefore the physical nature of the interface between the rhizosphere and the bulk soil is crucial for plant development. It is well known that the soil microbial community play a significant role in the evolution of the rhizosphere and some studies have shown that it is structurally a different environment to the bulk soil. However how this evolution or genesis is influenced by the underlying soil physical properties and how this interacts with different plant species is less well understood. Examining the undisturbed rhizosphere has represented a major obstacle to research, due to its microscopic size and often fragile nature. Here we have employed X-ray Computed Tomography (CT) to successfully map the physical architecture of the developing rhizosphere in natural soils for the first time. We compared the temporal changes to the intact porous structure of the rhizosphere during the emergence of a developing root system, by assessing changes to the soil porous architecture across a range of soil textures and plant species. Our results indicate the physical zone of influence of a root at an early stage is more localised than previously thought (at the µm rather than mm scale). Soil porosity increases at the immediate root surface due to localised crack formation in both fine and coarse textured soils. As such the soil porous architecture at the root interface is enhanced and not compacted as previously considered. This 'rhizosphere structure' and associated dynamics have important consequences for several important root-soil processes including water uptake efficiency and gaseous exchange between individual aggregates and subsequently our efforts to model their behaviour. 159 Effects of crop rotation on pathogen-suppressive activity and shifts in antibiotic activity of soilborne actinomycete communities Patricia Vaz*1, Nora Altier2, Carlos Pérez3, Linda Kinkel4 1 INIA- Las Brujas, Uruguay, 2Sección Bioinsumos, INIA- Las Brujas, Uruguay, 3Facultad de Agronomía, Universidad de la República, Uruguay, 4Department of Plant Pathology, University of Minnesota, United States Indigenous soil microbes have intrinsic potential to protect plants from pathogens, yet our understanding of the factors that determine the dynamics of antagonistic populations remains limited. Bacterial antibiotic production may change in the proximity of another organism, thus not only the ability of the bacterial community to produce antibiotics but also interactions with neighbors are likely to determine community antagonistic potential. We explored the relationships between crop rotation and edaphic characteristics with community antagonistic activity, and with induction of changes in inhibition among sympatric actinomycetes. Soil samples were taken from an experiment established 13 years earlier, with 10 treatments in a randomized block design (n = 3 blocks). Bacterial, actinomycete, and inhibitor densities were measured for all plots. Mean inhibition of actinomycetes against 203 Engineering the Rhizosphere Tuesday 23 June – Poster session target organisms was also determined. Three plots were selected for isolation of a random collection of sympatric actinomycetes using a 1 cm diameter soil corer. Ten isolates from each community were tested for their abilities to induce changes in inhibition among all possible sympatric isolate pairs. Soils having different rotation histories had significantly different bacterial, actinomycete and inhibitor densities. Soil communities from longer rotations had significantly higher microbial densities than communities from shorter rotations. Moreover, communities with higher inhibitor densities had better inhibitors. Soil nitrate, organic matter and soluble potassium were significantly correlated with microbial community characteristics. Inhibition among sympatric isolates was both increased and decreased by the presence of a partner. However, no significant differences in the frequencies of such shifts were observed among communities. Our work suggests that agronomic practices that contribute to increasing total bacterial densities are likely to enhance the potential for microbial communities to suppress plant pathogens. In this study, longer rotations, with higher plant diversities over time, were most effective in achieving high bacterial densities and strong inhibitory populations in soil. 160 Managing replant disease of apple and sweet cherry with compost Tristan Watson*1, Louise Nelson1, Tom Forge2 1 The University of British Columbia - Okanagan Campus, Canada, 2Agriculture and Agri-Food Canada, Canada Replant disease refers to the poor growth of fruit trees planted into soil previously used for tree-fruit production. The plant-parasitic nematode, Pratylenchus penetrans, and a diverse array of root-associated fungi have been implicated in replant disease worldwide. Recent restrictions on soil fumigants have generated interest in alternative management strategies, particularly those associated with promotion of beneficial microorganisms in the rhizosphere. This study aimed to investigate: (1) the impacts of amending soil with biocontrol agents and composts on plant growth and recovery of root pathogens, and (2) differences in N mineralization between two different composts. A greenhouse pot experiment was performed, consisting of apple and sweet cherry seedlings grown in old apple orchard soil amended with either a biocontrol agent, an agricultural or municipal waste compost, granular chemical fumigant (Basamid®), or nothing (untreated control). After 19 weeks growth, the abundance of P. penetrans was determined for each pot, and colonization by root-associated fungi assessed through isolation and molecular identification (ITS region). Differences in soil microbial communities were determined using Biolog EcoPlatesTM combined with real-time PCR analyses of total bacteria. Differential nutrient mineralization was assessed using freeliving nematode community indices of nutrient enrichment as well as analysis of NO3-N content. Fumigation and compost amendments decreased the abundance of P. penetrans and colonization by root-associated fungi, however only fumigation and agricultural waste compost improved root volume. Fumigation and compost amendments selected for distinct microbial communities, with agricultural waste compost supporting a greater abundance of total bacteria relative to non-compost amended treatments. Similarly, agricultural waste compost also supported a greater nematode enrichment index, lower channel index, and higher soil NO3-N content relative to municipal waste compost, suggesting enhanced Nmineralization through the bacterial decomposition pathway. Overall, agricultural waste 204 Engineering the Rhizosphere Tuesday 23 June – Poster session compost shows potential as a non-fumigant option for control of replant disease impacting apple and sweet cherry. 205 Engineering the Rhizosphere Wednesday 24 June – Poster session Symbiosis 1 Effect of Sinorhizobium sp. and phosphorus on growth, nodulation, and nitrogen fixation of fenugreek (Trigonella foenum-graecum L.) Abdolreza Akhgar*, Sara Larki Vali-e-Asr University of Rafsanjan, Iran The Legume-rhizobia symbiosis is the basis of biological nitrogen fixation and improving soil fertility. Inoculation of legumes with rhizobia is an important practice to maximize biological N2 fixation capacity in legume crops. Phosphorus is also an essential element for Rhizobia to convert atmospheric nitrogen into an ammonium form usable by plants. In this study, a greenhouse experiment was carried out to investigate effects of simultaneous application of Sinorhizobium sp. isolates and phosphorus levels on growth, nodulation, and nitrogen fixation of fenugreek. The experiment studied four Sinorhizobium isolates combined with four phosphorus levels (0, 20, 40 and 60 mg kg-1soil). The Sinorhizobium isolates were isolated from root nodules of fenugreek grown in farms located in Khuzestan and Kerman Province. The greenhouse experiment showed that simultaneous application of Sinorhizobium and phosphorus significantly increased shoot dry weight and nodule number and significantly enhanced N, P, Ca, Mg, Fe and Mn uptake. 2 Plant Growth Promoting Rhizobacteria (PGPR) to benefit root architecture and nutrition of nursery crops Shelby Berg*, Stephen Mudge, Susanne Schmidt, Yun Kit Yeoh The University of Queensland, Australia A healthy root system is fundamental for plant growth and development. Plant growth promoting rhizobacteria (PGPR) form complex symbiotic relationships with plants and benefit plant growth in the rhizosphere, on the root surface, within intercellular spaces and root cells. Here we investigated microbes for their ability to act as PGPR and colonise root systems of young maize and tomato plants in the presence of native soil biota. A number of putative PGPR isolates with strong phytate degrading and phosphorus solubilising activities were identified of which two isolates (an Enterobacter sp. and Burkholderia sp.) were selected for further testing. Tomato and maize seedlings inoculated with only Burkholderia or both Enterobacter and Burkholderia, cultivated in low P soil did not show improved plant growth. However Enterobacter inoculation led to increased shoot and root biomass, root surface area, branching and lateral root growth of both maize and tomato seedlings. Such changes in root architecture are desirable for nursery container production whereby roots inoculated with PGPR remain confined within the pot and do not proliferate from the base. These results also suggest that the Enterobacter strain, although originating from the sugarcane rhizosphere, has a wider host range. We are currently using genetically-engineered Enterobacter expressing green fluorescent protein (GFP) to study microbial colonisation of root systems and survival post-inoculation. The effects of Enterobacter inoculation on pre-existing rhizosphere microbial populations is also being analysed using bacterial community profiling technologies. We discuss the findings in context of formulating efficient PGPR inoculums to specifically target crops of interest for growth promotion. 206 Symbiosis Wednesday 24 June – Poster session 3 Nutritional value of tomato and strawberry fruits is affected by plant inoculation with soil bacteria Elisa Bona*1, Elisa Gamalero2, Guido Lingua2, Simone Cantamessa2, Nadia Massa2, Valeria Todeschini2, Paola Manassero2, Giorgia Novello2, Andrea Copetta2, Giovanni D'Agostino3, Graziella Berta2 1 Università del Piemonte Orientale, Italy, 2Università del Piemonte Orientale, DiSIT, Italy, 3Mybasol, srl, Italy Soil bacteria can stimulate plant growth; however, little information is available about the impact of these bacteria on the nutritional value of fruits. Therefore, we considered two economically relevant crops: tomato for the evidence of reduced risk of cancer and cardiovascular diseases and strawberry because they are an important source of antioxidants and anti-inflammatory phytonutrients. Tomato plants were inoculated with Pseudomonas sp. 19Fv1T, or Pseudomonas fluorescens C7 and grown in open field condition. Strawberry plants were inoculated with P. fluorescens Pf4 or Pseudomonas spp. 5Vm1K and grown in pot under glasshouse condition. Both plant species were subjected to reduced fertilization. The impact of the bacterial strains on the fruit nutritional value was assessed by measuring the concentration of soluble sugars, organic acids, vitamins, anthocyanin, lycopene and b-carotene together with nitrate and nitrite amount in fruits. The size and biomass of tomato fruits were unaffected by the bacterial strains. However, the sugar content was modulated by the microorganisms (i.e. fructose increase and glucose reduction). 19Fv1T induced an enhancement of malic, tartaric, ascorbic and glutamic acids, beta carotene and a reduction of oxalic acid in the fruits compared to controls. The strain C7 induced reduction of the glutamic, oxalic and ascorbic acid, and decrease of nitrite level in fruits compared to those produced by uninoculated plants. Both strawberry fruit size and yield were increased by the bacterial. The concentrations of sugars were differently affected according to the microorganism. Both bacteria enhanced the amount of ascorbic acid in fruits, but only Pf4 increased folic acid concentration. Fruit yield and quality can be improved by plant inoculation with soil bacteria; this can have a real world application, leading to economical, ecological and human health benefits in relation to the reduced chemical input and to the increased quality of the fruits. 4 Roots and nodules bacterial diversity of Scorpiurus muricatus in western Algeria Zoulikha Bouchiba*1, Zineb faiza Boukhatem1, Zohra Ighilhariz1, Mustapha MIsbah el Idrissi2, Abdelkader Bekki1 1 University of Oran 1 Ahmed Benbella, Algeria, Algeria, 2University Mohamed V Agdal,Rabat - Morocco, Morocco Algeria is facing chronic forage deficit, in order to improve the production and contribute to increasing animal production for sustainable development, the use of spontaneous forage legumes of pastoral interest as Scorpiurus muricatus ssp. sulcatus seems interesting. The 207 Symbiosis Wednesday 24 June – Poster session diversity of microorganisms associated with this plant is very little studied worldwide. The purpose of this study is to investigate the genotypic characteristics of microorganisms associated with this plant. Fifty nodular and root bacterial strains respectively were isolated from plants sampled in six regions in western Algeria. Nodular isolates are grouped into different clusters according to profiles obtained by repPCR by ERIC and BOX primers. NodC amplification allowed us to select renodulant strains. Nodular and root endophytes are studied by PCR-RFLP of DNAr16s and compared with references strains, a representative of each group is sequenced. The endophytes ability to solubilize phosphorus and IAA production is determined. 5 Biological Nitrogen Fixation Book Volume I and II Frans J. de Bruijn* INRA/CNRS Laboratory of Plant Microbe Interactions, France Nitrogen is arguably the most important nutrient required by plants. However, the availability of nitrogen is limited in many soils and although the earth’s atmosphere consists of 78.1% nitrogen gas (N2) plants are unable to use this form of nitrogen. To compensate, modern agriculture has been highly reliant on industrial nitrogen fertilizers to achieve maximum crop productivity. However, a great deal of fossil fuel is required for the production and delivery of nitrogen fertilizer. Moreover carbon dioxide (CO2) which is released during fossil fuel combustion contributes to the greenhouse effect. Biological nitrogen fixation is one alternative to nitrogen fertilizer. It is carried out by prokaryotes using an enzyme complex called nitrogenase and results in atmospheric N2 being reduced into a form of nitrogen diazotrophic organisms and plants are able to use (ammonia). It is this process and its major players which will be discussed in the “Biological Nitrogen Fixation” Books. The best known and most extensively studied example of biological nitrogen fixation is the symbiotic interaction between nitrogen fixing “rhizobia” and legume plants. Here the rhizobia induce the formation of specialized structures (“nodules”) on the roots of the legume plant and fix nitrogen which is directly assimilated by the host plant;. It is this symbiotic interaction which will be highlighted in the Book. While legumes are important as major food and feed crops, cereals such as wheat, mays and rice) do not have this symbiotic nitrogen fixing interaction with rhizobia. It has thus been a “holy grail” to transfer the ability to fix nitrogen to the cereals and different timely approaches towards this goal are also discussed in the Books. 208 Symbiosis Wednesday 24 June – Poster session 6 Phosphorus use efficiency in N2-fixing legume-rhizobia symbioses Jean Jacques Drevon*1, J. Abadie1, L. Amenc1, A. Bargaz2, O. Dommergue3, M. Lazali4, M. Zamanallah5 1 INRA Ecologie Fonctionnelle & Biogéochimie des Sols & Agroécosystèmes, France, 2Swedish University of Agricultural Sciences, Department of Biosystems and Technology, Sweden, 3Laboratoire des Symbioses Tropicales et Méditerranéennes, Campus International de Baillarguet, France, 4Université de Khemis Miliana, Algeria, 5CIMMYT, Southern Africa Regional Office, Zimbabwe Low phophorus availability in about 40% of the world’s arable land limits crop yield, most particularly for leguminous crops when their growth depends upon symbiotic N2-fixation (SNF). Therefore, our work aims to increase the phosphorus use efficiency (PUE) for SNF, and its contribution to a more effective coupling between the P and N bio-geochemical cycles. Myo-inositol hexakisphosphate (phytate) constitutes the main source of organic P in soils, but is unavailable to plants. Phytases are the only phosphatases able to hydrolyse phytate efficiently into inorganic Pi, thus increasing P bio-availability for plants. In this work we demonstrate the existence of phytase activity in root-nodules, and show the expression of a purple acid phytase within nodule inner-cortex by in situ RT-PCR. Also histidine acid- and bpropeller- phytases (HAP and BPP) were found among legume-nodulating bacteria, i.e. rhizobia, with major expression in nodule infected-cells. Moreover, phosphoenol pyruvate phosphatase and trehalose 6P phosphatase were discovered in nodules with puzzling localization. The nodular expression of all these genes and the enzymatic activity of their products increased significantly under P-deficiency, and varied among recombinant inbred lines of Phaseolus vulgaris that are contrasting in their PUE for SNF. It is concluded that the differential expression of bacterial and plant phosphatase-genes in nodules offers a new understanding of the N2-fixing legume physiology and its specific P requirements. The potential of these phosphatase-genes for a virtuous circle of P and N soil-fertility will be addressed in relation with the interdisciplinary research strategy of the FABATROPIMED federative project of Agropolis Montpellier. 7 Diagnosis of common bean inoculation needs and selection of effective bacterial biofertilizers adapted to soil conditions of the Skhirate region in Morocco Imane El Attar*1, Jamal Aurag2, Imane Thami Alami3, Mohamed El Khadir3 1 Laboratory of Microbiology and Molecular Biology (LMBM), Faculty of Science, Mohammed V University, Morocco, 2Laboratory of Microbiology and Molecular Biology (LMBM, LMI), Faculty of Science, Mohammed V University, Morocco, 3National Institute for Agricultural Research (INRA), Morocco The Skhirate region (North-West part of Morocco) provides about 20% of of common bean production in the country under an intensive production system based on the uncontrolled use of chemical fertilizers. In this context our research project aims to reduce the amount of fertilizers used during the cultivation of common bean causing negative environmental impacts. For this purpose, we plan to develop bacterial biofertilizers that can contribute efficiently to the nutrition of common bean and its growth by replacing, partially, the intakes of mineral fertilizers. This approach is based on the establishment of a sustainable 209 Symbiosis Wednesday 24 June – Poster session agrobiological system for the nutrition of common bean using mostly symbiotic nitrogen fixation and inorganic phosphate solubilization. During the growing season of common bean, a general survey was carried out in the Skhirate region in order to diagnose the nodulation status of this legume and determine if there is a need for inoculation with bacterial inocula. Common bean nodulation was found to be very low or even absent in most of the visited fields, thus we decided to carry out additional surveys in other production areas in Morocco (Beni Mellal, Moulay Bouselham and Berkane). Nodules collected during these surveys have enabled the creation of a collection of 143 bacterial isolates which were subjected to a physiological characterization, especially their tolerance to salt and pH stresses and their potential plant growth promoting activities (PGP activities). The results of this characterization revealed a large number of isolates with interesting PGP activities and tolerance to both types of stress studied. The symbiotic properties of these isolates are being characterized under controlled conditions. The strains able to establish a symbiotic relationship with the host plant (Phaseolus vulgaris) will be identified by sequencing the 16S rRNA gene. 8 Does mycorrhizal phosphate uptake influence the root-associated microbiome? Nina Gerlach*, Martin Willmann, Eva Koebke, Alexander van Burgeler, Marcel Bucher University of Cologne, Botanical Institute, Cologne Biocenter, Cluster of Excellence on Plant Sciences (CEPLAS), Germany The plant microbiome is a key determinant of plant growth and fitness. Arbuscular mycorrhizal fungi are part of the fungal microbiome of most terrestrial plants and function as an extension of the root system facilitating bi-directional exchange of soil-born nutrients and plant-derived carbon between both symbionts. Maize plants colonized by arbuscular mycorrhizal fungus Rhizophagus irregularis exhibited enhanced growth under nutrient-deficient conditions. Systemic metabolic alterations included anthocyanin and lipid metabolism likely in response to an improved P-status of mycorrhizal maize leaves. An overall increase in leaf C versus N metabolism highlighted changes in metabolic fluxes. A parallel induction of defense gene expression and accumulation of secondary metabolites suggested priming of mycorrhizal maize leaves. Phosphate uptake in plants is mediated by transporters of the Pht1 protein family. A pht1;6 transposon insertional maize mutant is strongly impaired in mycorrhizal phosphate uptake and thus exhibited reduced biomass accumulation in agricultural soil poorly fertilized with phosphate. Arbuscular mycorrhizal fungal colonization of isolated pht1;6 plants was strongly diminished. However, when cultivated together with wild type plants, fungal colonization of pht1;6 was restored but mutant roots exhibited increased arbuscule degeneration and formation of strongly septated stunted arbuscules. In consideration of the role of Pht1;6 in maize root and shoot physiology, our current interest is on the impact of Pht1;6 transporter activity on the root-associated fungal microflora 210 Symbiosis Wednesday 24 June – Poster session including mycorrhizal but also other fungi. To this end, the fungal microbiome of pht1;6, heterozygous and wild type plants was analyzed by a PCR-based automated ribosomal intergenic spacer analysis. Trans-complementation assays highlighted the impact of mycorrhizal neighboring plants on the mutant root-associated microbiome. Moreover the taxonomic structure of fungal assemblages as a function of Pht1;6 will be resolved by a culture-independent molecular approach. 9 Endofungal bacteria in plant symbiotic fungi Anton Hartmann*1, Dan Li1, Michael Rothballer1, Jochen Blom2, Peter Kämpfer3, Stefanie P. Glaeser3, Karl-Heinz Kogel4 1 Helmholtz Zentrum München, German Research Center for Environmental Health, Department Environmental Sciences, Research Unit Microbe-Plant Interactions, Germany, 2Bioinformatics and Systems Biology, Justus Liebig University, Germany, 3Institute of Applied Microbiology, Research Centre for BioSystems, Land Use and Nutrition, Justus Liebig University, Germany, 4Institute of Phytopathology, Research Centre for BioSystems, Land Use and Nutrition, Justus Liebig University, Germany Endofungal bacteria occur in some ectomycorrhizal as well as arbuscular mycorrhizal fungi; a mycorrhizal helper function was attributed to these bacteria. In the order of Sebacinales (Basidiomycota), also endofungal bacteria have been demonstrated. In many cases, the endofungal bacteria are not cultivated yet, but in the case of Piriformospora indica, the endofungal bacterium Rhizobium radiobacter F4 (RrF4) was cultured and its genome and physiology could be investigated. A better understanding of the role of R. radiobacter in this tripartite plant-fungus-bacterium symbiosis is of high relevance, because P. indica has a wide range of plant beneficial applications in agriculture. The genome of RrF4 has been sequenced by second generation pyro-sequencer (454 GS FLX Titanium). It is organized in a circular and a linear chromosome and two plasmids (pTiF4 and pAtF4) in the same manner as in R. radiobacter C58 (formerly Agrobacterium tumefaciens). The average amino acid identity (AAI) of RrF4 to biovar I strains (e.g. C58) is 99.8%. While RrF4 and C58 showed a high degree of similarity based on the circular and linear chromosomes, the plasmids were more diverse. In addition to some rearrangements between the two plasmids, RfF4 most importantly lacks – in contrast to C58 - the complete transferred DNA (T-DNA) region and some adjacent genes belonging to the nopalin catabolic region. Like in C58, strain RrF4 contains the structural gene traI for N-acylhomoserine lactone (AHL)synthesis and traR for the AHL-receptor traR. Detailed chemical analysis showed that RrF4 produces a range of oxo- and hydroxyl-AHLs with C8-, C10-, or C12 alkyl side chains. These autoinducer signaling molecules are able to induce systemic resistance in different plant hosts as well as having plant growth promoting effects. We therefore hypothesize that the endofungal bacterium RrF4 of P. indica contributes to its plant growth promoting action in the tripartite plant-fungal-bacterium symbiosis. 211 Symbiosis Wednesday 24 June – Poster session 10 An investigation on the colonisation of bacterial endophytes in oilseed rape, grown in the presence of entomopathogenic nematodes Mary Jo Hurley*, Dina Brazil, Thomais Kakouli-Duarte Institute of Technology Carlow, Ireland Pathogenic microorganisms and pests are a predominant threat in agriculture worldwide and control of these has relied heavily on chemical pesticides. The overuse of pesticides has been directly related to the intensification of agriculture over previous decades, but with substantial environmental costs. The overall aim of this project focuses on the biocontrol of economically important insects utilising a combination of entomopathogenic nematodes and bacterial endophytes, to help reduce pesticide input in the environment. Both entomopathogenic nematodes and endophytic bacteria have widespread applications as biocontrol agents, however the potential of synergism between these nematodes and endophytic bacteria has yet to be explored. Investigating the effects of various endophytic bacteria on nematode biology and behaviour will not only contribute to knowledge on these but also increase nematode efficiency and predictability within a dynamic environment. Results are presented here from experiments designed to (a) investigate the effects of bacterial endophytes on nematode virulence and (b) examine the potential of increased bacterial-plant colonisation in the presence of entomopathogenic nematodes. Nematode dose response experiments were carried out to determine negative or stimulatory effects of endophytes on nematode infectivity. In all bacterial treatments Heterorhabditis bacteriophora infected in lower numbers than Steinernema feltiae and Steinernema carpocapsae. Following bacterial exposure fewer S. feltiae (e-nema) infective juveniles were recovered, when compared to the control. Moreover, for all nematodes species fewer numbers were recovered from insects exposed to bacterial strain L228. Bacterial colonisation of oilseed rape was determined via bacterial plate counts and the polymerase chain reaction, using green fluorescent protein specific primers. To date gfp labelled bacteria have been isolated from root, rhizosphere and stem samples, in the presence and absence of nematodes. This work is currently on-going. 11 Symbiotically fixed N substitutes fertilizer N by enhancing C and N assimilation during reproductive stages of soybean Jian Jin*1, Yansheng Li2, Xiaobing Liu2, Guanghua Wang2, Zhenhua Yu2, Mathesius Ulrike3, Judong Liu2, Herbert Stephen4 1 Northeast Institute of Geography and Agroecology, Chinese Academy of Sciences, China, 2Key Laboratory of Mollisols Agroecology, Northeast Institute of Geography and Agroecology, Chinese Academy of Sciences, China, 3Division of Plant Science, Research School of Biology, The Australian National University, Australia, 4Stockbridge School of Agriculture, University of Massachusetts, USA Excessive fertilizer has been commonly applied in the soybean cropping system in fertile Mollisols in northeast China. It is necessary to understand the mechanisms of how reducing N fertilizer application impacts the plant N acquisition and remobilization, which is associated 212 Symbiosis Wednesday 24 June – Poster session with photosynthetic C assimilation and seed yield. Thus, the aim of this study was to investigate the origin of plant N, i.e. N2 fixation-, fertilizer- and soil-derived N under two different N application levels, and the subsequent influence on C assimilation. A pot experiment was conducted with soybean grown in a Mollisol supplied with 5 (N5) or 100 (N100) mg N kg-1 soil. Nitrogen was applied as 19.83% of 15N atom excess in urea before sowing, and 13CO2 labeling was performed at the R5 (initial seed-filling) stage. Plants were harvested at R5 and R8 (full maturity) stages to determine the 15N and 13C abundance in plant tissues. Seed yield and N content were not affected by different N rates. The symbiotically fixed N accounted for 64% of seed N in N5, while fertilizer-derived N dominated seed N in N100, resulting in 58% of seed N. The proportion of soil-derived N in seed showed no difference between the two N treatments. The enhanced N2 fixation in N5 significantly increased the assimilation of N and C during the seed-filling period compared to N100. The nodule density (nodule number per unit root length) and the amount of photosynthetically fixed 13C in roots in N5 were greater than those in N100. These results indicate that prolonging N2 fixation to increase assimilated N during the seed-filling period is likely to meet N demand for maintaining soybean yield when fertilizer N supply is reduced. More allocation of photosynthetic C to roots and enhanced nodulation would greatly contribute to the alteration of N acquisition pattern. 12 Selection of specific Rhizobium leguminosarum genotypes by different Fabeae legume hosts: A Pool-Seq mesocosm study Beatriz Jorrin*1, Amalia Soenens1, Juan Imperial2 1 Centro de Biotecnolgia y Genomica de Plantas. Universidad Politecnica de Madrid, Spain, 2Centro de Biotecnolgia y Genomica de Plantas. Universidad Politecnica de Madrid. Centro Superior de Investigaciones Cientificas, Spain Rhizobium leguminosarum bv viciae can establish effective symbioses with members of the Fabeae legume tribe (Pisum, Lathryrus, Lens and Vicia). Previous studies have suggested that different Fabeae select specific genotypes of rhizobia from those available in soil. We extended these observations at the genomic level by applying a Pool-Seq analysis to isolates selected from a soil population by pea, lentil, fava and vetch plants, and showed that plantselected sub-populations differ at the single nucleotide polymorphism level. The nature and extent of plant-specific genotypic preference were further studied by performing controlled mesocosm assays. Sterilized seedlings were planted in soil:vermiculite 1:1, and 21-day roots were collected, sterilized superficially, homogenized and centrifuged at low speed. This supernatant was used as inoculant for the next cycle of selection in the same soil with new seedlings. After 5 cycles, roots were collected, and rhizobia (25 from each host) isolated from nodules. Each of the isolates was grown separately, all cultures from the same host were pooled, and their pooled DNA was isolated and subjected to Pool-Seq analysis. Sequence reads were aligned against the R. leguminosarum bv viciae 3841 reference genome and both coverage and polymorphism analyses were performed for specific regions. No genotypic selection by the host was observed in the rDNA region, whereas increased genotypic selection was very clear along the mesocosm experiment for the nod cluster, but only in the pea and vetch subpopulations. In contrast, no further polymorphism profile changes were observed in the nod region for the fava and lentil subpopulations after the first 213 Symbiosis Wednesday 24 June – Poster session round of plant host selection. These results suggest the existence of differences in plant host selection of rhizobial genotypes along the mesocosm experiment for different hosts and genomic regions. 13 Effect of different biochars on the establishment of the symbiosis between Rhizophagus irregularis and leek grown in peat-based substrate Vicky Levesque*1, Hani Antoun1, Martine Dorais2, Noura Ziadi2, Martin Trépanier3 1 Laval University, Canada, 2Agriculture and Agri-Food Canada, Canada, 3Premier Tech, Canada Arbuscular mycorrhizae are affected by the use of peat-based substrates and inhibition of mycorrhization was observed with Allium species. We hypothesized that the use of biochar could alleviate the observed inhibition of mycorrhization of leek transplants grown in a soilless mix containing 75 to 85% sphagnum peat moss and inoculated with Rhizophagus irregularis. Since biochar physicochemical properties are influenced by feedstock nature and the pyrolysis temperature used, five different biochars were produced (maple bark 400˚C, 550˚C and 700˚C, pine chips 700˚C and willow chips 400˚C) and applied at three different rates (5, 10 and 15% in volume). The experiment was realized during nine weeks in a greenhouse with 5x3 factorial design for biochar types and rates. A control (0% of biochar) was also included for comparison purpose. Biochars were added to potting mix and inoculated with monoxenic culture of Rhizophagus irregularis DAOM 197198. Leek (Allium ampeloprasum var. Lancelot) was used as host plant. The effect of biochar amendment on leek root colonization by mycorrhizae, plant biomass and phosphorus uptake were evaluated at the seventh leaf stage. Results indicated that the type of biochar and the level of amendment have an effect on root colonization by mycorrhizal fungi. Indeed, in comparison to the control, the addition of 15% of maple biochar increased (ρ < 0.05) the percentage of root colonization (ranging from 23% to 46%). Nevertheless, the results have shown a reduction (ρ < 0.05) in shoot dry matter biomass and phosphorus uptake with maple biochar. Observed decreases could be attributed to the early stage of plant development. In conclusion, biochar could alleviate peat inhibition in soilless mix and improve mycorrhization, which will produce more robust leek transplants. 14 Nitrogen transfer in soybean/maize intercropping system inoculated arbuscular mycorrhizal fungi and rhizobium Shumin Li*1, Aiyuan Zhang2, Lingbo Meng3, Xiaoguang Han2, Fei Wang2, Dejiang Wang2 1 Northeast Agricultural University, China, 2Resource and Environmental College, Northeast Agricultural University, China, 3Department of Life Science, Harbin University, China The tripartite symbiosis between legumes, rhizobia and mycorrhizal fungi are generally considered to be beneficial for nitrogen (N) uptake of legumes, but the facilitation of the symbiosis in legume/non-legume intercropping system is not clear.Therefore, the aims of the research are 1) to certify if the dual inoculation can facilitate the N uptake and N transfer in 214 Symbiosis Wednesday 24 June – Poster session maize/soybean intercropping system, 2) to calculate how much N will be transferred from soybean to maize. A pot experiment with different root separations (solid barrier, mesh(30µm) barrier and no barrier) was conducted and 15N isotopic tracing method was used to calculate how much N transferred from soybean to maize inoculated arbuscularmycorrhizal fungi (AMF) and rhizobium in soybean (Glycine max L.cv. Dongnong No.42)/maize (Zea mays L.cv.DongnongNo.48) intercropping system. In comparison with the inoculatingGlomus mosseae(G.m.), Rhizobium SH212 and no inoculation, both inoculating Glomus mosseaeand Rhizobium SH212 increased the N uptake of soybean by 28.73%, 39.62% and 93.07% in solid barrier system. N uptake of maize inoculated both Glomus mosseae and rhizobium was 1.20, 1.28 and 1.67 times greater than that ofinoculating Glomus mosseae, Rhizobium SH212 and no inoculationrespectively. In addition, the amount of N transferred from soybean to maize of dual inoculation with mesh barrier was 7.25 mg, 7.01 mg and 11.21 mg greater than that ofinoculating Glomus mosseae, Rhizobium SH212 and no inoculation, and similarly 6.40 mg, 7.58 mg and 12.46 mg increased in no barrier system. Inoculating both AMF and rhizobium in soybean/maize intercropping system could improve the N fixation efficiency of soybean, and promote N transfer from soybean to maize, which result in the improvement of yield advantage in legume/non-legume intercropping. 15 Insect vectors efficiently convey complex endophytic communities across grapevine plants Sebastian Lòpez-Fernàndez, Valerio Mazzoni, Pier Luigi Bianchedi, Ilaria Pertot, Andrea Campisano* Fondazione Edmund Mach, Italy Microbial endophytes colonize the inner tissues of plants. It is commonly held that most endophytes invade the host tissues through the roots or through discontinuities on the plant surface, including wounds and stomata. Microorganisms can also be transferred through root anastomoses, as it occurs for instance with some pathogenic mollicutes, such as the phytoplasmas. Some insects able to penetrate the plant surface are also vectors of phytoplasmas. Very little is known about the ability of such vectors to harbour and transfer other microorganisms. To unravel the ecological role of insects for endophytic microorganisms, we used freshly hatched nymphs of the sap-feeding leafhopper Scaphoideus titanus (vector) to transport microorganisms across grapevine plants. We used adult, greenhouse-grown (donor) plants with an established endophytic fauna, and micropropagated (acceptor) grapevines hosting no detectable bacteria. We used 454 pyrosequencing of the bacterial 16S rDNA gene to estimate the composition of bacterial endophytic communities in donor plants, vector insects and acceptor plants, and to track microbial communities along the insect-plant-microbe network. After contact with the vector, acceptor plants were colonized by a complex endophytic community dominated by Proteobacteria, highly similar to that present on donor plants. Interestingly, a similar bacterial community, but with a higher ratio of firmicutes, was found on S. titanus. Insects feeding only on acceptor plants transferred an entirely different bacterial community dominated by Actinobacteria, where the opportunistic human pathogen Mycobacterium abscessus played a major role. Despite the fact that insects dwelled mostly on 215 Symbiosis Wednesday 24 June – Poster session plant stems, the bacterial communities in plant roots resembled more closely those inside and on insects, when compared with above-ground plants. We establish here for the first time the potential of insect vectors to transfer entire bacterial communities across plants. We also define the probiotic role of plants and microbial endophytes in establishing microbial communities in plant-feeding insects. 16 Inoculation with selected strains of Azospirillum spp. replaces nitrogen in pearl millet (Pennisetum glaucum (L.)) in Brazilian cerrado soil Ivanildo Evodio Marriel*1, Izabelle Melo2, José A. Rodrigues1, Christiane A Oliveira1, Eliane A. Gomes1, Francisco A Souza1 1 Embrapa Maize and Sorghum, Brazil, 2UFSJ, Brazil The cultivation of pearl millet has received increasing attention as an alternative crop for mulch formation in no-till systems and as forage for livestock production, mainly in the Brazilian Cerrado fields. There are evidences of the benefic contribution of the inoculation of diazotrophic bacteria to agronomic crops as a source of nitrogen (N), including pearl millet, but there is no strains recommended for this crop in the country. Here we report the effect of inoculation of Azospirillum strains on pearl millet (BRS 1501) growth and nitrogen fertilizer replacement. We evaluated 20 Azospirillum strains (CMS01 to 20) under three doses of N (0, 30 and 60 kg N.ha-1 on field condition at the Municipality of Sete Lagoas, MG State, Brazil. The crop was cultivated in plots at the field; the experimental design was randomized blocks with four replications. A basic fertilization was applied at sowing and consisted of 40 kg ha-1 of urea, 400 kg.ha-1 of superphosphate, 100 kg ha-1 of potassium chloride and 20 kg ha-1 FTE. At the flowering stage, plants were collected for determination of dry matter accumulation, concentration and content of macronutrients in the shoots. There were significant differences (p <0.05) between treatments for the variables analyzed, except magnesium content. The dry matter accumulation ranged from 163 to 367g for 3 plants and N content between 3.1 to 6.84g for 3 plants. The strains CMS 7 and 11 provided shoots growth and nitrogen uptake similar to those observed with 60 kg / ha of N. Our results also suggested the role of these bacteria as plant growth promoters. We highly the importance of screening of several different strains to obtain promising bacteria for inoculant formulation, seeking replacement of the nitrogen in the pearl millet crop. 17 Microbes from Inner Space: II. Diversity of seed inhabiting endophytes in white clover (Trifolium repens) across continents and their plant beneficial potential Jana Monk*1, Richard Johnson2, Rhys Jones3, Katharine Adam3, Damien Fleetwood4, Nigel Bell3 1 AgResearch Ltd, New Zealand, 2AgResearch Ltd, Grasslands Research Centre, New Zealand, 3AgResearch Ltd, Ruakura Research Centre, New Zealand, 4Biotelliga Ltd, Institute for Innovation in Biotechnology, New Zealand Endophytes are microbes that can be found inside plant tissues, where they can live commensally or execute beneficial functions for the host. It is believed that every plant 216 Symbiosis Wednesday 24 June – Poster session species harbors endophytes, and seeds of many plant species have been reported inhabiting endophytes. This study investigates whether seed endophytes in white clover are conserved at a global scale and what beneficial traits are associated with the culturable fraction. Seed from 18 countries across the globe was provided by the Margot Forde Forage Germplasm Centre (New Zealand). The culturable microbial fraction as well as DNA was extracted from seed followed by identification of endophytic microbes and DGGE fingerprinting analysis. No fungal endophytes were detected in clover seed. DGGE analysis of Gammaproteobacteria showed that the seed community composition varied across samples with only a few shared bands. The richness of bands was low with only 2 to 5 bands per seed sample. In contrast, DGGE fingerprints of Betaproteobacteria showed greater level of richess and some common bands across most samples. Gammaproteobacteria dominated the culturable fraction with Pantoea spp., Erwinia spp. and Pseudomonas spp. prevalent. Around 200 isolates are currently being evaluated for their ability to influence rhizosphere functions such as producing auxins, catabolizing the precursor of ethylene (ACC), solubilizing phosphorus, sequestering iron and antagonizing pathogens. For isolates with desirable effects on clover plant growth, in-depth analysis of the interactions with the host will be performed to determine the spatial distribution within the host plant and whether the endophyte(s) can be transmitted to new seed following artificial inoculation. The most promising isolates will be used to test for effects on root pathogens and invertebrate root pests. 18 Comparative effects of TRP and TSP on the development of two multi-purpose trees in the presence of mycorrhizae Sacko Ousmane*1, Kane Aboubacry2, Sanon Kadidia Bibata3, Yattara Inamoud Ibny1 1 Faculty of Sciences and Technics, University of Sciences, Technics and Technologies of Bamako, Mali, 2Common Laboratory of Microbiology IRD / ISRA / UCAD, Senegal, 3INERA, Burkina Faso West African soils are structurally deficient in nitrogen (N) and phosphorus (P). P is one of the nutrients that affect agricultural and forestry production. Its supply is mainly provided through the provision of agricultural inputs. There are large deposits of rock phosphate (RP) in most countries of West Africa and among these deposits, Tilemsi rock phosphate (TRP) coming from northern Mali, is one of the best and most receptive. Unfortunately, it is underutilized because of its slow dissolution compared with triple superphosphate (TSP) which cost is very expensive for farmers. Mycorrhizal fungi allow to plants which they are associated a better absorption of water and phosphorus (P). Experiments were conducted under greenhouse and laboratory to highlight the effects of the TRP on development, growth and nutrition of two multipurpose trees, Gliricidia sepium and Sesbania sesban, inoculated with an endomycorrhizal fungus (Glomus aggregatum). The effects of the TRP were compared to those of TSP used as fertilizer of reference. The soilof Nioro (Senegal) was used as the culture substrate for the trees. 217 Symbiosis Wednesday 24 June – Poster session The results showed that the TRP has been the preferential phosphate; used at the rate of 75 mg P.kg-1, it was better used by S. sesban in which the height, biomass, and the absorption of the major elements (N, P, K) have been improved, especially in the presence of mycorrhizal fungus. 19 Expression analysis of two novel lipid transfer proteins during the symbiotic interaction between Medicago truncatula and Sinorhizobium meliloti Chiara Santi1, Barbara Molesini1, Youry Pii*2, Tiziana Pandolfini1 1 University of Verona, Italy, 2Free University of Bolzano, Italy Lipid transfer proteins are small basic proteins that constitute a large family characterized by the ability to transfer phospholipids between a donor and an acceptor membrane. MtN5 is a non-specific lipid transfer protein expressed at a very early phase of the rhizobial symbiosis in the epidermal cells, root hairs and root nodules. By manipulating MtN5 expression in the roots, we demonstrated that this rhizobial protein positively regulates the nodulation process. To get more information about MtN5 function in the symbiosis we have now obtained stably transformed plants overexpressing and silencing MtN5. MtN5-silenced plants were impaired in nodulation, showing a 40% reduction in the number of nodules compared with wild type plants, while transgenic plants overexpressing MtN5 developed 34% more nodules with respect to wild type plants. These evidences indicate a crucial role for MtN5 in controlling bacterial invasion and nodule organogenesis in M. truncatula. We have identified two novel putative lipid transfer proteins of Medicago truncatula, Medtr3g055250 and Medtr7g052640, showing high similarity to MtN5. We have observed that the expression pattern of these lipid transfer proteins is modulated during the rhizobial symbiosis. In roots bearing nodule primordia (at 6 days post inoculation) the transcript level of Medtr7g052640 was almost 6 fold higher than in non-inoculated roots. At 14 days post inoculation, both genes were highly expressed in the root nodules. These data suggest that the different M. truncatula lipid transfer proteins might be involved in the regulation of root nodule development. 20 Microbial improvement of compost and biochar products by combination with arbuscular and ericoid mycorrhizal fungi Areg Poghosyan*, Henning von Alten Leibniz University Hanover, Germany The EU funded project REFERTIL has the mission to transform organic waste from a costly disposal process into an income generating activity. The output products will be safe, economical and standardized compost and biochar products containing phosphorous and nitrogen that can be economically and beneficially used by farmers. Within this framework LUH has the objective to combine inocula of arbuscular and ericoid mycorrhizal fungi with biochars and composts. The joined application shall combine the positive effects on plant nutrition and soil properties the latter have with the known beneficial effects of mycorrhizal fungi. 218 Symbiosis Wednesday 24 June – Poster session Isolates of arbuscular mycorrhizal fungi (AMF) and ericoid mycorrhizal fungi (ERMF) can be produced in vitro or in living plants and be used to inoculate crops. In the REFERTIL project LUH has shown that these isolates can be successfully applied together with biochar made from plant material or animal bones (ABC) and with composts. Aim was the microbial improvement of these soil amendments and organic fertilizers. The AMF isolates differed in the amplitude of their effects; they transported P from ABC to the plant roots and improved plant growth in combination with composts. The mixed application of biochars and composts together with AMF on expanded clay as carrier material is problem-free from a technological point of view. AMF/compost mixtures can even be stored for a certain time. That is of importance for the practical use of the micro-biologically improved new products. Aim of the combined application together with ERMF was the microbial improvement of the composts with the objective of a combined or synergistic suppression of the soil-borne disease caused by the Oomycet Phytophthora cinnamomi. The ERMF isolates in combination with composts were able to reduce the disease. 21 Functional diversity of the arbuscular mycorrhizal symbiosis Sabine Ravnskov*1, John Larsen2 1 Aarhus University/Department of Agroecology, Denmark, 2Universidad Nacional Autónoma de México/Laboratorio Agroecologia, Mexico Arbuscular mycorrhiza is known to alter growth and nutritional status of plants, and to enhance plant tolerance to pathogens. However, functional diversity of the arbuscular mycorrhizal symbiosis has been revealed in studies focusing either on plant phosphorus uptake or on plant tolerance to pathogens. The objective of the present study was to examine the functional diversity of arbuscular mycorrhiza in symbioses between three arbuscular mycorrhizal fungi, Funneliformis mosseae, Rhizophagus irregularis and Claroideoglomus claroideum and six varieties of Cucumis sativus (cucumber) with respect to plant growth and nutritional status, and to the tolerance of cucumber cv Tiffany to Pythium ultimum. The results showed a phenotypic variation of each of the 24 combinations of plant/fungal genotypes involved in the arbuscular mycorrhizal symbioses both with respect to root colonization by the arbuscular mycorrhizal fungi, to plant growth, to nutrient composition in shoots and to plant tolerance to the pathogen, but the functional diversity between the measured parameters varied and was not correlated. As the measured functions are related e.g. by the uptake of phosphorus could be influenced by uptake of ions of other nutrients, growth of plants could be influenced by the nutrient uptake by the plant, and both plant growth and nutrient content is determinant for the quality of the plant as habitat and carbon source for the pathogen, functional diversity of arbuscular mycorrhizal symbiosis seems to be more complex than earlier understood. This underlines that arbuscular mycorrhizal fungi are an integrated part of plant functioning across different plant functional parameters, and it calls for more research within the complex functional diversity of the arbuscular mycorrhizal symbiosis. 219 Symbiosis Wednesday 24 June – Poster session 22 Drought tolerance of rhizobia from nodules of Indigofera argentea Guiling Ren*, Leo van Overbeek, Henk Franssen, Mirjam Schilder, Jan Verver Wageningen University, Netherlands Rhizobium bacteria are able to form nitrogen fixing root nodules on legume plants. In agriculture rhizobia are used as inoculum to obtain optimal nodulation on the root system of legume crops. Seed inoculation with effective strains of rhizobia provide the nitrogen requirements of the legume to achieve increased yields. However, rhizobia are sensitive to desiccation, and inoculum have a short half-life during seed coating and subsequently storage. We questioned whether phenotypic variation in desiccation tolerance can be observed in different rhizobial species. How rhizobia in crucial dry desert environment keep active in inducing nodule formation in desert legume. Up to date, all literatures investigating the activity of desiccated rhizobia in the laboratory has used broth-grown, planktonic cells. In this study, a diverse range of 78 rhizobia strains from desert soils were isolated. And we developed a quantitative bioassay to illustrate desiccation tolerance on planktonic cells as well as biofilm-formed colony cells. By comparing fitness of desert isolates to elite strains that are used as inoculum under low humidity conditions, we found that colony formation improves the fitness of rhizobia dramatically when compared to bacteria grown in liquid cultures. Bacteria inside a micro-colony survive as long as 4 weeks at 28% relative humidity, whereas individual bacteria that were grown in a liquid culture do so for only 1-2 days. We noted a difference in colony structure of desert rhizobia when compared to elite rhizobial strains, as colonies formed by dessert strains are more compact and the extracellular matrix undergoes different physical changes upon rehydration. This difference in colony structure may contribute to the fitness of the bacterium under drought stress conditions. Our results demonstrate that the micro-colony structure protects rhizobia during drought stress. This finding may be exploited to improve rhizobial inoculum used in agriculture. 23 Deciphering the role of plant growth-promoting rhizobacteria in the tolerance of Spartina densiflora to physicochemical properties of marshes soils Ignacio D Rodriguez-Llorente*1, Enrique Mateos-Naranjo1, Mesa Jenifer1, Alfonso Pérez-Martín2, Miguel A Caviedes1, Eloisa Pajuelo1 1 University of Sevilla, Spain, 2IRNAS-CSIC, Spain In the salt marshes of the SW Spain, Spartina densiflora grows on contaminated sediments under unfavourable physicochemical conditions. 22 different cultivable bacterial strains were isolated from the rhizosphere of S. densiflora grown in those soils. 70% of the strains showed one or more plant growth promoting (PGP) properties, including phosphate solubilisation and siderophores or indolacetic acid production. Among them, a bacterial consortium, composed by Aeromonas aquariorum SDT13, Pseudomonas composti SDT3 and Bacillus sp. SDT14, was selected for further research. A glasshouse experiment was designed to investigate the role of this consortium in plant growth and physiological tolerance to the physicochemical properties of marshes soils. Plants of S. densiflora were randomly assigned to two inoculation treatments (with and without inoculation) in combination with two soil types with different physicochemical characteristics (from Piedras and Odiel marshes, SW Spain) for 50 days. Plant responses were examined using growth analysis, combined with measurements of gas exchange, efficiency of PSII biochemistry, total content of 220 Symbiosis Wednesday 24 June – Poster session photosynthetic pigments and leaf water content. In addition, the accumulation of nutrients and trace elements in roots and leaves were determined. The results confirm that inoculation improved growth of S. densiflora through a beneficial effect on its photochemical apparatus due to its impact on chlorophyll concentration. This growth enhancement happened under both soil conditions and was mainly reflected in a greater length and diameter of roots. Moreover, this consortium was able to stimulate ions uptake in roots and leaves. Rhizospheric bacteria appear to play a significant role in S. densiflora growth response and tolerance to the physicochemical properties of soils, through diverse protective effects on the photosynthetic apparatus, water-use efficiency and mineral nutrient balance. 24 The high efficiency species arbuscular mycorrhizal fungi play the main function for plant growth in symbiont Ning Shi*, Gu Feng College of Resources and Environmental Sciences, China Agricultural University, China Different species Arbuscular mycorrhizal fungi (AMF) would affect each other when they lived together in one root system. This study used two experiment designs to determine the interaction of AMF on contribution for Maize. One pots experiment was randomized in complete with three AMF species (Rhizophagus intraradices, Funneliformis mosseaeand Gigaspora margarita) either separately or in mixtures and two harvest times. The real-time PCR was used to determine the composition of AMF communities in roots by the number of large ribosomal subunit (LSU) genes. The other was used the root compartment design and 32 P labeled to explain the P uptake contribution for plant of three species AMF when they existed in one strip root. The fungi communities were advantage for symbionts although the fierce competition happened between the different species fungi. R. intraradices significantly promote the growth and had the highest radioactivity of 32P, but with less amount of LSU in the communities, it was "high efficiency" fungi. Gig. Margarita and F. mosseae emerged benefit for plant at 8 week but with larger number of LSU at single inoculation, and were decreased significantly when combined with R. intraradices, they were "low efficiency" fungi. Used the real-time PCR and 32P labeled, direct evidence is provided for competition among species within the AMF community existed in one single root system. 25 Mycorrhiza as a biotic elicitor in in vitro developed dual root culture system of Ocimum basilicum Shivani Srivastava*1, David Cahill2, Alok Adholeya3 1 TERI, India, India, 2Deakin University, Australia, 3TERI, India Arbuscular mycorrhizal fungi (AMF) are the most abundantly found symbionts of the plants. They colonize plant roots which are rich source of secondary metabolites having medicinal importance. Rosmarinic acid is a caffeic ester derivative obtained from Ocimum basilicum (Sweet basil) and is used as an anti-inflammatory, antiviral and anti-cancer agent widely. This study reports selection of best cultivar of Ocimum basilicum and development of an in vitro dual culture system between pRi derived transformed roots of Ocimum basilicum and CMCCROC3 for the production of mycorrhiza and rosmarinic acid. Developed co-culture 221 Symbiosis Wednesday 24 June – Poster session system was studied to provide insights into interaction mechanism between transformed roots and AM fungi. The developmental stages were studied by light, confocal and SEM microscopy.Potential of mycorrhiza as a biotic elicitor in comparison to non-mycorrhized root cultures was also studied. It was found that mycorrhization significantly affected RA level shown by HPLC and enzyme activity studies. Detailed results obtained through these studies will be presented and discussed. 26 Population genetic diversity and structure of symbiotic rhizobia nodulating lentil (Lens culinaris) in Morocco Kaoutar Taha*1, El bekkay Berraho2, Gilles Béna3, Jamal Aurag2 1 Laboratoire de Microbiologie et de Biologie Moléculaire - Laboratoire Mixte International (LMBM-LMI), Faculté des Sciences, Université Mohammed V, Morocco, 2Laboratoire de Microbiologie et de Biologie Moléculaire - Laboratoire Mixte International (LMBM-LMI) - Faculty of science, Mohamed V University, Morocco, 3Laboratoire des Interactions Plantes Microorganismes Environnement, IRD, France Lentil (Lens culinaris L.) is a food legume appreciated for its richness in protein, vitamins and minerals and also for its ability to fertilize soil. In Morocco, cultivated area by this culture is ranked in third position among food legumes, after bean and chickpea; however the mean yields remains low. To improve the production of this crop, it is necessary to identify the best bacterial symbiotic partners to use as inoculums. To attain this objective, both nodules and soils were sampled from the four major Moroccan producing regions: Rabat Zemmour Zaer, Chaouia Werdigha, Azilal and the Eastern region. A collection of 400 isolates was constituted from root nodules collected either directly from the field or by trapping in the laboratory. Isolates were characterized by sequencing housekeeping genes (partial 16S rRNA, recA and gln II) and the nodulation nodC gene. The analysis based on the 16S rRNA gene sequencing showed that all our isolates belong to the Rhizobium genus, with a genetic similarity with R. leguminosarum specie ranging from 97 to 99%. Maximum likelihood phylogenetic trees constructed based on analysis of recA and gln II genes showed that we recovered 21 distinct multilocus haplotypes most of which are clustered with isolates of R. leguminosarum bv. vicia recovered previously at a high frequency in Eastern and Central Europe. The nodC gene analysis also revealed a high diversity, with 34 different haplotypes where 10 of them include 186 isolates over the 228 sequenced. 41% of our isolates are closely related to strains previously identified in Mediterranean countries such as Tunisia, Morocco and Spain; others being related to strains isolated from different regions of the world. Phenotypic and symbiotic characterizations of isolates are in progress in order to select the most effective strain to be used as inoculums in Moroccan soil and climatic conditions. 222 Symbiosis Wednesday 24 June – Poster session 27 Streptomyces-elicited actions of rhizosphere microbes and pedunculate oak Mika Tarkka*1, Florence Kurth2, Markus Bönn2, Lasse Feldhahn2, Sylvie Herrmann3, Liliane Ruess4, Silvia Schrey5, François Buscot2 1 Helmholtz-Centre for Environmental Research, Germany, 2Dept. Soil Ecology, Helmholtz-Centre for Environmental Research, Germany, 3Dept. Community Ecology, Helmholtz-Centre for Environmental Research, Germany, 4Institute of Biology, Ecology Group, Humboldt-Universität zu Berlin, Germany, 5IMIT-Dept. Physiological Ecology of Plants, University of Tübingen, Germany The establishment of mycorrhizal symbiosis can be stimulated by bacterial isolates termed Mycorrhiza Helper Bacteria (MHB). It has been suggested that certain MHB do not only promote mycorrhization but also reduce damage by phytopathogens. Using a culture system in which the mycorrhizosphere isolate MHB Streptomyces sp. strain AcH 505 is grown in a soil-vermiculate substrate with a defined microbial starting community and pedunculate oak as plant host (www.trophinoak.de), we investigated the interactions of AcH 505 with microorganisms, nematodes and oak. Using specific primers, we observed that the AcH 505 PCR signal increased in the presence of the mycelium of the mycorrhizal fungus Piloderma croceum. Oak rhizosphere inoculation with AcH 505 counteracted the damage of the oak by the root parasitic nematode Pratylenchus penetrans. This antagonistic effect of AcH 505 was linked to shifts in the rhizosphere microbial community fostering fungi. Using plant RNA sequencing, we estimated how the oak responds to AcH 505. In the Streptomyces treatment, oak genes encoding microbe-associated pattern recognition-related leucine-rich repeat receptor protein kinases and xyloglucan cell wall transglycolases/ hydrolases were up-regulated in both roots and leaves. Co-inoculation of the rhizospheres with the mycorrhizal fungus Piloderma croceum attenuated AcH 505-elicited defense gene expression. In contrast, the defense gene expression in oak leaves was, in part, enhanced upon the challenge with oak powdery mildew Microsphaera (Erysiphe) alphitoides, and this response resulted in reduced powdery mildew symptoms, indicative of priming of plant defenses by the bacterium. Our studies suggest that the mycorrhizal fungus can enhance MHB growth, and that AcH 505 treatment affects the structure of the microbial community. It offers novel insights into the mechanisms of priming by actinobacteria and highlights the diversity of services provided by this Streptomyces strain. 28 Exudation of acid phosphatase from extraradical hyphae of arbuscular mycorrhizal fungus Rhizophagus clarus Keitaro Tawaraya*1, Takumi Sato1, Tatsuhiro Ezawa2, Weiguo Cheng1 1 Yamagata University, Japan, 2Hokkaido University, Japan Arbuscular mycorrhizal (AM) fungi enhance uptake of available phosphorus (P) from soil. The mechanism underlying this P uptake enhanced by AM fungi is the increase in surface area for absorption of available P. Little is known about utilization of unavailable P by AM fungi. We 223 Symbiosis Wednesday 24 June – Poster session investigated whether extraradical hyphae of AM fungi exude ACP and whether ACP activity responds to phosphorus condition. Sterilized Andosol was packed in pots that were separated into the mycorrhizal and hyphal compartments with a nylon net of 30 μm pore size. Seeds of Allium fistulosum L. were inoculated or uninoculated with the AM fungus Rhizophagus clarus in P fertilized soil (0, 0.15, 0.30, and 0.50 g P2O5 g-1 soil). Mullite ceramic tubes were buried in the soil of each compartment, and soil solution was collected. A. fistulosum L. and Linum usitatissimum L. inoculated with R. clarus were grown in sand culture and in vitro monoxenic culture, respectively. The soil solution, hyphae extracts, root extract, and root exudates were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. Shoot P concentration, shoot P content, and shoot dry weight were higher in the inoculated treatment than in the uninoculated treatment. Activity staining of the gel revealed that ACP activity at 187 kDa was observed in the soil solution in the inoculation treatment and hyphal extract collected from sand culture and in vitro monoxenic culture, but neither in the exudate of non-mycorrhizal roots grown in the hydroponic culture nor in the root extracts irrespective of mycorrhizal status. ACP activity in soil solution of inoculated treatment was higher in P0 than in P3 but there was no deference between P0 and P3 in uninoculated treatment. These findings suggest that extraradical hyphae of AM fungi exudes ACP into the hyphosphere and its activity is induced by Pdeficiency of host plant. 29 Ectomycorrhizal and soil enzyme activity profiles at the tree line Lixia Wang*1, Burenjargal Otgonsuren2, Douglas Godbold1 1 University of Natural Resources and Life Sciences, Vienna/forest ecology institute, Austria, 2Mongolian University of Life Sciences/School of Agroecology, Mongolia At a tree line site (1700m) and a lower elevation site (1100m) in the Austrian Alps, the ectomycorrhizal community structure was determined on Picea abies and Pinus mugo. The activity of a number of enzymes was determined on two of the dominant ectomycorrhizas for each tree species. In soil, the activity of a range of enzymes was determined from soil under the tree species and under Rhododendron hirsutum. The ectomycorrhizal community structure differed between Pinus mugo and Picea abies at the higher elevation site, but also between the higher and lower elevation Picea abies sites. At the higher altitude Picea abies site, the ectomycorrhizal community was dominated by Cortinarius sp, whereas at the lower elevation site the community was dominated by Russula. In Pinus mugo the dominant species were Amanita muscaria and Russula orchroleuca. In soil, the activity of the enzymes protease, exoglucanase, phenol oxidase, glucosidase, phosphatase, peroxidase were highest at the lower altitude Picea abies site. At the high altitude site, with the exception of phosphatase, the enzyme activities increased in the order Picea abies > Pinus mugo> Rhododendron hirsutum. In roots, the enzyme activities of βglucosidase, N-acetyl-β-D-glucosaminidase, acid phosphatase, leucine aminopeptidase were higher in mycorrhizal root tips than non-mycorrhizal tips. Between ectomycorrhizal species, the activity of acid phosphatase, averaged for all species of mycorrhizas investigated, was higher in both tree species at the high elevation site, compared to Picea abies at the lower 224 Symbiosis Wednesday 24 June – Poster session elevation. No clear relationship was shown between enzyme activity in the soil and that of the dominant ectomycorrhizal species, and also between the enzyme activity and rate of net nitrogen mineralization. 30 Modelling resource allocation in the legume - rhizobium symbiosis Annet Westhoek*1, Neil Dalchau2, Lindsay A. Turnbull3 1 University of Oxford, United Kingdom, 2Computational Science Laboratory, Microsoft Research, United Kingdom, 3University of Oxford, Department of Plant Sciences, United Kingdom The symbiosis between legumes and the nitrogen-fixing bacteria, rhizobia, is of major global importance, with symbiotic nitrogen fixation accounting for a third of the total nitrogen input in agricultural systems. Increasing demands for protein are only likely to increase reliance on legumes, which already form a key source of protein in human diets. Rhizobia provide legumes with nitrogen in return for carbon in the form of photosynthates. It is well established that investment by legumes in rhizobial symbionts depends both on the external nitrogen environment and on the rhizobial strain. However, we lack a quantitative understanding of how legumes allocate resources to symbionts of different nitrogen-fixing efficiency under a wide range of external nitrogen concentrations. In this study, we develop a model which investigates how a plant can maximise its fitness through optimising its resource allocation. The model simulates the growth of a plant infected with one or more rhizobial strains. We assume that growth is limited either by carbon or by nitrogen. The plant obtains carbon via investment in leaves while nitrogen is obtained by direct uptake via roots (which depends on the soil nitrogen concentration) and/or via nitrogen fixation in nodules (which depends on the nitrogen fixation rate of rhizobial strains). We then use the model to test alternative hypotheses about resource allocation strategies under different conditions. In particular we investigate whether the optimal allocation strategy depends on the assumptions made about the way inorganic nitrogen is supplied in soil. The model will be able to answer questions such as: 1) under what conditions investment in inefficient strains could be beneficial for legume growth; 2) and what would be optimal allocation for maximum growth? 31 Symbiotic effectiveness and host range of indigenous Rhizobia isolates nodulating Sesbania sesban Wassie Haile Woldeyohannes*1, Elias Dogiso Dagne2 1 Hawassa University, Ethiopia, 2Bureau of Agriculture, Ethiopia Exploitation of biological N-fixation as a cheap and sustainable source of N for plants requires identification of Rhizobia strain capable of inducing nodulation in plants and effective in fixing atmospheric nitrogen. Experiments were conducted to evaluate symbiotic 225 Symbiosis Wednesday 24 June – Poster session effectiveness of forty indigenous Rhizobia isolates on Sesbania sesban and to determine their host-ranges. Each isolate was grown on YEMB for 3-5 days in the Laboratory and inoculated to pre-germinated Sesbania seeds in modified Leonard Jars. -Ve and +ve-N control treatments were also included. The experiment was laid out in CR design with three replications. Data on nodulation, plant growth and yield parameters; and plant tissue N content were collected and subjected to ANOVA. Cluster analysis of data was also done. The results revealed that all isolates except AC100e induced nodulation on S. sesban. The isolates varied significantly in their effects on nodulation and plant growth parameters, biomass yield and tissue N contents of S. sesban. Accordingly, 70% of the isolates produced significantly higher growth and biomass yield of the plant than that produced in –N control. But, only 25% of the isolates produced growth and biomass yield of the plant similar to that produced in +N treatment. Cluster analysis data revealed that the 40 isolates were grouped in to six clusters. Those isolates in cluster-VI were AC50b, AC51C, AC61a, AC61d and AC100c resulted in the highest yield and N content of S. sesban and were best. The results of cross inoculation study revealed that out of 10 isolates tested only 40 and 90% of the isolates induced nodulation and growth on soybean and haricot bean respectively. It is concluded that there is a high potential to isolate infective and effective Rhizobia strains capable of inducing nodulation and N-fixation in S. sesban, Soybean and haricot bean from indigenous sources in Ethiopia. 32 Arbuscular mycorrhizal fungal hyphal exudates prime bacterium mediated phytate mineralization in hyphosphere Lin Zhang*1, Minggang Xu2, Yu Liu3, Fusuo Zhang1, Angela Hodge4, Gu Feng1 1 China Agricultural University, China, 2Chinese Academy of Agricultural Sciences, China, 3Zhejiang University, China, 4University of York, United Kingdom Ecology and evolutionary biology seeks to understand how cooperative strategies evolve and are maintained in species networks. Here, we focus on the three-partner relationship between plants, arbuscular mycorrhizal fungi (AMF) and hyphosphere bacteria to ask if the interaction between AMF and bacteria can pay back an essential resource (in this case phosphorus) to their associated host plant by consuming plant derived carbon (C). A microcosm and two Petri plate experiments which separate the plant roots, AMF hyphae and bacteria were conducted to demonstrate the direct effects of hyphal exudates on the growth and activity of bacteria in organic phosphorus (P) mobilization and, the reciprocal impact of the bacteria on growth and activity of the AMF in P uptake and transfer to the plant. Results showed that AMF released substantial C to the environment, triggering bacterial growth and activity resulting in enhanced organic P mineralization and turnover. While, in return, bacteria enhanced AMF hyphal proliferation which in turn, resulted in enhanced capture of the available P released. Under low soil C:P conditions, the AM fungi-bacteria interaction improved aboveground plant P nutrition. Our results suggest a C-P tradeoff occurs in plantAMF-bacteria systems. AMF and bacteria share the photosynthate of the plant, and as reciprocation, the AMF -bacteria interaction repays the plant with P by jointly mobilizing soil organic P forms and P acquisition for the plant through division of labour between these two microbial groups. 226 Symbiosis Wednesday 24 June – Poster session 33 Specificity and resilience of the arbuscular mycorrhizal fungal community in intensive agroecosystems Jiachao Zhou*, Xiaojing Wang, Yang Chen, Gu Feng China Agricultural University, China The composition of arbuscular mycorrhizal (AM) fungi communities can have a large effect on the performance of their plant hosts. The dynamic changes of AMF in ecosystem is hotspot recently. It is generally believed that the growth period of plants influenced the AMF community composition obviously. In agroecosystem, whether AMF communities can keep stable and be helpful for crop growth under traditional fertilization and pesticide is yet unresolved. We have carried out indoor pot and field test, to study the effect od benomyl and N fertilization on AMF communities and crop development. Changes in the community were characterized by root colonization, cloning, sequencing, tRFLP and DNA copy. While we used biomass and shoot P to measure crop development. Benomyl inhibited AM fungi infection while N fertilization increased abundance of AMF, some new batchs of fungi emerged after nitrogen added AMF diversity after nitrogen added. Community composition changed at different development stages of crop growth. The study found that after using benomyl G. intraradices appeard, the reason might be that G. intraradices can produce rich exogenous hyphae network and has strong restorability, thus adapt to benomyl. In vegetative growth period, AMF abundance was larger than reproductive period and different groups of AMF respond to differennt period altered. In vegetative period, mycorrhizal colonization rate was positive correlation with shoot biomass, nitrogen and phosphorus absorption. During reproductive growth stage, mycelium density was positive correlation with phosphorus absorption. In addition, during reproductive period mycorrhizal colonization has linear negative correlation with nitrogen nutrient absorption. In maturation phase, mycelium density had certain relevance with corn biomass, grain yield and harvest index. AM fungi in this study differed greatly in their response to perturbation and can be helpful to plant growth. In agroecosystems, AMF communities was important to crop even if apply traditional fertilization and pesticide. 227 Symbiosis Wednesday 24 June – Poster session Towards Application 36 ToT (transfer of technology) needs for promoting farmers on IPNM in agricultural production system in Indian sub-continent Manas Mohan Adhikary*, Anandamoy Puste, Kalyan Jana Bidhan Chandra Krishi Viswavidyalaya (State Agricultural University), India Modernizing agriculture is essential for meeting challenges of reducing hunger and poverty, increasing & sustaining productivity. Agriculture is the backbone of growth for developing countries like India, may achieve how well and how fast be able to manage a rural transformation (ToT) on integrated plant nutrient management (IPNM). Keeping this, field research on IPNM was emphasized during consecutive summer seasons at farmers’ field in different agro-zones of this sub-tropics. Improvised field demonstrations were undertaken on rice cv. Satabdi (IET 4786) in medium (S1) and medium-lowland situation (S2) to standardize production trend with IPNM (both organic and inorganic sources), which comparable with control and farmer’s practice [T1 - Control, T2 - Farmer’s practice, applied generally N:P2O5:K2O @ 50:30:30 as inorganic sources, T3 - N:P2O5:K2O @ 120:60:60 as 100% inorganic and T4 and T5 - 25% of N through FYM (farm yard manure) as organic, and 25% of N through green manuring + 75% of N:P2O5:K2O @ 120:60:60 kg ha-1, respectively as inorganic sources]. Results showed that grain yield of rice were significantly increased with IPNM in balanced form over farmers & control plot. Among organic sources, highest yield exhibited with green manuring along (25% N) with 75% of N, P2O5 and K2O through fertilizers. Gaining yield of rice is almost double in respect to farmers’ practice of the zones (Cluster I to IV). It reveals that benefit-cost ratio had gone in favour of highest productivity of rice grain (3.58 t ha-1), using maximum and balanced use of plant nutrients (IPNM), which is more compatible with the nutrients available to the crop plants and this was economically viable (B-C ratio 2.28) to the rural farming communities, it may be concluded that ToT field-based technological intervention on IPNM is imperative for boosting up crop productivity and promoting livelihoods of the rural farming community. 37 Combined use of Kosakonia radicincitans and Trichoderma harzianum in the northern and southern hemisphere: First results Beatrice Berger*1, Hernan Paillan Legüe2, Eduardo Donoso3, Silke Ruppel1 1 Leibniz Institute of Vegetable and Ornamental Crops, Germany, 2Universidad de Talca, Chile, 3Bio Insumos Nativa, Chile Microorganisms originating from nature are promising candidates to keep high yield gain and quality standards of agricultural products. However, successful use of microorganisms in bio-economy presupposes elucidation and understanding of microbial behavior in various environmental niches. Our bilateral project, funded by the BMBF (Project Management Jülich), between Germany (IGZ, Grossbeeren) and Chile (University of Talca) aims to study mechanisms of plant-microbe interaction using the plant growth-promoting bacteria Kosakonia radicincitans (German partner) and the fungal antagonist Trichoderma harzianum strain Queule (Chilean partner) in interaction with various horticultural plants. In cooperation with our Chilean partners we investigate the microbial mechanisms on the plant when grown 228 Towards Application Wednesday 24 June – Poster session under different nitrogen- and phosphor nutrition regimens, salinity stress under natural occurring soil conditions in the southern and northern hemisphere. In detail, we will measure changes in primary and secondary plant metabolism, moreover the nutrient up-take by the plant and we will evaluate the growth -promoting potential of bacteria and fungi as well as their colonization ability. Our results are intended to support the use of microbial products worldwide. Here we present first results on the combination of K. radicincitans and T. harzianum in greenhouse and field studies. 38 Efficacy evaluation of seed coated plant – root associated plant growth promoting rhizobacteria in microcosm system Rita Choudhary*, Alok Adholeya The Energy and Resources Institute (TERI), India Plant growth promoting Rhizobacteria (PGPR) are the important group of microorganisms which play a major role in stimulating plant growth through mobilizing nutrients in soils, producing numerous plant growth regulators, protecting plants from phytopathogens by controlling or inhibiting them and by improving soil fertility. This study was conducted with a view to isolate bacteria associated with roots of wheat, maize and soybean from different locations of Punjab, Rajasthan and Madhya Pradesh, India. A total of 130 bacterial isolates were screened biochemically for their plant growth promoting traits like phosphate solubilization, production of Indole Acetic acid (IAA), hydrogen cyanide (HCN) and siderophore. It was found that 58.5% of them showed IAA production, 22% showed phosphate solubilization, 48.3% siderophore production, 70% showed HCN production, whereas 9.7 % isolates showed all the plant growth promoting characteristics. Under polyhouse conditions, the efficacy of these biochemically characterized isolates was assessed. Formulations were prepared and an uniform <0.1mm thickness one layer coating was done in NIKLAS (W5/0.1) seed coating machine for 30 seconds on maize seeds. Results amply proved that by using plant beneficial rhizobacteria and by delivering viable number of cells on seed surface, promote better germination, enhanced dry weight, plant height and root length. Thus, these isolates have the potential not only to be used as biological seed coating but are also effective in decreasing the global dependency on hazardous agriculture chemicals which destabilize the soil health. 39 Soil protection against concentrated flow erosion with Arabidopsis roots S. De Baets*1, T. Denbigh2, T. Liverpool3, I. V. Chenchiah3, B. Higgins3, T. A. Quine1, C. Grierson2 1 College of Life and Environmental Science, University of Exeter, United Kingdom, 2Biological Sciences, University of Bristol, United Kingdom, 3Department of Mathematics, University of Bristol, United Kingdom Soil resources are under more pressure than at any time in human history. The current extent of global soil degradation, of which soil erosion and soil salinization are the dominant processes, is estimated at ~2 × 109 ha. Soil erosion is responsible for a global contemporary agricultural sediment flux on cropland of about 22 Pg year-1 and an additional approximately 229 Towards Application Wednesday 24 June – Poster session 11 Pg year-1 is mobilized on pasture- and rangelands. This corresponds with a total agricultural soil organic carbon erosion rate and hence soil fertility loss of 0.47 to 0.61 Pg C year−1. Due to increased rainfall intensity, climate change will lead to a significant increase of soil loss by 2050. Soil protection measures are therefore of great importance for future soil conservation. Plant roots have proven to be very effective in stabilizing the soil and protecting the soil against erosion. However, no clear insights are yet obtained into the root traits that are responsible for root-soil cohesion in order to better select the best species for soil protection. Research using Arabidopsis mutants has made great progress towards explaining how root systems are generated by growth, branching, and responses to gravity, producing mutants that affect root traits. In this study, the relative importance of root traits (e.g. root hairs, ratio primary/lateral roots) will be ranked by comparing the performance of selected Arabidopsis mutants in root-soil cohesion assays. Our first results show that wild type Arabidopsis (Col-0) is very effective in reducing concentrated flow erosion rates compared to other previously tested soil protection crops and that other mutants, containing less root hairs or having less laterals, are less effective. The results of this study can be used to rank root traits in order of importance for reducing erosion and to define molecular markers to look for in crop plants, optimizing soil protection. 40 Screening of biological and chemical treatments to control blackleg disease in potato Mout De Vrieze*1, Laure Weisskopf1, Santiago Schaerer1, Wolfgang Vogt2, Brice Dupuis1 1 Agroscope, Switzerland, 2Sourcon Padena, Germany In Switzerland, blackleg caused by Dickeya spp. in potato is the main cause of seed potato lot rejection during field inspections for seed potato certification. In order to elaborate a control strategy against this disease, nine Pseudomonas strains were evaluated for their potential to reduce the development of blackleg symptoms on potato plants in the field. Their inhibitory capacities were compared to those obtained for several biological and chemical treatment products including disinfectants, plant extracts, elicitors, essential oil and fertilizer. Initially, all candidate strains and products were tested in vitro for their ability to inhibit D. dianthicola’s growth and pectinolytic activity. The three best strains and the two best candidates of each of the other product categories were then selected for a greenhouse trial. A field trial was also conducted for the three selected Pseudomonas strains and completed with the already commercialized biological and chemical products. In both trials, treatment with the candidates consisted of tuber treatments, as well as an additional soil treatment for the antagonistic bacteria. Experiments performed in vitro and on potato tubers revealed that all nine strains, as well as the tested disinfectant, essential oil and two plant extracts, were capable of inhibiting the growth of Dickeya dianthicola. In the greenhouse, tuber and soil treatment with the bacteria did not lead to symptom reduction. The disinfectant sodium hypochlorite showed the most promising results, reducing the number of rotten stems per plant by 67 % in comparison with untreated plants, while the elicitors Bion® and chitosan showed slight symptom reduction, with reductions of rotten stems per plant of 20 and 15 % respectively. However, in the field, the best protection was obtained with the bacterial 230 Towards Application Wednesday 24 June – Poster session antagonist Pseudomonas DSMZ 13134, the active ingredient of Proradix®, which reduced disease incidence by 40 % in comparison with untreated plots. 41 Identification of a new cheap carrier for rhizobium inoculant Soumaya Tounsi Hammami1, Sana Dhane Fitouri*1, Salah Rezgui1, May Granier2, Faysal Benjeddi1 1 INAT, Tunisia, 2ATAE, Tunisia Production and quality of rhizobial inoculants in several developing countries are frequently limited that are attributed to inaccessibility of suitable carriers. Experiments were conducted to evaluate the potential of a new affordable and widely available carrier material in Tunisia. The carrier is a compost produced from the mixture of two native plant species: Arundo donax L and Medicago arborea L. Survival of Rhizobium sullae RSU9 was monitored over a period of 6 months at 4 ° C in sterile and non-sterile conditions. Compost maintained rhizobial population during this period. There was over than 109 and 107 rhizobia per gram of inoculant, in sterilized and unsterilized conditions. Throughout the storage period, compost maintained rhizobial population similar or higher than that observed in peat independently of the production conditions . The effect of this carrier material on nodulation and growth of Sulla coronarium L. plants was also investigated in glasshouse experiment. The sterilized compost showed the highest number and dry weight of nodules, respectively 30 nodules per plant and 0.13 mg per plant. It increased also shoot dry weight (0.31 gram per plant) compared to sterilized peat (0.25 gram per plant). Compost can produce high quality, inexpensive inoculants, maintaining high bacterial populations for at least 6 months. Further testing of this carrier material should be confirmed to assess its efficiency with other rhizobial species or other microorganisms and under field conditions. 42 Overcoming barriers for the microbial suppression of Rhizoctonia root rot on wheat Christopher Franco*1, Stephen Barnett2, Sophia Xue Lian Zhao3, Ross Ballard2 1 Flinders University, Australia, 2South Australian Research and Development Institute, Australia, 3Flinders University, Australia Root rot caused by Rhizoctonia solani AG8 is the major fungal root disease of cereals in low to medium rainfall areas in southern Australia. The application of microbial inoculants is being explored as a potential control option, because chemical and agronomic control options have had limited success. Attempts at obtaining strains effective in the field have been incosistent due to a lack of competence of the selected strains in the rhizosphere, and because the strain taken into commercial development do not always possess optimal traits. Success was achieved though collaboration of mycologists and bacteriologists (including 231 Towards Application Wednesday 24 June – Poster session actinobacteriologists) in testing a wide range (>2300) from a number of sources including the ecto and endorhizosphere of native plants, cereals, legumes and included endophytes and rhizosphere colonising microorganisms. The first phase of the high-throughput in planta screening consisted of field soil with added R. solani in 50 ml tubes and test strains added as a suspension directly to seeds. The 4.3 % of strains that reduced disease symptoms were then screened in more rigorous disease bioassays and characterised for properties important for the commercial development for such products. The 43 effective strains represented a diversity of microbial genotypes that included fungi and 4 phyla of bacteria-Actinobacteria, Firmicutes, Proteobacteria and Bacteroidetes. Further rigorous evaluation of minimum effective inoculum reduced the number to 6 strains for field trails. A range of inoculum delivery methods –from seed coating, drenches at different locations around the seed at sowing- and testing methods to account for the patchiness of the disease- resulted in the selection of three strains which were effective in a range of soil types in more than a single season. This holistic approach coupled with a team that understood each class of microorganism showed how inoculants can be developed to be effective in the rhizosphere. 43 The effect of nitrogen fertilization on rooting patterns and nitrogen recovery of catch crops Dina in 't Zandt*1, Alison Arico2, David Lehnert2, Christian Fritz1, Florian Wichern2 1 Radboud University Nijmegen, Netherlands, 2Rhine-Waal University of Applied Sciences, Germany In agriculture, nitrogen is typically applied in excess to increase crop yield. However, crops only take up 30-50% of the applied nitrogen leaving large amounts of nitrogen behind causing nitrate leaching and subsequent pollution of ground, surface and coastal waters. To reduce nitrate leaching, residual nitrogen can be immobilized in plant biomass by cultivating so called catch crops after harvesting the main crop. The objective of our study was to establish the relationship between root distribution and residual nitrogen immobilization by catch crops. In addition, we studied the effect of nitrogen addition to stimulate plant growth and nitrogen uptake. Since soil microorganisms compete with plants for nitrogen and immobilize nitrogen, microbial biomasses were also estimated. In a pot experiment, three catch crops, Raphanus sativus oleiformis L., Brassica rapa oleifera L. and Phacelia tanacetifolia Benth., were grown in a loamy sand soil at three fertilizer levels: 0 kg ha-1, 40 kg ha-1 and 80 kg nitrogen ha-1. Catch crops decreased nitrate concentrations up to tenfold compared to soils without catch crops. Interestingly, Brassica produced almost twice as many roots as Raphanus, but recovered comparable amounts of nitrate. Phacelia, on the other hand, captured a smaller fraction of the applied nitrogen, which was consistent with a decrease in root length and an increase in root diameter. For both Brassica and Raphanus, leftover nitrogen was comparable between all three nitrogen levels indicating that these plants respond to higher nitrogen availability by increasing their nitrogen uptake. Furthermore, nitrogen fertilization increased microbial carbon, but not nitrogen biomass, whereas catch crop cultivation increased only microbial nitrogen biomass. In conclusion, catch crops did not only contribute to reduced nitrogen losses by nitrogen uptake, but also by stimulating microbial nitrogen immobilization. 232 Towards Application Wednesday 24 June – Poster session 44 Effects of microbial fertilizer on soil chemical and biological properties of soil cultivates with wheat, barley and corn plant in different climatic conditions Nurgül Kitir*, Meti̇ n Turan Yeditepe University, Turkey This study was conducted to determine effectiveness of microbial fertilizers on soil chemical, biological properties and biodiversity of soil cultivated with wheat, barley and corn plant under different climatic conditions in Turkey. The Microbial Fertilizer was applicate in field conditions to wheat, barley and corn plant 30 L/ha. Each treatment were conducted three region that Kayseri, Urfa and Erzurum in Turkey. Plant and soil samples were taken at the end of the growing period each plant and each region. Some soil enzymes such as acid and alkaline phosphate, urease and dehydrogenase, amino acid exhausted from soil and plant roots, and macro and micronutrients elements of plant, soil microbial type and amount of the microorganism in the soil rhizospere were determined. The results obtained have shown that amino acids, plant and soil enzymes exhausted from root parts significantly affected the LifeBac NP microbial fertilizer. Results show that efficiency of LifeBac NP microbial fertilizer was affected to plant species and region climatic conditions. All plant species and region that studied LifeBac NP microbial fertilizer increased yield and yield parameters, and this increasing value has been very significantly as statical. 45 Development of an atlas of fine roots of European tree species Tanja Mrak*, Jožica Gričar, Hojka Kraigher Slovenian Forestry Institute, Slovenia Tree fine roots are an active component of belowground carbon cycle because of their fast turnover rates, and the exchange sites for nutrients and water. In this part of the rhizosphere, multiple interactions with various soil organisms occur, mycorrhizas being one of the central for the functioning of the forest ecosystems. Identification of tree fine roots is needed to elucidate the role of different tree species in belowground functional traits. Furthermore, tree root identification may serve in the field of cultural heritage protection as tree roots may cause structural damages to historical buildings. Since the molecular tools for identification of tree fine roots are costly and do not allow for quantification of species occurrence, the identification with anatomical-morphological approaches is a good alternative, especially for routine work. In our study ten temperate European tree species were investigated and for each species 3-5 individuals sampled. Roots of 5, 3 and 1 mm in diameter, as well as the most distal fine roots, were embedded into paraffin, longitudinal and radial sections prepared and studied with a light microscope. Morphology of fine roots was observed under the dissecting microscope and photographed. Compared to the stem wood of the same species, root wood differs in several characteristics, such as wedging growth rings, pattern of porosity, smaller size of vessels etc. Some characteristics not present in stem wood occur in fine roots – e.g. central channel in Abies alba Mill. roots. As the bark and primary tissues represents a high proportion of tissues in roots of smaller diameter, their anatomical characteristics can be used for identification purposes. The most important morphological characters for identification are the colour and texture of the bark, pattern of ramification and type of mycorrhiza. 233 Towards Application Wednesday 24 June – Poster session 46 Analysis of secondary metabolites produced by different strains of Pseudomonas chlororaphis isolated from halophytes, mesophytes and xerophytes Salma Mukhtar*1, I Shahid1, Muhammad R2, Deeba Baig1, Rahman Saleem3, Kauser Malik1 1 Forman Christian College (A Chartered University), Pakistan, 2Pakistan Council of Science and Technology, Pakistan, 3School of Science and Engineering, LUMS, Pakistan Several bacterial strains have been isolated from plants growing in diverse environments, such as halophytes, mesophytes and xerophytes (paragrass, sugarcane, cotton, cactus), of Pakistan. Among these, eight isolates were identified as strains of Pseudomonas chlororaphis subspecies chlororaphis and P. chlororaphis subspecies aurantiaca, based on 16SrRNA gene sequence. Four strains, RP4 (paragrass), ARS38 (cotton), FS2 (cactus) and PB-St2 (sugarcane), representing isolates from three different habitats, were selected for detailed study. These strains were screened for phosphate solubilization, indole acetic acid production, activity against plant pathogens, phenazine O (phzO) and pyrrolnitrin A (prnA) genes. All strains were positive for antifungal activity, indole acetic acid production, phzO and prnA genes. Secondary metabolites produced by these strains were analyzed and compared with each other by using MS technique. Phenazines, cyclic lipopeptides, homserine lactones, pyoverdin, pyrolnitrin and derivatives of lahorenoic acid have been detected in variable amount in these strains. Screening of these compounds against fungal pathogens is going on. These strains have great potential to be used as biocontrol agent due to phenazines and antibiotic production. 47 Studies on the bio-ecological characteristics and control methods of melon necrotic spot nepovirus (MNSV) and its soil fungal vector, Olpidium spp. Jin-woo Park*1, Kyung-Seok Park2, Se-weon Lee3 1 Agricultural Microbiology Division, National Academy of Agricultural Science(NAAS), South Korea, 2National Academy of Agricultural Science(NAAS), South Korea, 3Technology Cooperation Bureau, South Korea Melon necrotic spot nepovirus(MNSV) transmitted by seed and fungal vector Olpidium spp. in soil, is a most serious viral disease on melon. This study aimed to analyze the biological characteristics of MNSV and the impact of soil environment on the outbreak of a disease, as well as to select chemical control agents against Olpidium spp. which transmits MNSV. For the detection of MNSV in plant and Olpidium spp. in soil, this study used the real-time PCR, and genetic analysis of capsid protein gene of MNSV and resting spore ITS region of Olpidium were performed, respectively. Based on the result, it was found that among MNSV 128 isolates, the homogeny of 34 isolates whose pathogenicity is strong was more than 96% while 88~95% variations were confirmed over 84 isolates whose pathogenicity was weak. In addition, the genetically homology was found to be higher related to Spanish and Israeli strain than Japanese strain. Genetically analysis result showed that the Olpidium isolated from MNSV occurring areas was identified to be Olpidium blassicae. As a result of the analysis of the correlation between MNSV outbreak and in-plastic house environmental factors, it was found that MNSV occurrence was severe in soil with alkalinity of more than 7.5 while moisture in soil had no big impact on MNSV occurrence. The results of the study showed that the proper soil temperature for MNSV outbreak was 25°C. The result of survey on the density 234 Towards Application Wednesday 24 June – Poster session of Olpidium spp. which transmits MNSV from the surface soil to 45cm on a 15cm unit by six regions was found to be high at the layer near the surface soil of the region with high Olpidium spp. From the seven kinds of agricultural chemicals selected for fungal vector control, the Benomyl and Chlorothalonil water-dispersible powders were found to have high control effect. 48 The interactions between plant, microorganism and soil affect Fe acquisition in cucumber plants Youry Pii*1, Alexander Penn1, Concetta Eliana Gattullo2, Ignazio Allegretta2, Roberto Terzano2, Carmine Crecchio2, Tanja Mimmo1, Stefano Cesco1 1 Free University of Bolzano, Italy, 2University of Bari, Italy Plants have evolved two different strategies (Strategy I and II) to cope with Fe shortage, based on the exudation of organic and inorganic compounds to favor its mobilization and the root uptake. The role of the soil biotic component in the nutritional processes in the rhiszophere needs to be elucidated, since plants inoculated with PGPR showed an increased content of nutrients and a stronger resistance to abiotic stresses. The aim of the present work is the evaluation of the physiological effects, induced by Azospirillum brasilense in a calcareous soil on cucumber plants. Plants were grown in hydroponic Fe deficient solution followed by a 7-day period of contact with the A. brasilense-inoculated calcareous soil. At sampling, biometrics measurements, quali-quantitative analyses of root exudates and analyses of the nutrients content in plant tissues were carried out. Variations in soil mineralogy were assessed by X-ray powder diffraction (XRPD). Our results showed that A. brasilense facilitates plant growth in calcareous soils due to an enhanced recovery from the micronutrient deficiency. A. brasilense increases most likely the Fe availability within the rhizosphere by a) affecting the solubilisation of Fe thanks to the siderophore release and b) up and down-regulating the exudation activity of plants with an effect also on its molecular complexity. Further studies are needed to better understand and highlight the interactions between these two mechanisms and microorganisms. In particular, the present study shed light for the first time on two AAs, namely Gly and Glu, which could be involved in the plant-microorganism-soil interaction for the retrieval of Fe within a calcareous soil. XRPD analysis revealed a slight decrease of calcite and an increase of smectite under Fe-deficiency conditions. 235 Towards Application Wednesday 24 June – Poster session 49 Growth and nutrient uptake of sugarcane inoculated with diazotrophs Veronica Reis*1, Valfredo Chaves2, Willian Pereira2 1 Embrapa Agrobiologia, Brazil, 2Universidade Federal Rural do Rio de Janeiro, Brazil Sugarcane is a important crop for Brazilian economy by the production of sugar, ethanol and energy. Technologies that favor growth and better use of nutrients using diazotrophic plant growth promoting bacteria (PGPB) can reduce costs, environment impact and improve crop yield. The aim of this work was to evaluate the effects of inoculation with selected PGPB in the sugarcane varieties RB867515 and IACSP95-5000. Germination was measure during 40 days in a sterile substrate sand/vermiculite and also growth and biomass accumulation was measured in pots containing a mixture of sand/soil during 50 days. Treatments used: uninnoculated control and inoculation with G. diazotrophicus (Gd - strain PAL-5T); H. rubrisubalbicans (Hr - HCC103); H. seropedicae (Hs HRC54); A. amazonense (Aa - CBAMc) and B. tropica (Bt - PPe8T) applied in as a mixture or individually by immersion using a sett composed of a single stem node. The assays used a randomized block experimental design with 8 replications. In general inoculation with PGPR improved germination, biomass accumulation and nutrient uptake specially phosphorus and potassium but the response was dependent by the variety and strain used, showing that plant-bacteria interaction was modulated by plant and bacteria genotypes. the magnitude of inoculation has a different response in each variety but the single inoculation of Gd, Hr and Hs presented better results in the two sugarcane genotypes tested. 50 Enhancing the effective use of rhizobium inoculants by legume growers in southeastern Australia Maarten Ryder*1, Judith Rathjen1, Matthew Denton1, Ross Ballard2 1 School of Agriculture, Food and Wine, University of Adelaide, Australia, 2South Australian Research and Development Institute, Australia Legumes growers in Australia commonly use rhizobium inoculants. However farmers can be unsure whether or how frequently inoculants should be used on a particular field. A nodulation assessment guide has been made available (online at http://www.agwine.adelaide.edu.au/research/farming/legumes-nitrogen/legumeinoculation/) that guides grain legume growers in determining whether or not the inoculation of a crop has been successful. Preliminary surveys have been conducted in several farming regions of southeastern Australia to test this approach. Although nodulation levels have often been rated as adequate, poor nodulation of an inoculated crop should be investigated, and a troubleshooting guide is being developed to help solve problems with nodulation. Poor nodulation of an uninoculated crop implies that inoculation is advisable in future, to help improve nodulation and nitrogen fixation rates. Farmers are also often uncertain about any negative effects of mixing inoculant with other treatments at sowing, for example fertilizers, trace elements or pesticides. The compatibility of faba bean rhizobia (R. leguminosarum bv. viciae WSM1455) with a liquid zinc preparation used by farmers for improved plant nutrition 236 Towards Application Wednesday 24 June – Poster session was tested. When mixed in proportions used by the farmer, the zinc sulphate preparation (final pH approx pH 3.2) was found to kill 80% of rhizobia within 10 minutes and no rhizobia were detected after 2 h of incubation. This result clearly shows an incompatibility between current farm practice and rhizobium survival, and has led to a change in method. Further data on the impact of Zn, low pH and water quality on survival of rhizobia at sowing time will be presented. 51 Two volatile organic compounds trigger plant self-defense against a bacterial pathogen and a sucking insect in cucumber under open field conditions Choong-Min Ryu, Geun Cheol Song* KRIBB, South Korea Systemic acquired resistance (SAR) is a plant self-defense mechanism against a broad-range of pathogens and insect pests. Among chemical SAR triggers, plant and bacterial volatiles are promising candidates for use in pest management, as these volatiles are highly effective, inexpensive, and can be employed at relatively low concentrations compared with agrochemicals. However, such volatiles have some drawbacks, including the high evaporation rate of these compounds after application in the open field, their negative effects on plant growth, and their inconsistent levels of effectiveness. Here, we demonstrate the effectiveness of volatile organic compound (VOC)-mediated induced resistance against both the bacterial angular leaf spot pathogen, Pseudononas syringae pv. lachrymans, and the sucking insect aphid, Myzus persicae, in the open field. Using the VOCs 3-pentanol and 2-butanone where fruit yields increased gave unexpectedly, a significant increase in the number of ladybird beetles, Coccinella septempunctata, a natural enemy of aphids. The defense-related gene CsLOX was induced by VOC treatment, indicating that triggering the oxylipin pathway in response to the emission of green leaf volatiles can recruit the natural enemy of aphids. These results demonstrate that VOCs may help prevent plant disease and insect damage by eliciting induced resistance, even in open fields. 52 From seed to whole plant: seed defense priming by rhizobacteria and its determinant dipeptide Choong-Min Ryu, Geun Cheol Song*, Hye Khung Choi KRIBB, South Korea Seed priming is a technique to be controlled hydration and drying of seeds resulting in more rapid germination when are re-imbibed. Defense priming that has been induced by pretreatment of certain beneficial microbes and natural/synthetic compounds can enhance defense responses more rapidly or aggressively to biotic- or abiotic stresses. In this study, we were newly developed a immerged technology of the two priming methods referred to as “seed defense priming (SDP)” that is by seed priming with its supernatant (secreted metabolites) from root-associated Bacillus spp. in order to induction of systemic resistance (ISR) seven after transplanting to field. Seed defense priming mediated by bacterial supernatants from the 1800 strains of Bacillus spp. isolated from various soil samples in South Korea were tested ISR against Pseudomonas syringae pv. lachrymans (PSL) in cucumber 237 Towards Application Wednesday 24 June – Poster session seedlings. Symptom development on the SDP with strains PB69 and 1628 was reduced 40 and 28% respectively in vitro. Under field condition, pretreated pepper plants were assessed their disease severity by infiltration of Xanthomonas axonopodis pv. vesicatoria (Xav) at 20, 30, and 40 days post-transplanting (DAT). SDP by Bacillus spp. strains PB69 and 1628 elicited ISR as compared to control treatment at 20 and 30 but not 40 DAT. Cyclo (leu - pro) was isolated by various column chromatography and NMR spectra from culture filtrates of PB69. SDP by cyclo (leu - pro) (0.1 ppm) induced systemic resistance in cucumber plant. Our results indicate that seed defense priming triggered by bacterial supernatants can be de novo method to induced ISR even under field condition and cyclo (leu - pro) involves in the activator of plant defense reactions, leading to induced resistance against PSL in cucumber. 53 Is thinking about a below-ground landscape useful for scaling up understanding to practice? Elizabeth Stockdale* Newcastle University, United Kingdom Below-ground processes result from the interaction of soil habitats and their associated populations where the structure, composition and flows between these components are critical in defining the outcome and rate of the processes observed at the soil scale. Soil can be conceptualised as a series of linked habitats, including the rhizosphere, rather than a single habitat for soil organisms. Habitat types in soil: Resources (places): root, root surface (rhizoplane), rhizosphere, organic matter (litter to old humus), mineral surfaces. Pores (spaces): storage (Air/water filled), transmission (AIR filled) and residual (WATER filled). A multi-habitat (landscape) conceptualisation has been shown to provide a useful representation of soil faunal populations. However, application of landscape ecology approaches to below-ground ecology is not easy. Above-ground landscape ecology is moving away from a simple patch-matrix view of landscape and consequently connectivity is considered as an aggregate property of the structural configuration of the landscape elements. Habitat characteristics must be defined from across a range of scales and pattern prediction is complex and multifactorial – interaction between access to resources and refuge from predators. Habitat elements defined below-ground should clearly differ in quality; in the Table each has distinctive physical and chemical characteristics together with distinguishable communities of soil organisms. The context and connectivity of these elements are then key. The plant or plant community integrates across the diversity of below-ground ecosystem functioning via the roots and rhizosphere; in some way this role can be compared to that of the top predator in above ground systems. 238 Towards Application Wednesday 24 June – Poster session This conceptual model is applied to an analysis of the impacts of agricultural management on the size, activity and diversity of soil organisms and highlights the key role of the rhizosphere in mediating plant-soil interactions and their resilience. 54 Innovative biochar (hydrothermal carbonization) as an additive to soil-substrate for sustainable plant production Maren Stollberg*1, Thorsten Kraska2, Ralf Pude2, Guido Dericks3, Ulrich Schurr1, Arnd Kuhn1 1 Forschungszentrum Jülich, Germany, 2Friedrich-Wilhelms Universität Bonn, Germany, 3Grenol GmbH, Germany Just as natives peoples thousands of years ago started the story about “Terra preta”, an anthropogenic fertile tropical soil, we would like to imitate this phenomenon today using hydrothermal carbonization (HTC-biochar). The advantage of HTC-biochar is the possible use of all kinds of wet organic material as a parent material. We used digestate originating from biogas production which was converted to charcoal in a reactor at 200°C and 20 bar within 6 hours. The better-known pyrolysisbiochar instead is based on dry woody material only. In first pre-experiments we found a depression of Lactuca sativa var. crispa. growth after adding HTC-Biochar to field soil to increase carbon content. Therefore our aim was to remove any growth-reducing substances (e.g. aromatic compounds) by different extra treatments of the HTC-Biochar. The biochar treatments were (1) drying at 80°C, drying and washing with (2) water or (3) with 10% Ethanol solution. 5% HTC-Biochar (dry matter) was mixed with a silty loamy soil (field-soil, 40 soil-points, sieving at 2 mm) in pots and either Zea maize, Lactuca sativa var. crispa. or Brassica rapa subsp. pekinensis were grown on this substrate for 14-49 days. We performed a sequence of 3 - 4 harvests per species. Growth parameters such as dry matter, leaf area and minerals were measured. Zea maize showed after 14 days a negative effect on plant growth with untreated HTC-Biochar and also of the pyrolysis variant. The biomass of these variants was increased up to 20% compared to the control variant (pure soil) after 28 and 35 days. All HTC-Biochar treatments in comparison to untreated HTC-Biochar -as well as compared to pure soil- had negative effects on plant biomass. Therefore our chosen extra treatment of HTC-Biochar did not lead to an improvement in plant growth. Our assumption is that we lost mineral nutrients during these treatments. 239 Towards Application Wednesday 24 June – Poster session 55 Tracing of Pseudomonas inoculants in root and rhizosphere samples Carla Mosimann, Sarah Symanczik, Thomas Oberhänsli, Paul Mäder, Cécile Thonar* FiBL (Research Institute of Organic Agriculture), Switzerland Plant growth-promoting rhizobacteria (PGPR) are able to facilitate plant nutrient acquisition and can act as biocontrol agents by suppressing soil-borne diseases. Efficient strains can be formulated as microbial inoculants and their successful use for field application often requires a certain ability of persistence in the soil where they are inoculated. In this respect, there is a need to create tools enabling the tracing of inoculated PGPR which can also serve to monitor their spread in space and time. Here we report the development and application of a molecular method allowing the quantitative detection of two Pseudomonas strains contained in commercial formulations. The method is based on a Taqman qPCR assay targeting two polymorphic regions of the bacterial genome in order to ensure the specificity of the detection. The assays have been used with several samples (root or rhizosphere DNA) originating from various pot and field experiments with maize as host plant. The first results achieved in pot experiments indicate that for one strain the survival is influenced by the soil management (organic versus conventional) and that in general the high abundance of native Pseudomonas strains will not prevent a reasonable persistence of the inoculated Pseudomonas strains. In field conditions, the method has shown that the strain survival was improved when inoculated in combination with compost amendments. More results using these detection tools will be presented and will highlight the set of conditions (e.g. soil, inoculation techniques and frequency, co-inoculation or combination with other amendments) associated with reasonable persistence of inoculated PGPR. 56 Altered carbon and nitrogen cycling in soils following biochar application Tess van de Voorde*1, Simon Jeffery1, T. Martijn Bezemer2, Jan Willem Van Groenigen1, Liesje Mommer1 1 Wageningen University, Netherlands, 2NIOO-KNAW, Netherlands Biochar, pyrolysed biomass, is being widely promoted as a means to improve soil quality, sequester carbon, and improve soil-based ecosystem services. However, large knowledge gaps remain and the majority of research has been performed in managed agricultural land or controlled pot experiments. We aimed to study the effects of biochar amendment under semi-natural conditions. To do so we utilise a field experiment set up in 2011 in a nature restoration area near Ede, The Netherlands. Biochar was produced from cuttings collected from a local semi-natural grassland and pyrolyzed at 400oC or 600oC. The field experiment consisted of 4 treatments in 6 replicate blocks, resulting in 24 plots in total. The four treatments are: incorporation of biochar produced at 400oC, biochar produced at 600oC, incorporation of the non-pyrolyzed cuttings from which the biochar was produced, and a control treatment in which no material was incorporated (Control). Biochar and residue were applied at a rate of 10 ton/ha and mixed through the top soil layer (~10 cm). 240 Towards Application Wednesday 24 June – Poster session Three years later, in autumn 2014, we collected soil samples in all plots, which were used for a series of experiments to investigate the functioning of the soil microbial community, focussing especially on carbon (C) and nitrogen (N) cycling. We hypothesised that microbial communities exposed to biochar (1) differ in their ability to utilise a range of carbon substrates, and that (2) these communities will be better at decomposing more recalcitrant substrates. Overall, net N mineralisation measurements, a denitrification inhibition assay, substrate induced respiration experiments and a MicroRespTM assay (both using multiple Csubstrates), showed increased CO2 production in the soils that received biochar and hay residue, and reduced N2O production in the biochar amended soils, as compared to the noamendment control. We will link these findings to functional gene activities using GeoChip 5.0. 57 Root traits and aboveground yield of silage maize varieties under field conditions Nick Van Eekeren*1, Natalie Oram2, Joachim Deru1 1 Louis Bolk Institute, Netherlands, 2Nature Conservation and Plant Ecology, Wageningen University and Research Centre, Netherlands Drought tolerance of agricultural crops is critically important in the face of water shortages caused by climate change and competition for drinking water. Silage maize is a major fodder crop for the dairy industry on dry sandy soils in The Netherlands. Root architectural traits play a key role in improving drought tolerance of crops. Our objective was to study root architecture of silage maize in relation to aboveground production under field conditions. An experiment with 14 commercial maize varieties was carried out on a sandy soil in Loosbroek, The Netherlands. The experiment was managed according to common agricultural practice. At harvest, aboveground nitrogen content (g N / kg dry matter) and yield (kg N / ha) were measured along with other yield components (dry matter, starch, calculated feeding value). Root traits were quantified in two ways: 1) using ‘shovelomics’: a practical in-field measurement of brace and crown root number, angle, and branching; 2) harvested brace and crown roots were dried, weighed and scanned. Root length was determined with image software, discerning lateral from main roots. Aboveground, results show that nitrogen content differed between varieties and nitrogen yield did not. In root architecture, we found differences in crown root branching and root weight between varieties but not in root biomass or angle. Variety effects were also found in the proportion of lateral root length in total root length. Aboveground nitrogen yields were significantly positively correlated with root traits: total root length (R2 = 0.29), lateral root length (0.28) and crown root branching (0.32). There was no correlation between root traits and nitrogen content. Equivalent responses were found for the other yield components. Our results show that root traits that are quickly assessed in the field could be used in maize breeding to improve drought tolerance without affecting yield components. 241 Towards Application Wednesday 24 June – Poster session 58 Root properties of dike vegetation and their effects on concentrated flow erosion Wouter Vannoppen*1, Jean Poesen1, Sarah De Baets2, Matthias Vanmaercke1, Patrik Peeters3, Bart Vandevoorde4 1 KU Leuven, Belgium, 2University of Exeter, United Kingdom, 3Flanders Hydraulics Research, Belgium, 4Research Institute for Nature and Forest, Belgium The predicted climate change and the associated sea level rise are major challenges for the near future. The Scheldt basin will be exposed to an increased risk of flooding due to wave overtopping. To safeguard the land from dike breakthrough both above-ground and belowground biomass are important to keep the erosion resistance of the dikes sufficiently high. Plant roots are more effective in reducing soil erosion by concentrated flow compared to plant shoots. Therefore, the main goal of this study is to determine the erosion-reducing potential of the belowground biomass of dike vegetation types. Root properties of five dike vegetation types were studied: i.e. species-rich grassland (SR), species-rich grassland dominated by Arrhenatherum elatius (SRAe), grassland dominated by Arrhenatherum elatius (Ae), grassland dominated by Arrhenatherum elatius with few nettles (AeN) and nettle- dominated vegetation (NDV). The erosion-reducing effect (RSD) was estimated using a Hill curve model linking RSD to root length density (RLD, km/m³) based on an analysis of a global database on the erosionreducing potential of plant roots. Results based on the measured RLD in the topsoil (0-5cm) indicate that erosion rates would be reduced by more than 80% compared to a bare soil for dike vegetation types without nettles (129 < RLD < 235 km/m³). Dike vegetation types with nettles (22 < RLD < 58 km/m³) were less effective in controlling erosion due to concentrated flow. The degree of overgrowth of grassland by nettles, which have thicker roots and therefore a lower RLD, explains this pattern. To maintain a high resistance of the topsoil against concentrated flow erosion it is important to avoid the overgrowth of grassland by nettles through an effective management of the dike vegetation. Preventing the enrichment of the topsoil by nutrients is therefore an important measure. 59 How do plant roots affect rill and gully erosion? Wouter Vannoppen*1, Jean Poesen1, Matthias Vanmaercke1, Sarah De Baets2 1 KU Leuven, Belgium, 2University of Exeter, United Kingdom An important ecosystem service of plant roots is their effectiveness in controlling concentrated flow erosion rates. However an extrapolation of model results from individual case studies to estimate the erosion-reducing potential of plant roots is not reliable as rootsoil interactions in different types of environments, with differences in both root and soil characteristics are not yet fully understood. The objectives of this study were therefore: i) to provide a state of the art on studies quantifying the erosion-reducing potential of plant roots in controlling concentrated flow erosion rates; and ii) to explore the overall trends in erosion reduction as a function of root density (RD, kg/m³) and root length density (RLD, km/m³), root system architecture and soil texture, based on a meta-analysis of published research results. We therefore compiled a dataset of measured relative soil detachment rates (RSD) for RD and RLD. The decreases in RSD as a function of RD and RLD could be best described by a Hill curve model. Taking into account root system architecture and soil texture improved the model accuracy, especially for root length density. Fibrous root systems are in general more 242 Towards Application Wednesday 24 June – Poster session effective in controlling soil erosion by concentrated flow as they have a larger root-soil contact. As there was little variation in soil types among the collected data, the effect of soil texture on the erosion-reducing potential of plant roots could not be proved. The remaining unexplained variance is attributed to measuring errors and differences in experimental set ups (e.g. plant species tested, soil characteristics) that could not be explicitly accounted for. By taking these uncertainties into account we were able to establish relationships to assess the likely erosion-reducing effects of plant roots which are reliable irrespective of differences in root and soil characteristics. 60 Mixtures of maize genotypes produce more biomass than monocultures when mycorrhizal Xinxin Wang*1, Ellis Hoffland1, Gu Feng2, Thomas Kuyper1 1 Wageningen University, Netherlands, 2China Agricultural University, China Arbuscular mycorrhizal fungi can play a key role in enhancing plant productivity in multispecies natural ecosystems; however, their role in enhancing crop productivity and P uptake efficiency in single species genotype mixtures is hardly known. Therefore, we grew monocultures (one genotype) and mixtures of two genotypes of maize in low P soils in a greenhouse (with Funneliformis mosseae) and in a field experiment with an unidentified species mixture. We measured P uptake, hyphal length density and plant biomass. Genotype mixtures showed overyielding and enhanced P-uptake when mycorrhizal but not when nonmycorrhizal. The increase in relative yield total and P uptake was largely due to complementarity effects, and not to enhanced competitive ability of the larger genotype. Genotype mixing increased hyphal length density. The effects occurred both in the greenhouse and in the field. Our results suggest that genotype mixing increases the extent and activity of the mycorrhizal network. So mixing maize genotypes may be beneficial for enhancing productivity and P uptake efficiency. 243 Towards Application Wednesday 24 June – Poster session Rhizo Remediation and Fate of Pollutants 62 On-site PAH removal and microbial diversity during six-years monitoring under plant-assisted attenuation Thierry Beguiristain*1, Amélia Bourceret1, Aurélie Cébron1, Noële Raoult2, Pierre Faure1, Emilie Tisserant3, Pascal Poupin1, Pascale Bauda1, Corinne Leyval1 1 LIEC UMR 7360 CNRS-Université de Lorraine, France, 2GISFI, France, 3IAM UMR1136, INRA, France During the XXth century, the intensive coal and steel industries contributed to the chronic pollution of soils. Their dismantling left large areas of wasteland soils highly contaminated by recalcitrant organic compounds such as polycyclic aromatic hydrocarbons (PAH). We are interested in studying the potential for soil bioremediation using plants assisted rhizodegradation combining the potential of plant with those of rhizospheric microorganisms. Our objectives were to evaluate the fate of PAH and characterize the microbial diversity of an aged contaminated soil using on-site natural attenuation in experimental plots. A 24 plots devices with 6 different treatments in 4 replicates allowed the comparison between bare soil and different rhizospheric soils (alfalfa, naturally colonising vegetation) during 6 years monitoring. Soil characteristics and fate of pollutants were analysed together with microbial parameters. Our work is one of the first to characterize the microbial diversity in such aged PAH-contaminated soil. The impact of plant rhizosphere on both bacterial and fungal densities and diversities was assessed using qPCR and pyrosequencing. During this 6 year period of time an increase of pH and C/N was observed and a 50 % PAH removal was estimated. Plants had a limited impact on these soil parameters. On the contrary, plants favoured a higher density and originally increased the diversity of microorganisms. The bacterial community was dominated by Proteobacteria, Actinobacteria and Bacteroidetes and the fungal community was mainly represented by Ascomycota. Moreover some OTUs were foster by plants, such as members of Arthrobacter, Fusarium, Bionectria, Acremonium genera. Similarly, the plant rhizosphere seemed to favour the PAHdegrading bacteria belonging to the Actinobacteria as shown by qPCR targeting PAH-ring hydroxylating dioxygenases. Plants seemed to favour the PAH-degrading functional community but, in such aged contaminated soils, the main limiting factor to their activity remained the low PAH-bioavailability. 63 Dark-septate endophytic fungi: a solution for trees on metal-contaminated sites? Damien Blaudez*1, Charlotte Berthelot1, Julie Foulon2, Michel Chalot2, Corinne Leyval1 1 Université de Lorraine, France, 2Université de Franche-Comté, France Dark-septate endophytes (DSE) are conidial or sterile ascomycetous fungi that colonize living plant roots without causing apparent negative effects. DSE comprise a heterogeneous group 244 Rhizo Remediation and Fate of Pollutants Wednesday 24 June – Poster session of root associated endophytic fungi, characterized by melanized inter- and intra-cellular running hyphae and microsclerotia within the epidermis and/or the cortex of plant roots. High tolerance of DSE to metal pollution and their relatively high abundance in contaminated habitats suggest that DSE might have an important function for host survival in these extreme conditions. Fungal endophytes could affect heavy metal uptake of their host plants and increase plant metal tolerance. Therefore, in the context of phytoremediation assisted by symbiotic fungi, we have first isolated a set of DSE strains from poplar roots from metal-polluted sites. A wide-range screening of the strains was performed to select the best promising symbionts. Fungal isolates were identified as members of the Phialophora, Cadophora, Leptodontidium, and Exophiala genera. They were characterized for their plant growth promoting abilities, through different tests such as production of indol-3 acetic acid (AIA), release of volatile organic compounds or production of antifungal compounds. Metal tolerance of the fungal isolates was also studied under axenic conditions. For the most promising strains, an inoculation experiment was performed to monitor the effects of the fungi on the growth of poplar and birch on metal-contaminated soils. Moreover, we also investigated the interaction between DSE and endomycorrhizal fungi through dual inoculations of host plants. 64 Degradation of iprodione, vinclozolin and propanil by an Arthrobacter spp. strain isolated from a pristine rhizospheric soil Marco Campos*1, Dimitrios Karpouzas2, Maria Cristina Diez3 1 Universidad de la Frontera, Chile, 2Department of Biochemistry and Biotechnology, University of Thessaly., Greece, 3Chemical Engineering Department, Universidad de La Frontera., Chile Biological degradation constitutes the major processes controlling the dissipation of pesticides in soil. In this way, rhizodegradation in cooperation with pesticide-degrading bacteria could be a suitable tool to avoid point source contamination by iprodione, vinclozolin and propanil pesticides. We aimed to test the degradation of iprodione, vinclozolin and propanil by a rhizospheric bacteria strain isolated from a pristine grassland. Rhizospheric soil samples collected from ryegrass (Lolium perenne) grassland with and without previous exposure to pesticides were used for the bacterial isolation. After several enrichment in mineral soil medium (MSM) amended with iprodione, the isolation of an effective iprodione-degrading culture (C2.7) was reached from the pristine soil enrichments. Molecular fingerprinting revealed that C2.7 was composed of two strains identified via cloning as Arthrobacter spp. (C1) and Achromobacter spp. (C2). Degradation studies with the purified strains C1, C2 and their combination in minimal and rich media showed that C1 was the key iprodione-degrader, whereas C2 was only able to slowly co-metabolize iprodione. After that, C1 capacity to degrade pesticides of similar chemical structure to iprodione (vinclozolin, procymidone, propanil, diuron and isoproturon) was investigated. This strain completely degraded vinclozolin, a chemical analogue of iprodione belonging to the dicarboxamide family, in 20 days, and this was accompanied by bacterial growth and the 245 Rhizo Remediation and Fate of Pollutants Wednesday 24 June – Poster session formation of 3,5-dichloraniline. In contrast, this only partially degraded propanil with the production of small amounts of 3,4-DCA, while no degradation of procymidone (the other member of the dicarboxamide fungicides family) diuron and isoproturon were observed. These results provided first evidence that the degradation of iprodione proceeds via cleavage of the carboxamide bond common in iprodione and vinclozoline. Additionally, degradation of propanil provide significant information about the degrading versatility of our Arthrobacter spp. strain for future application in rhizodegrading systems. 65 Short-term effect of plants on the identity, abundance and activity of phenanthrene-degrading microorganisms slows down bioremediation in a contaminated soil François Thomas, Aurelie Cebron* LIEC, UMR7360 CNRS-Universite de Lorraine, France Microbial degradation is a promising soil remediation strategy for polycyclic aromatic hydrocarbons (PAHs) frequently polluting post-industrial environments, but the effect of plants on PAH dissipation rates and overall microbial community diversity and activity, is still unclear. Here, we compared the short-term dynamics of pollution, microbial communities and PAH-degraders in bare or ryegrass-vegetated aged-contaminated soil, using phenanthrene as a model PAH. Actively growing roots were allowed to colonize phenanthrene-recontaminated soil for 2 to 10 days in compartmented microcosms. Samples were collected from rhizospheric soil directly adherent to roots, bulk soil from vegetated microcosms and bare soil from non-vegetated controls. Phenanthrene concentrations were significantly lower after 10 days in bare soil than in rhizospheric or bulk vegetated soils. Measurements of total dissolved organic carbon, organic acids and carbohydrates showed that root exudation provided labile substrates that might be preferentially consumed instead of phenanthrene, therefore impeding its dissipation. Although the abundance of 16S rRNA genes and transcripts increased throughout the time course for both Bacteria and Archaea, these communities were more active in rhizospheric than in bulk vegetated soil after 8 days. In contrast, while plants favored the abundance of PAH-degrading genes compared to bare soil, their transcription level was similar in all conditions except after 10 days where the Actinobacteria activity was enhanced in bulk vegetated soil. To specifically target metabolically active PAH-degraders, similar microcosms were spiked with 13C-labelled phenanthrene in a stable isotope probing experiment. After 10 days, 16S rRNA gene fingerprinting revealed that 13C-phenanthrene was metabolized by different bacterial taxa depending on the presence of plants. Metagenomic characterization of the 13C-DNA fractions is underway. Together with the analysis of variations in total and active microbial community composition over the 10 days, it will provide an unprecedented view of the effect of plants on the identity of PAH-degraders in aged-contaminated soils. 246 Rhizo Remediation and Fate of Pollutants Wednesday 24 June – Poster session 66 Characterizing zinc tolerance genes in Suillus luteus, an ectomycorrhizal fungus with properties promising for use in phytostabilization applications Laura Coninx*1, Joske Ruytinx2, Michiel Op De Beeck3, Vangronsveld Jaco1, Colpaert Jan1 1 Universiteit Hasselt, Belgium, 2INRA, France, 3Lund University, Sweden Pyrometallurgical industry and mining activities have led to the contamination of vast areas with heavy metals. In these areas, biodiversity of plants and micro-organisms is often greatly reduced. Surviving organisms are subjected to a high selection pressure for metal tolerance, often resulting in the evolution of metal tolerant ecotypes of plants, fungi and bacteria. Zn contamination in the northern part of Limburg (Belgium), has led to the evolution of Zn tolerant ecotypes of Suillus luteus (L.) Roussel, an ectomycorrhizal basidiomycete that forms symbiotic associations with Pinus sylvestris L. These Zn tolerant ecotypes thrive in heavily contaminated soils and in the meantime protect their hosts from metal toxicity. This protective feature combined with the fact that S. luteus is a pioneer species common to sandy soils in temperate climate regions make S. luteus a suitable candidate for use in phytostabilization applications. However, to fully exploit the potential of such applications, a better understanding of the Zn tolerance mechanism is crucial. Previous investigations have shown that the basis of the tolerance trait is a mechanism promoting Zn efflux. Yet, since Zn is an essential nutrient, many homeostatic pathways are expected to be involved in maintaining an optimal Zn concentration in all cell compartments. This makes it a challenging task to characterize the tolerance mechanism. Hence, we first focused on establishing which Zn homeostatic pathways are present in S. luteus. Here we report 7 Cation Diffusion Facilitator proteins and 4 Zrt- Irt- like Proteins that have been identified in the S. luteus genome. Further characterization of these proteins and their response to increased zinc concentrations in zinc tolerant and sensitive isolates may lead to a better characterization of the zinc tolerance mechanism in S. luteus. 67 Bacterial resources for assisted phytostabilization of acid mine drainageaffected mountain stream bank Anna Corsini*1, Sarah Zecchin1, Milena Colombo1, Nicoletta Guerrieri2, Giorgio Lucchini1, Gian Attilio Sacchi1, Lucia Cavalca1 1 University of Milano, Italy, 2CNR-ISE, Italy Several areas of the Italian Alps have an arsenic content that exceeds the Italian law limit (20 mg kg-1, D.Lgs 152/2006), due to mineralogy of bedrock and to mining activities. Rio Rosso mountain stream (Anzasca Valley, Piedmont), is affected by arsenic (1015 mg kg-1) and iron (22 g kg-1) contaminated sediments that leach from an abandoned gold mine. In view of a bacterial-assisted phytostabilization action, rhizospheric bacteria and root endophytes of liverwort, fern and willow inhabiting stream bank were isolated and characterized. A total of 240 colonies were isolated and screened for arsenic-related features and plant growpromoting traits. Most of rhizobacteria and endophytes were resistant up to 7500 mg L-1 arsenate (21 and 20 respectively) and up to 750 mg L-1 arsenite (17 and 24 respectively), regardless plant species. More than 80% of the isolates were capable to reduce 75 mg L-1 of arsenate, while arsenite oxidation rarely occurred (<5% of the isolates), reflecting the prevalence of As(V) in the sediments. The endophytes of fern and willow mostly exhibited 247 Rhizo Remediation and Fate of Pollutants Wednesday 24 June – Poster session indole acetic acid and exopolysaccharide production, while rhizobacteria primarily possessed P solubilizing activity. This evidenced that the two different plant compartments exerted an effect on plant grow-promoting traits. These were homogenously distributed among rhizobacteria and endophytes of liverwort. A super imposed selective pressure was exerted by arsenic on the plants of the mountain stream bank, thus resulting in the distribution of arsenic resistance in plant rhizobiomes. The bacterial resources characterized in the present study will be exploited for the development of a bacterial-assisted phytostabilization of the arsenic contaminated stream. 68 Evaluation of the bioremediating ability of bacterial endophytes using nematodes as bioindicators in bioenergy crop Aoife Egan*, Thomais Kakouli-Duarte Institute of Technology Carlow, Ireland Nematodes are representative of their habitat, they respond quickly to disturbance in the soil composition and are therefore well suited as bio-indicators for environmental monitoring. Bioenergy crops, hosting certain bacterial endophytes, can be both utilised for bioenergy production and in bioremediation efforts, if they are grown on contaminated land. The work presented here utilises nematodes as indicators of soil health to evaluate the effectiveness of the bioremediation process of oilseed rape. This work is currently ongoing. The plants were grown in nickel contaminated soil in the presence of endophytic bacteria. Two approaches were adopted to evaluate nematodes as indicators of bioremediation efficiency: (1) Nematode assemblages in the rhizosphere of the bioenergy crops were characterised morphologically. In addition, a molecular analysis was performed using polymerase chain reaction of the 18S gene, followed by denaturing gradient gel electrophoresis. So far, results in the control samples (nickel free soil) were indicative of favourable conditions free from stresses. The most commonly occurring coloniser persister value was 4, with a maturity index of 4. The Shannon wiener index varied from 1.73-2.54 and the Simpson index varied from 0.80-0.99. (2) Transgenic Caenorhabditis elegans biosensors are utilised to assess the nickel bioremediating capacity of the bacterial endophytes. Both approaches aim to indicate the level of soil health and evaluate the phytoremediating capacity of oilseed rape in the presence of endophytic bacteria. The ability of nematodes as indicators of soil health in this system will be discussed. 69 Impact of diesel on the symbiotic partners Medicago truncatula/Rhizophagus irregularis in in vitro conditions Mónica Garcés*1, Maryline Calonne2, Stephan Declerck2 1 Université catholique de Louvain UCL, Belgium, 2Université catholique de Louvain, Belgium The tropical rain forest of Ecuador located in the Amazon region, in particular the Yasuni Reserve, is a hotspot of diversity. It also contains a major reservoir of oil. Since 1970, the oil 248 Rhizo Remediation and Fate of Pollutants Wednesday 24 June – Poster session industry development has become a serious threat to the environment, resulting in pollution of soil. The remediation of polluted soils by crude oil has become a priority for Ecuador. Currently, there is an increased interest in phytoremediation methods assisted by arbuscular mycorrhizal fungi (AMF). Indeed, these fungi colonize the majority of plant roots and are considered essential for the survival of many plants in polluted environments. In the present study, we investigated in monoxenic culture conditions the direct impact of oil on the symbiotic partners. The host plant Medicago truncatula and the AMF Rhizophagus irregularis MUCL 41833 were cultivated in absence or in presence of diesel which was added to the culture medium at the concentrations of 0.05; 0.1; 0.5 and 1%. The results demonstrated that the fungal spore germination as well as plant growth were inhibited in the presence of the two highest concentrations of diesel. As a consequence, this beneficial association could be hampered under such polluted conditions. These preliminary results open the door to investigate the mechanisms behind this inhibition. The in vitro system also allows the study of the impact of diesel on one of the major function of the mycorrhizal symbiosis, i.e. the nutrient transport from the fungus to the roots. 70 Ecopiling: A combined rhizoremediation and passive biopiling system for remediating hydrocarbon impacted soil Kieran Germaine*1, Xuemei Liu2, David Ryan1, David Dowling1 1 Institute of Technology, Carlow, Ireland, 2MicroGen Biotech Ltd, Ireland Remediation of large tracks of hydrocarbon contaminated land can be labour intensive and extremely expensive remediation projects. Biopiling is an ex situ bioremediation technology that has been extensively used for remediating a wide range of petrochemical contaminants in soils. Biopiling involves the heaping of contaminated soils into piles and stimulating the biodegrading activity of microbial populations by creating near optimum growth conditions. Rhizoremediation is another very successful bioremediation technique and involves the use of plants and their associated microbiomes to degrade, sequester or bioaccumulation pollutants from contaminated soil and water. The objective of this study was to investigate the effectiveness of a combined phytoremediation/biopiling system to remediated hydrocarbon impacted industrial soils. The effectiveness of the Ecopiling process was investigated at two former industrial sites. One site was a food manufacturing site and held ~4800m3 of soil contaminated with waste engine oil. The second site was a former wood preservation facility with soil heavily contaminated with creosote. At both sites the contaminated soil was amended with chemical fertilisers, inoculated with petroleum or polycyclic aromatic hydrocarbon degrading bacterial consortia and used to construct passive biopiles. Finally, a phyto-cap of Perennial rye grass (Lolium perenne) and White Clover (Trifolium repens) was sown on the soil surface. Monitoring of important physico-chemical parameters and hydrocarbon concentrations were carried out at regular intervals throughout the trials. The Ecopile system is a multi-factorial bioremediation process involving biostimulation, bio-augmentation and phytoremediation. One of the key advantages to this system is the reduced costs of the remediation process, as once constructed, there is little additional costs in terms of labour and maintenance (although the longer process time may 249 Rhizo Remediation and Fate of Pollutants Wednesday 24 June – Poster session incur additional monitoring costs). The other major advantage is that the aesthetics and many ecological functions are rapidly restored to the site, with concurrent bioremediation processes taking place within the piles. 71 Impact of aerobic-anaerobic interfaces established in the rice rhizosphere on the fractional distribution of pentachlorophenol and the microbial community Malik Hayat* COMSATS University, Pakistan Rhizoremediation is an emerging technology for remediating organic pollutants such as pentachlorophenol (PCP) in soils. Wetland plants like rice contain aerenchyma tissue, which channel air transports from the leaves to the roots, and uniquely develop aerobic-anaerobic interfaces in the rhizosphere. To investigate the rice rhizosphere effects on the dynamic changes of various extractable fractions of PCP and the microbial community, a glasshouse experiment was conducted by using a rhizobox in which rice seedlings were grown for 45 days. The soil was spiked with 20 and 45mg kg-1 PCP. Soil in the rhizobox was divided into five compartments at various distances from the roots. Sequential PCP extractions were conducted with three extractants: CaCl2 (0.01molL-1), butanol (99%), and DCM (99%). Butanol extractable form of PCP showed a significant difference in the rhizosphere at 3mm distance from the roots at both PCP levels. Thirty four phospholipid fatty acids (PLFAs)in the rice rhizosphere were identified in soils given the two different PCP concentrations. The total soil PLFAs concentration in the planted soils ranged from 29 to 52 nmol g-1. The highest concentration of PLFAs was at 3 mm distance from the roots at both PCP concentrations. The results suggested that the aerobic-anaerobic interface established by the root in the rhizosphere of rice and microbial community in the rice rhizospher sheowed significant role in the dissipation of PCP. 72 Improving phytoremediation of chlorendic acid by exploiting plant-associated microorganisms Inge Jambon*, Francois Rineau, Nele Weyens, Robert Carleer, Jaco Vangronsveld Hasselt University / Centre for Environmental Sciences, Belgium The possibility to remediate an industrial site contaminated with chlorendic acid, a fire retardant and expected carcinogen, by applying phytoremediation using poplar (Populus deltoides x (trichocarpa x deltoides) cv. Grimminge) is investigated. To improve phytoremediation efficiency, plant-associated (including rhizospheric and endophytic) microorganisms can be exploited. Bacteria can use organic pollutants as a carbon source and metabolize them to a greater extent than plants. Moreover they possess plant growthpromoting characteristics. Fungi can produce extracellular enzymes that act on a broad array of organic compounds, thereby degrading them. However, no biological degraders were yet described for chlorendic acid. Therefore, the aim of this research is to isolate chlorendic aciddegrading plant-associated microorganisms, which can later be applied together with poplar, to obtain an efficient phytoremediation of chlorendic acid whereby the plants bring the contaminant into close contact with the degrading microorganisms. 250 Rhizo Remediation and Fate of Pollutants Wednesday 24 June – Poster session To find suitable microorganisms, the cultivable plant-associated bacterial and fungal community of the chlorendic acid-contaminated site was isolated and screened for their degradation capacity. Of the 75 isolated fungi, 4 were able to significantly lower the concentration of chlorendic acid after 2 weeks, one even to 29% of its original concentration. After selective enrichment of a soil sample, 1 bacterial consortium was found to significantly decrease the concentration of chlorendic acid, but only to 94% of its original concentration. The role of the isolated fungi in the degradation of chlorendic acid therefore seems more important than that of the bacteria. However, plant-associated bacteria can still improve phytoremediation by promoting plant growth. Therefore, the isolated bacteria are screened for different plant growth-promoting characteristics, after which a consortium will be selected, consisting of chlorendic acid-degrading fungi and plant growth-promoting bacteria. The effect of inoculation of poplar with this consortium on the phytoremediation efficiency will then be evaluated in a greenhouse experiment. 73 Influence of silicon on phytotoxicity and uptake of antimony(V) in maize Miroslava Vaculikova1, Ying Ji*2, Susan Tandy2, Rainer Schulin2 1 Slovak Academy of Sciences, Slovakia, 2ETH Zürich, Switzerland With the increasing antimony emissions on a worldwide scale, Sb-polluted soil are receiving more and more attention. Of particular concern is the potential transfer of this toxic element into the human food chain, as the mechanisms of Sb uptake by plants are not sufficiently understood. On the other hand, silicon (Si) has been reported to protect plants from various types of stress including heavy metal toxicity. Thus, we addressed the question how Si would affect the uptake and phytotoxicity of Sb. In a hydroponic experiment, we investigated how Sb(V) at concentration varying between 0 and 30 mg L-1 affected root growth and anatomy of maize seedlings in presence and absences of 70 mg L-1 Si. Increasing Sb concentration led to a decrease in root length, root surface and aerenchyma. The addition of Si diminished these effects. It also increased the distance from root tip at which the Casparian bands formed, possibly explaining why Si increased the uptake of Sb into the shoots at low to medium Sb concentrations (5 to 10 mg L-1). The Casparian bands are known to interrupt the apoplastic pathway from the root cortex to inner root cylinder and thus inhibit the transfer of solutes in the apoplast from entering the xylem and being translocated into shoots. At the highest Sb concentration (30 mg L-1), at which the plants showed the most severe toxicity symptoms, the addition of Si greatly reduced Sb uptake into the shoots. The mechanism behind this is unknown and needs further research. 251 Rhizo Remediation and Fate of Pollutants Wednesday 24 June – Poster session 74 Root elongation responses of a drought tolerant shrub species Acacia chisholmii to elevated concentrations of cobalt and nickel in solution Sebla Kabas*1, Longbin Huang1, Peter Kopittke2 1 The University of Queensland / Sustainable Minerals Institute, Australia, 2The University of Queensland / School of Agriculture and Food Sciences, Australia Sulfidic base metal mine tailings such as the Cu/Pb-Zn tailings at Mount Isa Mines, Australia, contain elevated concentrations of copper (Cu), cobalt (Co), nickel (Ni), lead (Pb), and zinc (Zn), their phytostabilization requires introduction of tolerant native plant species. Acacia chisholmii, a drought tolerant shrub native to north-eastern Australia, has been found to colonize weathered Cu/Pb-Zn tailings under semi-arid climatic conditions. Despite past focuses on Cu, Pb and Zn-tolerance in plants, little attention has been paid to the toxicity of Co and Ni, which are present at high concentrations (ca.14 and 512 (mM) respectively) in pore water of the tailings. The present study investigates the growth responses of A. chisholmii to elevated Co and Ni concentrations in solution examining root growth and plant metal accumulation/distribution, initially under laboratory conditions. Young (5 days) and old (3 months) seedlings were treated with different Co, Ni or Co+Ni concentrations. Root elongation was limited at 25 µM in the young after 2 day exposure and growth reduction was observed at 60 µM in the old after 7 day. Individual exposure of young seedlings to the same concentrations of soluble Ni and Co showed similar root growth reductions, but the seedlings accumulated ca. 4 times more Ni (418 µg g-1 d.w.) than Co (114 µg g-1 d.w.). In 60 µM Co-only treatment, Co concentration in the roots of the old seedlings was up to 472 µg g-1 d.w.), but low as 28 µg Co g-1 d.w. in the shoots. This tentatively suggested Co exclusion in this species. Nevertheless, in the Co+Ni mixture, the Ni competed against Co uptake, resulting in lowered Co bioconcentration factor ([metal]root/[metal]solution). In conclusion, A. chisholmii showed Co/Ni-exclusion behaviour compatible with the phytostabilization purposes. Further investigations are required to elucidate differential rhizosphere mechanisms involved in the Co/Ni root uptake. 76 Influence of biochar and compost on microbial community in Technosols Ali Kanso*1, Catherine Sirguey2, Emile Benizri2, Antoine El-Semrani3, Ahmad Kobeissi3, Guillaume Echevarria2 1 Universite de Lorraine, France, 2Université de Lorraine, France, 3Lebanese University, Lebanon The LORVER project, supported by Lorraine Region (France) and European Union (FEDER), aims to create industrial chains for plant biomass production, from derelict lands and industrial by-products. In this work, phytoextraction was used as a sustainable restoration technology allowing metals recovery from constructed Technosols with industrial soil and byproducts. The effect on the microbiological and soil biochemical properties, which are known to play a key role in soil functioning and fertility, was assessed. Technosols were constructed according three formulations: a Biocentre® treated soil artificially contaminated in trace elements by additions of a mix of industrial sludges (BTS), 252 Rhizo Remediation and Fate of Pollutants Wednesday 24 June – Poster session BTS amended with biochar (B2TS), and BTS amended with a green waste compost (BTSC). Planted (two hyperaccumulators plants: Noccaea caerulescens and Alyssum murale) and nonplanted soils were transferred to a growth chamber for three months. Measured parameters were: soil physicochemical properties, plant growth, microbial biomass carbon (MBC) and nitrogen (MBN), soil enzyme activities and bacterial and fungal genetic structure. MBC and MBN were significantly higher in BTSC soils without any plant effect, whereas MBN was significantly lower in cultivated B2TS soils. Mean microbiological biomass C:N ratios were not significantly different among non-cultivated Technosols and reached 19.7. In planted BTS and B2TS soils, this value significantly increased up to 41.5 and 106 respectively whereas it was not affected in cultivated BTSC soils. Fluorescein Diacetate and enzyme (urease, phosphatase and beta-glucosidase) activities were systematically higher in BTSC soils when measured per soil mass unit, whereas urease and phosphatase activities were higher in B2TS soils when values were normalized to MBC. Technosols may have a low potential for microbial activity due to soil nutrient microbial limitations. If biochar strengthened nutrient limitations, green waste compost alleviated them. Compost amendments could thus improve Technosols fertility and microbial activities linked with biogeochemical process at both structural and functional levels. 77 Rhizoremediation of soils contaminated with petrogenic hydrocarbons using Australian native plants Navjot Kaur*, Paul Greenwood, Andy Whiteley, Suman George, Megan Ryan University of Western Australia, Australia Most industries require hydrocarbon-based fuels; however routine activities such as transportation, storage and refilling pose considerable spillage risk on land. We conducted a glasshouse experiment to examine the mechanism of breakdown of persistent hydrocarbons by native Australian plants, and associated microbial communities, in crude oil contaminated soil. A concentrated mix of oil 12% (w/w) and white sand was aged for 6 months in the glasshouse. The mix of soil and sand, or sand only (control), was added to dried, sieved (2 mm), low-nutrient field soil. Fresh soil was then applied to achieve 1% oil contamination. Eight species (7 native, 1 exotic) belonging to five families were grown for 3 months. There were 5 replicates of each species/treatment combination. We hypothesized that the level of oil biodegradation would depend on the populations of degrader communities selected by the plant species on the basis of type and/or amount of low molecular weight root exudates released under contamination stress. At harvest, total plant biomass, root area and plant height differed little between oil-contaminated and control pots. Community-level physiological profiling of bulk soil showed microbial communities from contaminated soil had an increased response for carbohydrates and carboxylates. The response was significantly higher for Viminaria juncea and Lolium perenne, possibly due to higher root mass. Unplanted controls were most effective in reducing soil total aliphatic and aromatic content, by 71% and 65%, respectively, which was marginally better than for Chloris truncata, Iseilema vaginiflorum, and V. juncea (aromatic) and Acacia holosericea. The least effective species were Trachymene pilosa, Rhagodia preissii and Rhodanthe chlorocephala. The results suggest a competitive interaction among plants and primary microbial communities. Total 253 Rhizo Remediation and Fate of Pollutants Wednesday 24 June – Poster session root exudates will be measured by high-pressure liquid chromatography and molecular techniques will be applied to understand quantitative changes in microbial communities after oil contamination. 78 Rhizosphere competence of atrazine-degrading Arthrobacter bacteria Dmitry Bazhanov1, Chengyun Li*1, Hongmei Li2, Jishun Li1, Hetong Yang2 1 Key Laboratory for Applied Microbiology of Shandong Province, Biotechnology Center of Shandong Academy of Sciences, China, 2Biology Institute of Shandong Academy of Sciences, China The occurrence and diversity of atrazine degraders in the rhizosphere of cogon grass and common reed growing near an atrazine factory and in the rhizosphere of maize from geographically distant fields were investigated. Atrazine degrading bacteria were isolated by direct plating on the specially developed SM agar. ERIC-PCR genotyping of the isolates followed by 16S rRNA gene phylogenetic analysis revealed a variety of atrazine-degrading Arthrobacter spp. in the rhizosphere of cogon grass and common reed, with a strong prevalence of one genomospecies related to the cluster comprising A. aurescens, A. ilicis, and A. nitroguajacolicus. Genetically similar A. ureafaciens bacteria which occurred as minor inhabitants of cogon grass roots in the industrial soil were predominant atrazine degraders in the maize rhizosphere at all the agricultural sites. The genetic structure of atrazine degrading communities of heavily contaminated industrial soils appeared to be governed mainly by the contamination rate. More complex factors seemed to influence the survival of atrazine degrading A. ureafaciens in agricultural soils exposed to low rates of atrazine. We found that the atrazine degrading strains of A. ureafaciens exhibited active movement in liquid and semisolid agar media, spread on the surface of solid agar along fungal hyphae and competitively colonized roots of maize, wheat and alfalfa after seed inoculation. The atrazine was not required for the root colonization. However, the highest population densities of the root-associated atrazine degrading A. ureafaciens were detected in the atrazinesupplemented soil. The root colonization resulted in the atrazine degradation and protection of the plants from atrazine injury. Our results demonstrate rhizosphere competence of atrazine-degrading Arthrobacter bacteria and their potential for use in rhizoremediation of polluted soils. 79 Ranking of mechanisms governing the phytoavailability of cadmium in agricultural soils by using a model analysis Zhongbing Lin1, André Schneider2, Thibault Sterckeman2, Christophe Nguyen*2 1 INRA UMR 1391 ISPA, France, 2INRA, France This study aimed at determining the influential mechanisms governing the phytoavailability of the toxic trace element cadmium (Cd) in agricultural soils using a mechanistic model. We built a phytoavailability model simulating the Cd2+ uptake by 1 cm² root surface and including the transport by diffusion and convection, the kinetics of sorption/desorption and of complexation with the dissolved soil organic matter. The model variables were ranked by 254 Rhizo Remediation and Fate of Pollutants Wednesday 24 June – Poster session performing a sensitivity analysis where a 30-days uptake of Cd2+ was simulated for 100 000 combinations of the parameters chosen within ranges consistent with the French agricultural soils. Generally, the most influential variable was the initial concentration of Cd2+, which was low, making the root capacity for absorbing Cd2+ of little influence. The convection was almost always negligible and the diffusion was the dominant process of Cd phytoavailability. The soil impedance factor, the water content and more particularly the soil buffer power for Cd2+ were therefore the following most influential variables. The kinetics of adsorption and desorption were of little Influence. The Cd-complex was shown to generally contribute little to the uptake due to a strong kinetic limitation for the dissociation. Consequently, the dominant process buffering the Cd2+ concentration at the root surface was desorption. Significant complex contributions were observed for very labile complex and high water flux toward the root surface. On the whole, the simulated uptake correlated very well with a simplified model considering only the transport and sorption at equilibrium of Cd2+. This model relied on the total soil Cd concentration, the soil pH, the soil organic carbon content, the concentrations of calcium and dissolved organic matter, and the partitioning of the dissolved organic matter between fulvic and humic acids; these variables determined the soil/solution partitioning of Cd2+. 80 Characterization of bacteria isolated from heavy metals contaminated soils of the region of Oujda (Morocco) Malika Oubohssaine*1, Laila SBABOU2, Jamal Aurag2 1 Faculty of Sciences, Mohammed V University, Morocco, 2Faculty of Sciences, Morocco After stoppage of mines activities, most of the exploitations are abandoned and the soils are heavily loaded with heavy metals. Toxic particles are transported in the ambient areas, causing a serious imbalance in the biogeochemical cycles and important changes in the functioning of ecosystems. These contaminated habitats are considered extreme regarding the abundant and chronic presence of heavy metals,. This combination of environmental conditions allows the development of pioneer microorganisms possessing the necessary biological mechanisms conferring them tolerance or resistance to toxic concentrations of metals and facilitating of the installation and colonization of plant species in these hostile environments. It is therefore interesting to study the diversity of this particular microflora and its interaction with the local plants metallophytes, in order to use them in the development of phytoremediation strategies for metalliferous sites. For this purpose rhizobacteria from metal contaminated soils will be selected and inoculated to Hedysarum spinosissimum plants under metal stress conditions, in order to study the usefulness of this biotechnological approach in phytoremediation programs.For the achievement of this objective, we have first realized a collection of 700 bacteria isolated from the rhizospheric and bare soils collected from three mining zones of Oujda (Sidi Boubker, Oued El Heimer and Touissit). The tolerance of the isolates to heavy metals (Pb, Zn, As) has been studied and multi-resistant bacteria to different concentrations of these metals has been identified. Qualitative and quantitative studies of 255 Rhizo Remediation and Fate of Pollutants Wednesday 24 June – Poster session potential plant growth promoting activities permitted the identification of 47 rock phosphate solubilizing isolates , 145 auxin producing strains and 54 siderophores producing bacteria . Sequencing the 16S rDNA gene of the most interesting bacteria allowed their identification at the genus and / or species levels. It was found that they belong to different families and clusters of bacteria and they are dominated by the genus Bacillus. 81 Effect of nitrogen addition on rhizospheric Zn, Pb and Cd mobility and plant uptake in contaminated technosol Bashar Qasim*1, Mikael Motelica-Heino2, Domenico Morabito3, Sylvain Bourgerie3, Arnaud Gauthier4 1 Université d'Orléans/ ISTO, France, 2ISTO, France, 3LBLGC, France, 4Université Lille 1, France Mining and smelting of metal ores activities have increased the spread and occurrence of potential toxic elements such as metal(loid)s in soils. However, they can be readily absorbed and enter into the food chain, causing serious threats to human health. Our study aimed at reporting the effect of the addition of two nitrogen form (NH 4+ and NO3−) on rhizospheric soil pore water pH, dissolved organic carbon concentration, potential toxic elements concentrations in SPW and their uptake by Populus euramericana Dorskamp grown in contaminated soils. Soil samples were taken from metallophyte grassland contaminated with Zn, Pb and Cd located at Mortagne – du –Nord (North France). A plant growth experiment with poplar woody stem cuttings was conducted with forty-five pots (3 soil samples × 3 treatments × 5 replicates for each treatment) for 35 days. The experiment consisted of two nitrogen treatments (NH4Cl and KNO3) and an untreated control soil (without fertilizer) was included for comparison. For all studied soils, rhizospheric pH decreased and increased gradually with NH 4+ and NO3– addition respectively, whilst, it decreased initially then increased at the end of the experiment for the untreated control soil. Dissolved organic concentrations increased gradually with time during the cultivation period. An increase in the rhizospheric total dissolved concentrations of Zn, Pb and Cd was observed with NH4 + addition associated with the lowest pH, whereas the opposite was observed with the addition of NO3 –. Nitrogen form addition enhanced plant roots metal uptake compared to plant shoots uptake for all studied metals. 256 Rhizo Remediation and Fate of Pollutants Wednesday 24 June – Poster session 82 Assessment of ecosystem services rendered by agromining of nickeliferous soils: association of leguminous and hyperaccumulator Ramez Saad*1, Guillaume Echevarria1, Ahmad Kobeissi2, Emile Benizri1 1 Université de Lorraine - UMR 1120 Sols Environnement INRA, France, 2Université Libanaise, Faculté des Sciences 1, Laboratoire Biologie Végétale et Environnement, Lebanon The objectives of Agromining are to decontaminate soils by extracting metals with high economic importance with the use of hyperaccumulators and then to improve their agronomic value for traditional agricultural use. Nevertheless, some hyperaccumulator species grow slowly and produce little biomass because of the soil’s low fertility, which could limit their use for phytoextraction, in terms of quantities exctracted. Thus, it is necessary to improve phytoremediation by the implementation of new and appropriate cropping systems dedicated to Agromining. To attain this goal, we propose to identify the effect of the combination of plants (hyperaccumulator and non-accumulator, i.e. legume) on the efficiency of phytoextraction (biomass production, remediation and restoration of soil quality). Indeed, legumes have shown to increase the soil-N pool which Brassica could benefit from. Intercropping or co-cropping with legumes has the potential to combine high economic performance and low environmental impact by reducing the amount of fertilizer supplied. Our aims are to evaluate the efficiency of associating Lens culinaris with Alyssum murale on an ultramafic Ni-rich soil in order to enhance biomass production and to achieve better Ni phytoextraction. However, it is known that abiotic stress, originating from the presence of metals in soil, causes the production of ethylene in plants, which in turn affects the development of roots and nodule formation in legumes. So, the purpose of the present study is to test if Lens can make nodules and if these are functional when it grows on Ni-rich soil. The soil used in this experiment naturally contains Rhizobium and was artificially contaminated with different concentrations (0 to 90 mg Ni.kg-1 soil) of nickel sulfate. Natural 15 N abundance will be used to estimate the percentage of plant N derived from N2 fixation (%Ndfa) and to estimate the capacity of Lens to grow and form functional nodules under Ni contamination. 83 A meta-model to predict the phytoavailability of cadmium in French agricultural soils André Schneider*1, Zhongbing Lin2, Thibault Sterckeman3, Christophe Nguyen1 1 INRA, Biogeochemistry of Trace Elements, UMR 1391 ISPA, France, 2School of Water Resources and Hydropower Engineering, Wuhan University, China, 3INRA, Laboratoire Sols et Environnement, UMR 1120, France Predicting the phytoavailability of toxic trace elements such as cadmium (M) in soils is an important issue for crop quality. A set of partial differential equations can be derived for 257 Rhizo Remediation and Fate of Pollutants Wednesday 24 June – Poster session simulating the root influx of M2+, the transport by diffusion and convection, the kinetics of sorption/desorption, and the kinetics of complexation in solution, for the free ionic species (M2+), for a mean ligand, and for the complex. The aim of this study was to develop a simple meta-model that would be easier to handle than these partial differential equations. d and are out of equilibrium in a reaction layer of thickness µ < d; in this reaction layer the concentration of the complex is assumed constant, and the association rate of the complex is neglected. Under these assumptions and for two extreme behaviors of the complex, i.e. when complex is totally labile (µ=0) or totally inert (µ=d), the analytical equation for calculating the concentration of trace element at the root surface, and therefore the uptake, was derived. For intermediate complex lability (0 <µ < d), the uptake was obtained by linear interpolation of the uptake between µ=0 and µ=d. The meta-model was evaluated for cadmium in the context of French agricultural soils. For a dataset of about 105 simulations covering the variability of soil characteristics, the metamodel estimates of the uptake fitted very well the values obtained by the numerical integration of the partial differential equations (r²=0.996). Indeed, for 95% of the simulations, the values of the relative error of prediction ranged between -14 and 27%, and the mean absolute relative error of prediction is quite excellent (5.9%). 84 Phenanthrene-induced alterations in Noccaea caerulescens roots and consequences for metal phytoextraction Ivan Zelko1, Catherine Sirguey*1, Stéphanie Ouvrard2 1 Université de Lorraine - UMR 1120 LSE, France, 2INRA - UMR 1120 LSE, France Industrial decline in recent decades has led to the emergence of thousands of acres of brownfields in North-East France. Some of them are multi-contaminated by heavy metals and organic pollutants and their remediation need to address both problems. A combined decontamination using simultaneous phytoextraction of metals and rhizodegradation of organic pollutants appears a promising solution. However, organic pollutants may adversely affect hyperaccumulator plants growth and extraction activity thus limiting their use in this context. The aim of this work was to study root morphological and structural features of the heavy-metal hyperaccumulator Noccaea caerulescens exposed to phenanthrene (PHE) in combination with cadmium (Cd). Seedlings of N. caerulescens were cultivated, during one week in a climatic chamber, on a nutritive agar medium non supplemented (control) or supplemented with 2 mM PHE and/or 5 µM Cd. At the end of the culture, we assessed plant growth and production parameters (elongation, biomass and plant water content), root architecture (WhinRhizo® software). Root structural features (suberin lamellae and peri-endodermal layer) were observed using epifluorescence microscopy. The tested PHE concentration caused serious inhibition (about 40%) of growth and biomass production of both roots and shoots, while Cd had no significant adverse effect. Moreover, Cd moderately impacted root architecture whereas PHE strongly reduced (about 50%) total root length and surface. Additionally, PHE promoted lateral root formation and inhibited root 258 Rhizo Remediation and Fate of Pollutants Wednesday 24 June – Poster session hair elongation. Moreover, endodermal cells with suberin lamellae appeared closer to the root apex when exposed to PHE compared to control and Cd treatments. This can be explained by modified lateral root formation. The stage with well-developed suberin lamellae was not influenced by PHE. However peri-endodermal layer development was impaired in PHE-treated plants. These morphological and structural root modifications in response to PHE exposition might in turn limit Cd phytoextraction by N. caerulescens in co-contaminated soils. 85 Nodulation and growth of Fabaceae in mine spoils as influenced by inoculation of resident rhizobacteria displaying multi-trait phytostimulation properties Souhir Soussou*1, Brigitte Brunel1, Marjorie Pervent1, Diederik van Tuinen2, JeanClaude Cleyet-Marel1, Ezekiel Baudoin1 1 Tropical and Mediterranean Symbiosis Laboratory, France, 2UMR1347 AGROECOLOGIE, France Heavy metals (HM) extraction by metallurgical activities generates highly toxic mine spoils that still contain noxious levels of HM. To limit further HM dissemination by leaching and eolian transport, rubble mines can be phytostabilised by engineering a dense and perennial plant cover. In particular, autochtonous legume ecotypes spontaneously growing in severely HM-polluted areas are attractive candidates in such vegetalisation scenarios owing to their tolerance mechanisms and their organic nitrogen-carbon inputs in such nutrient-depleted substrates. In this background, inoculations of rhizobia and plant growth promoting rhizobacteria (PGPR), isolated from the rhizosphere of such metallophytes growing in mine spoils, are to be tested so as to facilitate the development of legumes. Here, we report on the rhizosphere of Anthyllis vulneraria ssp. carpatica, a local legume ecotype grown in highly Zn-Pb-Cd contaminated mine ponds (St. Laurent-le-Minier, France), as a reservoir of diverse fluorescent Pseudomonas species displaying multi-trait phytostimulation properties. 11% of the strains harboured the acdS gene and were ACCdeaminase positive. Development and nodulation patterns of the metallophyte legumes A. vulneraria ssp. carpatica and Lotus corniculatus grown in the same mine substratum were observed following individual inoculations of 11 selected PGPR strains together with the relevant rhizobium strains. Plant development was highly dependent on the identity of both metallophyte and PGPR strain. A single strain proved the most efficient on both legumes and two strains systematically induced a strong inhibition of plant development, irrespectively of their in vitro PGPR potentials. These data suggest that successfull inoculations of resident PGPR strains as a tool to facilitate resident metallophyte development in mine tailings first rely on the integration of the metallophyte-PGPR genotypes interaction. 259 Rhizo Remediation and Fate of Pollutants Wednesday 24 June – Poster session 86 A radio-isotopic dilution technique for functional characterisation of the associations between organic and inorganic contaminants and natural nanoparticles Ehsan Tavakkoli*1, Albert Juhasz2, Enzo Lombi2 1 NSW Department of Primary Industries, Wagga Wagga Agricultural Institute, Australia, 2Centre for Environmental Risk Assessment and Remediation, University of South Australia, Building X, Mawson Lakes Campus, Australia Despite evidence that the fate and behaviour of organic and inorganic contaminants are influenced by their interactions with water-dispersible naturally occurring soil colloids, our understanding of the mobility and bioavailability of contaminant–colloid associations has been hampered by the limitations of common operationally defined analytical techniques. In this paper, an isotopic dilution method was developed to quantify the isotopically exchangeable and non-exchangeable forms of zinc, phosphorus and phenanthrene in filtered soil-water extracts. In addition, the effect of filter size on the determination of contaminant exchangeability was investigated. The results showed that the isotopically non-exchangeable Zn and P in filtered soil-water extracts respectively ranged between 5 and 60 % and 10 and 50 % and was associated with water-dispersible colloids. Filter pore size had a significant effect on Zn and P exchangeability. Whereas the <0.1-µm filtrates contained isotopically exchangeable Zn and P fractions equal to the total Zn and P concentrations (i.e. 100 % isotopically exchangeable Zn and P), the filtrates obtained from larger filter sizes (0.22, 0.45 and 0.7 µm) contained increasing proportions of non-exchangeable Zn and P. Our findings also suggest that the exchangeability of phenanthrene in sodium tetraborate is controlled by both inorganic and organic colloids, while in aqueous solutions inorganic colloids play the dominant role (even though coating of these by organic matter cannot be excluded). 87 Regulation and stabilisation of physical, chemical and microbiological alteration processes by plants’ exudates and antioxidants in Callovo-Oxfordian argillite Dimitri Ubersfeld*1, Corinne Leyval1, Paul-Olivier Redon2, Christian Mustin1 1 LIEC UMR 7360 CNRS-Université de Lorraine, France, 2Andra, Direction Recherche et Développement, Centre de Meuse/Haute-Marne, Route départementale 960, France This study is focused on vegetalisation of Callovo-Oxfordian clay rock stockpiles excavated from the underground Andra research laboratory (CMHM, Bure, France). The aim was to understand the effect of planting hardy and resilient plants (i.e. lavender) on the physical and chemical weathering processes (erosion, oxidation, leaching) of clay materials, made of smectites, sulphides and carbonates. A bioweathering experiment was conducted on sieved and ground clay material samples inoculated with a microbial inoculum coming from the site’s topsoil. Percolation devices were watered once a week and incubated at 28°C for three months. Plant effect was simulated by the addition of antioxidant molecules (linalool and thymol) and synthetic root exudates to eluents. Both physico-chemical and biological transformations of clay materials were assessed by quantifying metal amounts in leachates. Bacterial density was estimated by DNA extraction from clay material and quantification of 16S rDNA gene copy number. Microbial inoculation and exudate addition lead to a significant pH decrease of the leachates and an important leaching of Al, As, Ca and Fe. With antioxidant inputs, the lixiviation of most important metals (Fe, S…) was slightly lower and the 260 Rhizo Remediation and Fate of Pollutants Wednesday 24 June – Poster session bacterial density concomitantly and significantly decreased with thymol. In an opposite way, exudates stimulated the growth of microflora. In conclusion, these results motivate the use of hardy plants producing antioxidants (essential oils) for phytostabilisation of clayed excavated residues. 88 Effect of glyphosate on rhizosphere microbial communities of Cortaderia selloana plants Mikel Anza1, Lur Epelde1, Unai Artetxe2, Julen Urra*1, Jose Maria Becerril2, Carlos Garbisu1 1 Neiker-Tecnalia, Spain, 2University of the Basque Country, Spain In the north of the Iberian Peninsula, Cortaderia selloana plants have invaded not only human degraded areas but also ecosystems with high ecological value such as marshes, dunes and riparian forests. As a consequence, in many areas, the high proliferation of C. selloana has become a matter of great ecological concern, as few strategies are available for its control. In our region, the control of this invasive species is attempted by spot treatment with a postemergence application of glyphosate (a non-selective systemic herbicide). The main objective of this work was to determine the impact of glyphosate treatment on rhizosphere microbial communities of C. selloana plants. To achieve this goal, a microcosm pot experiment was established with previously grown C. selloana seedlings. After two months of growth, pots were treated with glyphosate. Three rhizosphere soil samplings were carried out: just before treatment, and one and two months after treatment. A variety of parameters that provide information on the abundance, activity and diversity of soil microbial communities (ergosterol concentration, enzyme activities, basal respiration, community-level physiological profiles, etc.) were determined in these rhizosphere soil samples. After one month of treatment, our results indicate a negative impact of glyphosate on soil microbial communities. In the longer term (after two months of glyphosate application), this impact was more than reversed with a significant increase in the values of the microbial parameters determined here. Most importantly, glyphosate-treated soil kept its capacity to properly support plant growth (in particular, the growth of Festuca rubra plants; in our region, this species is a good candidate for revegetation after glyphosate-treatment of C. selloana plants). Finally, soil microbial properties have proven to be good indicators of soil ecosystem health. 261 Rhizo Remediation and Fate of Pollutants Wednesday 24 June – Poster session 89 Spatial pattern of bacterial diversity in a site with mixed and uneven contamination, and assessment of rhizoremediation potential F. Mapelli1, L. Vergani*1, R. Marasco2, B. Chouaia1, M. Fusi2, A. Di Guardo3, G. Raspa4, E. Zanardini3, C. Morosini3, S. Armiraglio5, S. Anelli6, P. Nastasio6, V.m. Sale6, D. Daffonchio1, S. Borin1 1 Defens, University of Milan, Italy, 2King Abdullah University of Science and Technology, Saudi Arabia, 3DSAT, University of Insubria, Italy, 4DCEME, University of Rome La Sapienza, Italy, 5Municipality of Brescia, Museum of Natural Sciences, Italy, 6ERSAF, Italy The SIN Caffaro is a large polluted site of national priority located in the Northern Italy, originated by the activities of the former Caffaro s.p.a. chemical factory. The soil in the site presents a mixed contamination of halogenated Persistent Organic Pollutants and heavy metals in variable concentrations, uneven distributed in the area and often exceeding the safety values. The use of plants to extract and modify the pollutants (phytoremediation) together with root associated microbes to i) degrade or modify the pollutants (rhizoremediation) or ii) support plant growth (plant growth promotion, PGP) has recently arose as a promising approach for bioremediation. In this context, we collected soil samples from three different areas in the site to a depth of 1 meter, which were chemically and microbiologically characterized. A DNA-based fingerprinting approach was applied to describe the bacterial community’s structure, which proved to be significantly different according to the area and depth of collection. We also tested the influence of selected environmental parameters, showing that the concentration of different classes of pollutants was significantly related to the microbiome structure. Furthermore, the rhizosphere of three autochthonous plant species was collected in the most contaminated area of the site. The overall bacterial community was studied by 16S rRNA pyrosequencing and a collection of bacterial strains was in parallel established and tested for PGP potential. The results showed that the rhizosphere-dwelling microbiome was highly similar between the plant species, in terms of both phylogenetic diversity and PGP potential, confirming the existence of a strong selective pressure given by the pollution profile rather than the plant species. Overall, this work highlighted the occurrence of distribution patterns in bacterial populations related to gradients in soil pollution, and showed the intrinsic potential of the highly contaminated soils at the Caffaro site for rhizoremediation potential. 90 Quantitative and qualitative synthesis of siderophores by Streptomyces sp. strains in the presence of Cd2+ Michał Złoch*, Dominika Thiem, Sonia Szymańska, Renata Gadzała-Kopciuch, Katarzyna Hrynkiewicz Nicolaus Copernicus University, Poland Microbial siderophores are low-molecular mass chelators which act as solubilizing agents for iron from minerals or organic compounds under conditions of iron limitation. Additionally, 262 Rhizo Remediation and Fate of Pollutants Wednesday 24 June – Poster session they can also form stable complexes with other heavy metals like Cd, Cu, Pb, Al or Zn. Moreover, stimulating effect of heavy metals on siderophore biosynthesis by some bacteria, which may indicate involvement of heavy metals in regulation of siderophore biosynthesis pathways, was suggested. We hypothesized that: (1) production of siderophores by 5 rhizosphere Streptomyces sp. strains can be stimulated or inhibited by increasing concentration of Cd2+ in the medium and (2) diversity of siderophores depend on the concentrations of Cd2+ in the medium (0, 0.5, 1, 2 and 3 mM Cd2+). In the analysis we used a spectrophotometric chemical assays, high performance liquid chromatography (HPLC) as well combined techniques - LC-Q-TOF/MS. We have revealed relevant differences between investigated strains both in type and amount of synthesized siderophores under Cd2+supplementation. In case of two investigated strains increasing concentration of Cd2+ in the substrate stimulated synthesis of hydroxamic, catechol and phenolic siderophores. In general, presence of Cd2+ stimulated synthesis of siderophores compare to the control variant (not supplemented with Cd2+), however effect of metal concentration differed between investigated strains. HPLC analysis allowed for detection and identification of ferrioxamine B in culture medium of 4 studied strains. 263 Rhizo Remediation and Fate of Pollutants Wednesday 24 June – Poster session New Methods and Concepts in the Rhizosphere 91 High-throughput amplicon sequencing for microbe identification in complex colony communities of culture collections Jaderson Armanhi*1, Laura Migliorini de Araujo1, Rafael Soares Correa de Souza1, Natalia Verza Ferreira1, Vagner Katsumi Okura1, Beatriz Jorrín Rubio2, Juan Imperial2, Paulo Arruda1 1 State University of Campinas (UNICAMP), Brazil, 2Centro de Biotecnología y Genómica de Plantas (CBGP), Campus de Montegancedo, Universidad Politécnica de Madrid, Spain Construction of a microorganism culture collections isolated from environment can be timeconsuming since individual colonies containing single microbes might demand several streaking steps for colony purification. In addition, the pursuing of pure colonies may restricts important insights on mutual association. Potentially, in extreme cases, microorganisms in consortia might depend strictly on beneficial microbe-microbe interactions. Here we present a method for construction and identification of a collection of microorganisms and consortia of microorganisms isolated by a single step of plating and colony picking. Complex microbial mixtures isolated from sugarcane roots and stalks were plated in defined media at proper dilution to obtain colonies. Colonies were picked, grown in 96-deepwell plates and stored at 80 °C. This collection comprises over five thousand wells containing individual microorganisms or consortia. In order to identify these microorganisms we have developed a method to sequence pools of amplicons in 96-well plates using barcodes to tag plates, rows and columns in two PCR steps. Pooled plates can be sequenced into a single run of PacBio and delivers high quality amplicons relying on circular consensus sequence (CCS) for error corrections. Together, the multi-tagging system and PacBio platform allows the identification of microbe composition of each well, regardless if it contains pure or consortia colonies. Results will be discussed in the context of cost-effective alternatives for large-scale identification of individuals in complex colony communities by sequencing pools of multiplexed 96-well plates. 92 Combination of X-ray micro tomography and soil solution studies to analyse root system development and soil chemistry in situ as a response to different Nforms Sebastian Blaser*1, Doris Vetterlein1, Enrico Thiel2 1 Helmholtz Centre for Environmental Research - UFZ, Germany, 2SKW Stickstoffwerke Piesteritz GmbH, Agricultural Application Research, Germany Only 30-50% of the applied N fertilizers are captured by crops. Therefore increasing both, N use efficiency and crop production is a major challenge for sustainable agriculture. Urea is the most widely used N fertilizer. Urea undergoes rapid hydrolisation applied at the soil, after which ammonium is oxidized to nitrate. Due to the use of nitrification inhibitors, the importance of NH4+ as N source has increased. NH 4+ is more protected against nitrification creating benefits for environment and plant growth. 264 New Methods and Concepts in the Rhizosphere Wednesday 24 June – Poster session Application of urea granules with or without inhibitors is expected to result in a spatially heterogeneous and temporally dynamic distribution of different N-forms in the soil matrix. Plasticity of root growth can be observed when roots are exposed to localized sources of nitrogen. Two general ways of response seem to be common including systemic repression of lateral root growth by high N status of the plant and local stimulation by initiation and elongation or inhibition of lateral root growth by availability of NO3 or NH4. These effects are related to the nutritional and signalling effect of the respective ion but dynamics are difficult to observe due to the opaque nature of soil. The aim of this work is to verify hypotheses, derived from studies with flow-through-systems or gel plates, for soil systems, using X-ray micro tomography to visualize and characterize dynamic root system development in soil as a response to different N-forms in situ. These analyses are combined with soil chemical studies in the same temporal resolution. Micro suction cups are installed to extract soil solution with known distance to roots and localized fertilizer. This setup is applied to answer the questions when, where and why do modifications of root system architecture occur in relation to soil chemical and N-nutritional conditions. 93 Microdialysis – an alternative approach to identifying plant-available nitrogen in soils Scott Buckley*, Richard Brackin, Susanne Schmidt The University of Queensland, School of Agriculture and Food Sciences, Australia Given the importance of nitrogen (N) availability in controlling N acquisition in plants and microbes, estimating soluble and exchangeable N forms in soil is vital to understanding these processes. However, we have relied on extraction methods that severely disrupt the soil environment. Microdialysis offers an alternative method of sampling soil N with minimal disturbance, but it is unclear whether this method better represents plant-available N in soil. We evaluated microdialysis and conventional salt (KCl) and water extraction methods in the context of a microcosm experiment, sampling N release (NO3 -, NH4+ and amino acids) from sugarcane litter (high C:N ratio) and soybean litter (low C:N ratio) decomposition over 30 days. Microbial activity (microbial biomass-N, protease activity, CO2 respiration) was also measured to assess microbial response to varying litter quality, and to predict N cycling processes. Diffusive fluxes (measured via microdialysis) provided a high-resolution snapshot of N availability at day 30, highlighting N cycling processes that were insufficiently resolved using salt or water extractions. Patterns of N immobilisation were observed in sugarcane treatments, whereas significant N mineralisation was found in soybean treatments. Each pattern increased with litter concentration, and was consistent with observed microbial activity to differing qualities of litter (microbial biomass-N, protease activity and CO2 respiration being greatest in soybean treatments). Such patterns were not apparent in KCl or water extractions, both showing uniform N concentrations and N species across treatments, with the exception of highly amended soybean treatments. These findings challenge the effectiveness of conventional soil extraction techniques to sufficiently estimate plantavailable N, and to resolve N cycling processes in soil environments. 265 New Methods and Concepts in the Rhizosphere Wednesday 24 June – Poster session 94 Water repellency and percolation across the rhizosphere Mohsen Zarebanadkouki, Eva Kroener, Andrea Carminati* Georg-August University of Goettingen, Germany The ability of plants to extract water from the soil is influenced by the hydraulic conductivity of roots and their rhizosphere. Recent experiments showed that the rhizosphere turned water repellent after drying and it remained dry after rewetting of the bulk soils. Our objective was to investigate whether and in what conditions rhizosphere water repellency is a limit to root water uptake. We used neutron radiography to trace the transport of deuterated water (D2O) in the roots of lupins experiencing local soil drying. The soil was partitioned in nine compartments. We let one of the upper compartments dry, while keeping the other compartments wet. We injected D2O in different locations. The radiographs showed that root water uptake in the soil region that was let dry and then irrigated was 4-8 times smaller than in the wet soil regions. In a parallel experiment, we monitored the rehydration of lupin roots that were irrigated after severe drying. Based on measurements of root swelling and xylem pressure, we found that the hydraulic conductivity of the rhizosphere was the limiting factor for root rehydration. Our hypothesis is that rhizosphere water repellency was caused by mucilage exuded by roots. We measured capillary rise in soils with varying mucilage concentration. We found that below a critical mucilage concentration water could easily cross the soil, while above the critical concentration water could no longer percolate through the soil. The critical mucilage concentration depended on soil particle radius. The results were well reproduced by a new pore-scale model of water percolation through the rhizosphere. Our studies suggest that the rhizosphere is near the percolation threshold, where small variations in mucilage concentration sensitively alter the soil hydraulic conductivity. It is tempting to conclude that mucilage exudation is a plant mechanism to efficiently control the rhizosphere conductivity and water uptake. 95 Effect of root exudates on soil mineralogy: possible implications on nutrient mobilization Concetta Eliana Gattullo*1, Ignazio Allegretta1, Giovanni Cuccovillo1, Luca Medici2, Tanja Mimmo3, Youri Pii4, Stefano Cesco4, Nicola Tomasi5, Roberto Pinton5, Roberto Terzano1 1 Department of Soil, Plant and Food Sciences - University of Bari, Italy, 2Institute of Methodologies for Environmental Analysis, CNR, Italy, 33Faculty of Science and Technology, Free University of Bolzano, Italy, 4Faculty of Science and Technology, Free University of Bolzano, Italy, 5Department of Agricultural and Environmental Sciences University of Udine, Italy Plants release in the rhizosphere flavonoids and organic acids which exert multifunctional roles. In a recent study, we observed that the flavonoid rutin, alone or combined with genistein or organic acids (citrate, malate, oxalate), usually present in plant exudates, mobilized Fe from a calcareous soil with a great efficiency. However, the effects of these 266 New Methods and Concepts in the Rhizosphere Wednesday 24 June – Poster session exudates on soil mineralogy, especially on Fe-bearing minerals and clays, remained unexplored. This work aims at evaluating the changes in the mineralogical composition of a calcareous soil after 24h-treatment with an aqueous solution of rutin (35 µM), alone or combined with other flavonoids (10 µM) or organic acids (1 mM). After centrifugation, the solid fraction was dried, homogenized with corundum (internal standard) and analysed by X-ray powder diffraction (XRPD), using the Rietveld method for mineralogical quantification. XRPD analysis of the natural soil revealed the presence of calcite (57.0%), illite (17.9%), smectite (5%), quartz (4.2%), heulandite (2.8%), and amorphous phases (13%). No Fe-oxides and oxyhydroxides were detected. In all the treatments with rutin, the sum of illite and smectite considerably increased, compared to the natural soil, especially when rutin was combined with organic acids or genistein. Conversely, the amorphous residue decreased. It can be assumed that the increase of illite+smectite was caused by the transformation of the amorphous phases into clay minerals. Rutin and its combinations with organics possibly mobilized Fe from soil along with other cations which could then find suitable conditions to recrystallize as new forming clay minerals. Na, K, and Ca could enter in the interlayer regions of phyllosilicates, while Al and Si could induce the formation of new sheets. Although the variation in soil mineralogy could not be easily related to Fe mobilization, it was evident that some treatments producing the higher Fe solubilization showed also the higher increase of the illite+smectite fraction. 96 A microfluidic device for imaging and asymmetric perfusion of Arabidopsis roots Claire Stanley1, Jagriti Shrivastava2, Rik Brugman2, Dirk van Swaay1, Andrew deMello1, Guido Grossmann*2 1 ETH Zürich, Institute for Chemical and Bioengineering, Germany, 2Universität Heidelberg / Centre for Organismal Studies, Germany Plant roots are highly sensitive to changing environmental conditions such as water and nutrient availability, biotic and abiotic stresses. Long-distance communication from root to shoot has recently been reported to be specific for salt stress perceived by roots. Open questions include how and why certain stimuli cause local, cell-autonomous responses, whilst others result in cell-cell communication and coordinated responses by tissues, organs or the entire plant. To understand how roots sense and process information about their environment we need tools that allow live imaging of roots and provide precise control over the root microenvironment. Over the past years, a number of microfluidic devices have been developed to cultivate and perfuse Arabidopsis roots at the microscope. These devices have substantially advanced experimental access to roots and some devices, e.g. the RootChip, allowed pulsed treatments. So far, in devices where roots grow in channels it has been difficult to locally apply treatments only to selected regions of the root, as the organ was usually perfused as a whole. Moreover, due to roots bending within observation chambers, an even perfusion was often challenging to achieve, thus affecting reproducibility of the treatments. 267 New Methods and Concepts in the Rhizosphere Wednesday 24 June – Poster session Here we present a new imaging and perfusion device for Arabidopsis roots that provides guidance of the root tip and centering of the root within the chamber to allow symmetric or asymmetric perfusion of the root. By applying treatments specifically to one side of the root we are able to distinguish between cell-autonomous responses and coordinated responses to changing environmental conditions. 97 Screening for allelochemical stress on cytoplasmic protein synthesis pattern of selected plants Aasifa Gulzar*, M.B. Siddiqui, Shazia Bi Aligarh Muslim University, India Determining the mode of action of allelochemicals is one of the challenging aspects in allelopathic studies. Recently, allelochemicals have been proposed to cause differential protein expression in target tissue and inhibit the early growth. Phenolic acids, one of the common allelochemicals emitted from rhizosphere soil is known for its growth-inhibitory activity. The aim of the present study was to determine the inhibitory effect of Phenolic acids on seedling growth, dry biomass, total protein content and expression levels of proteins. Effects of allelochemicals on early seedling growth and dry biomass were studied in three test species, Glycine max Willd., Lycopersicon esculentum L. and Lantana camara L. Total protein content and their differential expression were studied in the leaves of test species. Rhizosphere soil was analysed for the detection of phenolic acids. The root length, shoot length and dry biomass were significantly reduced in rhizosphere soil as compared to control soil. SD-PAGE analysis showed different protein expression in three species on exposure to rhizosphere soil. Protein banding pattern not only differ between the control soil and rhizosphere soil, but also among test species. The protein content was increased in Glycine max, and esculentum while it was decresed in L. camara in response to rhizosphere soil. Rhizosphere soils contained significantly higher amount of phytotoxic phenolics as the putative allelochemicals, which were ferulic acid, vanillic acid, p-coumaric acid and benzoic acid. The study concluded that C. procera roots and rhizosphere soil exerted allelopathic effect on test species by releasing water-soluble phenolic acids as putative allelochemicals in soil which will act as a structural substitute for synthetic herbicides. 268 New Methods and Concepts in the Rhizosphere Wednesday 24 June – Poster session 98 Quantification of C- and N- rhizodeposition of peas under field conditions Anke Hupe*1, Florian Wichern2, Hannes Schulz3, Franziska Näther3, Rainer Georg Jörgensen1 1 University of Kassel, Soil Biology and Plant Nutrition, Germany, 2Rhine-Waal University of Applied Sciences, Faculty of Life Sciences, Germany, 3University of Kassel, Department of Organic Farming and Cropping, Germany Quantification of the C- and N- rhizodeposition under field conditions is difficult as labelling plants with stable or radioactive isotopes and separating labelled and unlabeled roots is difficult and often creates unnatural conditions. As a consequence experiments are often done under controlled conditions. To estimate a realistic amount of C and Nrhizodeposition, experiments have to be conducted under field conditions without influencing the root system or the water and nutrient budget. The presentation will focus on the results of a study, which quantified the C- and N- rhizodeposition of peas in a field experiment in 2013. In particular, the spatial and temporal distribution of rhizodeposition and their transfer into different soil compartments was measured. Furthermore, the percentage of the microbial biomass C and N deriving from rhizodeposition was quantified. For this, the pea (Pisum sativum cv. Santana) was labeled fortnightly (beginning at BBCH 13) with a solution of 2% 13C-glucose (99 atom%) and 0.5% 15N urea (95 atom%) using the cotton wick method. Sampling took place on four dates depending on the growth stage (beginning at BBCH59). Soil samples were taken at 0-30 and 30-60 cm depth. The sampling of the soil always took place in three defined sectors of the microplots (directly on the plant; between two plants in the row; between two rows). In order to calculate the complete root biomass and rhizodeposits of one pea plant and to reduce errors, the weighted mean of the three sectors in 0-30 cm and 30-60 cm depth was calculated. In the presentation the following hypotheses are discussed: 1. the amount of C and N rhizodeposition under field conditions is substantially higher compared to pot experiments; 2. the development of rhizodeposition depends on the development of above-ground plant biomass. 99 Density-based approaches to interface models and data in root biology Dimitris Kalogiros*1, Michael Adu2, Xavier Draye3, Mariya Ptashnyk4, Glyn Bengough4, Lionel Dupuy2 1 University of Dundee / The James Hutton Institute, Scotland, 2The James Hutton Institute, Scotland, 3Earth and Life Institute, Université Catholique de Louvain, Belgium, 4University of Dundee, Scotland Understanding the development of root system architectures is difficult, because they consist of numerous interconnected roots which develop in parallel in response to many intrinsic and environmental cues. Root system architecture models have proved useful to deal with that complexity, yet they suffer from the lack of quality growth data for roots grown in soils. 269 New Methods and Concepts in the Rhizosphere Wednesday 24 June – Poster session We present recent advances in the construction, calibration and validation of root system architecture models. Mathematical models use density distributions and time-delay partial differential equations to describe root developmental mechanisms. We also present methods to estimate root density distribution from experimental image data (kernel-based estimators) and test different optimisation algorithms to parameterise the mathematical model on such data. Results show that density-based models can be parameterised accurately from experimental density distribution function, paving the way for new approaches to understand growth responses to environmental cues. Plants develop diverse root system architectures to capture soil water and nutrients. This study shows that density-based models provide a simpler link with experimental data, thereby complementing existent root system architecture models and setting the stage for characterisation of root properties and investigation of root functions. 100 Application of laser capture microdissection for assessing pyoverdine synthesis in planta by endophytic bacteria Laure Avoscan1, Christine Arnould2, Aline Bonnotte3, Jeannine Lherminier2, Philippe Lemanceau*1, Sylvie Mazurier1 1 INRA/UMR Agroécologie, France, 2INRA/UMR Agroécologie/Plateforme DimaCell, Centre de Microscopie INRA, France, 3Université de Bourgogne/UMR Agroécologie/Plateforme DimaCell, Centre de Microscopie INRA, France Previous studies have shown that some isolates of fluorescent pseudomonads are able to colonize root tissues and are detected in root intercellular spaces and root cells. These bacteria are also known to scavenge iron from the soil and from the rhizosphere environment, thanks to the synthesis of secondary metabolites called siderophores. Meanwhile a bacterial siderophore produced by Pseudomonas fluorescens C7R12 (pyoverdine) has been shown to be incorporated in planta and to contribute to plant iron nutrition. Taken together, these observations raise the question of whether siderophores may be produced in planta by endophytic bacterial cells. To address these questions, laser capture microdissection (LCM) and gene expression studies have been combined. More precisely, the model plant Medicago truncatula has been grown axenically under controlled conditions of iron supply and inoculated or not with P. fluorescens C7R12. Firstly, root zones colonized by the bacteria have been described by transmission and scanning microscopy observations. Then, labeling and fixation procedures for microscopy have been adapted to allow the selective laser capture microdissection of endophytic bacterial cells and nucleic acid microextraction prototocols were adjusted to extract RNAs from the dissected samples. PCR primers targeting genes of P. fluorescens C7R12 implied in the pyoverdine synthesis were designed and RT-qPCR protocols were developed to compare the level of expression of these genes ex planta and in planta. The results of this comparison will be presented together with further plans to evaluate the influence of the pyoverdine on the plant iron homeostasis. 270 New Methods and Concepts in the Rhizosphere Wednesday 24 June – Poster session 101 Plant water-uptake effects on salt distribution at the root-soil interface Adi Perelman*1, Helena Jorda Guerra2, Jan Vanderborght3, Andreas Pohlmeier2, Shimon Rachmilevitch4, Naftali Lazarovitch4 1 Ben-Gurion University of the Negev, Israel, 2Forschungszentrum Jülich, Germany, 3Forschungszentrum Jülich, KU Leuven, Belgium, 4Ben-Gurion University of the Nege, Israel In recent years, increased salinization of soils, along with the depletion of available water, has become a major threat for agriculture. The rhizosphere plays an important role in connecting processes at the soil-plant- atmosphere interface. A better understanding of these connections and their mutual influences can help in improving crop yield. New technologies provide better tools with which to study root structure and function in non-destructive ways. These techniques also provide a refine resolution regarding processes occurring where the roots meet the soil, e.g., water uptake and salt accumulation. Bell peppers are considered to be sodium excluders, whereas carrots take up sodium. Therefore, we hypothesize that salt accumulation around the roots will create a distinction between them under the same irrigation regime. The initial results from rhizoslides (capillary paper growth system) show the salt concentration gradient is decreasing with distance from the root, compared with the bulk that remained more stable. At the microscopic scale, magnetic resonance imaging (MRI) will be implemented to observe root structures, water content and sodium concentration distributions around single roots. At the macroscopic scale, root systems and solute and water distributions within the root zone of plants grown in lysimeters will be monitored using a combination of electrical resistivity tomography (ERT) and local soil sensors. Data will be used to calibrate a model that is expected to predict root water uptake in saline soils for different climatic conditions and different soil water availabilities. Sensitivity analyses from a simulation study with this detailed model for different soil salinities, irrigation regimes and weather scenarios will be presented. 102 Dissecting root secretion using novel lux biosensors Joshua Roworth*, Philip Poole University of Oxford, United Kingdom Plant roots secrete a complex mixture of metabolites, which are key to determining the structure of the microbial community (microbiome) in the rhizosphere. The microbiome in turn can alter plant growth both positively (plant growth promotion) or negatively (pathogenesis). Lux biosensors were developed in Rhizobium leguminosarum biovar viciae 3841, which respond to metabolites secreted by plant roots both specifically (e.g. phenylalanine) and generally (e.g. expression is dependent on available carbon). By using a highly sensitive, cooled CCD camera these biosensors can be used for non-invasive, temporal and spatial mapping of metabolite secretion. Using this novel system it is possible to screen plant mutant libraries of rice for both increased and decreased metabolite secretion. Once secretory mutants have been identified the nature of a mutation can be determined (e.g. increased formate secretion) and phenotypic characterisation of the effects these mutations have on plant growth, productivity and rhizosphere colonisation can be assessed. 271 New Methods and Concepts in the Rhizosphere Wednesday 24 June – Poster session Identifying the roles that key metabolites play in plant health and the rhizosphere is an important step in understanding how plants manipulate the rhizosphere to their advantage. Identifying metabolites that play a key role in the rhizosphere are key targets for genetic manipulation to aid in plant growth, health and productivity. In the case of rice, this in the future will hopefully lead to increased yields in one of the most important food crops. 103 Fungi as food source for rhizosphere bacteria Max-Bernhard Rudnick*1, Wietse de Boer2, Hans van Veen2 1 Leibniz Institute of Vegetable and Ornamental Crops (IGZ), Germany, 2Netherlands Institute of Ecology (NIOO-KNAW), Netherlands Fungi and bacteria are primary consumers of plant-derived organic compounds and, therefore, considered as basal members of soil food webs. Trophic interactions among these microorganisms could, however, induce shifts in food web energy flows. Given increasing evidence for a prominent role of fungi as primary consumers of root exudates, we propose that fungus-derived carbon may be an important resource for rhizosphere bacteria. To test this assumption, two common rhizosphere-inhabiting saprotrophic fungi, Trichoderma harzianum and Mucor hiemalis, and a plant-pathogenic fungus, Rhizoctonia solani, were confronted in microcosm systems with bacterial communities extracted from the rhizospheres of two plant species, Carex arenaria and Festuca rubra. Bacteria that associated with hyphae were screened for their ability to feed on growing fungal hyphae. This screening revealed a widespread ability of rhizosphere bacteria to feed on energy resources provided by living fungi. The identity of the fungi had a strong effect on the composition of these mycophagous bacteria, whereas the effect of rhizosphere origin (sedge versus grass) was small. Our results suggest that secondary consumption of fungal resources instead of primary consumption of root exudates can be an important nutritional strategy among rhizosphere bacteria. Food web models should take this possibility into account as this has important consequences for carbon fluxes with more carbon dioxide released by microbes and less microbial carbon available for the soil animal food web. Another, interesting finding is that several of the “fungus-feeding” bacteria appeared to be closely related to plant-pathogenic bacteria. 272 New Methods and Concepts in the Rhizosphere Wednesday 24 June – Poster session 104 Use of high throughput sequencing to study oomycete communities in soil and roots Rumakanta Sapkota*, Mogens Nicolaisen Aarhus University, Denmark Many of the plant diseases caused by oomycetes such as cavity spot and damping off involve a complex of several species emphasizing the need to use a community approach when studying these organisms. Despite the economically importance of plant pathogens such as Phytophthora and Pythium, we have limited understanding of the diversity of oomycetes in symptomatic plant tissue as well as in root zones. The aim of this study was to improve and validate techniques for using high throughput sequencing as a tool for studying oomycete communities. Primer sets ITS4, ITS6 and ITS7 that have been used earlier in similar studies but with limited success, were used in this study with an improved protocol. Our result shows that the proportion of retrieved oomycete sequences dramatically increased, mainly by increasing the annealing temperature during PCR. The optimized protocol was validated using mock communities, DNA extracted from carrot tissue samples with symptoms of Pythium infection and soil samples collected from agricultural fields. Sequence data from Pythium and Phytophthora mock communities showed that our strategy successfully detected all included species. Taxonomic assignments of operational taxonomic units from symptomatic lesions in carrot resulted in 94% of the reads belonging to oomycetes with a dominance of species of Pythium that are known to be involved in causing cavity spot. Moreover, soil samples showed that 95% of the sequences could be assigned to oomycetes including Pythium, Aphanomyces, Peronospora, Saprolegnia and Phytophthora. A high proportion of oomycete reads was consistently present in all symptomatic lesions and soil samples showing the versatility of the strategy and thus demonstrating the usefulness of the method in plant and soil DNA background. 105 Quantifying and comparing AM fungi biomass in three soils colonized by maize using signature fatty acids and microscopic methods Mahaveer P Sharma*1, Jeffrey S Buyer2 1 ICAR-Directorate of Soybean Research, India, 2USDA-ARS, SASL, BARC-West, United States The arbuscular mycorrhizal (AM) fungi association of the Phylum Glomeromycota is one of the most common associations colonizing the roots of more than 80% terrestrial plants. AM fungi are well-known plant symbionts which provide many benefits to plants such as enhanced growth, mineral nutrition particularly phosphorus and zinc uptake, protection against soil pathogens, and improved response to moisture stress. There is increased interest in the area of mass production and application of AM fungi which necessitates selection of efficient AM strains. Selection of superior AM candidates for mass production and growth promotion is being done based on quantification of AM fungi, hence the quantification of AM fungal biomass in soil and roots is essential to every experimental setup involving the AMFplant interaction. Quantification of mycorrhizal biomass and root colonization has traditionally been performed by root staining and microscopic examination methods which are time-consuming, laborious and rely on utilizing specific stains to visualize the AM-roots 273 New Methods and Concepts in the Rhizosphere Wednesday 24 June – Poster session and then quantify based on person’s skills in microscopic observations and often lacks reproducibility of quantified data by different laboratories. In this study we compared two microscopic methods, spore counts and root colonization, to three biomarker methods: neutral and phospholipid fatty acids (16:1ω5 PLFA and NLFA), and 16:w5 ester-linked fatty acid analysis in three different soils raised on maize. Irrespective of soil type, consistently the 16:1ω5 ester-linked fatty acid (EL-FAME) was found to be positively correlated both with spore count and root colonization whereas neutral lipids correlated well only with spore count. Overall, analysis of 16:1ω5 ester-linked fatty acid was the most useful biomarker for assessing AMF biomass, while 16:1ω5 neutral lipid fatty acid analysis was useful for estimating mycorrhizal spore counts. 274 New Methods and Concepts in the Rhizosphere Wednesday 24 June – Poster session Rhizosphere Microbiome 107 Effects of four plant functional types on rhizosphere bacterial community composition in a subarctic Sphagnum peatland Sylvain Monteux*1, James T. Weedon2, Frida Keuper3, Ellen Dorrepaal4 1 Climate Impacts Research Centre, Department of Ecology and Environmental Sciences, Umeå University, Sweden, 2University of Antwerp, Department of Biology, Belgium, 3French National Institute for Agricultural Research, Laon, France, 4Climate Impacts Research Center, Department of Ecology and Environmental Sciences, Umeå University, Sweden Northern peatlands store about one third of the world’s soil organic carbon, the future stability of which is a global concern that remains poorly understood. Bacterial communities play a major role in decomposition, and can be affected by many environmental factors. Of these, rhizodeposition is particularly interesting as its possible effects on bacterial community structures and priming effects could greatly affect the rate of soil organic carbon decomposition. While effects of hydrology and microclimate on microbial communities have recently been investigated in peatlands, little is known about the impacts of rhizodeposits from different vascular plants. We therefore investigated whether bacterial communities are affected by differential rhizodeposition by vascular plant species in Sphagnum peatlands. We hypothesized that plant species belonging to different plant functional types with different rhizodeposits harbour different bacterial communities in their rhizosphere than those observed in bulk Sphagnum peat. Alternatively, it may be that Sphagnum peat composition is sufficiently homogeneous to overcome rhizodeposits effects. To test this hypothesis we sampled Sphagnum peat soils from a subarctic peatland in northern Sweden (close to Abisko, 68°21′N). We compared Sphagnum peat from areas with low vascular plant cover (bulk peat) with the rhizosphere of four common species of vascular plants representing four functional types: sedges (Eriophorum vaginatum), forbs (Rubus chamaemorus), deciduous shrubs (Betula nana) and evergreen shrubs (Empetrum nigrum). We present bacterial communities’ profiles obtained through Illumina sequencing of 16S rRNA gene amplicons and test the extent to which community composition in bulk peat differs from those in the rhizosphere, and whether plant species from different functional types harbour distinct bacterial communities in their rhizosphere. 108 Plant traits related to nitrogen uptake influence plant-microbe competition Delphine Moreau*, Barbara Pivato, David Bru, Hugues Busset, Florence Deau, Céline Faivre, Annick Matejicek, Florence Strbik, Laurent Philippot, Christophe Mougel INRA, France Plant species is an important driver of soil microbial communities. However how plant functional traits are shaping these communities has received less attention while linking plant and microbial traits is crucial for better understanding plant-microbes interactions. Our 275 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session objective was to determine how plant-microbes interactions were affected by plant traits. Specifically we analyzed how interactions between plant species and microbes involved in nitrogen-cycling were affected by plant traits related to nitrogen nutrition in interaction with soil-nitrogen availability. Eleven plant species, selected along an oligotrophic-nitrophilic gradient, were grown individually in a nitrogen-poor soil with two levels of nitrate availability. Plant traits for carbon and nitrogen nutrition were measured and the genetic structure and abundance of rhizosphere microbial communities, specifically the ammonia oxidizer and nitrate reducer guilds, were analyzed. The structure of the bacterial community in the rhizosphere differed between plant species and these differences depended on nitrogen availability. The results suggest that the rate of nitrogen uptake per unit of root biomass and per day is a key plant trait, explaining why the effect of nitrogen availability on the structure of bacterial community depends on the plant species. We also showed that the abundance of nitrate reducing bacteria always decreased with increasing nitrogen uptake per unit of root biomass per day, indicating that there was competition for nitrate between plants and nitrate reducing bacteria. This study demonstrates that nitrate reducing microorganisms may be adversely affected by plants with a high nitrogen uptake rate. Our work puts forward the role of traits related to nitrogen in plant-microbe interactions whereas carbon is commonly considered as the main driver. It also suggests that plant traits related to ecophysiological processes, such as the nitrogen uptake rate, are more relevant for understanding plantmicrobe interactions than composite traits, such as nitrophily, related to a number of ecophysiological processes. 109 Soil microbial diversity effects on primary production and symbiotic interactions Christophe Mougel*1, Clémentine Lepinay2, Julie Aubert3, Sébastien Terrat4, PierreAlain Maron2, Christophe Salon2, Thierry Rigaud5 1 INRA, France, 2INRA Agroécologie, France, 3INRA Mathématiques et Informatique Appliquées, France, 4INRA Agroécologie & Plateforme GenoSol, France, 5UMR CNRS 6282 Biogéosciences, France The consequences of biodiversity losses on ecosystem functioning are now frequently addressed in ecology and biological conservation, mainly because of the importance of ecosystem functioning in providing goods and services to human activities. Most studies on the consequences of biodiversity losses are on plants. However, the importance of telluric microorganisms linked together by trophic exchanges with plants, that sustain all ecosystems through primary production, is known. However, the role of soil microbial diversity for primary production remains controversial. We test the hypothesis that decreased soil microbial diversity may impact negatively plant primary production and symbiotic associations. For this, we manipulated the soil microbial diversity by experimental erosion, and measured the changes in microbial community diversity using massive sequencing. We then measured the effect of diversity loss on plant/symbionts association and on plant growth and fitness. The results showed that the impact of microbial diversity on plants depends on their reliance on symbionts. On the whole, Medicago. truncatula was negatively affected by erosion, Brachypodium distachyon was positively affected and there was no significant effect on Arabidopsis thaliana. These results are of interest to predict the consequences of soil microbial diversity loss, on 276 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session crop plants productivity under low nutrient inputs, according to microbial communities and symbiotic interactions. 110 Taxonomic and metabolic diversity of Pseudomonas spp. fluorescents isolated from the rhizosphere of the southern Algerian palm groves Ben Oussaid Nacera*1, Benchabane Messaoud2, Phillipe Thonart3 1 University Blida 1, Departement of Biology, Algeria, 2University Blida1, Departement of Agronomy, Algeria, 3University of Liege, Walloon Center for Industrial Biology, Belgium Pseudomonas spp. fluorescents characterized by genetic and phenotypic diversity in relation to their positive impact on the functioning of the rhizosphere, exerting direct and / or indirect actions on the development of the plant. However, the exploitation of microbial resources occupies a prominent place in applications preservation and restoration of soil fertility. The effectiveness of some strains of Pseudomonas spp. fluorescents organic plant protection has been demonstrated in numerous studies in experimental conditions and practices. The intended benefits are essentially on biocontrol of plant pathogens, the nutritional status of the plant and its defensive natural abilities. Our work presents the characterization of a collection of strains of Pseudomonas spp. fluorescent, isolated from the rhizosphere of different palm of Bechar, Ghardaia and Ouargla province southern of Algeria and their potential use as biological means. The selection of effective isolates was carried out on the basis of exhibition of substantial involved in the processes of biocontrol and phytostimulation (siderophores, HCN, phenazines, AIA...). In addition to the basic bacteriological tests (morphological, physiological and biochemical tests, Biolog), 16S rRNA was amplified by PCR. Sequencing was performed using an ABI Prism® 3130XL Genetic Analyzer (Applied Biosystems, Foster City, CA, USA) and the phylogenetic analysis was performed using the software package BioNumerics (Applied Maths, Belgium).). Phenotypic and genotypic identifications showed that the predominant species is P. fluorescens (50%), in addition to the presence of P.putida, P.chlororaphis, P.aureofasciens and intermediate strains. The results indicate the presence of a taxonomical diversity and the particular metabolic faculties in the group of Pseudomonas spp. fluorescent and particularly in type species P. fluorescens and P. putida. The selection of microbial taxa, in palm groves, can be evoked. 111 Diversity and abundance of genes for the synthesis of osmoprotectans in the rhizosphere of the halophyte Arthrocnemun macrostachyum Salvadora Navarro-Torre*1, José María Barcia-Piedras1, Eloisa Pajuelo1, Miguel Ángel Caviedes1, Susana Redondo-Gómez1, María Camacho2, Ignacio D. Rodríguez-Llorente1 1 University of Seville, Spain, 2IFAPA, Centro Las Torres-Tomejil, Spain Arthrocnemum macrostachyum is a halophyte naturally growing in salt marshes and salt steppes of the south of Spain. Our interest is focused in the utility of this plant and the microorganisms colonizing its rizosphere to desalinate coastal agricultural soils. In this context, the aim of this work was to study the diversity of bacteria able to produce osmoprotectants related with salt tolerance (ectoine and betaine) in the rhizosphere of the plant. Based on their salt tolerance and requirements, several halophilic and halotolerant 277 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session bacteria were isolated and identified. All of them showed the presence of ectC (ectoine synthase) and betB (betaine aldehyde dehydrogenase) genes. Three of this isolates (Cs16b, Cs25 and Hvs18) have been identified as halophilic strains, being Hvs18 able to tolerate up to 20% NaCl. Cs16b, Cs25 and the halotolerant strain named Hv16, have been described as novel species within the genera Microbulbifer, Labrenzia and Kokuria, respectively, indicating that rhizosphere of halophyte plants are an excellent ecosystem to isolate new halophilic/tolerant species. In order to study the biodiversity of bacteria with ectC and betB genes, an internal fragment of each was amplified from total soil DNA isolated from the rhizosphere of A. macrostachyum growing in two different soils. Amplified ectC fragment was cloned into pGEM-T easy vector and 100 positive clones were sequenced. 70% of the sequences corresponded to ectC and at least 17 different sequences were identified. On the other hand, betB fragment was used use as template for a DGGE analysis of the abundance and diversity of betB in those rhizospheres. Our results show a great biodiversity of bacteria harboring genes for the synthesis of osmoprotectants, most of them described for the first time in this work. 112 Rhizosphere bacterial communities from wheat field in Brazil and the occurrence of wheat blast Vanessa Nessner Kavamura*1, Ana Gabriele Barbosa-Casteliani2, Rodrigo Taketani1, Angelo Sussel3, Itamar Melo1 1 Embrapa Environment, Brazil, 2University of São Paulo, Brazil, 3Embrapa Cerrados, Brazil Brazil is the second largest wheat producer in South America. However, wheat blast disease outbreaks caused by Magnaporthe grisea result in high yield losses. Recent studies have shown that this fungus has the ability to infect through the roots as well as the foliar system. Understanding the changes in the structure of rhizosphere microbial communities of wheat plants in areas with different levels of infection might help unveil which groups of microorganisms could be acting to supress M. grisea. Rhizosphere and bulk soil samples were collected from a wheat field in Distrito Federal (DF), with disease levels of 10% and 100%. DNA was extracted and bacterial community profiling by 16S rRNA (V6-V7) amplicon sequencing was achieved through the Ion Torrent platform. Analyzes were performed using QIIME, PAST and STAMP software. Nonmetric multidimensional scaling (NMDS) plots revealed three major groups (stress value=0.1218), one comprised bulk soil samples and the other two were made up of rhizosphere samples, which were separated by disease incidence. Similarity percentage (SIMPER) analysis revealed that bacterial communities from rhizosphere samples were more different from each other (overall dissimilarity=96.16%) than the equivalent bulk soil samples (overall dissimilarity=88.12%). Besides, at class level, STAMP analysis revealed that Cytophagia, Deltaproteobacteria and Rubrobacteria are more abundant in both bulk soil and rhizosphere samples with higher incidence of disease (P<0.05). Future work will comprise more samplings to obtain a better understanding of the structure of bacterial communities associated with wheat in different fields from Brazil, with different levels of disease incidence. This, combined with isolation of antagonists to M. grisea, might help in the development of biological control methods. 278 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session 113 Microbiome of Vitis vinifera cv. Pinot Nero in two vineyards with different soil managements Giorgia Novello*, Elisa Gamalero, Elisa Bona, Lara Boatti, Flavio Mignone, Patrizia Cesaro, Nadia Massa, Guido Lingua, Graziella Berta Università del Piemonte Orientale, DiSIT, Italy Soil is a complex microhabitat where microorganisms regulate plant productivity and the maintenance of biogeochemical cycles. Due to the release of carbon-rich exudates (rhizodeposition), bacterial densities increase in the rhizosphere. The rhizosphere is a site of intense interactions among the plant roots, the soil and the microflora. The composition, the activity and the density of microbial cells living in the rhizosphere is stimulated by the rhizodeposition: as a result, the community structure of the rhizosphere is different by that found in bulk soil. Both the density and the biodiversity of bacteria in soil are affected by the soil condition and by its management. The bulk soil and the rhizospheric soil of the grapevines were sampled before and after grape production. DNA was extracted from 1 g of soil (bulk soil and rhizospheric soil) using the UltraClean™ kit and amplified with primers for the V1 and V4 regions of 16S rDNA according to the methods optimized for 454 Roche pyrosequencing. Preliminary data indicated that the sampling time is an important factor regulating biodiversity. Actinobacteria was the dominant group followed by Sphingobacteriales. Among Proteobacteria, alpha Proteobacteria were prevalent with the Rhizobiales as the most representative group. Burkholderiales was the most prevalent among the beta Proteobacteria, while members of gamma Proteobacteria were not detected in bulk soil. The abundance of firmicutes was low in all sites; however, the species Desulfosporinus meridiei was dominant among the firmicutes found in the bulk soil. 114 Molecular identification and functional diversity of microorganisms isolated from the rhizosphere of plants grown in a suppressive soil substrate Iakovos Pantelides*, Stella Papageorgiou, Αnastasis Antoniou, Eleftheria Demetriou, Maria-Dimitra Tsolakidou, Dimitris Tsaltas Cyprus University of Technology, Cyprus In this study we investigated the effectiveness of a compost amendment, consisting of recycled plant material, against the wilt pathogens of tomato Verticillium dahliae and Fusarium oxysporum f. sp. lycopersici. The plants grown in sterile compost (SC) mix exhibited significantly more severe symptoms and fungal biomass in their vascular tissues compared to the plants grown in non-sterile compost (NSC) substrate. Plant height, total leaf surface and fresh weight of tomato plants grown in the NSC mix were significantly more compared to the plants grown in the SC substrate. Therefore, the microorganisms inhabiting the compost are likely responsible for the suppressive properties observed and may act as growth promotion inducers. In order to identify the microorganisms responsible for these phenotypes we first characterized the compost’s microbial population and then we identified the microbes that colonized the rhizosphere of tomato plants grown in the NSC substrate. 132 bacterial and 79 fungal compost isolates and 143 bacterial and 50 fungal rhizosphere isolates were identified 279 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session by sequencing of the 16S and ITS region respectively. It was found that the incidence of the rhizosphere microorganisms was different compared to that of the compost. The isolated rhizobacteria were evaluated for their biological activity against 4 soilborne pathogens (V. dahliae, F. oxysporum f. sp. lycopersici, F. oxysporum f. sp.radicis- lycopersici and Sclerotinia sclerotiorum) by the dual culture technique and tested for hydrogen cyanide and indole-3acetic acid production, phosphate solubilization, protease, chitinase, pectinase and cellulose activity. The present study revealed a high degree of functional and genetic diversity among the microorganisms isolated from the rhizosphere of plants grown in a suppressive soil. These isolates are considered to play a vital role in disease suppression and plant growth promotion due to their potential of producing an array of antifungal metabolites, hormones and plant growth promoting enzymes. 115 Bacterial inoculation on maize crops seen under invasion ecology perspectives – is the best invader the best plant growth promoter? Pedro Beschoren da Costa*1, Samanta Bolzan de Campos2, Paul Dirksen2, André Dresseno1, Volker Wendisch2, Luciane Passaglia1 1 Universidade Federal do Rio Grande do Sul, Brazil, 2Bielefeld University, Germany Bacterial inoculants applied to crops are very important for sustainable agriculture, but might not be always effective under field conditions. As interaction between the inoculants and the native microbial community plays a key role in effective root colonization and plant growth promotion, classical invasion ecology might be applied for inoculants on crops to improve microbial management effectiveness. In this work, we apply hypothesis based on invasion ecology to crop fields under bacterial inoculation. We evaluate the associated microbial community composition of inoculated and non-inoculated plants to test if: (I) an inoculant with a higher invasion potential will provide a higher plant growth promotion effect;(II) if lower diversity/resource ratio in the environment facilitates invasion. To do this, we analyzed 3 different maize crops locations in south Brazil, which received the same 4 bacterial inoculants (one Azospirillum, one Achromobacter, and two Pseudomonas). Metagenomic samples were taken from bulk soil immediately before planting, and from rhizospheric soil of 10 day old plants. The V4 region of the 16S rDNA gene was then sequenced on the MiSeq platform. Principal Coordinate Analysis shows that samples clustered by the 3 locations. Principal Component Analysis, based on the Similarity Percentage test results of the controltreatment pairs, was used as a proxy for inoculant invasion ability. Data from this approach suggests that 2 inoculants may induce very different shifts in the community upon inoculation, acting differently in different locations. These different shifts that suggest higher invasion ability, however, were not associated to greater crop productivity. The diversity/resource ratios are still to be calculated, so invasion difficulty across environments must yet be estimated. Our preliminary conclusion is that it is not possible to predict the most effective inoculants by looking at shifts on the rhizospheric community 10 days after planting – what would be extremely useful in field trials. 280 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session 116 Soil microbiomes are different in healthy and declined Quercus suber (cork oak) Diogo Pinho*1, Cristina Barroso1, Paula Gomes1, Miguel Pinheiro1, Conceição Santos Silva2, Conceição Egas1 1 Next Generation Sequencing Unit of CNC/Biocant, Portugal, 2Coruche Forest Landowners Association (APFC), Portugal Cork oak is an evergreen tree of utmost importance in Portugal because of the social and economic value of cork and also in the preservation of forest biodiversity. However, cork oak forests have been declining, due to a complex series of biotic and abiotic factors. The recognition of plant-associated microbiomes as key determinants of plant health and productivity led us to deeply characterize the cork oak soil microbial community and test the hypothesis that a specific soil microbial community is associated with Quercus suber health. The bacterial and fungal soil communities were surveyed by pyrosequencing and the microbial community associated with healthy and diseased cork oaks compared. Results unveiled a rich bacterial and fungal soil community in the cork oak forest, with Proteobacteria and Basidiomycota as the most abundant phyla. When compared, soils from healthy cork oaks were significantly enriched in ectomycorrhizal fungi while saprobes were enhanced in soils from diseased trees (p<0.05). Interestingly, the healthy cork oaks fungal community was dominated by a single ectomycorrhiza, Cortinarius, while several saprobes and other ectomycorrhiza prevailed in soils from declined cork oaks. A global comparison of the bacterial and fungal communities was performed by Principal Coordinates Analysis and significant differences were observed between the tree health conditions (RANOSIM=0.344, p=0.002; SIMPER analysis: group dissimilarity of 43.7%), with Cortinarius, Steroidobacter and Mycobacterium as the largest fungal and bacterial contributors for the differences. This study provided a deep description of the soil microbiome of cork oak rhizosphere, supporting the hypothesis that tree health condition has a specific microbiome associated. In the future, the combination of rhizosphere microbiome characterization with Q. suber genomic resources (transcriptome and genome) will enhance the understanding of cork oak biology, and promote strategies leading to improved resistance to biotic and abiotic stresses, growth promotion and increased cork production. 117 Changes in rhizosphere microbial community association with different parental types of Arabidopsis thaliana MAGIC lines Carla Porges*1, Tesfaye Wubet1, Jörg Overmann2, Boyke Bunk2, Guillaume Lentendu1, Johannes Sirkorski2, Francois Buscot1 1 Centre for Environmental Research UFZ, Germany, 2Leibniz Institute DSMZ, Germany Plants can interact with microbes through changes in the composition and concentration of root exudates. The composition of bacterial communities in the rhizosphere and within the roots are distinct between plants, developmental stages and even ecotypes and differ from the microbial community in bulk soil. However, while studying the microbial community in the rhizosphere of A. thaliana most works are focusing on the bacteria, disregarding the fungal part. 281 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session In this study, which is part of the joint project “Chemical Communication in the Rhizosphere”, 19 parental lines of the model plant species A. thaliana were cultivated in pots under controlled phytochamber conditions in two different soils. When the plants had reached an early flowering stage rhizosphere soil and bulk soil were collected for DNA extraction. Afterwards the genomic DNA of the rhizosphere of the 19 parental lines was analyzed for potential bacterial communities by paired end Illumina 16S rDNA sequencing. We found that the main driver of the rhizosphere bacterial communities was the soil origin. Furthermore the presence of plants on the soil as well as the accession type had an influence on the rhizosphere bacterial community due to the release of root exudates. Those differences where significant in relative abundance and richness. In addition we identified a dominance of the Alphaproteobacteria in the rhizosphere among the parental lines. Further investigation, including sequencing of the rhizosphere fungal community via paired end Illumina sequencing targeting the ITS2 region, will aim to elucidate the selection effect of A. thaliana accessions on fungal taxa. 118 Effect of plant domestication on the rhizosphere microbiome of common bean (Phaseolus vulgaris) Juan Esteban Pérez Jaramillo*1, Rodrigo Mendes2, Jos M. Raaijmakers1 1 Netherlands Institute of Ecology, Netherlands, 2Brazilian Agricultural Research Corporation, Embrapa Environment, Brazil Plant domestication was a pivotal achievement for human civilization and subsequent plant improvement increased crop productivity and quality. However, domestication also caused a strong reduction in the genetic diversity of modern cultivars compared to their wild relatives. It is known that plants rely, in part, on the rhizosphere microbial community for growth, development and tolerance to (a)biotic stresses. Hence, plant domestication events may have adversely affected recruitment of and interactions with their beneficial microbial partners. In here, we investigated the bacterial diversity of the rhizosphere of two wild relatives, three landraces and three modern cultivars of common bean (Phaseolus vulgaris). These different lines belong to the Mesoamerican bean gene pool of Colombia and were selected amongst more than 37,000 accessions kept in the Genetic Resources Program of the International Centre for Tropical Agriculture (CIAT, Colombia). The eight accessions were grown in both native and in agricultural soils collected in the province of Antioquia (Colombia). At different plant growth stages, DNA was extracted from rhizospheric soil and bacterial taxonomic diversity was analysed by metagenomic sequencing of the V3-V4 region of the 16S rRNA. Our approach of going “back to the roots” using native soils together with wild relatives provides new fundamental insights in host genotype-mediated recruitment of beneficial microbes and in the functional and metabolic potential of the rhizosphere microbiome of native soils and wild relatives of modern crop cultivars. 282 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session 119 Description of tomato root colonization by Bacillus subtilis EA-CB0575 using Fluorescence in situ hybridization (FISH) and Scanning Electron Microscope (SEM) Luisa Fernanda Posada Uribe*1, Valeska Villegas Escobar2, Javier Correa Álvarez2, Magally Romero T.3, Luz Estela Gonzáles de Bashan4, Yoav Bashan4 1 Universidad EAFIT Medellin, Colombia, 2Universidad EAFIT Medellín, Colombia, 3Universidad Nacional de Colombia, Colombia, 4CIBNOR México, Mexico Different Bacillus species have potential applications on important agricultural crops as Plant Growth Promoting Bacteria (PGPB). Studying PGPB colonization is an important subject to understand plant-microbe interactions; nevertheless, few studies have been conducted with Gram positive bacteria. Techniques such as culture-based detection, PCR (polymerase chain reaction), gfp insertion, immunodetection, FISH (Fluorescent in situ hybridization), have been used for detecting colonization of microorganisms in plant roots or rhizospheric area. FISH combines precision of molecular techniques, visual information of microscopy, and does not generate genetic instability issues with transformed cells, therefore it is highly recommended for Gram positive colonization studies. Bacillus subtilis strain EA-CB0575 isolated from the rhizosphere of a banana plant in Urabá, Colombia was reported as PGPB of different crops such as pepper, coffee, maize and banana. In this study, the colonization of Bacillus subtilis EA-CB0575 was evaluated on tomato roots using FISH and SEM techniques and the effect on plant growth promotion was also determined. Probes Bs575-FAM and Bs575-Cy3 were designed based on 16s rDNA sequence and their specificity was evaluated with different strains of Bacillus genus and other different genus. It was determined that Bs575 probe was specific to Bacillus subtilis species, stable in time and it was found in roots of tomato grown in MS medium, Hoagland nutrient solution and arid soil. Additionally, when using the probe to determine the colonization patterns of the strain in tomato roots, it was found that B. subtilis EA-CB0575 forms microcolonies at the intersection between main and secondary roots after one month of inoculation. Moreover, exogenous application of this strain to tomato plants promoted plant length and total dry weight with significant statistical differences respect to control without bacteria. 120 Effect of tillage systems on the prevalence of antifungal genes implicated in fungal disease suppression of wheat within the microbiome of winter oilseed rape Ridhdhi Rathore*1, Kieran Germaine1, Dermot Forristal2, John Spink2, David Dowling1 1 Institute of Technology Carlow, Ireland, 2Teagasc Crop Research Centre, Oak Park, Carlow, Ireland Plants grow in intimate association with complex microbial communities, which live in the rhizosphere, endosphere and phyllosphere. The relationships between this microbiota and its host plant can vary from pathogenic to beneficial. This microbiota has been shown to promote plant growth and productivity, improve carbon sequestration and enhance phytoremediation of pollutants. In addition, some plant associated microorganisms play an important role as biocontrol agents against pathogens. 283 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session Winter oilseed rape (wOSR) is very important crop in Europe for biodiesel and edible seed oil production. This crop is widely used as a break crop for Wheat to reduce ‘take all’ a major fungal disease. Conventional cultivation of Wheat requires the use of chemical fungicides to reduce losses resulting from this disease while significantly increasing the cost of crop production. Since the crop production sector is facing a major profitability problem due to continuing high input costs in its cultivation, novel technologies and practices are being investigated to help reduce these costs. The implementation of conservation tillage practices can significantly reduce the cost of cultivation and has huge environmental benefits associated with it. However, there is currently very limited knowledge on how conservation tillage practices impact the microbial communities associated with wOSR. We have investigated the impact that conservation and conventional tillage practices have on the prevalence of bacterial genes involved in fungal disease suppression within the microbiome associated with OSR. The prevalence of genes involved in two of these antifungal systems (PhlD and PhzB) was determined using qPCR on DNA extractions isolated from rotational trials of Wheat and wOSR. 121 Effect of Azospirillum inoculation on the abundance and genetic diversity of key phytobeneficial microbial functional groups in the maize rhizosphere Sébastien Renoud*, Jordan Vacheron, Jacqueline Haurat, Claire Prigent-Combaret, Laurent Legendre, Yvan Moënne-Loccoz, Daniel Muller UMR CNRS 5557 Microbial Ecology, France Cereal seed inoculation with plant growth-promoting rhizobacteria (PGPR) can lead to enhanced root system development, thereby allowing seedlings to rapidly explore deeper soil layers and gain access to larger supplies of water and mineral nutrients. In addition, PGPR inoculation can modify rhizomicrobial community structure, at least in the first stages of plant development, which may result from inoculant effects on root physiology and rhizodeposition, as well as more direct inoculant interactions with other rhizosphere microorganisms. These microbial interactions between PGPR inoculants and the indigenous microbiota may, in turn, have an impact on root and rhizosphere functioning, but this is hardly documented. Microbial functioning of the rhizosphere relies on individual functions carried out by functional groups e.g. nitrogen fixers, phytohormone producers, etc., which often are comprised of multiple taxa, with taxa-specific contributions to a given ecological function. Therefore, it is not feasible to infer the impact of PGPR inoculation on rhizosphererelevant microbial functional groups based on our knowledge of inoculant impact on the taxonomic composition of the rhizomicrobial community. To address this issue, we assessed the effect of seed inoculation with the phytostimulatory PGPR Azospirillum lipoferum CRT1 on the size and diversity of microbial functional groups driving N dynamics or promoting plant growth in the rhizosphere of field-grown maize. The first results with quantitative PCR showed that inoculation resulted in small reductions in the size of microbial groups involved in plant hormonal modification in the three fields studied. Sequencing data will be shown in order to evaluate whether diversity modifications also took place in the maize rhizosphere. 284 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session 122 Dynamics of the microbial community structure in the rhizosphere of narrowleafed lupin and tomato as related to nitrogen form provided Ana Alejandra Robles Aguilar*1, Oliver Grunert2, Vicky Temperton1, Stephan Blossfeld1, Dirk Reheul3, Emma Hernandez-Sanabria2, Nico Boon2, Nicolai Jablonowski1 1 Forschungszentrum Jülich GmbH, Institute für Bio- und Geowissenschaften, Germany, 2Laboratory of Microbial Ecology and Technology (LabMET), Ghent University, Belgium, 3Department of Plant Production, Ghent University, Belgium Ammonia-oxidizing bacteria (AOB) and nitrite oxidizing bacteria (NOB) are the most important organisms responsible for ammonia and nitrite oxidation in agricultural ecosystems and growing media. Ammonia and nitrite oxidation are critical steps in the soil nitrogen cycle and can be affected by the application of mineral fertilizers or organic fertilizers. The microbial community and structure associated with the growing medium will also be influenced by the plant species identity. The functionality of the microbial community has a major impact on the nutrient turnover and will finally influence plant performance. In our study, we used two different plant species from different functional groups. Lupin, a legume that makes a nitrogen-fixing symbiosis with the microbial community and tomato, both with the availability to exudate nutrient mobilizing acids. We studied plant performance in rhizotrons (a phentotyping system for imaging roots), including an optical method (planar optodes) for non-invasive, quantitative and high-resolution imaging of pH dynamics in the rhizosphere and adjacent medium. The horticultural growing medium was supplemented with organic-derived nitrogen or ammonium derived from struvite. The possible differences in the root structure between treatments is compared with the total root length. Destructive growing medium sampling and high throughput sequencing analysis of the bacterial abundance of the communities present in the rhizosphere and the bulk soil will be used in order to study the growing medium-associated microbial community structure, and this will be related to pH changes in the rhizosphere and the bulk soil. Our hypothesis is that the growing medium-associated microbial community structure changes depending on the nitrogen form provided, the plant species used, the rhizosphere and bulk soil. We expect a higher abundance of bacteria in the treatment with organic fertilizer and a higher abundance of AOB and NOB in the rhizosphere in comparison to the bulk soil. 123 Use of 14C labelled model exudates to evaluate plant species effects on carbon use efficiency in rhizosphere soil Harriet Robson*1, Tom Sizmur1, Andrew Neal2, Liz Shaw1 1 University of Reading, United Kingdom, 2Rothamsted Research, United Kingdom Rhizodeposition, defined as the flow of organic carbon from plant roots to soil, plays an important role in the soil carbon cycle: it represents a major flux of photosynthetic C from plant to soil and also helps shape the structure of the rhizosphere microbial communities that are responsible for the rhizodeposit decomposition. Growing evidence suggests that belowground plant inputs make a large contribution to the stable soil organic carbon pool which is composed mainly of organic carbon that has been processed by the microbial biomass rather than selectively preserved plant litter C. Therefore, it follows that the efficiency by which 285 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session rhizosphere microbial communities process rhizodeposited carbon to microbial products (as 286 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session opposed to CO2; i.e. the carbon use efficiency) will be important for the long-term stabilisation potential of plant-derived C in the rhizosphere. Here we examine how carbon use efficiency in rhizosphere soil varies with respect to plant species with a focus on both legume and non-legume grassland species. We determine carbon use efficiency through tracking the fate of 14 C-labelled model exudates (glucose, succinate, cysteine, salicylic acid) in sampled rhizosphere soil and examine the relationships with rhizosphere microbial community structure. This poster presents results from this ongoing experiment. 124 Effects of bacterial inoculation on sugarcane growth, physiology and assembly of rhizosphere bacterial community Mauricio Rocha Dimitrov*1, Raquel de Paula Freitas2, Adriana Parada Dias da Silveira2, Eiko E. Kuramae1 1 Netherlands Institute of Ecology, Netherlands, 2Instituto Agronômico, Brazil Sugarcane is one of the most important agricultural crops in the world, being source of sugar, renewable energy (biofuel) and biomaterials. Brazil is the largest sugarcane producer in the world and the leader in production of bio-ethanol, a renewable energy source that has been presented as a great potential replacement for gasoline. However, to maximize production, high rates of fertilizers are often applied to the field, which may lead to nutrient leaching, increase in greenhouse gases emissions and reduction in biodiversity. Therefore, a key challenge is to intensify agriculture practices while minimizing harmful environmental effects of intensive fertilization. The use of beneficial bacteria, such as Plant Growth Promoting Bacteria (PGPB), has been shown to enhance plant growth and health in controlled conditions, even with low levels of fertilization. Effects of six bacterial isolates, which had been previously isolated from sugarcane stalks, on sugarcane growth (shoot and root dry mass), physiology (total sugar, sucrose, starch and nitrate concentration, as well as activity of enzymes related to the nitrogen cycle) and assembly of rhizosphere bacterial community were assessed in greenhouse conditions for six weeks. Bacterial isolates and controls were tested in two different treatments regarding nitrogen content (0 and 350 mg N kg-1 of soil). All six isolates promoted a significant increase on shoot and root dry mass in both nitrogen treatments, when compared to controls that did not receive any inoculation. Results of physiological endpoints as well as determination of rhizosphere bacterial community are currently under analysis. Such results will contribute to a better understanding of the effects promoted by bacterial isolates used here on sugarcane plants. Furthermore, result may provide a glimpse of the sugarcane rhizosphere microbiome, as well as indicate whether previous inoculation of beneficial bacteria may affect the formation of rhizosphere bacterial community. 287 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session 125 Microbial community of Bacillus in the rhizosphere, an alternative to improve growth and plant health Ligia Sanchez*, Lucia Corrales, Graciela Lancheros, Yesid Ariza, Estefania Castañeda, Lina Mendez, Andres Zarate, Paola Barrera, Adriana Diaz, Solange Benitez, Cristian Layton, Luisa Monroy, Edna Maldonado, Jenny Celis, Milena Cordero Colegio Mayor de Cundinamarca University, Colombia In Colombia, agriculture plays an important role in the development of the country, geographical position, biological diversity and climatic zones, related to the variety of plants as food for its people and open markets throughout the world. Factors such as technological development, cultural roots and the use of agrochemicals have not allowed high production rates, reason why universities and research centers are focusing their projects towards environmentally friendly alternatives that enable the farmer improve the growth and health their crops. The research group has dedicated its efforts to determine the functional potential from bacteria of the genus Bacillus from indigenous rhizosphere. The aim of this study was to determine if some native gram positive bacilli from rhizosphere of Ornithogallum umbelatum, Thymus vulgaris and Rosmarinus officinalis had capacity to fix nitrogen, solubilize phosphate and produce antibiotics to promote growth and development in plants. The methodology included functional tests with agar Ashby, Pikovskaya, evidence of antagonism against Fusarium sp, and coding presence of genes for several antibiotics. Taking into account the results, it was obtained the most effective antibiotic against the fungus by aerobic fermentation. Finally, the Bacillus was genotyped as potential biochemical antibiotic. It was found that several bacilli grew in culture media Ashby; most microorganisms showed phosphate solubilization and in vitro antagonism against Fusarium sp. All bacillus showed the presence of some gene encoding antibiotic. Even though, one of the rods showed presence of the secondary metabolite Iturin A. They were identified by 16s all bacillus Biochemical potential. It was concluded that the genus Bacillus can be a good alternative for growth and development in plants by their functional versatility and be an environmental friendly option. Currently, a master student of research group is developing a project to microencapsulate bacilli and use it as biofertilizer. 126 Seasonal observation of prokaryotic and root exudate diversity in pneumatophore rhizosphere microhabitat (PRM) of Avicennia marina in Vellar estuarine Dinesh Sanka Loganatha Chetti*, Sundararaman Muthuraman Bharathidasan University, India Pneumatophores are aerial roots originating from the primary roots of mangrove plants. It produces numerous fine roots, and creates a zone around them, called as the pneumatophore rhizosphere microhabitat (PRM). Metagenomic DNA was isolated from PRM associated composite soil samples (n=2) of Avicennia marina, Vellar estuarine with the aim of studying the prokaryotic diversity and root exudates during different seasons. The variable regions of 16s rRNA, V5-V9 (bacteria) and V3-V5 (archaea) regions were amplified from metagenomic DNA. The amplicons were sequenced using a Tag Encoded FLX Amplicon Sequencer (TEFAP). The predominant phylum for bacterial domain was Proteobacteria, irrespective of the seasons. Cyanobacteria was dominant during the monsoon, whereas 288 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session Acidobacteria was dominant in summer. In archaea domain, Thaumarchaeota phylum was dominant in both seasons, while Crenearcheota was higher in summer and Euryarcheota dominant in monsoon. Thermoprotei (class) was three times higher in summer compared to monsoon. Highest bacterial species richness and diversity was observed in summer at 97% sequence similarity. Meanwhile, highest archaeal richness and diversity was observed in monsoon at 97% sequence similarity. Ethanolic extracts (70% v/v) of pneumatophores were identified using gas chromatography-mass spectrometry. Fifteen metabolites during monsoon, fifty two metabolites during summer and twenty one common to both the seasons were observed. PCoA analysis revealed the effect of seasons by placement seasons at opposite ends for archaeal and bacterial phyla. Prokaryotic community structure variation and differential expression of metabolites observed in this study might be attributed to prevailing conditions during seasons. To the best of our knowledge, it is identified that this is the first study to reveal the PRM prokaryotic diversity and metabolites. Further studies needed to affirm the role of these root exudates in shaping prokaryotic diversity in PRM. 127 Abundance and functional traits of rhizosphere yeasts in maize Marcela Sarabia*1, Cecilia Madrigal2, Yazmin Carreon3, Sylvia Fernandez Pavia3, John Larsen2 1 Universidad Nacional Autonoma de Mexico, Mexico, 2Universidad Autonoma de Mexico, Mexico, 3Universidad Michoacana de San Nicolas de Hidalgo, Mexico Yeasts are common soil inhabitants, but little is known about their ecological functions in this environment. In this study we characterized natural populations of yeasts in the maize rhizosphere in nine maize fields with contrasting production systems (low, medium and high input) and soil physico-chemical characteristics (pH, CEC, Base saturation, N, P, K, Ca, Mg, Cu, Na, organic matter and texture). Rhizosphere soil samples were collected at the vegetative, flowering and senescence plant growth stages from which a culture collection of 190 isolates was obtained. Pure yeast isolates were examined for different traits in relation to plant nutrition (P solubilization) and root health (biocontrol against the root pathogens Pythium sp. and Fusarium verticilloides). The results showed that yeasts were abundant (104 - 105 CFU g-1 soil) and diverse (10 morphotypes) in all maize fields during the complete growing cycle. The highest abundance of yeasts was obtained in fields without use of agrochemicals during the florescence plant growth stage. On the other hand, the agrosystem managed intensively with high input of agrochemials presented the highest number of morphotypes. Among the soil physico-chemical characteristics examined the yeast population density correlated positively and negatively with organic matter and percentage of clay, respectively. In terms of function, P solubilization seems to be a common trait in rhizosphere yeasts (45% of the 190 yeast isolates examined solubilized CaPO4 in in-vitro assay), which in some cases coincided with plant growth promotion in corresponding plant bioassays. Another interesting trait of rhizosphere yeasts was a strong antagonistic potential against the maize root pathogens Pythium sp. and Fusarium verticilloides as examined in terms of in-vitro confrontation tests and plant bioassays. In conclusion, yeasts seem to be a common part of the maize rhizosphere microbiome with important functional traits related to improved plant nutrition and root health. 289 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session 128 SYMBIO BANK - the collection of beneficial soil microorganisms Lidia Sas Paszt*, Paweł Trzciński, Beata Sumorok, Anna Lisek, Edyta Derkowska, Sławomir Głuszek, Eligio Malusa, Krzysztof Weszczak, Michał Przybył, Mateusz Frąc The Research Institute of Horticulture, Poland An important part of the project called EcoTechProduct, which is carried out at the Research Institute of Horticulture in Skierniewice (Poland), is to establish and maintain a Bank of Symbiotic Microorganisms, called SYMBIO BANK. The collected material of isolated spores of mycorrhizal fungi and PGPR bacteria comes from organic orchards and plantations in Central Poland, the Bieszczady and Białowieża areas. Results of studies to date have shown that there are large differences in the occurrence of mycorrhizal fungi depending on the species and plant cultivation method. Plant-soil microorganisms can modulate the uptake of mineral nutrients through feedback processes that reflect plant responses to environmental conditions. The intimate interrelation between the root and symbiotic arbuscular mycorrhizal fungi and the resulting enhancement in the uptake of N and P by the plant are further expanded by the interactions between the fungus and bacteria present in both the rhizosphere and mycorrhizosphere. Numerous species of plant growth promoting bacteria form biofilm when colonizing roots, which can affect bio-geochemical processes and can result in increased availability of poorly available mineral nutrients. The soil biotic communities are formed by several kinds of microorganisms that can live symbiotically or in association with roots. Four major groups of microorganisms are considered as beneficial to plants: arbuscular mycorrhizal fungi, plant growth promoting rhizobacteria, nitrogen-fixing rhizobia, which are usually not considered to be PGPR, and microbial biocontrol agents, which are composed of bacteria, yeasts and fungi. The collection in SYMBIO BANK contains over 53 thousands of arbuscular mycorrhizal fungi spores isolated from the soil of the different plant species and over 1500 isolates of rhizosperic bacteria and filamentous fungi. The most effective strains and species of microorganisms will be registered in Poland as bacterial and mycorrhizal inocula to be used in fruit production and in phytoremediation. 129 Characterization of plant growth promoting activities of bacteria in the rhizosphere of barley and tomato Marina Scagliola*1, Patrizia Ricciuti1, Youry Pii2, Tanja Mimmo2, Stefano Cesco2, Carmine Crecchio1 1 University of Bari "A. Moro", Italy, 2Free University of Bolzano, Italy Plant growth promoting rhizobacteria might be an alternative to chemical fertilizers and pesticides as they influence several mechanisms such as enhancing nutrient uptake, plant pathogens suppression and phytohormone production. This work aims at isolating and characterizing rhizobacteria and their beneficial activities like siderophores and indole 3-acetic acid (IAA) production and phosphate solubilization. 290 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session Microorganisms were isolated from rhizosphere of barley and tomato, which were grown in the RhizoTest-System first hydroponically (either in a full nutrient solution or in an iron deficient solution) and then in soil. Two hundred isolates for each soil sample were screened and eighty isolates were selected on the basis of their ability to produce siderophores. Siderophores were further purified by chromatographic analysis and characterized by spectrophotometric analysis. To investigate their capability to produce IAA, isolates were tested by colorimetric method using Salkowski reagent; the presence of IAA was confirmed and quantified by HPLC. Their capability to solubilize phosphates was evalutated on Pikovskaya agar; quantitative estimation was performed in liquid medium by spectrophotometric method using ascorbic acid. Isolates were also characterized by molecular analysis. A region of the 16S rRNA gene of about 1 kb was amplified and sequenced from both ends. Sequences were aligned by BLASTn to 16S rRNA sequences available on NCBI. Molecular evolutionary and phylogenetic analysis was performed using Seaview 4; phylogenetic trees were constructed according to the maximum likelihood method. Phylogenetic analysis of the selected population was not able to differentiate the microbiome evolved by different plant species in function of the nutritional status (Fe deficiency vs sufficiency). Forty-seven isolates, mostly belonging to genus of Pseudomonas, Azotobacter and Rhizobium, exhibited high levels of the three activities simultaneously. Further studies are needed to characterize the role of the selected microorganisms on the mechanisms underlying the nutrient acquisition in plants. 130 Exploring the mycorrhizosphere of urban trees Angelika Schartl*1, Josef Valentin Herrmann1, Arthur Schuessler2 1 Bavarian State Institute for Viticulture and Horticulture, Germany, 2SYMPLANTA Laboratory, Ghana Urban trees grow in an unnatural environment. Limited tree pits restrict their rooting zone, degraded, compacted soil disturbs water and air balance. Multiple pollutants as well as frequent mechanical damage affect the tree vitality. Additionally, climate change has worsened these stressful conditions. Under such adverse circumstances mycorrhizal associations play an even more significant role for plant health. To select urban trees for the future, a long-term project was started in 2010. Different tree species were planted at three sites with differing climate conditions. Some, representing different mycorrhizal preferences, were chosen (Carpinus betulus, Fraxinus pennsylvanica ’Summit’, Magnolia kobus, Ostrya carpinifolia, Parrotia persica, Quercus cerris, Tilia tomentosa ’Brabant’) for root monitoring, especially to scrutinize their mycorrhization status. At planting half of the trees were inoculated with a commercial mycorrhiza product (INOQ). 291 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session Since 2011 root monitoring has been performed twice a year. Besides microanalysis of mycorrhizal fungi, molecular biological analyses were performed for species identification by BLAST against the UNITE and NCBI databases, including a deep sequencing by SYMPLANTA in 2012. In all trees tested both ecto- and endo mycorrhiza could be detected, except for Magnolia, where only endomycorrhiza was present. The frequency was always higher than 50% and displayed seasonal changes. Intensity of endomycorrhiza colonization averaged 15%. A difference between inoculated and non-inoculated trees could not be detected. Sequence analyses revealed a complex community of mycorrhiza on all trees with individual patterns. Nevertheless, some characteristic associations between tree and fungal species could be detected. Conventional sequencing rarely identified endomycorrhizal fungi. A connection of mycorrhizal patterns to nursery origin and/or cultivation site has to be discussed. 131 Effect of plant genotype and soil type on the rhizosphere microbiome structure of Sorghum bicolor (L.) Moench and Strigolactone production Thiago Roberto Schlemper*, Eiko Kuramae, Jos Raaijmakers, Hans van Veen Netherlands Institute of Ecology - NIOO-KNAW, Netherlands In terrestrial systems, rhizosphere microbes are important players in plant growth, plant health and nutrient acquisition. Here we determined if different Sorghum bicolor cultivars select different microbial assemblages in the rhizosphere. Originated from Africa, S. bicolor is a valuable staple crop mainly used for feed and food. S. bicolor is the major cereal crop in the semi-arid regions of the world, in particular in sub-Saharan Africa. To determine the effect of the Sorghum genotype on the rhizosphere microbiome structure, a greenhouse experiment was conducted with 7 different cultivars, planted in two different soils. Bulk soil and rhizosphere samples were taken at 4 different time points representative of different developmental stages of Sorghum. The rhizosphere and bulk soil samples were subjected to total genomic DNA extraction and the bacterial/archaeal communities were assessed by 16S rRNA gene sequencing and the fungal community by 18S rRNA gene sequencing. We will present results on soil - and host - genotype specific effects on the rhizosphere microbiome of S. bicolor and will provide insight into the correlation of microbiome structure and Strigolactone exudate profiles. 132 Diversity and spatial distribution of diazotrophs associated with microenvironments of wetland rice Hannes Schmidt*, Dagmar Wöbken University of Vienna, Department of Microbiology and Ecosystem Science, Division of Microbial Ecology, Austria Rice is one of the world’s most important crop plants. The production is strongly limited by nitrogen (N), which is typically supplied by industrial fertilizers that are costly and hazardous 292 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session to the environment. It is known that Biological Nitrogen Fixation through N2-fixing bacteria and archaea (diazotrophs) can alleviate the N-shortage in rice cultivation. However, our knowledge on the micro-sites of N2 fixation, as well as the diversity and in situ N2 fixation activity of diazotrophs in the soil-microbe-plant interface (i.e. rhizosphere) of flooded rice fields is still rudimentary. A greenhouse study was performed to identify key factors that control rice-diazotroph association and related N2 fixation activities. Paddy soils were cultivated with two genotypes of wetland rice. Samples were taken from the bulk soil, the root-associated rhizosphere soil, the rhizoplane, and the endosphere at flowering stage of rice plant development. These samples were subjected to functional assays and various molecular biological techniques including high-throughput sequencing to identify the diazotroph community. Based on Illumina sequencing of 16S rRNA and nifH genes and transcripts, we will present first insights into the diversity of bacterial/diazotroph communities with emphasis on assessing the potential influence of (a) the soil microbial “seed bank” or (b) plant genotype in shaping the microbiomes in each micro-environment. These data will be combined with N2 fixation activity measurements as assessed by incubation assays using acetylene-enriched atmospheres. The localization of selected diazotrophs on the rhizoplane of rice roots via fluorescence in situ hybridization and confocal laser scanning microscopy is currently underway to identify areas of potential N-transfer between diazotrophs and rice roots. 133 Evaluation of strategies for the separation of root-associated microbial communities Tim Richter-Heitmann1, Thilo Eickhorst2, Michael W. Friedrich1, Hannes Schmidt*3 1 University of Bremen, Microbial Ecophysiology, Germany, 2University of Bremen, Soil Microbial Ecology, Germany, 3University of Vienna, Department of Microbiology and Ecosystem Science, Division of Microbial Ecology, Germany The analysis of plant-microbe interactions has emerged as an important topic in rhizosphere research. Based on comparative studies applying high-throughput sequencing, it has been postulated that plants may shape distinct root microbiomes consisting of microbial populations attracted from the soil seed bank. A main premise for the value of such investigations is that microorganisms living near, on, and inside roots were well separated.We applied different protocols to plant roots of two soil-grown plant species (Oryza sativa, Vicia faba) and assessed their potential to detach rhizoplane colonizing cells by washing, sonication, and treatment with NaOCl. Sonication was performed with (i) sonication bath and (ii) sonication probe at low and high intensity, respectively. The number of microbial cells attached to the rhizoplane after each treatment was evaluated via SYBR Green-staining and fluorescence microscopy. Accordingly, cell numbers of rhizoplane-detached cells and the estimated level of plant cell destruction were recorded. Community fingerprinting and next generation sequencing of root samples is applied to assess the effect of each treatment on the composition of microbial populations associated with roots. First results showed that washing alone yielded in the majority of cells still being attached to root surfaces. The highest amount of cells detached from rice roots was found with the 293 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session sonication bath system at highest energy. However, the intensity of plant cell destruction increased as well, leading to a potential release of endophytes into the rhizoplane compartment. Our results also suggest that presence of root hairs - as found in Vicia faba interfered with all procedures as the numbers of cells detached from the rhizoplane did not increase with increasing intensity of the applied treatments. Hence, we conclude that strategies to separate root associated bacterial communities need to be carefully evaluated for each plant genotype, which can be easily performed by fluorescence microscopy. 134 A fragrant neighborhood: The role of volatiles in the formation of bacterial communities in soil Kristin Schulz*1, Wietse De Boer2, Paolina Garbeva1 1 Netherlands Institute of Ecology, Netherlands, 2Netherlands Institute of Ecology, Wageningen University, Netherlands Soil bacterial species living in the rhizosphere interact in numerous and versatile ways. It is increasingly recognized that volatile organic compounds (VOCs) produced by bacteria can function as bioactive growth-promoting or growth-inhibiting agents as well as info-chemicals for inter- and intra-specific communication. Due to their physical properties, VOCs can act on a wide scale in soil and, consequently, may play a key role in interspecific bacterial interactions and in the development of microbial soil communities. Soil microcosm experiments with bacterial model strains growing on artificial root exudates are performed to assess (1) the influence of volatiles on the formation of bacterial communities and (2) the ability of VOCs to stimulate non-active bacteria. Pilot microcosm experiments with monocultures and mixture of bacterial isolates (Burkholderia, Dyella, Janthinobacterium, Pseudomonas, and Paenibacillus) revealed that each strain produced a different set of volatiles and that the production of different volatile compounds was triggered by interspecific interactions in the mixture. Interestingly, the poorly-growing Gram-positive Paenibacillus strain had a strong effect on the growth-performance of the other strains as well as on the production of volatiles in the mixture. We are currently testing how trapping (removal) of microbial volatiles from the soil microcosms will affect the development of the microbial community. Furthermore, we are testing if VOCs produced by an active bacterial community can stimulate the growth of nonactive (starving) bacteria. In conclusion, with the obtained results we aim to reveal new insights into the ecological role of volatiles in microbial interactions and community dynamics in soil. 294 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session 135 Deciphering plant growth promoting rhizobacteria from rice rhizosphere soil and plant growth promotion in rice Lalan Sharma*, Dipak T. Nagrale, Dhananjaya P. Singh Indian Council of Agricultural Research - National Bureau of Agriculturally Important Microorganisms, India Sampling of rhizospheric soil samples have been done of different rice varieties of IndoGangetic plain of Uttar Pradesh. Soil samples have analysed for physico-chemical properties like pH, electric conductivity and organic carbon. It recorded that soil pH ranges 7.0 to 8.4, electric conductivity ranges 1.3 to 1.8ds/m and organic carbon in soil medium to medium high and free living rhizospheric bacteria were isolated by using different inoculation techniques on various culture media. 143 rhizobacteria have isolated and characterized by Gram staining, Streo-microscopically, Scanning Electron Microscope and plant growth promoting attributes like HCN production, Siderophore production, and Phoshphate solubilisation. 16s rDNA amplification has been done by using universal primers and also phenolic chromatogram has prepared by High Performance Liquid Chromatography for secondary metabolites detection. Seed bio-priming has done with potential rhizobacterial isolates using Gnotobiotic system. The data are recorded and calculated that some isolates either individually or in consortium form like MAU143, MRT84 and MRT92 are promising microbial inoculants to enhance rice seed vigour and antioxidants properties in rice seedlings. 136 Characterisation of medium-term selection of Rhizobium leguminosarum bv viciae populations by different legume plant hosts Amalia Soenens Martinez de Murguia*1, Beatriz Jorrin1, Juan Imperial2 1 Universidad Politécnica de Madrid, Spain, 2Universidad Politécnica de Madrid / Centro Superior de Investigaciones Cientificas, Spain Rhizobium leguminosarum bv viciae is a nitrogen-fixing soil bacterium able to establish specific root-nodule symbioses with legumes of four different genera: Pisum, Vicia, Lens and Lathyrus. Available evidence suggests that although Rhizobium leguminosarum bv viciae strains are able to nodulate the four plant genera, plant hosts select specific rhizobial genotypes from those present in the soil. This has been shown previously in our laboratory at the genomic level following a population genomics approach (Pool-Seq). Pool genomic sequences from 100 isolates from each of four plant species: P. sativum, L. culinaris, V. faba and V. sativa show different specific profiles at the single nucleotide polymorphism (SNP) level for relevant genes. In this work we characterised the extent of plant host selection on Rhizobium leguminosarum bv viciae genotypes both in nodules and in the soil. For that purpose we proceed to do a mesocosm experiment, where we started with a wellcharacterised soil population, applied plant host selection and those rhizobia selected by the different legume hosts: P. sativum, L. culinaris, V. faba and V. sativa were used as the inoculum for the next plant growth. This was done for 5 cycles. Direct soil and nodule individual strains from each of these mesocosm studies have been isolated and initially tested for specific rhizobial genes (glnII and fnrN) and symbiotic genes (nodC and nifH). The selected populations were further characterised by means of Sanger sequencing of both the rpoB phylogenetic marker gene and the symbiotic genes nodC and nifH. The distribution and composition of the different rhizobial populations showed changes for each legume host 295 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session during the mesocosm study. We hypothesize that these changes mimic those occurring under real field conditions and should help reveal the underlying bases of rhizobial genotype selection by the plant host. 137 Changes of the bacterial community in the rhizosphere of wild soybean (Glycine soja) at different growth stages Youngmi Lee1, Jae-Hyung Ahn1, Hang-Yeon Weon1, Jung-Hoon Yoon2, Jaekyeong Song*1 1 Agricultural Microbiology Division, National Academy of Agricultural Science, RDA, South Korea, 2Department of Food Science and Biotechnology, Sungkyunkwan University, South Korea We analyzed bacterial community of rhizosphere at four growth stages of soybean, Glycine soja using pyrosequencing based on the 16S rRNA genes. In the bulk soil and the rhizosphere of wild soybean the predominant phylum was Proteobacteria (32-42%) and followed by Firmicutes (7-27%), Actinobacteria (7-12%) and Bacteroidetes (3-16%). The rhizosphere of wild soybean had higher abundance of the phyla Proteobacteria and Bacteroidetes than bulk soils. In the class level, Gammaproteobacteria was much more abundant in bulk soils, while Alphaproteobacteria and Betaproteobacteria were more abundant in the rhizosphere of wild soybean. The predominant genus was Bacillus in the rhizosphere of the wild soybean, followed by Flavobacterium. There were some changes in bacterial community structure of the rhizosphere of wild soybean during growth unlike those of bulk soils. Of which, the abundance of the genus Bacillus changed markedly between growth stages, full seed stage and maturity stage of wild soybean, indicating that Bacillus responds preferentially to change of materials such as root exudates of wild soybean. 138 Defining the microbiome of a sugarcane plant Rafael Souza*1, Vagner Katsumi Okura1, Nuria Lozano García2, Jaderson Silveira Leite Armanhi1, Laura Migliorini de Araujo1, Natalia Verza Ferreira1, Manuel GonzálezGuerrero2, Homayoun Bagheri3, Marcio José da Silva1, Juan Imperial1, Paulo Arruda1 1 State University of Campinas (UNICAMP), Brazil, 2Centro de Biotecnología y Genômica de Plantas (CBGP), Campus de Montegancedo, Universidad Politécnica de Madrid, Spain, 3Repsol Technology Center, Spain Studies on beneficial microorganisms associated to crops have traditionally used culturebased techniques, which are known to sample only a minute portion of the microbial community. Sugarcane has been a target of such researches, since there are repeated evidences of a microbial community supporting plant development and sustaining high yields under conditions of little or no fertilization. However, none study has taken into consideration the diversity or the qualitative/quantitative microbial communities in the biology of the plant. We intend to unravel the microbial community associated to sugarcane plants cultivated for four cuts without fertilization. Exophytic and endophytic microbial communities were evaluated in root, stalk, leaf and young shoot. We also accessed the microbial communities of bulk soil. Illumina platform was used to sequence 16S and ITS 296 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session amplicons. Our data revealed that factors driving the community composition differ between prokaryote and fungi. Although the microbial diversity is huge for any tissue, a core microbiome comprised of less than 1% of the diversity represents 65% to 96% relative abundance of the total microbial diversity. Interestingly, the most abundant microbial groups are not the ones that have been isolated and evaluated over the past decades in sugarcane. The data open a new window for the investigation of untargeted microbes potentially beneficial for sugarcane growth and development. 139 Deciphering the cross-talk between host genotype, microbiota structure, and plant growth in barley Rodrigo Alegria Terrazas*1, Katharin Balbirnie2, Eric Paterson3, Elisabeth Baggs4, Davide Bulgarelli2 1 The University of Dundee at The James Hutton Institute, United Kingdom, 2University of Dundee, United Kingdom, 3The James Hutton Institute, United Kingdom, 4University of Aberdeen, United Kingdom The rhizosphere microbiota is a potential resource of plant probiotic functions. However, its deployment in agriculture is impaired by the limited knowledge of the mechanisms regulating plant-microbiota interactions. Wild barley (Hordeum vulgare subp. spontaneum) populations from marginal soil areas are considered a genetic resource for crop improvement. To investigate the contribution of the wild barley-microbiota interactions to plant growth, we have grown 14 wild populations, representing the three major ecotypes in Israel (i.e., ‘north’, ‘coast’ and ‘desert’), and a modern variety in a reference agricultural soil under greenhouse conditions. At early stem elongation, rhizosphere specimens were collected and stem dry weight measured. We adopted an llumina MiSeq protocol to generate high-resolution 16S rRNA gene profiles of the rhizosphere and unplanted soil controls. We used QIIME to quality-filter the reads and to identify Operational Taxonomic Units (OTUs) at a 97% sequence similarity. Consistent with previous investigations, we observed a taxonomically coherent barley microbiota, dominated by members of the phyla Proteobacteria and Bacteroidetes, whose enrichment discriminates rhizosphere and soil profiles. Beta diversity calculation revealed distinct host-dependent substructures of the rhizosphere microbiota. In particular, the microbiota retrieved from the desert ecotype accessions clustered separately compared with those from other ecotypes and the modern variety. Non-parametric statistical analyses and supervised learning classification indicated that the recruitment of members of the families Comamonadaceae, Cytophagaceae, Hyphomicrobiaceae and Micrococcaceae support the observed diversification, several of which have previously been reported as plant growth promoting rhizobacteria. Remarkably, host-dependent microbiota profiles correlated with a biomass gradient among the tested plants, underlying a potential cross-talk among the host genotype, the establishment of substructures of the microbiota and plant growth. To further explore these relationships, we are currently investigating the contribution of the barley rhizosphere microbiota to plant adaptation to soil limiting nutrient supplies. 297 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session 140 Rhizobacterial community structure and function in a dryland agroecosystem Linda Thomashow*1, James Parejko2, Melissa LeTourneau3, Olga Mavrodi4, Dmitri Mavrodi4, Robert Bonsall3, David Weller5 1 USDA-ARS Washington State University, United States, 2University of Wisconsin La Crosse, United States, 3Washington State University, United States, 4Southern Mississippi State University, United States, 5USDA-ARS, Washington State University, United States Certain members of the Pseudomonas fluorescens species complex produce phenazine antibiotics inhibitory to soilborne root pathogens. Included among these bacteria are the model strain 2-79 and representatives of three other Pseudomonas species that produce phenazine-1-carboxylic acid (PCA) and comprise up to 10% of the culturable rhizobacteria on the roots of wheat grown in dryland soils of the inland Pacific Northwest, USA. PCA production on roots of field-grown spring wheat occurred mainly during the first half of the growing season but remained detectable for at least 130 days after planting. In vitro, the compound persisted with a half-life of 3 or 4 days in dry or moist soils, respectively, and remained detectable throughout the 20-day experiment. These data indicate that most PCA synthesis in the field occurs early in the season, but suggest that some synthesis continues to occur even as soil water potentials approach as low as -400 to -500 kPa. We suggest that PCA producers (Phz+) survive desiccation on roots in biofilms. PCA influenced biofilm formation by some, but not all, Phz+ strains from dryland wheat and its impact varied with matric and osmotic stress levels. Structural differences also were observed in colony biofilms grown from strains of different species under control and stress conditions, and distinct differences were observed between some wild-type Phz+ strains and their Phz- mutants. 141 High-throuput detection of plant pathogens using next generation sequencing Andrzej Tkacz*, Philip Poole Plant Sciences, University of Oxford, United Kingdom The soil microbial community has an enormous impact on crop yields. There has been extensive research conducted using cultivation and environmental DNA and RNA sequencing methods. However, only recently with the advent of the new high-throughput sequencing methods are we starting to understand the delicate structure of the soil microbiota, both in its spatial and temporal scale. Here we present our recent findings on the soil community inter- and intra-kingdom structure caused by the rhizosphere influence of different plants. We are detecting pathogen build-up in the soil of plants grown in monoculture. Using selected plant species and secretion mutants, we are correlating pathogen invasion of the rhizosphere with plant host genetics. We are especially interested in wheat lines and their progenitors, rice secretion mutant lines and signalling mutants of Medicago. 298 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session 143 Comparison of the diversity, structure and function of the fungi and bacterial rhizosphere community of main rice production areas from Colombia (SA) Daniel Uribe*1, Giovanna Landazabal2, Catalina Camelo2, Javier Vanegas3 1 Universidad Nacional de Colombia/ Biotechnology institute, Colombia, 2Universidad Nacional de Colombia/ Biotechnology Institute, Colombia, 3Universidad Nacional de Colombia / Biotechnology Institute, Colombia The deterioration of agricultural soils, mainly represented by the erosion, loss of fertility and environmental contamination with different types of substances, has resulted in decrease of productivity and sustainability of many agroecosystems. This phenomenon, has increased the interest to assess the impact of agricultural practices on soil microbial communities, in relation to soil ecological function in specific agroecosystems. Studies carried out for this purpose, have shown that the activity, diversity and structure of microbial communities are affected by different management practices and land use. In Colombia, the intensive cultivation of rice, the third largest crop in terms of planted area, has led to a progressive deterioration of the soil, which is reflected in the reduction in rice production levels, especially in Tolima and Meta, the two largest rice production areas of Colombia. This study compared the structure, diversity and enzymatic activity of rhizosphere bacterial and fungal communities of 16 rice crop farms, from Tolima and Meta, using denaturing gradient gel electrophoresis. Sixteen farms located in four areas contrasting in terms of fertility, physicalchemical characteristics and the rice production system in terms of water management were studied. Cluster and principal component analysis (PCA), showed that the structure, diversity and function of rhizosphere bacterial and fungal communities were influenced by the physical and chemical properties of soils and rice production system. Most bacterial and fungal OTUS identified belongs to unculturable species. 144 Rhizosphere bacterial community composition of the native potato Solanum tuberosum group phureja associated with nitrogen and phosphorus cycle Daniel Uribe*1, Nathalia Florez-Zapata2, Anyela Rodriguez2 1 Universidad Nacional de Colombia/ Biotechnology institute, Colombia, 2Universidad Nacional de Colombia/ Biotechnology Institute, Colombia Soils where Solanum tuberosum group phureja is cultivated are characterized as being highly soil phosphate binders. Besides, it is well known the important role of nitrogen in agricultural production. Therefore, to meet the nutritional demands of the crop, high phosphorous and nitrogen based fertilizers are applied, which increase production costs, as well as undesirable environmental effects. Because of that, some farmers apply organic amendments as an alternative to ameliorate such negative effects. However, the effects of such practices, as well as the soil type on S. tuberosum group phureja P an N related microbial communities are still unknown. The aim of this study was to analyze the edaphic communities related to phosphorous and nitrogen metabolism in spatially independent rhizosphere soil samples from S. tuberosum group phureja crops, contrasting in terms of soil structure and fertilization strategy. Such analysis was done through a polyphasic approach, which includes physicalchemical soil analyses, enzimatic activity quantifications, and culture-dependent and independent microbial communities’ composition analysis, in order to identify the factors that regulate P and N cycling in this ecosystem. Statistical significant differences were found 299 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session in microbial counts, community composition and enzimatic activities in the four soil samples. Besides, differences were found in terms of the effect of soil type on the structure and function of N and P associated bacteria, suggesting differences in the susceptibility of those two functional groups to the soil physical-chemical environment. 145 Forest tree species shape their soil microbiome Stephane Uroz*1, Oger Phil2, Tisserand Emilie1, Cébron Aurélie3, Turpault Marie-Pierre1, Marc Buée1, Wietse De Boer4, Johan H Leveau5, Pascale Frey-Klett1 1 INRA, France, 2CNRS, France, 3CNRS/Univ. Lorraine, France, 4NIOO, Netherlands, 5University of California, USA The impact of plant species on the soil microbial communities and the physico-chemical characteristics of the soil begins to be well documented especially for non-perennial plants. However, our understanding of the diversity and structure of the tree-associated microbial communities as well as the of tree species effect remains limited. Here, we investigated the archaeal, bacterial and fungal communities in replicate soil samples, using 16S rRNA, 18S rRNA and fungal ITS sequences, in the long term experimental site of Breuil-Chenue. We showed significant difference in abundance, composition and structure of the microbial communities associated to two contrasted tree species, Fagus sativa and Picea abies developed on the same soil. Our results highlighted a strong host effect on the soil microbial communities, with a stronger effect on the fungal communities. Although the pyrosequencing approach showed a limited rhizosphere effect, quantitative PCR revealed a significant enrichment of specific bacterial genera known for their ability to weather minerals in the tree root vicinity. At last, co-occurrence analysis revealed very different networks below the two tree species, suggesting a modification of the structure and abundance of the microbial communities, but also modification of the interactions established between microorganisms. In all the microbial communities considered we observed a host effect with variable intensity, suggesting that the tree host shapes its soil microbiome. 146 Organic amendment effects on lettuce production and rhizosphere soil Julen Urra*1, Carlos Garbisu1, Jose Maria Becerril2, Unai Artetxe2, Fernando Blanco1, Iker Martin1, Mikel Anza1, Iker Mijangos1 1 Neiker-Tecnalia, Spain, 2University of the Basque Country, Spain Intensive farming practices, and in particular the utilization of synthetic chemical fertilizers, lead to the degradation of the environment, including the soil resource. In consequence, there is growing interest in the development of sustainable agricultural practices which protect the integrity of the soil ecosystem while producing healthy and abundant crops. In this respect, fertilization with organic amendments not only provides a wide range of nutrients but also improves soil physicochemical properties and, most relevantly, might have a beneficial effect on soil microbial communities. Here, the effect of organic amendments from chicken and horse manure (i.e., fresh manure, compost, Bokashi compost), as well as the effect of a liquid amendment obtained from forest 300 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session leaf litter and Biovin®, on lettuce production and rhizosphere soil was studied under controlled conditions in a growth chamber. The effect of organic amendments on lettuce rhizosphere soil was investigated through the determination of a variety of physicochemical (pH, organic matter, nutrient concentrations, cation exchange capacity, texture, etc.) and microbial properties. Pertaining to these latter microbial properties, we determine a great deal of parameters which provide information on the biomass (e.g., microbial biomass C, substrate-induced respiration, RT-qPCR for bacteria and fungi), activity (e.g., RNA/DNA ratio, basal respiration, potentially mineralizable N, and enzyme activities such as β-glucosidase, urease, alkaline phosphatase and arylsulphatase) and diversity of soil microbial communities. The application of organic amendments resulted in optimal lettuce yields while stimulating the activity of soil microbial communities. 147 Microbiomics of cormosphere and rhizosphere of Crocus sativus, Saffron Jyoti Vakhlu*, Deepika Trakroo, Sheetal Ambardar University Of Jammu, India Microbial associations with the roots have been explored in a great detail but other belowground plant parts like corm, bulb and onion etc have been ignored so far. Crocus sativus, Saffron, is world’s costliest spice and has interesting corm-root cycle. Our laboratory for the first time has initiated the study on the microbial (bacterial and fungal) association of below ground parts of Crocus sativus i.e. corm and root and comparison of their specific associations. The study was initiated by cataloguing the bacterial diversity during vegetative stage and fungal diversity during flowering stage by cultivation-independent metagenomic approach, involving deep sequencing of phylogenetically relevant genes amplicons i.e 16S rRNA gene for bacteria and ITS for fungi. Bacterial diversity of cormosphere was high as compared to rhizosphere which was also represented by higher Alpha diversity in cormosphere (11) than in rhizosphere (9). Proteobacteria was the dominant bacterial phylum in both rhizosphere and cormosphere with greater abundance in cormosphere. In rhizosphere Lutteibacter (10.4%) was found to be predominant genus followed by Rhizobium (6.77%) and Sphingomonas (5.68 %). However Chryseobacterium (7.6%; Bacteriodetes phylum) was abundant genus in cormosphere followed by Sphingomonas (5.5%) and Burkholderia (3.1%). In fungi, Zycomycota was the dominant phylum in both rhizosphere and cormosphere but its relative abundance was higher in cormosphere (54%) than in rhizosphere (43%). The Shannon alpha diversity was found 6 and 7 for cormosphere and rhizosphere, respectively, indicating higher number of fungal species in rhizosphere than cormosphere. At genus level Rhizopus (Zygomycota) was dominant in rhizosphere. In cormosphere though Zycomycota was dominant phylum but Cryptococcus (Basidiomycota) was dominant genus. In this preliminary investigation there is clear indication of organ specific microbial– association. However what is the influence of such association, especially microbe–corm association is matter of further investigation. 301 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session 148 In depth exploration of the rhizosphere and endosphere of maize Tom Viaene*1, Raul Tito2, Hilde Nelissen1, Jeroen Raes2, Sofie Goormachtig1 1 VIB, UGENT, Belgium, 2VIB, KU Leuven, Belgium We have initiated an in-depth characterization of the microbiome composition in the rhizosphere and endosphere of maize in a Flemish soil type. Using 16S rRNA pyrosequencing on the Illumina Miseq platform, relative abundance values of the different bacterial phyla show a gradual transition from bulk via rhizosphere to endosphere samples, with the endosphere being most different. At the family level, we are able to identify bacterial families that are significantly upregulated in the endosphere (and/or) rhizosphere, compared to bulk soil samples. These are families (e.g. Rhizobiaceae, Comamonadaceae and Streptomycetaceae) to which well-known plant growth promoting bacteria belong. To understand the functionality of the microbiome data, we have developed two plant growth promoting assays in maize to evaluate the effect of selected microbes on maize growth. These assays are currently used to screen plant growth promoting effects of individual corn rhizosphere bacteria. 149 Impact of soil heat treatment on bacterial community reassembly in the rhizosphere Menno van der Voort1, Rodrigo Mendes*2, Jos M. Raaijmakers3 1 Wageningen University, Netherlands, 2Embrapa Environment, Brazil, 3Netherlands Institute of Ecology (NIOO-KNAW), Netherlands The rhizosphere microbiome offers a range of ecosystem services to the plant, including nutrient acquisition, tolerance to abiotic stress and protection against diseases. Here we studied how heat treatment of soil disturbs the reassembly of the bacterial community in the rhizosphere and how this affects tolerance to pathogen infection. Using PhyloChip-based community profiling, we assessed the impact of 1-hour heat treatments of 50ºC or 80ºC on the bacterial community composition in the rhizosphere of sugar beet seedlings grown in a soil that is naturally suppressive to the soil-borne fungus Rhizoctonia solani. The heat disturbance caused significant increase of alpha diversity and led to a partial (50ºC) or complete (80ºC) loss of protection against fungal infection. The bacterial families Bacillaceae, Comamonadaceae, Paenibacillaceae and Alcaligenaceae showed a significant increase in relative abundance with increasing temperatures. The Pseudomonadaceae and Burkholderiaceae showed higher abundance only when the soil was heat-treated at 80ºC. Conversely, the bacterial families Streptomycetaceae, Micrococcaceae, Solibacteraceae and Mycobacteriaceae showed a reduction in relative abundance when the soil was heat-treated at 80ºC. Based on these results, we propose a reassembly model where bacterial groups that are most heat-tolerant and with high growth rates increase in relative abundance after heat disturbance, while temperature-sensitive and slow growing bacteria have a disadvantage. The results also point to a potential role of slow growing bacterial families from Actinobacteria and Acidobacteria phyla in protection of plants against fungal infection. With this study we showed that heat disturbance in soil results in a rearranged rhizosphere bacterial community, which in turn leads to changes in the ecosystem services of the soil. 302 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session 150 The potato microbiome and its potential impact on late blight resistance Mout De Vrieze*, Aurélien Bailly, Christian Ahrens, Tomke Musa, Laure Weisskopf Agroscope, Switzerland Late blight caused by the oomycete Phytophthora infestans is a major threat for potato production worldwide. In organic farming, control is based on the use of copper-based fungicides, which are the only efficient control products currently available. However, due its toxicity towards the environment, the use of copper compounds for agricultural purposes is being questioned. In Europe, the current approval expires in 2018 and the EU aspires at reducing the use of copper in the future. The need to develop alternative organic control methods is therefore evident. In natural and agro-ecosystems, plant roots and shoots are colonized by a diverse community of microorganisms. In the model plant Arabidopsis thaliana, a protective role of this plant’s microbiome against a number of phytopathogens has been demonstrated. However, the composition of the potato leaf microbiome is so far unknown, as is its putative role in protecting the plant against pathogens such as P. infestans. In this project, the microbiome of non-infected and P. infestans-infected potato plants of various cultivars will be characterized using next-generation sequencing techniques with the aim to reveal its composition and the shifts occurring when the plants are infected with P. infestans. Furthermore, the genomes of selected Pseudomonas strains recently isolated from field-grown potatoes and showing antagonistic potential against P. infestans will be sequenced to identify the genomic determinants of the anti-Phytophthora activity. These findings should provide better understanding of the mechanisms involved in the antiPhytophtora activity of the bacteria and reveal how these mechanisms are regulated at the molecular level. The ultimate goal of this study is to improve our use of bacteria as biocontrol agents through a deeper understanding of their metabolic possibilities and of their needs. 151 Exploring the root-associated bacterial community turnover along a primary succession in salt marsh Miao Wang*, Joana Salles University of Groningen, Netherlands The root-associated microbiome represents the bridge between soil and plant hosts, being influenced by biotic and abiotic factors. Although soil characteristics provide a stronger selective force than plant species, i.e. the same plant species growing in different soils select for distinct bacterial communities, these comparisons are mostly performed in soils with distinct characteristics, origins and from different environments. Primary succession fields provide a perfect setting to test the relative influence of soil and plant type on plantassociate microbiome, as the same plant species grows along a gradient of soil development, under similar environmental conditions. Here, we used an undisturbed salt marsh chronosequence, spanning more than 100 years of primary succession, to study the bacterial communities associated with the soil, rhizosphere and the root endopshere of Limmonium vulgare using 454-pyrosequencing. Our hypotheses were that soil characteristics would determine community composition and that selective force exert by the endosphere would be stronger. The soil and rhizosphere bacterial communities were phylogenetically more diverse than those in endosphere, which could be explained by plant “filtration”. Moreover, this diversity remained stable over the chronosequence. Multivariate analyses showed that 303 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session endopshere communities were distinct from the others and did not cluster by soil age. Conversely, rhizosphere and soil-associated communities were similar and grouped according to succession stage, confirming our first hypothesis for rhizosphere communities. The variation in the bacterial community assembly, analyzed by co-occurrence network matrices, indicated bacterial species turnover along the succession to be the highest in the endosphere and lowest in the rhizosphere. Contrary to the perspective of more stable community composition in the endosphere due to less competition compared to the soil or rhizosphere, we showed that the rhizosphere performed higher selective force than endosphere, the latter being under the control of stochastic mechanisms and colonized by passenger endophytes. 152 Dilution of microbial soil community selects for K-strategists (serial dilution approach) Simone Weidner*, George Kowalchuk, Alexandre Jousset, Joost Keuskamp Utrecht University, Netherlands Soils are hot spots of diverse microbial communities. Soil microbial communities are important for plants in terms of being the species pool from which the plant recruits its rhizosphere microbes. However, there is not much known about how possible diversity loss might be linked to changes in microbial community structure. We created a diversity gradient in a microbial soil community using a serial dilution approach. Briefly, the microbial community from a natural soil was extracted, diluted, and γ irradiated soil was inoculated with these microbial communities. Although it is known that dilution decreases the diversity by out-selecting rare microbes, much less is known about structural changes in the soil microbial community which result from competitive interactions during the regrowth period after inoculation. In order to secure equal microbial biomass across the treatments, soils were incubated in the dark at 20°C for nine weeks before first analyses. Potential growth rates of the communities were determined via substrate induced growth respiration and used as proxy for the dominance of r vs K-selected species. Biolog plates were used to assess whether changes in community structure affected carbon resource use patterns. This study shows that dilution causes a bias towards r-selected species upon regrowth with moderate dilutions, whereas strong dilutions lead to K-selected communities. We suggest that the connectivity during the regrowth period determines the dominance of K vs rselected species after the regrowth period. Detected changes in carbon resource use patterns across dilution might result in effects of root exudate composition on the community and recruitment of rhizosphere microbes. As a next step we will investigate how these changes in community structure affect plants’ recruitment of certain microbes from bulk soil to the rhizosphere. Focus will be lying on effects of dilution on the role of the recruited microbial community for plant protection. 304 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session 153 Analysis of community structure of metabolically active bacteria in a rice field subjected to long-term fertilization practices Jae-Hyung Ahn, Min-Young Choi, Jaekyeong Song, Hang-Yeon Weon* Agricultural Microbiology Division, National Academy of Agricultural Science, RDA, South Korea To estimate the effect of long-term fertilization on metabolically active bacterial communities in a rice field, RNA was extracted from endosphere (rice root), rhizosphere, and bulk soil that had been subjected to different fertilization regimes for 59 years and the 16S rRNAs were analyzed using the pyrosequencing method. The richness and diversity of metabolically active bacteria were higher in bulk soil than in the endosphere and rhizosphere, and showed no significant difference between non-fertilized and fertilized plots. Weighted UniFrac analysis showed that each compartment had characteristic bacterial communities and that the effect of long-term fertilization on the structure of bacterial community was more pronounced in bulk soil than in the endosphere and rhizosphere. The 16S rRNAs affiliated with Alphaproteobacteria and Firmicutes were more abundant in the endosphere than in bulk soil while those affiliated with Chloroflexi and Acidobacteria were more abundant in bulk soil than in the endosphere. Several dominant operational taxonomic units (clustered at a 97% similarity cut-off) showed different frequencies between non-fertilized and fertilized plots, suggesting that the fertilization affected their activities in the rice field. 154 Metagenomic insight into the plant-microbe rhizosphere interaction under Rehmannia glutinosa consecutive monoculture regime Linkun Wu*, Juanying Wang, Hongmiao Wu, Jun Chen, Xianjin Qin, Wenxiong Lin Fujian Agriculture and Forestry University, China Under consecutive monoculture regime the biomass and quality of Rehmannia glutinosa, an important Chinese medicinal plant suffers from significant decline. The objective was to evaluate the response of soil bacterial and fungal communities to consecutive monoculture using omics technique and assess the positive and negative effects of root exudates on the key microbes associated with consecutive monoculture problems. The results showed that consecutive monoculture led to a great shift in rhizospheric microbial community. In details, consecutive monoculture led to significant reduction of populations assigned to genus Pseudomonas. However, Fusarium oxysporum or Fusarium sp. belonging to Ascomycota was significantly higher in consecutively monocultured soil than in newly-planted soil. Real-time PCR assay confirmed the reduction in abundance of Pseudomonas sp. and an increase in F. oxysporum in consecutively monocultured soil. Furthermore, the relative abundance of Pseudomonas sp. with the antagonistic activity against F. oxysporum was greatly lower in consecutively monocultured soil than in newly-planted soil. We aslo found that phenolic compounds mixture in an ratio as detected in the soil could greatly promote the growth of pathogenic F. oxysporum, but inhibit the beneficial antagonistic bacteria. The isolated hostspecific F. oxysporum caused wilt disease on the tissue culture seedlings of R. glutinosa. This study demonstrated that consecutive monoculture resulted in the alteration of rhizospheric microbial composition with fewer microorganisms providing beneficial functions and more microorganisms with pathogenicity, which is mediated by root exudates and had a negative impact on R. glutinosa growth and development. 305 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session 155 Selection of microbial community by plant and soil from diverse inocula Yan Yan*1, Eiko Kuramae1, Peter Klinkhamer2, Hans van Veen1 1 Netherlands Institute of Ecology, Netherlands, 2Leiden University, Netherlands The experimental approach applied is based on the assumption the biodiversity of the microbial community in soil can be altered by inoculation of diluted suspensions in a sterilized soil. Although this method has frequently been used earlier, little is known how the assemblage of bacterial communities in soil and rhizosphere proceeds after inoculation of more or less diluted suspensions. For that purpose, serial dilutions of a soil suspension were made, and re-inoculated into the original soil previously sterilized by γ-irradiation. We determined the structure of the microbial communities by using 454-pyrosequencing of the 16S rRNA gene. Upon dilution, the community diversity at the species level in the suspensions reduced dramatically. The structure of microbial community in soil was changed drastically after incubation as compared to the structure of the community in the inoculum suspension. Although the microbial community was more homogenous in the rhizosphere, differences between the original bulk soil and rhizosphere soil for all dilutions were only apparent at the genus or species level. The selection power by soil after incubation was stronger than that by the plant. Assemblage processes in soil of the high-diluted community followed clearly a niche type model, but the assemblage in the rhizosphere appeared to be a neutral, random, selection process. Network analysis showed a more complex system in the rhizosphere than in the bulk soil, which, in contrast to the assemblage rules models, indicates the importance of plant related mechanisms operating in the assembly of microbial communities in the rhizopshere. 156 Evasion and suppression of root immunity by beneficial microbes Ke Yu*1, Roeland Berendsen1, Chiel Pel2, Corné Pieterse1, Peter Bakker1 1 Plant-Microbe Interactions, Utrecht University, Netherlands, 2University of Toulouse, France Plants can detect pathogenic microbes by recognizing microbe-associated molecular patterns (MAMPs) through cell-surface pattern-recognition receptors (PRRs), which can induce MAMP-triggered immunity in hosts. Plant-beneficial microbes living in the rhizosphere possess similar MAMPs, however, this does not lead to massive activation of plant defenses. This indicates that these beneficial microbes actively evade or suppress local immune responses. Plants recognize flagellin as a MAMP through the PRR FLAGELLIN SENSING 2. For pathogenic pseudomonads it was shown that an alkaline protease A (AprA) that degrades flagellin monomers, limits recognition and thus virulence. The plant growth promoting rhizobacteria Pseudomonas simiae WCS417 and P. capeferrum WCS358 can both suppress the immune responses in Arabidopsis roots elicited by flagellin. An AprA homolog was also identified in the genome of WCS417. However the genome of WCS358 did not comprise an AprA ortholog, suggesting WCS358 deploys a different strategy to suppress the root immune responses. Site-directed mutagenesis and transposon mutant library screening have been initiated to explore mechanisms involved in the evasion and 306 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session suppression of root immune responses elicited by MAMPs during plant-beneficial microbe interaction. 157 Pyrosequencing reveals the structure and functional traits in a coastal halophyte rhizosphere microbiome Zhilin Yuan*1, Yuan Qin2, Yicun Chen2, Rusty Rodriguez3, Regina Redman3, Chulong Zhang4, Fucheng Lin4 1 Chinese Academy of Forestry, China, 2Institute of Subtropical Forestry, Chinese Academy of Forestry, China, 3Adaptive Symbiotic Technologies, United States, 4Zhejiang University, China Soil and root-associated microbes are always, if not all, of great importance for improving host fitness. Current knowledge of microbiome in salt circumstances, however, is largely lacking. As a step in this direction, the microbial community capable of prospering in the bulk soils (BS), rhizosphere soils (RS), root endosphere (R) of Suaeda salsa, a widely distributed halophyte in China's east coast, was investigated using platform of 454 pyrosequencing. Rarefraction curves documented an impressive decreasing trend of α-diversity from soils to endophytic compartments. Proteobacteria was the abundant phyla in all communities. The genus Acinetobacter dominated the roots. Many groups of bacteria showed high phylogenetic affinities to known halo-tolerant species. We used membership and phylogenybased Venn diagram to reveal the core bacterial colonizers. In addition, the relatively high abundance of unclassified fungal and bacterial OTUs was prevalent at different taxonomic levels, indicating the novelty of microbes in saline environment. For fungi, OTUs assigned to Pleosporales were frequently found. Beauveria bassiana, Leptosphaeria sp., Retroconis sp., Monosporascus sp. were heavily enriched in BS, RS, R respectively. All of them displayed a moderate to high level of genetic diversity. We hypothesize that such apparent phylogenetic conservatism (redundancy) would be responsible for plants to be well adapted to salt stress. Tajima's D test and the star-like structure of haplotype networks further revealed the population expansion of above taxa. Beyond this, Montagnulacea sp. and Monosporascus sp., much more common in roots than BS and RS, were also easily isolated, and inoculation test confirmed their ability to confer beneficial effects to plants under organic nitrogen or salt stress condition. Overall, our data yields some evidence that rhizosphere microbiome of S. salsa readily evolved to enable plants to survive in adverse conditions and opens new avenue for generating symbiotically-modified salt-tolerant plants. 158 Plant breeding and its effect on the rhizosphere microbiome - a sugar beet example Christin Zachow*1, Henry Müller1, Ralf Tilcher2, Gabriele Berg1 1 Graz University of Technology, Austria, 2KWS SAAT AG, Germany Worldwide crop yields are lost due to abiotic and biotic stresses like drought, salinity, pests and diseases. Breeding of stress-tolerant and pathogen-resistant cultivars has a long history and approaches targeting plant beneficial microbes are developing. We used omics and microscopic technologies analysing a naturally composed microbiome as seed treatment for sugar beet plants against abiotic stresses and the late root rot caused by the soilborne pathogen Rhizoctonia solani. Two sugar beet cultivars (BERETTA/JENNA) were used, which are 307 Rhizosphere Microbiome 2 Wednesday 24 June – Poster session characterized by R. solani sensitiveness/tolerance and by different abundance, enrichment and interaction of a beneficial, sugar beet-specific Pseudomonas genotype. In the Rhizoctonia-tolerant cultivar JENNA, the Pseudomonas genotype was enhanced in seed, endorhiza and rhizosphere independent of the surrounding soil as example of naturally occurring biocontrol. In contrast, the Rhizoctonia-sensitive cultivar BERETTA contains no detectable number of this genotype. One strain of the genotype was compared to other Pseudomonas strains of the same clade and characterized in detail; the genome information of the endophyte P. poae RE*1-1-14 explains the endophytic lifestyle and the antagonistic effect in 191 unique genes. Additionally, we analysed the impact of genetic variation in ten cultivars on the composition of the microbiota. The overall results will contribute to integrated management strategies in modern sustainable agriculture. 159 Iron-reducing bacteria in rice rhizosphere contribute to arsenic mobilization under flooded conditions Sarah Zecchin*1, Anna Corsini1, Raffaella Zanchi1, Maria Martin2, Gian Maria Beone3, Marco Romani4, Lucia Cavalca1 1 University of Milano, Italy, 2University of Torino, Italy, 3Università Cattolica del Sacro Cuore, Italy, 4Ente Nazionale Risi, Italy Rice is among the crops mostly affected by arsenic contamination. Whereas in arsenic contaminated soils (> 40 mg kg-1) rice farming contributes to population exposure, in rice field soils with low arsenic content agronomic conditions represent an issue for arsenic bioavailability Iron-reducing bacterial populations (Geobacteraceae and Shewanellaceae) inhabiting rice rhizosphere were assessed in box plots (total arsenic 18.4 mg kg-1) cultivated with different water regimes, in order to evidence their role in the release of iron and consequently arsenic from root ferric iron plaques. Geobacteraceae and Shewanellaceae 16S rRNA genes significantly increased in rice rhizosphere from the order of 105 to 106 copies (g dry weight)-1 under flooding, while no significant increase occurred in aerobic rice. Fluorescence in situ hybridization in rhizoplane evidenced a similar trend related to the active populations. This increase was concomitant with the release of arsenic in soil solution under continuous flooding: from 1.40 μg L-1 to 190 μg L-1 over the cropping cycle, whereas in aerobic rice no significant release occurred. Analogously, ferrous iron increased from 0.75 mg L-1 to 51.1 mg L-1 and dissolved organic C from 8.9 to 79.9 mg L-1 until late reproductive stage. In aerobic rice soluble iron remained almost negligible and dissolved organic C never exceeded 30 mg L-1. Arsenic content in rice grains was 237 μg kg-1 under flooding and 4.67 μg kg-1 in aerobic rice, thus reflecting this solubilisation trend. These outcomes evidence that iron-reducing bacteria promote the arsenic release from iron plaques under flooded conditions in non-contaminated rice fields thus contributing to grain contamination. 308 Rhizosphere Microbiome 2