Following text represents the selection of materials compiled

Transcription

IRP identification code
AV0Z50380511
Following text represents the selection of materials compiled
for the international evaluation of the
Institute of Experimental Botany AS CR
in 2004.
1
AV0Z50380511
Institutional Research Plan (IRP) proposal
Provider‘s code
Research plan title
Applicant 1
Institution 2
Principal investigator 3
A
A1.
AV0
Z50380511
Mechanisms of regulation of plant growth and development on the
level of cells, organs and whole organisms: physiological, genetic
and molecular bases
Institute of Experimental Botany AS CR
RNDr. Ivana Macháčková, CSc.
General information on the applicant
Organisational scheme of the applicant, including the number of employees in the
applicant’s units
Governing bodies of the apllicant are listed in part A2.
Number of employees at the IEB (including part-time jobs): in total 196 employees
(consisting of 63 researchers, 34 research assistants, 42 technicians, and 57 other
employees).
IEB is located in six centres in Prague and Olomouc. There are 16 laboratories in IEB
(names and heads of the laboratories are listed below).
CENTRE 1:
Rozvojová 135
Lysolaje
165 00 Prague 6
RNDr. Radomíra Vaňková, CSc.
(420) 220 390 427
fax: (420) 220 390 446
e-mail: [email protected]
Laboratory of Plant Morphogenesis
(since 2004 RNDr. Jan Martinec, CSc.)
(420) 220 390 453
fax: (420) 220 390 456
Ing. Miroslav Kamínek, CSc.
Laboratory of Hormonal Regulations in
Plants
(since 2004 RNDr. Eva Zažímalová,
CSc.)
(420) 220 390 445
fax: (420) 220 390 446
RNDr. Milena Cvikrová
(420) 220 390 409
fax: (420) 220 390 419
Laboratory of Biologically Active
Compounds
1
2
3
Legal name of the applying organisation, legal entity
Name of the department or applicant’s principal organisational unit, which will carry out the research
according to the proposal, if different from “applicant”. Applies only if more than one proposal of IRP was
submitted by the “applicant”.
Person in charge, who is responsible, on behalf of the applicant/institution, in scientific and financial
matters of IRP
2
AV0Z50380511
RNDr. Věra Čapková, CSc.
(420) 220 390 452
fax: (420) 220 390 461
RNDr. Viktor Žárský, CSc.
(420) 220 390 457, 220 390 458
fax: (420) 220 390 461
Laboratory of Pollen Biology
Laboratory of Cell Biology
CENTRE 2:
Na Karlovce 1a
Dejvice
160 00 Prague 6
RNDr. Milada Šindelářová, CSc.
secretariat: (420) 224 310 108
fax: (420) 224 310 113
RNDr. Milada Šindelářová, CSc.
: (420) 224 310 109
fax: (420) 224 310 113
RNDr. Karel J. Angelis, CSc.
(420) 224 322 603
fax: (420) 224 310 113
RNDr. Jana Pospíšilová, CSc.
(420) 224 320 198, (420) 233 331
032
fax: (420) 224 310 113
RNDr. Tomáš Gichner, DrSc.
(420) 224 310 109
fax: (420) 224 310 113
RNDr. Noemi Čeřovská, CSc.
(420) 224 320 338
fax: (420) 224 310 113
Ing. Jaroslav Tupý, DrSc.
Laboratory of Pathological Plant
Physiology
Laboratory of DNA Repair
Laboratory of Stress Physiology
Laboratory of Mutational Genetics
Laboratory of Virology
CENTRE 3:
Na Perníkářce 15
Dejvice
160 00 Prague 6
(since 2005 RNDr. Miloslav Juříček,
CSc.)
(420) 233 336 791
fax: (420) 233 339 412
Laboratory of Pollen Embryogenesis
(since 2005 RNDr. Miloslav Juříček,
CSc.)
(420) 233 336 791
fax: (420) 233 339 412
(420) 485 177 430
fax: (420) 233 339 412
Experimental Station Střížovice
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AV0Z50380511
Josef Holík
(420) 241 032 381, (420) 241 062
728
fax: (420) 241 062 150
Josef Holík
(420) 241 032 381, (420) 241 062
728
fax: (420) 241 062 150
CENTRE 4:
Vídeňská 1083
Krč
142 00 Prague 4
Isotope Laboratory
CENTRE 5:
Sokolovská 6
772 00 Olomouc
Doc. Ing. Jaroslav Doležel, DrSc.
secretariat: (420) 585 228 521-2
fax: (420) 585 228 523
Doc. Ing. Jaroslav Doležel, DrSc.
secretariat: (420) 585 228 521-2
fax: (420) 585 228 523
Doc. Dr. Jiří Vagera, CSc.
secretariat: (420) 585 228 521-2
fax: (420) 585 228 523
Laboratory of Molecular Cytogenetics and
Cytometry
Laboratory of Genetic Manipulations in
vitro
Laboratory of Plant Cytoskeleton and Cell
Cycle
Doc. RNDr. Pavla Binarová, CSc.
(420) 585 228 521-2,
(420) 241 062 130
fax: (420) 585 228 523
e-mail: [email protected],
[email protected]
CENTRE 6:
Joint Laboratory of IEB ASCR and
Faculty of Life Sciences Palacký
University in Olomouc
Šlechtitelů 11
783 71 Olomouc
Prof. Ing. Miroslav Strnad, CSc.
(420) 585 634 850
sekretariat: (420) 585 634 851
fax: (420) 585 634 870
Laboratory of Growth Regulators
Prof. Ing. Miroslav Strnad, CSc.
(420) 585 634 850
secretariat: (420) 585 634 851
fax: (420) 585 634 870
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AV0Z50380511
A2.
Governing bodies of the applicant and names of their personnel
DIRECTOR:
Rozvojová 135
Lysolaje
165 00 Prague 6
(4202) 220 390 453
- secretariat (4202) 220 390
455
fax: (4202) 220 390 456
DEPUTY DIRECTOR:
Rozvojová 135
Lysolaje
165 00 Prague 6
RNDr. Eva Zažímalová, CSc.
(4202) 220 390 429
fax: (4202) 220 390 446
SCIENTIFIC SECRETARY:
Rozvojová 135
Lysolaje
165 00 Prague 6
RNDr. Martin Vágner, CSc.
(4202) 220 390 414
fax: (4202) 220 390 419
CHAIRMAN OF THE SCIENTIFIC BOARD:
Rozvojová 135
Lysolaje
165 00 Prague 6
RNDr. Jan Martinec, CSc.
(4202) 220 390 416
fax: (4202) 220 390 419
MEMBERS OF SCIENTIFIC BOARD:
RNDr. Lenka Burketová, CSc.
RNDr. Noemi Čeřovská,CSc.
RNDr. Věra Cenklová, PhD.
RNDr. Věra Čapková, CSc.
Mgr. Lucie Perry, PhD.
RNDr. Radomíra Vaňková, CSc.
ADMINISTRATION:
Rozvojová 135
Lysolaje
165 00 Prague 6
Naděžda Pulcová
(4202) 220 390 475
fax: (4202) 220 390 474
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AV0Z50380511
B
B1.
Information on research and development activities
of the applicant/institution
Specification of the principal research and development (R&D) activity of the
applicant/institution
The aim of setting up this working site is to perform the scientific research in the field
of experimental botany and searching for possibilities of exploitation of its results.
The subject of business of IEB AS CR is the scientific research in the field of plant
physiology, genetics, plant biotechnologies, biochemistry and molecular biology with the
focus on the regulation of growth and development, physiology of photosynthesis, adaptive
mechanisms against stress induced by the surroundings and pathogenic agents, molecular
genetics of pollinic and somatic cells and mechanisms of mutagenesis and synthesis of
marked bioactive substances. Further activities include the participation in programs of
development of plant physiology, genetics and biotechnologies directed towards generation
of new genetic sources, improvement of new genotypes resistant against biotic and abiotic
factors, reproducing of plant material in vitro and its application in cultivation, as well as
development of new silvicultural technologies in agriculture and horticulture. Within the scope
of its subject of business, the Institute contributes to increasing the level of knowledge and
understanding, exploiting the results of research activities and its promotion. The Institute
receives, processes and spreads the scientific information by publishing magazines, textbooks, monographies and regular reviews of its research activities. The Institute provides
scientific references, standpoints, recommendations and searches. In co-operation with
universities, the institute organizes post-graduate studies and raises scientists. The Institute
develops international co-operation projects and, in the scope of the subjected activities,
organizes scientific conferences, symposia and seminars in the Czech Republic, with
international participation. The Institute obtains national and international grants as well as
other forms of support of its scientific activities. In order to exploit the results of scientific
research together with having the right of their registration and maintaining of patent
protection locally and abroad, the Institute administers the property rights to the patents
originated in its working site and enters into license agreements with local and foreign
subjects. The Institute executes its projects in co-operation with other scientific and
professional institutions.
A comparison of this Foundation document with the description of the results gained in the
Institute of Experimental Botany (IEB) in years 1999-2003 and the research proposal for
years 2005-2010 clearly shows that IEB fulfills its research and other tasks. IEB is active in
research in the fields of plant physiology, genetics, biochemistry and molecular biology,
namely regulation of plant growth and development (metabolism, transport and mechanisms
of action of phytohormones and other regulatory substances), regulation of cell cycle, signal
transduction, stress physiology, physiology of photosynthesis and interaction with pathogens,
DNA damage and repair and mutagenesis, gene mapping and functional genomics,
chromosome sorting and use of chromosomes in modern genomics. IEB participates in the
programmes preparing new materials for cereal breeding gained by in vitro manipulations or
transgenosis, and introduces new apple tree varieties resistant to scab. Researchers in IEB
have also started research in the field of potential production of pharmaceutically important
proteins in transgenic plants and they have developed new substances on the basis of the
structure of phytohormones cytokinins, which inhibit cell division and are tested as
medicaments against proliferative diseases. The Izotope Laboratory of IEB synthetises many
substances for research, in which colleagues abroad are also interested.
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AV0Z50380511
The researchers in IEB publish their research results in scientific journals (see the list, pp.
71-102 ) and also help the popularisation of these results and science as such (part 4.3). IEB
issues two international journals: Biologia Plantarum and Photosynthetica. Researchers of
IEB are also authors or coauthors of a number of textbooks and scripta (part 4.3) and many
of them actively participate in lecturing at several universities and supervise the diploma and
doctoral theses (part 4.1).
IEB has actively developed and supported international collaboration as may be seen from
the list of workers from abroad who have worked in the IEB for long periods (see part B4)
and from the list of publications (some of them come from collaboration) and grant projects.
IEB has organised several symposia and taken part in the organisation of some others. IEB
is the representative of Czech plant biologists in the EPSO - European Plant Science
Organisation.
IEB has several patents and many licences, the list of which may be found in D4, pp. 101102.
B2.
Contribution of the applicant/institution to the development of knowledge in the
disciplines referred to in B1, in national and international context
The Institute of Experimental Botany (IEB) is the only institution in the Czech Republic (CR)
whose research covers a wide area of plant physiology and genetics and interconnects and
integrates these two areas. Similar in orientation is the Institute of Plant Molecular Biology
(IPMB) in České Budějovice, but its research in plant physiology is limited to photosynthesis
and phytopathology. Earlier the research in the individual laboratories of the IEB was
relatively independent, but in the last 10 years we have interconnected and integrated the
research of individual teams and different teams participate in the work on common projects.
Collaboration with the Universities has also been successfully developed (with Departments
of Botany and Genetics at the Palacky University in Olomouc - common "Laboratory of Plant
Growth Regulators", the Department of Plant Physiology at Charles University in Prague common laboratory is being prepared, the Institute of Biochemistry and Microbiology at the
Institute of Chemical Technology in Prague, the Institute of Botany and Plant Physiology at
the Mendel University of Agriculture and Forestry in Brno and the Department of Botany at
the Czech University of Agriculture in Prague) as well as collaboration with colleagues
abroad. The workers of the IEB take an active part in teaching at the Department of Plant
Physiology in Prague and the Departments of Botany and Genetics, respectively, in
Olomouc. The researchers of the IEB supervised and supervise a number of diploma and
doctoral theses (in the frame of official acreditations or agreements with universities). In the
last 5 years we have integrated our research in the field of studies of signal transduction
pathways in plants, in which the IEB is the coordinator of a prestige Research Centre, which
integrates the best laboratories in the Czech Republic in this field of research. During this
period the number of foreign researchers working in the IEB increased - they were
colleagues from the Belorussia, Bulgaria, France, Great Britain, Guatemala, Japan, India,
Italy, Russia, Serbia, Ukraine and USA, Poland. The Institute has a good tradition in
organising international symposia on auxins and cytokinins - the last one was in Prague in
1999. We also participated in the organisation of the 17th Conference of the International
Plant Growth Substances Association (IPGSA) in 2001 in Brno.
The Institute also produces two major scientific journals: Biologia Plantarum and
Photosynthetica.
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The level of research performed in the IEB is high and the IEB has a leading position within
the CR in research on phytohormones and other regulatory substances, signal transduction
pathways, pollen biology, cytogenetics, cell cycle and DNA repair. The IEB together with the
IPMB were the first in the Czech Republic to study and evaluate the possibility of producing
pharmaceutically important proteins in transgenic plants (edible vaccines etc.). The
Laboratory of Genetic Manipulations in vitro contributed significantly, with its haploid and
other plant materials, to the breeding of new varieties of cereals and grasses. At the
Research Station in Střížovice new apple tree varieties resistant to scab were bred - licenses
for these varieties are now requested all over the world.
The number of publications continues to increase as does their quality and impact (see figure
bellow).
Several research areas in the IEB also have a high level in an international context fully
comparable with that of other European Institutions:
The Laboratory of Growth Regulators (Palacky University and IEB) together with the Isotope
Laboratory of the IEB have developed groups of substances based on the structure of
phytohormones of the cytokinin type, which inhibit the cell cycle and in collaboration with
hospitals in CR and abroad were tested as potential medicaments against cancer and other
proliferative diseases. These substances are patented and the Institute has a number of
license agreements for testing and potential use. This research has received several
international awards.
Phytohormone research has a long tradition and a high level, particularly research on auxins
and cytokinins. The Institute is known as a centre for analytical methods for the
determination of phytohormones and some other regulatory substances and for studies on
hormone metabolism and transport. In recent years the Laboratory of Molecular Cytogenetics
and Cytometry was very successful - they managed to localise some sequences on sorted
chromosomes and for the first time described isolation of pure high-molecular weight DNA
from sorted chromosomes.
Changes in the structure of the cytoskeleton during the cell cyle and division of plant cells
and function of some regulatory proteins were also described.
In collaboration with the University of Leicester (UK) gene expression at the level of
transcriptome in one developing cell (pollen tube) was described for the first time.
In collaboration with the Department of Plant Physiology (Charles University, Prague) first
indications of the occurrence of an Exocyst complex in plants were found and its possible
role in cell morphogenesis proposed.
In research on the phosphoinositide signalling system a method of in situ determination of
activities of different types of phospholipases was elaborated and a new type of
phospholipase cleaving phosphatidylcholine previously unknown in plants was described.
The regulatory role of excretion of chloride ions was shown in pollen tube growth and a new
model for this regulation proposed.
The occurrence of the hormone melatonin was shown in higher plants and a previously
unknown rhythm in its level, which depends on the photoperiod was described.
The Comet assay was elaborated for studies of DNA damage and is being used in the study
of DNA repair and the effect of mutagens.
Our research on the role of polyamines in cell division, studies of the role of various signals
and PR (pathogenesis related) proteins in the interaction of plants with pathogens as well as
the molecular characterisation of plant viruses also has a good European standard.
A significant contribution to the research level is made by the Isotope Laboratory of IEB,
where they synthetise compounds which inhibit the cell cycle mentioned above as well as
substances, both radioactive and non-radioactive, necessary for the research of other teams,
which either cannot be bought or are very expensive. A number of foreign colleagues are
interested in these substances.
The priority results described obtained in the last 5 years document a high quality of research
performed in IEB, both at local and international levels. Specificity of the research in IEB
resides mainly in the areas of phytohormones and signalling pathways (and their integration),
studies of the genome at the level of chromosomes, pollen biology and DNA repair. In these
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areas the research in IEB is the leading one in the CR. When compared with other European
institutions, the quality of research in IEB is good and it contributes to and complements
European research. The equipment at IEB for research in molecular biology, biochemistry,
cytology and physiology of plants is at a standard European level. Many institutions
concentrate in narrow research fields; the specifity of IEB is integration of several research
areas of plant biology in one moderate-sized institution and make them complementary.
Scientific publications of IEB
*
impacted papers
average impact factor
60
2.2
2.0
50
1.8
40
30
1.6
20
average impact factor
number of papers in impacted journals
70
1.4
10
1.2
0
1995
*
1996
1997
1998
1999
2000
2001
2002
2003
year
only papers published until November 2003 (currently at least 14 other papers are "in press",
some of them will be printed in 2003)
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B3.
Major R&D results achieved and implemented by the applicant/institution in the
disciplines referred to in B1 within the last five years (overall characteristics)4
Main results of IEB AS CR 1999 - 2003
The Research Programme of the Institute of Experimental Botany (IEB) for the period 1999 2003 was Physiological and Genetic Basis of Plant Development, Cell Cycle,
Morphogenesis, Reactions to Stress Conditions and Biotechnologies. Organisation
and Function of the Genome (No.: AV0Z5038910).
The programme was aimed at elucidating the basic molecular and cellular mechanisms in
the following topics:
• The integrated action of phytohormones and growth regulators in plant development
control;
• The regulation of cell morphogenesis and the role of cytoskeleton and relevant signalling
pathways;
• The regulation of the cell cycle and the use of synthetic regulators for its inhibition and
the inhibition of tumour growth;
• The regulation of ontogenesis of the male gametophyte: gene expression,
proteosynthesis and signalling; somatic and pollen embryogenesis, anther cultures,
haploid production;
• Genome organisation and function at the chromosome level;
• The detection of regulatory genes, dynamics of damage and repair of genomic DNA
under the influence of stress and mutagens;
• The factors affecting photosynthesis and water relations in stress conditions;
• The reaction of plants to infection and molecular characteristics of viruses.
This research has resulted in an improved understanding of the above topics and their
biotechnological applications.
The tasks of the programme imply that the IEB covers a relatively broad field of research in
the physiology and genetics of plants. The main aim was to integrate the individual parts of
the programme and the grant proposals were composed and the projects run accordingly.
One of the significant steps in the integration of this research was the award to the Research
Centre of the project „Signalling Pathways in Plants“ by the Ministry of Education, Youth and
Sports of the Czech Republic, No.: LN00A081), co-ordinated by IEB. Some of the results
reported in the area of signalling were performed within the framework of this project and are
included marked with sign ◘ at the end of the paragraph within this report.
The research running in the IEB during the period 1999-2003 in the frame of the Research
Programme, the Research Centre and grant projects, can be divided into two main parts:
•
The regulation of plant growth and development
•
The structure and function of the genome.
In the area of „regulation of plant growth and development“ research was focussed on growth
regulators (namely phytohormones: auxins and cytokinins and, to a lesser extent, ethylene
and abscisic acid). Some aspects of metabolism, transport and mechanism of action of
phytohormones and polyamines were described and characterised. In this area an important
achievement was reached in the investigation of auxin carriers and their regulation, and in
the characterisation of the role of cytokinin oxidase in the regulation of the level of active
cytokinins in cells and tissues. The derivatives of N6-substituted adenine were prepared and
4
Implemented results are those R&D results which have been published, applied in practice and/or protected
as an intellectual property according to a specific law (e.g. publications, patents, trademarks, and newly
applied technologies).
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tested as inhibitors of key reactions of the cell cycle and as a medicine against serious
diseases. One of the compounds designed, prepared and characterised in the IEB is now in
the second phase of clinical tests as a potential drug against cancer in several European
countries. Attention has also been aimed towards the participation of plant growth regulators
in the stress reactions of plants to high levels of radiation, high and low temperatures, water
deficits and some viruses.
Attention was also focussed on the investigation of signalling pathways and their coupling
with cell structures (cytoskeleton) and of cell cycle control. A new type of plant
phosphatidylcholine-hydrolysing phospholipase C was characterised and the complex
Exocyst (previously unknown in plant cells) was identified.
The distribution of gamma-tubulin and F-actin was described during mitosis, when centrioles
were not formed and cyclin B2 expression was monitored in the course of cell division.
In the area of genomics, the investigation was focussed on:
•
Characterisation of the genome structure and its variability in some plant species,
•
Localisation of some sequences and molecular markers using sorted chromosomes,
•
The preparation of high-molecular weight DNA from sorted chromosomes and
chromosome-specific DNA-libraries.
In the male gametophyte the expression of genes and regulation of transcription and
translation were investigated. In the male gametophyte, the expression of genes and
regulation of transcription and translation were investigated; it was the first time when the
expression has been analysed on the level of transcriptom in the range of the whole genome
of a single developing cell. The expression of genes encoding proteins involved in the
regulation of cell cycle was described, and the mechanisms of DNA repair and plant viruses
were characterised. Transgenosis has been characterised in relation to the plant protection
and for the food industry (potato) and for crop improvement (cereals). Preparation of
transgenic plants with controlled expression of genes related to metabolism of cytokinins was
started with the aim of increasing longevity and productivity. Research work was also
concentrated on androgenesis in vitro and haploid and dihaploid plants were prepared for
breeding. Varieties of apple tree resistant to fungal diseases were prepared and an
investigation of the possibilities of production of bioactive proteins in plants started.
In the following text, a more detailed description of results achieved is divided into two main
research areas and these are sub-divided into smaller thematic parts for better
understanding. Numbers in parentheses relate to the papers published, as listed on page 71102.
List of contents:
1
REGULATION OF PLANT GROWTH AND DEVELOPMENT
1.1
Plant growth regulators
1.1.1
Metabolism and mechanism of action of phytohormones
1.1.2
Transport of auxins and cytokinins
1.1.3
Hormonal regulation of some life processes in plants
1.1.4
Other plant growth regulators
1.1.5
The role of plant growth regulators in plant stress reactions
1.1.6
New methods in the investigation of phytohormones
1.1.7
Potential practical applications of research on plant growth regulators
1.2
Signalling and signal transduction
1.2.1
Signalling mechanisms
1.2.2
The relationship between signalling and the cytoskeleton
1.2.3
Signalling in plant defence mechanisms
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2
STRUCTURE AND FUNCTION OF THE GENOME
2.1
Functional genomics
2.2
Studies on genome structure using sorted chromosomes
2.3
Genotoxicity and DNA repair
2.4
Molecular aspects of plant virology
2.5
Potential practical applications of genetic research
1
1.1
REGULATION OF PLANT GROWTH AND DEVELOPMENT
1.1.1
Metabolism and mechanism of action of phytohormones
Using a newly developed method of large-scale synchronisation of tobacco cell suspension
(line BY-2), which allows synchronisation up to 30 g FW of cells, the dynamics of cytokinins
and activity of cytokinin-degrading enzyme, cytokinin oxidase/dehydrogenase (CKX), was
determined during the cell cycle progression. Results have confirmed a significant and
transient accumulation of cytokinins at the beginning of the S-phase and mitosis and brought
new knowledge about fast metabolic regulation of levels of physiologically active cytokinins.
It is based on their conversion to storage cytokinins of cis-zeatin and zeatin-O-glucoside type
in the premitotic phase of the cell cycle and on the degradation of isoprenoid cytokinins by
CKX at the beginning of the S-phase (229).
Novel N6-substituted derivatives of adenine were isolated and identified: In the
photoautotrophic cell culture of Chenopodium rubrum 6-[2-(β-D-glucopyranosyloxy)benzylamino]purine (oTOG), 6-[2-(β-D-glucopyranosyloxy)benzylamino]-2-methylthiopurine
(2MeS-oTOG) a 6-benzylamino-9-β-D-glucopyranosylpurine (BAP9G) were found. Their
cytokinin activity was confirmed in the Amaranthus bioassay and their endogenous origin
was proven using a novel method based on the incorporation of deuterium in situ from the
cultivation medium enriched by D2O into the compounds studied. The appearance of a
range of other new cytokinins, e.g. zeatin-9-glucoside-O-glucoside, and aromatic cytokinins
was detected in other species (224, 230, 353, 375). For the first time the new group of
naturally occurring plant hormones structurally derived from 6-(2- a 3methoxybenzylamino)purine was isolated. The identification was carried out in A. thaliana,
poplar leaves, and selected strains of A. tumefaciens using LC-MS. The high biological
activity of the compounds isolated was confirmed in three cytokinin biotests (364).
In co-operation with laboratory of Prof. Schmülling (Berlin, Germany) the effects of
expression of four different genes encoding the cytokinin oxidase/dehydrogenase (CKX) from
Arabidopsis thaliana on the development of transgenic tobacco plants were determined. A
high increase in CKX activity resulted in a significant lowering of the cytokinin content and,
consecutively, to pronounced changes of the phenotype of transformed plants. Cytokinindeficient plants were characterised by dwarf shoots with smaller apical meristems and
prolonged plastochron. In contrast, root meristems of transgenic plants were bigger; roots
were more branched and grew more intensively. This finding represents the first direct
experimental evidence of the physiological relevance of endogenous cytokinins in the control
of morphogenesis and meristematic activity in plants based on down-regulation of cytokinin
levels (195). The dependence of CKX activity on the level of endogenous cytokinins was
found in a cytokinin-autonomous line of A. thaliana, which thus represents a promising model
for studies of CKX induction by its substrates (89). The regulatory effects of cytokinins on
CKX activity were proved in tobacco cell suspension where increases in cytokinin levels
following either the application of exogenous cytokinin or the expression of IPT gene,
encoding enzyme for cytokinin biosynthesis, resulted in a significant increase in the activity of
CKX, namely of its glycosylated form, and its preferential secretion outside the plasma
membrane (336).
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By a combination of affinity chromatography with immobilised zeatin and liquid
chromatography with mass spectrometry detection, the adenosine kinase was isolated from
suspension culture of the tobacco cell line BY-2 and characterised (327). Adenosine kinase
is one of the enzymes responsible for the formation of cyclic nucleotides in cells. This is the
first report of this enzyme in plants and it shows the significance of this enzyme in the
metabolism of cytokinins.
The existence of the isopentenyl adenosine-5´-monophosphate- (iPMP)-independent
biosynthetic pathway for zeatin-type cytokinins was proven (82). This pathway was active in
both IPT-transgenic A. thaliana and wild type plants. The investigation of cytokinin
biosynthesis de novo in the IPT-transgenic A. thaliana using labelling with deuterium in vivo
and mass spectrometry showed that the rate of biosynthesis of zeatin riboside-5’monophosphate was ca. 66-times higher than that for iPMP, which was supposed to be the
primary product of isopentenyl transferase from A. tumefaciens. By the double-labelling
method, using [2H6]-isopentenyl adenosine and deuterium oxide, the existence of an
alternative, iPMP-independent biosynthetic pathway for zeatin and its derivatives was
discovered. This pathway operates in both IPT-gene-expressing and wild type plants. A
decrease in the activity of the alternative biosynthetic pathway after the application of
mevastatin, inhibitor of 3-hydroxy-3-methylglutaryl CoA reductase, indicates the terpenoid
origin of the side-chain precursor for this iPMP-independent pathway.
A second gene encoding cis-zeatin specific O-glucosyltransferase (cisZOG2) was isolated
from maize in co-operation with the laboratory of Profs Moks (Corvallis, Oregon, U.S.A.). The
gene is significantly expressed in roots and, in contrast to cisZOG1, its expression in kernels
is very low. Interestingly, high levels of zeatin riboside O-glucoside were found in kernels
using HPLC/MS. Results confirm the existence of specific metabolic pathways regulating
levels of cis-zeatin type cytokinins in plants (373).
Chloroplasts from tobacco plants (SR1) carrying the gene for β-glucosidase Zm-p60.1 under
the CaMV35S promoter (kindly provided by Dr. B. Brzobohatý, Institute of Biophysics, Brno)
contained almost unmeasurable levels of cytokinin O-glucosides (5). This implies that the
active product of the gene Zm-p60.1 is present in chloroplasts. Chloroplasts from tobacco
plants carrying the gene for isopentenyltransferase (IPT) under the promoter of the small
subunit of RUBISCO from pea (Pssu) (kindly provided by Dr. Valcke, Diepenbeck, Belgium)
had significantly higher cytokinin content than those from control plants. As the Pssu
promoter is a nuclear one, it is probable that cytokinins are transported to chloroplasts from
the cytoplasm. Chloroplasts from tobacco plants carrying the gene for cytokinin oxidase from
Arabidopsis (AtCKX3) under the CaMV35S promoter (received from Prof. T. Schmülling,
Berlin, Germany) had lower cytokinin content than those from control plants. For the
investigation of cytokinin biosynthesis in chloroplasts, these were immobilised into alginate
gel to prolong their longevity at the temperature necessary for metabolic studies (25oC).
Immobilisation prolonged the longevity of chloroplasts but, on the other hand, decreased the
rate of metabolism, due to diffusion through the gel. Therefore, the usage of immobilised
chloroplasts had not proved to be advantageous for metabolic studies. ◘
Cyclin-dependent kinase inhibitors developed in IEB (roscovitine, olomoucine and bohemine)
were proved to also inhibit N-glucosylation of cytokinins. They affected especially
exogenously applied cytokinins in a species-specific manner, depending on the prevailing
activity of individual cytokinin-down-regulating pathways (cytokinin oxidase/dehydrogenase
or N-glucosylation) (379). Their short-term effect on the endogenous physiologically active
cytokinins was balanced by the mechanisms involved in the maintenance of cytokinin
homeostasis, especially by the stimulation of the formation of physiologically inactive ciszeatin derivatives (300, 379). The long-term effect resulted in cytokinin accumulation,
however, only of the non-active forms. The growth of seedlings was significantly impaired.
Cytokinin-binding proteins which exhibit high binding activity to aromatic cytokinins were
isolated and characterised in oat and wheat grains. Although they have very similar
molecular weight to native proteins and their subunits, they exhibit very different
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immunological properties. Their accumulation in maturating wheat grains correlates with an
accumulation of aromatic cytokinins, including N6-(3-hydroxybenzyl)adenosine, which was
identified in wheat grains using mass spectrometry. On the basis of this correlation and
because of the release of proteins bound during germination the physiological function of
these proteins was proposed: These binding proteins temporarily immobilise aromatic
cytokinins during grain maturation and thus prevent their stimulatory effect on cell division
and, consequently, the premature germination. In contrast, cell division during germination of
mature grains is stimulated by the release of the bound aromatic cytokinins during
degradation of the binding protein (322).
1. 1. 2
Transport of auxins and cytokinins
Using model auxin-dependent and cytokinin-autotrophic tobacco cell lines BY-2 and VBI-0,
the action of auxins and cytokinins was characterized from the point of view of regulation of
their internal and external levels and their transport across the plasma membrane. It was
proved that auxin in the cultivation medium functions as an “external mitogen”, which
regulates the internal auxin level. The significance of the “fine-tuning” of the internal auxin
level for standard progress of cell division was confirmed in studies of auxin carriers.
Changes in the activity of the auxin efflux carrier in relation to the commencement of cell
division were monitored. The studies of the mechanism of action of a specific inhibitor of the
auxin efflux carrier (1-N-naphthylphthalamic acid, NPA) (Fig. 1) resulted in the proposition of
a mechanism controlling orientation (polarity) of cell division, based on regulatory NPAbinding-protein controlled targetting of the auxin efflux carrier to a specific region of the
plasma membrane (253). Kinetic studies of NPA effects on auxin transport out of the cells
revealed much higher effectiveness of NPA than the inhibitors of vesicle-mediated protein
traffic in inhibiting auxin efflux. Experimental data were also provided about the behaviour of
cytoskeletal structures and endoplasmatic reticulum, arguing against the idea proposed
earlier, that the action of NPA and other inhibitors of auxin efflux is more general and based
on the impairing of vesicle-mediated protein traffic (344). Some data on the regulation of
auxin action on plant development were summarised in the review (377), and data related to
auxin transport in papers (394, 395, 401). On the same experimental material, the excretion
of cytokinins from the cells into the cultivation medium within the growth cycle was also
characterised. Cytokinins were excreted from cells to the cultivation medium in relation to
their internal concentration during the whole subcultivation period. It was suggested that,
besides metabolic reactions, transport of cytokinins across the plasma membrane may
represent another mechanism involved in the regulation of internal cytokinin levels (252).
Fig. 1: The effect of 1-N-naphthylphtalamic acid (NPA) on the phenotype of suspensioncultured VBI-0cells. a – cells grown in control medium, day 9; b – cells grown in control
medium supplemented with NPA (final concentration 10 µM), day 9. Note abnormal cell
division planes. Scale bars = 50 µm. Modified from Petrášek et al. (253).
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The hypothesis has been tested that more erect leaves in the modern maize hybrid 3394,
than in the older variety 307, might be a cause of tolerance of the modern maize to
neighbours and, ultimately, the high yield in dense planting. It was found that light controlled
leaf angle development in maize via regulation of polar auxin transport and auxin sensitivity
in leaf tissues (314). The tissue of a new hybrid differed more in sensitivity (number of
receptors) than in affinity (of receptors) towards auxin and it was less sensitive to treatment
with antiauxin p-chlorophenoxyisobutyric acid (PCIB). In the maize leaf, at the junction of the
blade and the sheath, a specialised structure is formed, called the auricle. The growth of the
auricle is controlled by light, probably via polar auxin transport. The mutants with auxinbinding proteins affected differed significantly in the leaf angle. The results suggest the
important role of both auxin-binding proteins and polar auxin transport in determining the leaf
angle in maize (314).
1. 1. 3
Hormonal regulation of some life processes in plants
The optimisation of cultivation protocols and image analysis was a prerequisite for the
investigation of morphological characteristics and measurement of internal levels of
endogenous phytohormones (IAA, cytokinins, ABA) during the process of somatic and
zygotic embryogenesis of Picea abies (36, 76, 399). From the point of view of hormonal
changes, the courses of both somatic and zygotic embryogenesis were very much the same
and characteristic alterations of hormonal levels occurred in the same phases of embryo
ontogenesis. The lower level of auxins and higher level of cytokinins was typical for the early
phase of embryo development. During cotyledon and primary root establishment the level
of IAA increased temporarily but remarkably. During embryo maturation and desiccation the
level of endogenous cytokinins temporarily increased. Somatic and zygotic embryos differed
in the relative proportion of individual cytokinins and in the total endogenous cytokinin
content, which is one order of magnitude higher in zygotic embryos. These differences
disappeared after germination. The dynamics of alterations in hormonal levels implies an
important regulatory role of hormones in the process of embryogenesis.
Expression of the homologue of the gene ABI3 (abscisic acid insensitive from A. thaliana)
was followed in both embryogenic and non-embryogenic lines of Norway spruce differing in
their embryogenic capacity. In the embryogenic lines the ABI3 homologue was expressed
early in the proliferative phase and its expression increased during maturation, while in the
non-embryogenic lines its expression was almost not detectable during all phases of
development. Thus, the expression of ABI3 homologue is proportional to the embryogenic
capacity of the line.
The application of cytokinin N6-benzyladenine (BA) on bean plants increased the water use
efficiency because of a higher stimulation of the photosynthetic rate than the transpiration
rate (116, 183). The positive effect of BA was also observed during the rehydration of waterstressed bean plants. In sugar beet plants, N6-(m-hydroxy-benzyl)adenosine (HBA) was
tested in addition to BA, however, HBA only rarely stimulated the gas exchange (179, 374).
BA applied simultaneously with ABA was able to reverse stomatal closing induced by ABA.
However, BA applied after ABA was not able to induce re-opening of stomata previously
closed by ABA (347). During development of water stress, pre-treatment with ABA markedly
decreased gas exchange parameters at the beginning of the experiment but, in its later
phase, the effect was compensated by delay in development of water stress. Pretreatment
with BA delayed development of water stress and increased photosynthetic rate in waterstressed leaves. At mild water stress BA also reversed the effect of ABA (347).
The rate of photosynthesis decreased in cytokinin-overproducing transgenic tobacco plants
(Pssu-IPT grafts of shoots on control roots and poorly rooted plants of F1 generation) by 20
- 50 % probably as a result of closed stomata. Activity of photosystem 2 was hardly affected
in both transgenic types, nevertheless, in grafts, the activity of photosystem 1 was inhibited
up to 70 % (37). Although the number of chloroplasts in cells did not change in transgenic
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plants, their ultrastructure was markedly changed. Beside distinctive peripheral reticulum
and membrane-bound protein bodies of various densities, large crystalloids of lamellar
structure were localised within chloroplasts (359). 3D-reconstruction of chloroplasts showed
that up to 20 % of total plastid volume could be occupied by crystalloids. Transgenic
tobacco plants were more resistant towards mild water stress, probably due to the
increased activity of antioxidant enzyme systems (186).
A spontaneous mutant of tomato 7B-1 was selected and physiologically characterised. It is
a recessive mutant producing higher amounts of ABA, where germination and hypocotyl
growth were resistant to mannitol and ABA. The mutant was also resistant to osmotic, salt
and low-temperature stresses (160, 161) and showed blue-light-specific resistance to
osmotic stress and abscisic acid (232). Therefore, the mutant 7B-1 is an excellent object for
study of the role of light in plant responses to osmotic stress.
1. 1. 4
Other plant growth regulators
The control of cell growth and differentiation is one of the possible functions of phenolic
compounds in plants. Content and degree of methylation of phenolic substances were found
to be the important factors for initiation of sessile oak somatic embryos and for their further
development. A higher content of some cell-wall-bound phenolic acids could restrict cell
expansion and, consequently, a normal development of somatic embryos (49, 305). Phenolic
compounds could influence also the content of some phytohormones and polyamines. The
application of the inhibitor of phenylpropanoid biosynthesis to alfalfa suspension cultures
resulted not only in the marked decline in the content of phenolic compounds, but also in the
decrease in the content of free IAA and IAA-oxidase activity (97). The formation of
conjugates of phenolic acids with polyamines had a significant impact on the regulation of the
levels of free polyamines in plant cells (304).
An experimental system has been developed which enabled comparison of the endogenous
levels of polyamines with cytological changes of alfalfa explants cultured on media inducing
either (i) the intensive cell division and formation of proembryogenic structures or (ii)
enlarged, highly vacuolated cells with limited cell division. Certain polyamines were shown to
participate in certain developmental processes. High spermidine and spermine levels might
be essential for the development of proembryogenic structures in dividing alfalfa explants,
while higher content of putrescine was characteristic for elongating cells (11). Further
experiments have also shown the involvement of polyamines in the initiation of sessile oak
somatic embryos. The inhibition of phenylpropanoid biosynthesis in sessile oak embryogenic
culture resulted in a decrease in the level of conjugated polyamines and in the increase of
the content of their free forms. The results indicated an important role for spermidine in the
initiation of somatic embryo formation (305). ◘
The dynamics of polyamine content was described in the course of the cell cycle in
synchronous meristematic tissue of Vicia faba roots with the aim of confirming the direct
participation of spermidine in the control of cell division. Polyamine content was determined
in the individual phases of the cell cycle, characterised by flow cytometric analysis. Cells
were released from hydroxyurea block in the G1-phase and progression to S-phase
coincided with a decrease in the level of free putrescine and spermidine, while the G2-phase
was characterised by an increase in free polyamines. The transient prolongation of the Sphase, i.e. the delay in DNA replication, was accompanied by an accumulation of free
spermidine and soluble spermidine conjugates (304). The investigation of the dynamics of
polyamine content during the growth cycle of tobacco BY-2 cell suspension culture resulted
in the finding that the maximum in free spermidine content coincided with the onset of cell
mitotic activity. The results obtained suggest the relationship between DNA replication and
metabolism of spermidine and extend the knowledge about the role of polyamines in the
regulation of plant growth.
The accumulation of free polyamines (namely putrescine) induced by aphid feeding on winter
wheat plants has also demonstrated the important role of polyamines in plant responses to
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biotic and abiotic stresses (94).
Melatonin has been studied as another plant regulatory compound. This substance is known
from animal cells as the regulator of rhythmical and photoperiodical processes. In plants it
has been found that the rhythms in melatonin level in the above-ground parts of
Chenopodium rubrum plants was dependent on photoperiod. The longer the dark period, the
later the maximum of the rhythm occurred. Interestingly, in the case of C. rubrum it always
appeared 6 h before the start of the photoperiod (196). The application of melatonin, its
analogs and agonists, inhibited flower induction in C. rubrum when applied in concentrations
higher than 10-5M 1 h before the start of the inductive darkness. Melatonin has had an
inhibitory effect only when applied after the 6th h of the dark period and it did not change
either the phase or the period of the rhythm of flowering (323).
1. 1. 5 The role of plant growth regulators in plant stress reactions
High irradiation (700 µM m-2 s-1) brought about a decrease in the maximal efficiency of
photosystem 2, in quantum yield and in photochemical quenching and stronger
photoinhibition in tobacco plants cultivated in vitro in closed vessels in comparison with
plants cultivated with a higher supply of CO2 in ventilated “Magenta” vessels. The positive
effect of higher CO2 concentrations was obvious not only during the in vitro cultivation (22),
but also after the transfer of plants into ex vitro conditions (31, 115). Increased CO2
concentration stimulated plant growth to a higher extent by increasing the velocity of
photosynthesis more than by slightly increasing pigment content. At the same time there
were increased activities of peroxidase, glucose-6-phosphate dehydrogenase and malic
enzyme. Application of abscisic acid (ABA) immediately after the transfer ex vitro weakened
the “transplantation shock” and probably decreased formation of active oxygen species. This
resulted in a decrease of activities of glutathione reductase, Mn-superoxide dismutase,
peroxidase, glucose-6-phosphate dehydrogenase and malic enzyme (263). However, wheat
plants growing under the increased CO2 concentration had lower photosynthetic capacity and
on the opposite side, plants growing under lower CO2 concentration had an increased
photosynthetic capacity. These changes persisted even after return the of plants to natural
CO2 concentration (128).
The role of antioxidative enzymes was studied in the sensitivity to chilling of some inbred
hybrid maize lines. Low temperature induced activities of glutathione reductase and
ascorbate peroxidase and, at the same time, increased concentration of carotenoids. Chilling
without preceding acclimatisation caused higher changes in enzyme activities than a gradual
decrease of temperature.
Photosynthetic capacity decreased during senescence (74). The nature of products of radical
reactions in lipids did not change during ageing of bean cotyledons, while their amount rose
significantly with age. Mass spectrometry showed that the C12 compounds were the most
abundant. Young and old cotyledons differed in their composition. Oxidative damage of
soluble proteins was studied using antibodies against carbonyl groups and it was found that
this damage increased during ageing. Oxidatively damaged proteins were rapidly destroyed.
Beside reactive oxygen species, reactive nitrogen species are active as well. The content of
nitrotyrosine, determined by specific monoclonal antibodies, increased in old cotyledons.
Both enzymatic and non-enzymatic antioxidative system was studied. Ratios of reduced and
oxidised ascorbate and glutathione diminished during ageing. Similarly, activities of
glutathion reductase, ascorbate peroxidase, SOD and catalase decreased as well as the
capacity of the enzymatic and non-enzymatic systems (349). These events resulted in
oxidative damage in the course of ageing.
Phenolic acids play an important role in the reaction of plants to abiotic stress as was
demonstrated in acclimation of soybean roots to low temperature. Low temperature in
soybean plants brought about a decrease in the concentration of phenolic acids bound in the
cell walls and an increase of free phenolic acids (99, 241). In spruce needles damaged by
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emissions increased lignification was found together with an increased concentration of
conjugated phenolic acids and a lower concentration of phenolic acids bound to cell walls
(119).
1. 1. 6 New methods in phytohormone research
A new rapid method was developed for parallel extraction, separation and purification of
phytohormones of the auxin type, abscisic acid and cytokinins, based on the use of new twoparameter sorbents with the functions of cation exchanger and reverse phase. The method
substantially shortens the time needed for phytohormone analysis while at the same time
increasing their recovery and enabling subsequent mass spectrometric determination (228).
A new method was developed for immunolocalisation of cytokinins by means of antibodies
against zeatin riboside and isopentenyladenosine (prepared in the laboratory of Prof. Strnad,
IEB) and different fixation procedures for free bases, ribosides and ribotides, respectively.
Specific detection is possible only in the case of free bases fixed by formaldehyde. ◘
A new sensitive ESI-MS (electrospray ionisation - mass spectrometry) method was
elaborated which enables the use of a cheap and simple (in comparison with MS/MS
techniques) mass detector with one quadrupole analyser for quantitative cytokinin analysis
(337).
Tritium-labelled cytokinins were prepared with very high molar radioactivity (higher than
1TBq/mmol), namely cis- and trans-zeatin, N6-isopentenyladenine, N6-benzyladenine,
dihydrozeatin and their ribosides. For trans-zeatin a new synthesis was worked out. Two
original syntheses of non-radioactive cis-zeatin were elaborated. The following standards,
trans-zeatin-9-glucoside-O-glucoside, glucose esters of 3-indoleacetic and 2,4dichlorophenoxyacetic acids, as well as putrescine and spermidine amide conjugates of
tyramic, p-coumaric and ferulic acids were prepared for HPLC/MS and metabolic studies.
1. 1. 7 Potential practical applications of research on plant growth regulators
A new series of inhibitors of cyclin-dependent kinases (CDK) was developed as well as a
system of modelling enabling targeting of the molecules of CDK inhibitors into the binding
site for ATP in the molecules of CDK2 and CDK1. New compounds with high inhibitory
activity to CDK1 were designed derived from purines, pyrazolo[4,3-d]pyrimidines and 8azapurines. The most active of these substances with IC5O 0.01µM for CDK2 was given the
name olomoucine II (6-(2-hydroxybenzylamino)-2-{[1-(hydroxymethyl)propyl]amino}-9isopropyl purine) and it has the same inhibitory activity towards CDK as purvalanol, until now
the most efficient inhibitor. However, olomoucin II is more easily synthesised, it is, as well,
soluble under physiological conditions and more effective in the tests of cytotoxicity in tumour
cell lines. The inhibitor of CDK of the first generation, roscovitine (6-benzylamino-2-[1-R(hydroxymethyl)propyl]amino-9-isopropyl purine) entered the second phase of clinical tests in
several European countries. Among the synthesised compounds there are also substances
that do not exhibit pronounced inhibitory activity towards CDK, but they show high
cytotoxicity in tumour cell lines. Metabolism of one selected CDK inhibitor, bohemine (6benzylamino-2-[3-hydroxypropylamino]-9-isopropyl purine) was described and it was shown
that the main metabolite is its O-glucoside. In addition, a stimulatory effect of CDK inhibitors
on production of monoclonal antibodies was described (103, 114, 164, 170, 236, 242, 243,
245, 246, 272, 273, 333, 334).
It was demonstrated that both roscovitine and bohemine induce the expression of nonmutated form of protein p53 in tumour cell lines. The protein actively binds to DNA and is
capable of activation of some genes, e.g. the one coding the protein p21WAF1. This finding
should start development of a new generation of anti-tumour agents the molecular target of
which should be the gene/protein p53, which is most often mutated in human tumours (173).
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A number of complexed substances - derivatives of benzylaminopurine (BAP) (aromatic
cytokinins) and selected transition metals (Ni, Cu, Fe, Pd, Pt) - were prepared. Testing
biological activity of these compounds showed that complex formation can result in a
substantial increase of cytotoxicity against selected tumour lines (33, 175, 192, 248, 269,
367).
1. 2 Signalling and signal transduction
1. 2. 1 Signalling mechanisms
The regulatory role of plant proteins dependent on GTP (so called small GTPases) was
shown in establishing the cell polarity and in regulation of morphogenesis (79, 199).
Microinjection of non-metabolised analogues of GTP/GDP into living polarised plant cells led
to the loss of their polarity while microinjection of ATP/ADP was without any immediate effect
(158). We suggest that the most important small GTPase in the processes described is the
Rop GTPase. Currently the Rab geranylgeranyl transferase complex (GGTasell) is being
characterised, which is responsible for posttranslatory prenylations of Rab GTPases. Its
subunit was described called "Rab escort protein" (REP), which is very close to the
homologue of Rab GDP dissociation inhibitor as described by us earlier. In position 195 of
this protein asparagine was detected as a specific amino acid for plant REP. ◘
In Arabidopsis plants three subunits were characterised (Sec6, Exo70G1 and Sec10) of
complex Exocyst, which is an effector of small GTPases in polarised/localised exocytosis
not yet described in plants. Using antibodies against the subunit Sec6 its localisation was
shown in the membrane-bound complex in the tips of growing tobacco pollen tubes (313)
(Fig. 2). ◘
Fig. 2: Indirect immunolocalisation of Exocyst Sec6 subunit in proliferating pollen tubes of
tobacco (Nicotiana tabacum, cv. Samsun).
Plant phospholipases D (PLD) were phyllogenetically analysed in detail (283) and their
possible role in regulation of cell expansion was studied. It was shown that the product of
PLD activity - phosphatidic acid (PA) - acts as an important signal in the regulation of polar
morphogenesis of plant cells and of their growth. The specific inhibitor of PLD - 1-butanol stopped the growth of the pollen tube and application of phosphatidic acid restored it. Growth
was partially restored also after taxol application, which suggests binding of PLD to
microtubules (348). ◘
Basic components of plant phospholipid signalling pathways were studied. The receptor for
inositol-1,4,5-trisphosphate was localised inside the cells at the endoplasmic reticulum. This
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finding supported the hypothesis on the existence of an alternative source of calcium ions
(besides the vacuole) in plant cells (108). For the first time, the regulatory function of
phosphorylation of membrane-bound phosphatidylinositol bisphosphate-dependent
phospholipase D, isolated from 5-day-old hypocotyls of Brassica oleracea was shown (340).
◘
Three types of signalling enzymes degrading phospholipids - phosphatidylinositol-dependent
phospholipase C (PI-PLC) and two types of phospholipase D, PIP2- dependent and
independent (PIP2, PLD and PLDα, respectively) - were studied during ripening, germination
and early growth of oilseed rape. Activities of membrane-bound forms of all three types of
phospholipases changed during ripening in a different way to the activities of soluble forms.
PIP2-PLD activity changed in an opposite way to the activities of PLDα and PI-PLC, both
during ripening of rape seeds and during germination and growth of seedlings. In fractions
isolated from hypocotyls of seedlings most phospholipases were detected in the form bound
to the plasma membrane (341). ◘
To monitor phospholipase activity in situ a new method was developed using fluorescencelabelled substrates. These substrates are incorporated into membranes (mainly into the
plasmalemma) of living cells (Fig.3). Fluorescently labelled products of phospholipase activity
are then quantified using TLC or HPLC. ◘
Fig. 3: Plasmolysed BY-2 tobacco cell with incorporated fluorescent BODIPY FLC5 –
phosphatidylinositol -4,5-bisphosphate (PIP2) to the plasma membrane (green colour).
Fluorescent shuttle PIP2 carrier stay in cell wall (red colour). Cells with incorporated
BODIPY-PIP2 is used for in situ determination of phosphatidylinositol specific phospholipase
C activity.
Treatment of plant cells with elicitors induced changes in phospholipase activity.
Phospholipase A activity was increased after treatment of tobacco cells with cryptogein
(elicitor from the pathogen Phytophthora cryptogea) and after treatment of parsley cells with
a glycoprotein elicitor isolated from Phytophthora sojae. Under these conditions, the
formation of diacylglycerol was decreased. Phosphatidic acid was detected only in minor
amounts, which suggests that diacylglycerol was formed from the added phosphatidylcholine
by the activity of phospholipase C hydrolysing phosphatidylcholine. These results are the first
description of a new plant phospholipase C hydrolysing phosphatidylcholine and suggest its
role in signal transduction (260).
Oscillations in the excretion (flux) of chloride ions at the tip of pollen tubes were shown to be
the main regulator of the growth rate of the pollen tube (197, 276). This excretion of chloride
ions is regulated by a harmonic oscillator, which is switched on before growth oscillations can
be observed (378). The chloride oscillator is tightly coupled to changes in the pressure
potential in the cell and correlates with osmoregulation and cell volume changes and reacts
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to the signal inositol-3,4,5,6-tetrakisphosphate (Ins3,4,5,6P4), which determines the
frequency of the oscillator (197, 276) (Fig. 4). It seems that this oscillator is able to convert
the pollen tube to a sort of "pneumatic hammer" which enables the cells to penetrate stigma
tissue. ◘
Fig. 4:
Proposed minimal model for
dynamic Cl− efflux oscillations from the
apex of pollen tubes. The Cl− pool at the
apex is supplied from Cl− influx along
distal regions of the tube and an
endogenous source. Spatial coupling of
efflux and influx sites predicts a closed
loop of flux vectorially traverses the apical
region. Cell volume sensing circuits (grey
nodes and flowlines) are hypothesized to
be concentrated in the apical dome. The
Ins(3,4,5,6)P4 cycle and putative Ca2+
cycle are hypothesized to be phaseshifted by 180°. Negative flowlines are
denoted as l⎯I , positive flowlines are
denoted as ´. The thick outline around the cell denotes the cell wall, the thin interior line
denotes the plasma membrane.
1. 2. 2 Coupling of signal transduction to the cytoskeleton
Structural and functional analysis of the cytoskeleton in the course of microspore
development showed that in critical developmental phases specific cytoskeletal structures
are formed, which determine further direction of development. It was shown that
microtubules are important in nucleus migration and polarisation of microspores, that actin
and tubulin structures take part in the course of microspore asymmetric mitosis and that actin
has a significant role in postmitotic differentiation of vegetative and generative cells of the
pollen (46, 152). During the induction of pollen embryogenesis the actin cytoskeleton shows
significant reorganisation, namely around and inside the vegetative nucleus. Phases of
gametophytic pollen development are coupled with the occurrence of developmentally
specific fucosylated and especially mannose or hybrid N-glycoproteins (23). Glycoprotein 92
kDa, specific and, at the same time, dominant in the phase of asymmetric microspore
mitosis, corresponds in its amino acid sequence to β-galactosidase. Glycoproteins 51 and 59
kDa, which accumulate during pollen maturation are thermostable and glycoprotein 59 kDa
belongs to the group of dehydrins. In microspore cultures of potato, certain concentrations
and forms of organic nitrogen induce a change of asymmetric mitosis into the symmetric type
of division and reorientation of gametophytic development into an early embryogenic one
(34).
Nucleation and cytoskeleton organisation was studied in plant cells with the absence of
centers for nucleation of microtubules, like are e.g. centrosomes in animals. In acentrosomal
plant cells the distribution of γ-tubulin was characterised; this protein is known to play a key
role in nucleation and organisation of microtubules in animals (85, 299). Beside the
occurrence of γ-tubulin in the nucleus, it also was found to be associated with membranes in
the form of protein complexes (312) (Fig. 5). Using immunoprecipitation association of γtubulin with the dimers of α- and β-tubulin was proved. Large γ-tubulin complexes, resistant
to cleavage by salts, were detected in the microsomal fraction; molecular mass of these
complexes was higher than 1 MDa. These complexes were active in microtubule nucleation.
Association of γ-tubulin complexes with dynamic membranes obviously enables flexibility of
acentrosomal nucleation of microtubules.
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Fig. 5: Immunofluorescence localisation of microtubules (green), Golgi membranes (red) and
DNA (blue) in mitotic cell of Vicia faba. Golgi membranes are accumulated in close vicinity of
spindle poles.
Specific changes of cytoskeleton during somatic embryogenesis in Norway spruce were
described. F-actin was detected in dividing cells of the embryo head in the whole course of
mitosis. Transitory co-localisation of actin microfilaments with the preprophase microtubule
bundle was observed. F- actin was not detected in kinetochore microtubule bundles in the
course of metaphase and anaphase. A high intensity actin signal was seen in the spindle in
the late anaphase in the equatorial plane between separating chromatids (303). ◘
Regulatory aspects important for cytoskeleton organisation were also studied. Activation of
the cell cycle in zygotic embryos in the course of imbibition and its relation to tubulin
expression and microtubule orientation were analysed in Vicia faba and Medicago falcata
(19). Gradual polymerisation of microtubules was seen coupled with reactivation of the cell
cycle in those cells, which entered the cycle from the G1 phase, in which the embryo cells
were present in dormant seeds. Cell cycle reactivation was similarly coupled with significant
reorientation of the microtubular cytoskeleton in the induction phase of somatic
embryogenesis of alfalfa (11). ◘
1. 2. 3 Signalling in plant defence mechanisms
The ability of a plant to withstand unfavourable external conditions can be demonstrated in
the synthesis of the so-called small heat-shock proteins (sHSP). In collaboration with the
Department of Plant Physiology (Faculty of Natural Sciences, Charles University, Prague) a
new type of their regulation by means of the level of ATP in the cells was described (118), in
which ATP inhibits interaction of the sHSP complex with the substrate. In the phase of acute
stress, when the ATP level was decreasing, the affinity of the protecting sHSP towards the
cell components endangered by stress was increased. After the end of the acute phase of
the stress the increased ATP level loosened these components from the complex and
renaturation of HSP70, occurred.
Defence mechanisms of plants against pathogens (especially viruses) were studied with
special attention to the induction of systemic acquired resistance (SAR) and induced
systemic resistance (ISR) by synthetic and natural inducers. Synthesis of pathogenesisrelated proteins (PR-proteins) was also studied (9, 189, 267, 301, 302). The composition
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and amount of induced PR-proteins (9) as well as their localisation in tissues (301, 302)
depended on the applied inducer. In sugar beet (9, 302) and wheat, the most effective
inducer was benzothiazol (BTH) and also glycinebetaine and salicylic acid were active
inducers. Isozymes of β-1,3-glucanases and chitinases were found both in leaves and in
roots after treatment with chemical inducers, infection with Polymyxa betae and beet mosaic
yellow vein virus (BNYVV) (Fig. 6). The results show the possible role of these enzymes in
resistance to rhizomania. BTH application induced resistance to tobacco mosaic virus (TMV)
and potato virus Y (PVY) (266). Nine different inducers including BTH were efficient in wheat
and strongly decreased growth of the fungus Blumeria graminis. In contrast to the inducers,
herbicides with auxin-like activity increased the content of TMV in tobacco leaf discs but did
not induce PR-proteins (121). Enzymes of the biosynthesis of viral RNA precursors, their
regulation, subcellular localisation and relation to plant resistance were also studied (9, 40,
41, 122, 265, 266, 267, 361). Correlation between the activities of the key enzymes of
metabolic pathways and the virus reproduction was shown for glucose-6-phosphate
dehydrogenase (G6PDH) (9, 40, 41, 265, 266, 267, 361) and ribonucleases (9, 122, 267,
361). In the synthesis of the virus chloroplastic isozyme of G6PDH was predominantly active
(265). Activities of these pathways also increased considerably after BTH application, which
suggests possible competition between virus synthesis and defence reactions (267). In
contrast antivirus factor (AVF) decreased infectivity of TMV by lowering activities of the
above mentioned enzymes (363).
Fig. 6: Immunohistological localisation of basic chitinase Ch4 in healthy (left) and
rhizomania-diseased (right) root tissues. In sections of infected roots (right) (P.betae
cystosori- Pb), Ch4 was found mainly in endodermis cells (arrowheads) and xylem vessels,
but in some rhizodermal cells the accumulation of Ch4 was also detected (arrowheads). In
healthy control plants (left), slight staining indicates the constitutive presence of Ch4.
Patterns were determined in at least 10 plants and were remarkably consistent.
Infection of control and transgenic tobacco plants carrying the Pssu-IPT construct with potato
potyviruses A (PVA) or Y (PVY) resulted in different responses. Infection with PVY caused in
control and F1 transgenic plants a decrease in photosynthesis and in parameters of
chlorophyll fluorescence kinetics and an increase in activities of phosphoenolpyruvate
carboxylase, NADP-malic enzyme and pyruvate dikinase. The effect of PVA was negligible
(348). Electron microscopy showed aggregates of virus particles in the vicinity of
chloroplasts. Pssu-ipt grafts were highly resistant to virus infection and infection with PVA
even stimulated their photosynthesis (348).
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2 STRUCTURE AND FUNCTION OF THE GENOME
2. 1 Functional genomics
Using methods of in vitro transcription, translation and N-glycosylation we positively
demonstrated that the model tobacco pollen-specific N-glycoprotein p69 is encoded by the
ntp303 gene (130). We proved the phylogenetic conservation of this gene at the levels of 1-D
and 2-D SDS-PAGE protein spectra of pollen tube-walls in 16 plant species of 15 genera
(162). The ntp303 transcript is synthesised during pollen maturation after the first haploid
mitosis, accumulated in the non-translatable form and was utilised after germination during
the progametic phase of male gametophyte development. We observed the kinetics of
ntp303 transcript synthesis and, for the first time in plants, we described a new form of
mRNA storage RNP complexes. ntp303 transcripts are stored in the form of translationally
silent EPP particles („EDTA-puromycin resistant particles“), high molecular weight RNP
complexes resistant to polysome-destabilising agents EDTA and puromycin (Fig. 7) (95, 96,
168).
Fig. 7:
Potential model of the developmental
regulation of ntp303 mRNA subcellular distribution.
Newly synthesized ntp303 mRNA is released from the
nucleus in the transport form of mRNPs. Between
stages 3 and 5 ntp303 mRNA is distributed evenly
between polysomes and EPPs. All polysomes
associated with ntp303 mRNA are formed at this stage
and are translationally silent. EPPs are proposed to be
the long term storage compartment formed by
aggregation of individual ntp303 mRNPs probably with
other proteins. Between stages 5 and 6, polysomes
associated with ntp303 mRNA are still present in the
vegetative cell but their amount does not increase.
ntp303 mRNA synthesized at this time remains in the
transient form of “free” mRNPs with only small portion
of them combining into EPPs. In the final period of
maturation between stages 6 and dry pollen the
synthesis of ntp303 mRNA is complete, but a massive
redistribution of ntp303 mRNA from “free” mRNPs and
polysomes to EPPs occurs.
For the first time in the plant kingdom, we performed a genome-wide analysis of the gene
expression of a single developing cell at the transcriptome level. Mainly, we focussed on
changes in the gene expression during ontogenesis. As a model system, we used the
vegetative cell of Arabidopsis pollen. The gametophytic gene expression was quantified and
actively transcribed genes were assorted to functional categories. Gametophytic and
sporophytic transcriptomes were compared and the extent of their overlap determined (321,
326). We have also described putative gametophytic transcription factors and clusters of coexpressed genes containing candidates for putative regulons, expression of which is
controlled by particular gametophytic transcription factors (388).
The dynamics of expression of another plant translationally regulated and constitutively
transcribed D1 protein was studied. The translation intensity of D1 protein, one of the key
proteins of photosystem II, during the first two days following stress treatment remained
unchanged and high. Substantial decrease of its synthesis was observed the third day after
stress application, when other physiological and biochemical parameters were stabilised at
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the control level (166, 171, 240).
For proteomic studies, new original methods were developed for mitochondrial isolation and
fractionation and protein extraction from small amounts of starting material (grams instead of
kilograms). We demonstrated that heat stress (40o C) immediately induced intensive
expression of 9 low molecular weight proteins: 8 sHSPs and 1 transcription factor. Qualitative
and quantitative representation of all these proteins, which form part of the membrane
fraction, remained stable during the first 12 hours of stress treatment
2. 2 Studies on genome structure using sorted chromosomes
Although many authors believe in intraspecific variation in nuclear genome size, the results
obtained in the genus Musa (26), Sesleria (107) and Agave (342) do not support this
assumption. The genus Musa was used as a model to study the evolution of polyploid
species. The size of the nuclear genome has been established in wild diploid species and in
diploid and triploid parthenocarpic cultivars for the first time (26). Systematic genome
analysis resulted in the isolation and characterisation of a number of repetitive DNA
sequences which constitute a significant part of the banana genome (271). These include a
new type of „monkey“ retrotransposon (83).
Fig. 8: Analysis of genomic distribution of five repetitive DNA sequences (pSc119.2, GAA
microsatellite, Afa family repeat, and 5S rDNA) on chromosomes of tetraploid wheat
(Triticum durum cv. Langdon; 2n = 4x = 28) using fluorescence in situ hybridization (FISH).
Labelled probes were detected either with fluorescein (yellow-green signals) or Cy3 (red
signals); chromosomes were counterstained with DAPI (red pseudolocour for single colour
FISH; blue pseudocolour for double colour FISH). For each chromosome type, two
representative examples are given.
In collaboration with the laboratory of Prof. B. Vyskot (Institute of Biophysics, Brno), the
evolution of sex chromosomes was studied in plants. Nuclear genome size and genomic
distribution of genes for ribosomal RNA have been established in species of the genus
Silene (190). Development of a method for flow cytometric sorting of sex chromosomes
facilitated physical mapping of MROS genes, expressed in male plants of S. latifolia, to X
chromosome and autosomes (172). Also, the Y chromosome has been found to harbour a
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MADS box gene, which has been duplicated from an autosome (331).
Analyses of crop plant genomes are hampered by the genome size and complexity. In order
to simplify the genome analysis, methods for chromosome sorting have been developed in
barley (27), wheat (129, 222, 244) and rye (325). The genomic distribution of some repetitive
DNA sequences was first established by the method of sorted chromosomes (104, 207, 325)
and, using the same method, the chromosomal localisation of molecular markers was
determined (27, 325) (Fig. 8).
The development of a method for preparation of high molecular weight DNA from sorted
chromosomes and the construction of chromosome-specific DNA libraries cloned in BAC
vectors (360) have been a significant success. Flow-sorted chromosomes have also been
used to map nuclear genomes in legumes. In field bean, new repetitive DNA sequences
were isolated and characterised (28) and new microsatellite molecular markers for
chromosome 1 have been isolated (255). Flow sorted chromosomes were used for physical
mapping and integration of genetic and physical maps in garden pea (251) and chickpea
(275).
2. 3. Genotoxicity and DNA repair
Fig. 9: Images illustrating the induction of DNA damage in tobacco nuclei as expressed in
the Comet assay. Control nucleus (A) and nuclei with different levels of DNA damage (B, C,
D).
For DNA repair studies, the Comet assay was modified (Fig. 9) to enable the detection of
various types of DNA damage and their localisation by the FISH method (fluorescence in situ
hybridization) in specific sequences, for example telomeric regions or repetitive sequences
(110). The development of the modified method of the Comet assay was necessary for
assessment of efficiency and kinetics of DNA repair and, when the Arabidopsis thaliana
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genome sequencing is completed and knock-out mutants are available, has aided studies
on particular repair pathways. The sensitivity to chemical mutagens causing DNA damage
was tested. The repair of these DNA damages can be achieved by various mechanisms
involving various repair enzymes. The effects of chemical mutagens (alkylation mutagens
N‘-methyl-nitrosourea a methyl methanesulfonate; radiomimetic bleomycin causing doublestrand breaks in DNA; mitomycin C inducing cross-link bonds in DNA; plant morphoregulator
maleinhydrazide) aid the verification of the mutant phenotype (177). These results led to
studies of mutants defective in various repair pathways, mostly the ones defective in genes
involved in homologous recombination: AtSpo3-11, AtT3B, AtARF1, AtTop6B, AtRad9,
AtRad17, AtBRCA1, AtFANDC2, or non-homologous end-to-end joining of double-strand
breaks (NHEJ): AtKu70, AtKu80. So far, the characterisation of AtTop6B topoisomerase
mutant has been published (239). Beside the analyses of Arabidopsis repair mutants, the
adaptation to genotoxic stress has been proved in Vicia faba and Arabidopsis after
application of various alkylation mutagens. The effect of alkylation mutagens is not solely
connected to O6-guanine alkylation as had been expected, based on analogy with other
organisms, until now (80).
Gamma-irradiation of tobacco seedlings induced a dose-dependent increase in somatic
mutations and was highly correlated (r = 0.99) with the increased DNA damage in the nuclei
of the leaves. 24 h after irradiation a complete repair of DNA damage induced by gammairradiation and measurable by the Comet assay was observed, whereas the yield of somatic
mutations increased in relation to the radiation dose (181). By contrast, DNA damage
induced by the monofunctional alkylating agent ethyl methanesulphonate persisted over a 72
h period (91).
The plant growth regulator and herbicide, maleinhydrazide (MH), induced, in sprouting
tobacco plants, a high frequency of somatic mutations and recombination events, but no
significant increase in DNA damage measured by the Comet assay. MH represents the first
chemical agent which has proved to be highly mutagenic but does not cause DNA damage
as measured by the Comet assay in the same experimental system (90, 318).
2. 4. Molecular aspects of plant virology
The influence of viral infection caused by two different Potyviruses, Potato virus Y (PVY) and
Potato virus A (PVA) on plant metabolism and photosynthetic apparatus of Nicotiana
tabacum L. cv. Samsun and cv. Petit Havana SR1 was studied. The main emphasis was
focussed on the activities of enzymes that catalyse anaplerotic metabolic pathways phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31), NADP-malic enzyme (NADP-ME,
EC 1.1.1.40), and pyruvate phosphate dikinase (PPDK, EC 2.7.9.1). PVY infected tobacco
plants responded with enhanced activities of all three enzymes. NADP-ME was the most
sensitive to viral infection; the enzyme activity was 5 times higher in Samsun and 6-fold
higher in SR1, compared with control plants. The activities of PEPC and PPDK were
enhanced 2-3 times in both cultivars. In contrast to PVY, PVA infection affected enzyme
activities insignificantly, although a moderate increase of activities was observed. No
significant difference between both tobacco cultivars was found (352).
The entire nucleotide sequence of coding regions of Potato mop-top virus (PMTV) isolate
named 54-15 was determined. The genome was found to be highly conserved when the
nucleotide sequences and their derived respective amino acid sequences were compared
with other known isolates of PMTV, even though the PMTV isolates exhibited different
symptoms in host plants and originated from different parts of Europe. However, several
changes in nucleotide and amino acid sequences in parts of RNA coding for “triple-geneblock” (TGB) proteins and in the “read-through” part of the coat protein (CP) RNA were
detected. The phenotypic differentiation of these isolates could be attributed to these
mutations. In order to confirm this hypothesis, the TGB and read-through CP regions from
several other isolates were sequenced, namely 54-19 and 54-10. The comparisons of
sequences obtained with sequences of other isolates available in databases imply high
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genetic stability and slow evolution within the PMTV population. The most remarkable
evolutionary event observed would be the disruption of the fourth open-reading frame (ORF)
on TGB-coding RNA, identified as a gene for cysteine-rich protein in other PMTV isolates
(308).
2. 5. Potential practical applications of genetic research
To test the concept of plant derived edible vaccine, an agroinfection system has been set up
in tobacco and tomato in order to characterise molecular and immunological features of viral
proteins of human papilloma virus HPV-16. In this system, the production of 0.5 – 1 % of total
protein was achieved in the case of oncoprotein E7, which can potentially be used as a
therapeutic vaccine after the onset of cervical cancer and in the case of coat protein L1, that
could serve as prophylactic vaccine. L1 protein is produced as structural capsomeres. The
conditions influencing formation of the pseudovirions are under study, as well as the
immunogenicity of produced proteins.
Gene coding for the coat protein of Potato virus A (PVACP) was cloned. This gene was also
modified, so that the epitopes of structural and nonstructural proteins (L2 and E7,
respectively) from human papilloma virus (HPV), those attached to the N-terminus and/or Cterminus of PVACP protein (modified genes: PVACP+E7, L2+PVACP and L2+PVACP+E7),
could be expressed.
New apple varieties bearing the Vf resistance against scab, the most widespread and
harmful disease caused by the fungus Venturia inaequalis, have been bred. Eleven
selections were applied for Plant Breeder’s Rights in the Czech Republic under the numbers
MAL 7121, 7123, 7125, 7983, 8137, 8138, 8139, 8140, 8641, 8642, 8643 and 4 varieties
were applied for Community Plant Variety Rights in the European Union under file numbers
1999/0804, 2001/0082, 2003/1163, 2003/1164. Plant patents / Plant Breeder’s Rights
granted and licence agreements for commercialisation of the varieties concluded are listed in
the section 13. In addition testing agreements with option were concluded in Australia
(5 var.), South Africa (5 var.), New Zealand (1 var.) and the USA (10 var.). Within the
breeding of resistant varieties with compact, columnar growth habit derived from Wijcik
McIntosh mutant, one selection was applied for Plant Breeder’s Rights in the Czech Republic
(MAL 07120) and option agreements were concluded for 3 selections with a German society
for the European Union and for 8 selections in the USA. The resistant varieties bred at the
Střížovice Station were commercialised mainly for organic fruit growing in Europe and
regular royalties from the licences became an important financial source for the Institute. In
the follow up breeding of varieties with multiple durable resistance, breeding material was
selected for testing Vf homozygosity and combination of Vf with polygenically encoded
resistance using molecular markers.
Using the method of microprojectile bombardment, the genes DAP A and PHYT (gene dapA
codes for dihydropicolinate synthase form insensitive to feedback inhibition and increasing
lysine content and gene PHYT codes for phytase, which affects phosphorus content) were
transferred to the cell genomes of immature zygotic embryos of barley, from which
transgenic plants were derived. The transformation frequency in anther cultures of these
primarily transformed plants was low (215). The presence of DAP A gene was confirmed by
molecular analysis in four androgenous regenerants, one of which was capable of producing
seeds. The gene PHYT was detected in more than 70% of androgenous regenerants. A
tetrahaploid transgenic line of barley, carrying a phytase gene, has been derived, which we
consider to be a significant success.
The analysis of transgenic potato plants transformed with the introduced bacterial gene
LbPFK (phosphofructokinase of Lactobacillus) was performed in field tests (yield
characteristics). Among the transgenic plants, phenotypically standard plants prevailed; sole
27.5% of transformed KIamýk cultivar and 21,4% of transformed Korela cultivar, respectively,
showed a deviation from normal phenotype in either growth or flowering. The transformed
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lines generally displayed a yield equal or higher (by 3 to 32%) compared with control lines.
The levels of reducing sugars in harvested tubers of transgenic plants was higher than that
observed in tubers of control plants. During the cold-storage of tubers (160 days), however,
the amount of reducing sugars in transgenic tubers decreased, while in control tubers it was
increasing. By the end of a storage period, the amount of reducing sugars was lower in
transgenic tubers than in control non-transgenic tubers. One of the transgenic lines, which is
considered the most interesting, contained only 0,17% of reducing sugars after a period of
160 days cold-storage.
LIST OF ABBREVIATIONS
ABA
ABI3
ABPs
BAP (BA)
BNYVV
BY-2
BTH
CaMV35S
CDK (cdc2)
CHN/O analysis
CKX (AtCKX3)
CP
2D-HPLC
DsRed
EDTA
ELISA
ESI-MS
EPP
EST
FPLC
FRET
GDP
GTP
GTPase
GFP
G6PDH
GC/MS
GUS
HPLC
HPLC/MS (LC/MS)
(s)HSP
IAA
IEB
IPT
IPMP
IR spektrometry
LC
MALDI-TOF
MH
MS
NMR
NPA
ORF
PAGE
abscisic acid
ABA- insensitive (3) A. thaliana mutant
auxin binding proteins
N6-benzylamino purine
beet necrosis yellow vein virus
tobacco cell culture
benzothiadiazole
promoter from cauliflower mosaic virus
cyclin-dependant kinase (2)
elementary analysis
cytokininoxidase/reductase (from A. thaliana)
coat protein
two-dimensional HPLC
Discosoma species Red (fluorescent red dye from species
Discosoma)
ethylenediamine tetraacetic acid
enzyme-linked immunosorbent assay
mass spectrometry with „electrospray“ ionisation
EDTA, puromycin-resistant particles
expressed sequence tag
fast protein liquid chromatography
fluorescence resonance energy transfer
guanosine bisphosphate
guanosine trisphosphate
enzyme cleaving guanosine trisphosphate
green fluorescent protein
glucose-6-phosphate dehydrogenase
gas chromatography with mass spectrometric detection
β-glucuronidase
high performance liquid chromatography
high performance liquid chromatography with mass spectrometric
detection
(small) heat-shock proteins
indolyl-3-acetic acid
Institute of Experimental Botany
isopentenyl transferase
isopentenyladenosine-5´-monophosphate
infrared spectrometry
liquid chromatography
„matrix-aided laser desorption ionisation-time of flight“- type of mass
spectrometry
maleinhydrazide
mass spectrometry
nuclear magnetic resonance
1-N-naphthylphthalamic acid
open reading frame
polyacrylamide gel electrophoresis
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PEPC
PCIB
PIP2
PLC
PI-PLC
PC-PLC
PCR
PLD
PMTV
PPDK
PR proteins
Pssu-IPT
PVA
PVY
R/FR
RNAi
RT-PCR
SAR
SDS-PAGE
SOD
TGB
TLC
TMV
VC
YFP
phosphoenolpyruvate carboxylase
p-chloroisobutyric acid
phosphatidylinositol-4,5-bisphosphate
phospholipase C
phospholipase C hydrolysing phosphatidyl inositol
phospholipase C hydrolysing phosphatidyl choline
polymerase chain reaction
phospholipase D
potato mop-top virus
phosphopyruvate dikinase
pathogenesis related proteins
gene IPTcoding for isopentenyl transferase under the control of
promoter of small subunit of Rubisco from pea
potato virus A
potato virus Y
red light 660 nm/ far-red light 730 nm
small interfering RNA
real time polymerase chain reaction
acquired systemic resistance
polyacrylamide electrophoresis with addition of sodium dodecylsulfate
superoxid dismutase
triple gene block
thin layer chromatography
tobacco mosaic virus
research center
yellow fluorescent protein
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C5.
Objectives of IRP
Plans for the period 2005-2010
The traditional areas of research in the IEB have been focussed previously on two main
topics:
(i)
plant growth and development;
(ii)
plant genomics.
Both these research areas have been intensively studied on the Institute’s grounds and
acknowledged in previous years.
For plant growth and development, future research will be aimed mainly towards:
•
studies on metabolism, transport and function of various regulatory metabolites
(mainly phytohormones)
•
signal transduction (phosphoinositide signalling system, small GTPases, Exocyst
complex and formin-associated proteins).
In plant genomics, priority will be given to the exploration of plant genome structure and,
namely, the determination of the function of genes of the male gametophyte, as well as of
some genes during the cell cycle, and interaction of plants and viruses.
Both these areas of basic research in the IEB will be conducted so as to contribute
significantly to the understanding the mechanisms of regulation of plant growth and
development, and, consequently, to practical utilisation of the results obtained as well.
The main goal of research in the IEB is to integrate the two above mentioned areas of
research and thus achieve a complex view on the mechanisms of the basic life processes
within a plant cell. The plant cell will be studied at various levels:
•
those comprising the structure of the genome,
•
the regulation of gene expression,
•
the functional organisation of signalling pathways,
•
the mechanisms of plant growth regulation,
•
the physiological behaviour of the plant cells studied (studies on the cell cycle,
endogenous rhythmicity, cell polarity etc.).
Such complex research forms the basis for understanding the principles of the plant cell and
subsequent plant organ development and, finally, the whole plant organism itself.
Promising areas of research will be preferred, and projects likely to lead to future
breakthroughs in given topics will be supported with high priority. These topics include:
•
molecular cytogenetics (structure and evolution of the genome, construction of
cytogenetic maps using sorted chromosomes)
•
characterisation of transcriptom of the male gametophyte
•
phosphoinositide signalling system in plant cells
•
mechanism of function and regulation of the activity of auxin and cytokinin carriers
•
various aspects of metabolism of cytokinins
•
molecular mechanisms of nucleation and reorganisation of microtubules during the
cell cycle
•
production of pharmacologically-active proteins by transgenic plants.
The coordination of the projects gives a basis for fruitful cooperation between the teams in
the IEB, where the genome studies are an information source for further studies on the
signal and regulatory molecules and, eventually, for a deeper understanding of the
development of plant cells. The process of signal transduction is closely tied to cell
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structures, which further affect translation and protein transport to respective effectors.
The experimental work will be focussed on a limited number of experimental materials
(above all Arabidopsis thaliana, model tobacco cell lines, and – in special cases – the
others), so that a complex view on the course and control of key developmental processes
is achieved. Close connection among the teams in the IEB, together with methodological
knowledge and the support of highly trained personnel guarantees a modern approach to
basic research into the life processes in a plant cell.
The coordination of the two main research topics is documented below. For the purposes of
easier understanding, the sections do not strictly correspond to the running projects of the
respective teams but the names of key researchers responsible for individual areas are
given in parentheses.
List of contents:
1
REGULATION OF GROWTH AND DEVELOPMENT
1.1
1.1.1
Metabolism of phytohormones and other plant growth regulators
1.1.2
Transport of phytohormones and other plant growth regulators
1.1.3
Mechanism of action of phytohormones and other plant growth regulators
1.1.4
Hormonal regulation of some processes of growth and development and
stress reactions
1.1.5
Potential practical applications of the research of plant growth regulators
1.2
2
STRUCTURE AND FUNCTION OF THE GENOME
2.1
Basics of structural and functional genomics
2.2
Molecular aspects of plant virology and phytopathology
2.3
Contribution to the organisation of international scientific activity at the A.
thaliana research
2.4
Potential practical applications
1
REGULATION OF GROWTH AND DEVELOPMENT
1.1
1.1.1
Metabolism of phytohormones and other plant growth regulators
The detailed understanding of the mechanisms of regulation of levels of plant growth
regulators is a necessary prerequisite for the understanding and for targetted control of
plant growth and development. This knowledge is indispensable for design of novel
synthetic growth regulators, for preparation of transgenic plants with controlled biosynthesis
and metabolism of plant hormones, and for their use in plant biotechnologies.
Investigation of control of levels and action of cytokinins will include studies of
(h) the role of hormonal interactions in the regulation of cytokinin levels,
(i)
mechanisms of hormonal homeostasis,
(j)
the targetting of enzymes involved in the biosynthesis and metabolism of cytokinins
and its effects on intracellular cytokinin levels and cytokinin secretion to the apoplast,
(k) selected cytokinin metabolic pathways involved in regulation of cytokinin levels in
plants (spatial and temporal localisation, physiological significance, reaction to
stresses),
(l)
identification and role of specific cytokinin-binding proteins in the formation of a
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storage pool of immobilised cytokinins.
For separation of plant hormones (auxins, cytokinins, abscisic acid), HPLC/MS, GC/MS and
2D-HPLC with on line UV- and fluorescence detectors will be used. Activities of enzymes
involved in cytokinin metabolism will be determined by conversion of radio-labelled
substrates and HPLC with flow-through radioactivity determination. LC and FPLC will be
used for purification of cytokinin-binding proteins; their binding activity will be determined by
equilibrium dialysis and ultrafiltration using radio-labelled ligands. (Motyka, Vaňková,
Dobrev, Kamínek)
The development and stability of chloroplasts is under the strong influence of cytokinins.
Changes in cytokinin levels in plastids can regulate functioning of plastids and delay their
senescence. Consequently, the effectiveness of photosynthetic apparatus is enhanced.
Therefore, the cytokinin content will be measured in chloroplasts from tobacco plants
carrying the gene for zeatin-O-glucosyl transferase (obtained from Prof. D. W. Mok,
Corvallis, Oregon, USA). Biosynthesis of cytokinins in chloroplasts isolated from tobacco
leaves will be studied using incubation with radioactive precursors (pyruvate, mevalonate,
D-xylose) and after treatment with inhibitors of both isoprenoid biosynthetic pathways,
lovastatin and fosfidomycin. Using immunocytochemical method changes will be followed in
the levels and localisation of cytokinins in leaves and isolated chloroplasts from control
plants and from plants transformed by genes, products of which affect cytokinin levels.
(Vaňková, Macháčková)
Recent findings about the appearance of cytokinins in animal cells imply that cytokinins may
have a function, which is not known yet, also in animal cells. To investigate this
phenomenon, the modern analytical methods developed in the IEB will be used. The
appearance of cytokinins in animal tissues will be studied using LC/MS, with special
attention to the production and differentiation of blood cells (preliminary results showed
kinetin as a native cytokinin in some types of blood cells). The biosynthetic pathway of
cytokinins in these cells will be elucidated using incorporation of deuterium in vitro. (Strnad,
Doležal)
In last years, new native cytokinins, some of them displaying high biological activity and
metabolic stability, have been isolated and characterised in the IEB. This very successful
research topic will be further elaborated so that new N6-substituted derivatives of adenine
will be isolated from plants (and animals) and identified, their internal concentrations will be
measured by HPLC/MS and their biological activity will be determined by cytokinin biotests.
The metabolism of these compounds will be studied in relation to their potential utilisation in
plant biotechnologies; synthetic analogues of these compounds will also be prepared. Other
groups of organic compounds, structurally derived from N6-substituted adenine, will be
synthesised, as well as their complexes with selected transition metals; their structure will
be determined by NMR, X-ray diffraction, CHN/O analysis, magnetic susceptibility, MS-, IRand UV-spectrometry. Final structures will be derived from quantal-chemical computations.
(Strnad, Doležal)
Besides phytohormones, plant growth and development and the resistance of plants against
stresses is influenced also by polyamines. The metabolism of polyamines will be
investigated at the level of activities of biosynthetic enzymes (ornithine and arginine
decarboxylases), of contents of putrescine, spermidine and spermine, and of determination
of diamine and polyamine oxidases (the degradation enzymes), during the growth cycle of
tobacco BY-2 cell suspension culture. The metabolism of polyamines will be mapped during
the cell cycle of synchronous tobacco BY-2 cell suspension culture using inhibitors of
various biosynthetic pathways. (Cvikrová, Gemperlová)
Even more than in most of the other research areas, the progress in the research of
phytohormones depends on the improvements of analytical methods for their determination.
Immunodiagnostics of the new generation will be prepared for determination of cytokinins,
as well as other phytohormones (namely abscisic, indole-3-acetic, and jasmonic acids, and
brassinosteroids). The aim is to obtain both high-specificity antibodies for individual
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phytohormones for ELISA assays and for immunochemical detection of these compounds at
the cell level, and generic broad-specificity antibodies for detection of metabolites of certain
groups of phytohormones (applicable especially with immunoaffinity chromatography).
(Strnad)
1.1.2
Transport of phytohormones and other plant growth regulators
Mechanism(s) of transport of phytohormones (auxins and cytokinins) and other growth
regulators (polyamines) will be studied at the cell level (translocation of these compounds
across cell membranes) in relation to (i) modulation of intracellular levels of these
compounds, and (ii) regulation of basic processes of cell development (cell and growth
cycles, establishment and maintenance of cell polarity). The investigation of auxin carriers
will be concentrated on:
(h) mechanism of regulation of their activity,
(i)
localisation of carriers and their dynamics,
(j)
connection(s) with cytoskeletal structures and the endomembrane system,
(k) mechanism of action of phytotropins (i.e. inhibitors of auxin efflux carriers),
(l)
characterisation of specific inhibitors of auxin influx carrier(s).
Experimental approaches will involve transformation of plant cells using vectors bearing
genes for auxin carriers under constitutive and controllable promoters, and in translation
fusion with reporter genes for fluorescence proteins (GFP, YFP, DsRed); fluorescence and
confocal microscopy in vivo; immunofluorescence techniques; determination of kinetic
parameters of transport of auxins across cell membranes.
All experimental data will be related to regulation of growth (or cell) cycle of the cells of
model tobacco lines cultivated in vitro. A. thaliana wild type and mutant plants will be used
as a reference material for transformation and transport assays. On the same experimental
material the possible cytokinin carriers will be searched for with the emphasis on
mechanism(s) of control of their activity (kinetic parameters, effects of anticytokinins,
screening of possible inhibitors, etc.). (Zažímalová, Petrášek, Perry) The accumulation of
polyamines and their transport across the plasma membrane will be characterised in
relation to the cell cycle. Next progression will be analogous to that for auxin carriers.
(Cvikrová, Zažímalová)
1.1.3
Mechanism of action of phytohormones and other plant growth regulators
Hybrids of maize differ in toleration to the density of growth. The degree of this toleration is
probably connected with the leaf angle and thus with their ability to utilise light energy. The
role of auxin-binding proteins (ABPs) in growth and development of leaf angle will be
characterised. The mechanism of how light regulates ABPs and polar auxin transport will be
elucidated. Experimentation will include growth and molecular analysis, with the aim to
determine whether differential development of leaf angle in modern and older maize hybrids
and maize abp mutants is caused by differential expression of the ABP genes and/or
amount of ABPs in leaf tissues. The questions of whether and how light affects levels of
ABPs, and how possible changes in ABPs influence auxin- and light-induced growth and
leaf angle development in maize seedlings will be addressed. A homologue of the ATHB2
gene (coding for a HD-ZIP protein) will be isolated from maize. The gene is specifically
expressed by changes in R:FR (red:far-red light) and by auxin, and it is believed that
ATHB2 is a bridge between light and auxin signalling. Consequently, the expression of the
gene as a function of light and auxin will be studied in the above mentioned maize hybrids.
Physiological characterisation of mutants abp15 and abm19 (T-DNA-mutants of
Arabidopsis) will be finished and the primary changes due to the mutation in the genes
ABP15 and ABM19 will be described. Two hypotheses will be tested: (a) mutants abp15
and abm19 are affected in transduction of the light signal, which results in changes in
hormone levels and signalling, and thus in different growth under light and dark conditions,
(b) the above mentioned mutants are primarily affected in growth in the darkness, and
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consequently also the growth in light and sensitivity to hormones are changed. The
sequences of genes ABP15 and ABM19 in the Arabidopsis genome will be identified and
mapped. Possible products of these genes will be searched for and their function in
regulation of growth and development of Arabidopsis will be investigated. (Fellner)
A range of plant hormone-like compounds, both naturally occurring and synthetic, affect
animal cells as well. To be able to use these compounds e.g. in pharmacy, their effects and
mechanism(s) of action must be well understood. Therefore, molecular mechanisms of
action of plant and synthetic growth regulators in animal tissue cultures will be
characterised, especially in relation to their potential anti-tumour capacity. Pharmacological
properties of these compounds as well as their effects on the cell cycle and its regulatory
proteins will be analysed. Highly effective compounds will be selected, co-crystallised with
selected enzymes or receptors, and X-ray crystallography will be used to determine the
binding mode. Both cellular and molecular effects of these compounds will be analysed and
proteins with high affinity to them will be characterised in detail (proteomic MS analysis) with
the aim of describing the molecular mechanisms of action of these compounds in cells.
(Strnad, Doležal)
1.1.4
Hormonal regulation of some processes of growth and development and
stress reactions
High concentration of cytokinins (resulting from the expression of the ipt gene under Pssu
promoter) gives rise to the formation of a range of structural and functional anomalies. The
elucidation of these anomalies may contribute to the understanding of cytokinin action.
Anomalies in ultrastructure of cell organelles (crystalloids and spherical protein formations in
chloroplasts, peripheral reticulum and interactions between chloroplasts, mitochondria and
peroxisomes) of transgenic Pssu-ipt tobacco will be described using electron microscopic
analysis of ultra-thin sections and 3D computer reconstruction of organelles. Isolation and
analysis of crystalloids will be done by spectrofluorometry, SDS-PAGE, and
immunodetection using specific antibodies. A complex study will be carried out involving
virological, physiological, ultrastructural, and biochemical aspects of viral infection of
transgenic tobacco with elevated content of endogenous cytokinins. Attention will focus on
the role of enzymes of antioxidant and anaplerotic pathways, expression of pathogenesis
related (PR)-proteins and endogenous cytokinins during viral infection. (Synková)
Hormones very often interact in the regulation of physiological processes. The action of
cytokinins is often antagonistic to the action of ABA. In the research of possible antagonistic
actions of ABA and cytokinins in the regulation of stomatal opening and, in consequence,
leaf gas exchange, the interaction between cytokinins and ABA on the activity of stomatal
guard cells and metabolic interaction during water stress-induced ABA synthesis will be
investigated. The research will continue on plants with elevated ABA levels as a
consequence of water stress and on transgenic plants with modified cytokinin content. The
role of pigments of the xanthophyll cycle will be determined in plant protection against
photoinhibition and as precursors in ABA synthesis induced by water stress under low and
high irradiance, as affected by cytokinins and in transgenic plants with an elevated cytokinin
content. During development of water stress changes in activities of antioxidative enzymes
and the effect on these activities of ABA and cytokinins will be followed. (Wilhelmová,
Pospíšilová)
The use of mutants with changed content of and/or sensitivity to hormones is advantageous
for elucidation of the function of individual hormones in growth or developmental processes.
Genetic analysis will be performed on the 7B-1 mutant of tomato, characterised by changed
sensitivity and reaction to ABA, increased resistance to osmotic stress, and decreased
ability to react to blue light. The locus will be mapped by classical methods or using
molecular markers. The 7B-1 gene will be cloned by the most suitable molecular methods
followed by functional analysis. (Fellner)
Cytokinins are well-known for their strong effect on the delay of senescence. The
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hypothesis that the induction of plant senescence is related to free radical reactions and
their products will be tested. The accumulation and characteristics of free radical products
and activities of the antioxidative system (enzymatic and low-molecular) will be investigated.
Further, regulation of leaf senescence induction will be studied using genetically
manipulated plants with changed concentrations of cytokinins. The changes in the formation
of the inductor of senescence – ethylene, will be tested in relation to the production of nitric
oxide and nitrated proteins. (Wilhelmová)
There are more, mutually interacting hormones taking part in the regulation of complex
developmental processes in plants. To elucidate the mechanism(s) of hormonal regulation it
is necessary to monitor changes in levels of all hormones involved in the duration of such a
process. New possibilities of modern analytical methods for endogenous phytohormones
(LC-MS, GC-MS) enabling the detection of a higher number of different compounds
(cytokinins) with higher sensitivity (IAA, ABA, cytokinins) will be utilised in studies of somatic
embryogenesis. Cellular and organ distribution of endogenous phytohormones will be
elucidated by immunolocalisation (auxins, cytokinins). The role of the homologue of the
ABI3 (abscisic acid insensitive 3) gene will be determined in embryogenic cultures of
spruce, and the possible control of its expression and its potential use as a marker of
embryogenesis will be followed. (Vágner, Fischerová)
1.1.5
Potential practical applications of the research of plant growth regulators
Cytokinins prolong the leaf vitality and longevity and increase the flow of assimilates into set
grains. This knowledge will be utilised for the preparation of transgenic plants with increased
level of cytokinins and enhanced productivity. Transgenic plants will be prepared with
controlled expression of genes involved in biosynthesis and metabolism of cytokinins,
suitable for increased crop productivity. Genes encoding the cytokinin biosynthetic enzyme
(isopentenyl transferase, ipt) and cytokinin degradation (cytokinin oxidase, Atckx3, in sense
and antisense orientation) under control of chemically and developmentally inducible
promoters (induction by alcohol [PalcA]; induction of leaf senescence [PSAG12]) will be used for
transformation of the model plant (A. thaliana) and wheat. The idea is to enhance plant
productivity and longevity by delaying leaf senescence, prolongation of leaf photosynthetic
activity and assimilation of nitrogen. Plants will be transformed using the method of floral
dipping in a suspension of transformed clones of Agrobacterium tumefaciens bearing the
above specified genes and promoters obtained from collaborating laboratories. (Hoyerová,
Kamínek)
For cultivation of animal cell cultures, bovine serum albumin (BSA) is frequently used. This
may be dangerous with respect to Creutzfeld-Jacob disease. The serum can be replaced
with hydrolysates of some plant proteins. Therefore, innovative “safe“ protein-less (BSAfree) cultivation media, containing hydrolysates of wheat proteins (mixtures of peptides), will
be prepared and optimised. Their use for stationary and mixed cultures of animal cell lines
in biopharmacy will be tested. (Franěk)
1.2
The regulation of morphogenesis is often connected with cell endomembrane system, and
with the transport of material from cytoplasm to membranes or between individual types of
membranes. With cell membrane system a range of signalling systems is coupled. The
analysis will be performed of factors regulating plant cell morphogenesis with the special
focus on small regulatory GTPases (Rab, Rho, Arf) and interacting regulators and protein
complexes. The plant geranylgeranyl transferase complex responsible for post-translational
Rab GTPase prenylation will be characterised. The characterisation will be continued of
subunits of the plant Exocyst complex, especially the Exo70 one which, in contrast to other
eukaryotes, is present in multiple isoforms in plants. The complex of formin-associated
proteins will be studied in relation to the dynamics of the plant cytoskeleton and the mobility
and localisation of endomembrane compartments. (Žárský, Hála)
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The participation of PLD and phosphatidic acid (PA) will be analysed with special emphasis
on the control of metabolism of components of endomembrane metabolism in the plant cell.
Special attention will be concentrated on PLD as a potential microtubule-associated protein
(MAP) regulating an interaction of microtubules with the plasmalemma and at the same time
intervening in the actin dynamics. A new topic will be an analysis of the influence of PLD on
membrane compartments – especially the Golgi apparatus. Mutants of Arabidopsis will
serve as a basic experimental material, recombinant plant proteins will be expressed in
bacteria and their interactions will be observed in vitro or in the yeast two-hybrid system.
Relevant antibodies will be used also for immunofluorescence localisation in cells. In
parallel, the expression of relevant genes for these proteins in fusion with GFP and
functional analysis will be performed on transgenic plants as well as using detailed
phenotype analysis of mutants (predominantly Arabidopsis). (Žárský)
Molecular and biochemical characterisation of a novel plant enzyme – phosphatidylcholinehydrolysing phospholipase C (PC-PLC) - will be performed in relation to its potential role in
plant signalling systems. Functional analysis will continue. Classical approaches to
molecular genetics (cloning of homologous sequences, expression and biochemical
characterisation of recombinant protein) will be used to describe proteins highly homologous
with bacterial PC-PLC. Intracellular localisation of the protein will be determined using PCPLC-GFP construct, promoter will be studied using GUS constructs and functional analysis
of protein(s) will be performed using RNAi. (Martinec)
The revealing of mechanisms of signal transduction is of key importance for the
understanding of the interactions between plants and environmental factors (physical,
chemical, or pathogens). The molecular basis of aluminium toxicity in plants will be
elucidated from the point of view of the role of phospholipid-cytoskeleton signalling. The role
of phospholipases will be studied in situ using fluorescence-labelled substrates and specific
inhibitors for individual phospholipases. Tobacco cell culture will be used for these
experiments. Interaction between phospholipases and the cytoskeleton will be observed
using purified plant actin and tubulin. Proteins showing differential binding with actin before
and after treatment with aluminium ions will be further tested for phospholipase activity and
identified using specific antibodies and MALDI-TOF mass spectrometry. Constructs with
GFP of genes for phospholipases detected as above will be prepared and fusion proteins
visualised in A. thaliana during stress reaction to aluminium ions. Direct protein-protein
interactions will be proven by fluorescence resonance energy transfer (FRET) technology
using GFP-fusion proteins. A similar experimental approach (namely measurement of
phospholipase activities in situ) will be used to understand the role of phospholipid signalling
systems in biotic stress, especially in the defence mechanisms of Brassica napus against
the fungal pathogen Leptosphaeria maculans. (Martinec, Burketová)
Plant defence mechanisms against plant pathogenic agents will be studied, with the
emphasis on their induction by synthetic and natural environment-friendly compounds, with
perspective of the application to crop plants with minimised impact on the environment. Both
the early response of plants to infection and/or inductor (the role of phospholipases) and the
involvement of signalling pathways (pathway of jasmonic and salicylic acids) and the
production of defence compounds (PR-proteins) in the process will be characterised.
(Burketová, Šindelářová)
2.
2.1
GENOME STRUCTURE AND FUNCTION
Basics of structural and functional genomics
Isolated chromosomes are an ideal material for investigation of structure and function of
genome, the knowledge of which is the prerequisite for understanding of regulation of
growth and development.
Three groups of plant species will be used for the analysis of plant genome structure and its
evolution.
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In Banana, construction of a fine physical cytogenetic map will be a main goal. The map will
help to unravel chromosome changes accompanying evolution and speciation within the
genus Musa. In the same genus, dynamics of repetitive DNA sequences accompanying
evolution of cultivated forms will be followed.
In Legumes, procedures based on the use of sorted chromosomes will be used for targetted
isolation of molecular markers and physical mapping with the aim of aiding the cloning of
agronomically important genes. In cereals, the main focus will be on the use of flow-sorted
chromosomes for high-resolution physical cytogenetic mapping and construction of unique
chromosome-specific BAC libraries. Clones selected from these libraries will be used to
develop physical maps from regions of interest and, in collaboration with other laboratories,
important genes will be isolated. New avenues for using sorted chromosomes in highthroughput physical mapping of ESTs and in vitro (HAPPY) mapping will be explored.
(Doležel, Šimková, Lysák, Valárik)
Pollen tube, as a single intensively growing cell, is one of the most suitable materials for the
investigation of transcription. The male gametophyte of tobacco will be used for studies on
regulation of transcription and translation. A detailed analysis of previously described EPP
particles containing stored ntp303 transcripts will be performed. The role of the cytoskeleton
on the developmentally induced translational regulation of gene expression in tobacco male
gametophyte will be characterised. Currently available bioinformatic tools and reverse
genetic resources will be exploited to reveal, on a genome-wide scale, the network of
gametophytic transcription factors that regulate the start and flawless progress of the male
gametophyte developmental programme during Arabidopsis pollen maturation. (Honys,
Čapková)
Studies on the mechanisms underlying regulation of pollen development will be oriented on
analysis of the properties and functions of glycoproteins specific for the critical
developmental stages. Emphasis will be placed on sequencing, regulation of expression
and on the role of thermostable and dehydrin-like pollen proteins as potential markers and
genetic sources of tolerance to abiotic stresses. (Hrubá, Tupý)
New genes that encode proteins associated with microtubules and microtubule organising
proteins will be cloned in order to describe molecular mechanisms of plant microtubule
nucleation and acentrosomal organisation, one of the, so far, not well explained phenomena
in cell biology. (Cenklová, Binarová)
The majority of processes in organisms proceed in a rhythmical way. Therefore, the
investigation of mechanisms of rhythmicity is nowadays in the centre of interest. Using real
time RT-PCR, we will study the expression of genes connected with rhythmicity and
flowering (especially the gene CONSTANS) in Chenopodium rubrum. The influence of light
regime, various phytohormones and melatonin on the expression of these genes will be
studied in connection with flowering. Some CONSTANS-like genes expressed in flower
buds will be studied as well. (Štorchová, Kolář, Macháčková)
Both mutagens in the environment and UV-irradiation often cause DNA damage. Majority of
organisms, including plants, possess mechanisms to eliminate this damage, so-called DNA
repair. The interactions among DNA repair pathways will be studied in A. thaliana by
characterisation of T-DNA knock-out mutants and various types of DNA damage repair in
mutants will be analysed by the method of Comet Assay. The kinetics of the DNA repair and
its connection to the cell cycle will be observed. The main goal of our research will be to
induce the adaptation to genotoxic stress and to induce programmed cell death by DNA
damage. (Angelis)
Together with the DNA repair in Arabidopsis, the DNA repair of DNA double-strand breaks
will be monitored in Physcomitrella patens, the moss, which has an extremely high level of
homologous recombination, compared with other plant species. The aim of this project is to
develop an expression system for desired proteins by using targetted integration into the
coding sequences („gene targetting“) into the plant genome. (Angelis)
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2.2
Molecular aspects of plant virology and phytopathology
There is a possibility that in the future plants could produce a range of pharmacologically
important proteins. For this purpose, it is necessary to elaborate reliable systems of
transgenosis and to optimise the expression of transgenes. Using the effective system of
agroinfection, the possibility of producing pharmaceutically useful proteins, such as scFv
antibodies, viral proteins for vaccination, and others will be tested. In cooperation with
specialised teams, the immunological features of these substances will be confirmed. The
existing experience with the production of the coat proteins of human papillomavirus will be
used in confirming the possibility of using pseudovirions as the means of targetting DNA
sequences to the plant genome (and the development of DNA vaccines). In „edible vaccine“
development, the expression of defined sequences in edible tissues of transgenic plants will
be tested in each step. Emphasis will be put on defining the yield and stability, as well as
storage possibilities and dispersal of the protein in a given tissue. In addition to production
tomato plants, other plants will be tested for their feasibility of production and edibility.
(Angelis)
Plant viruses harm a range of crops. The knowledge of mechanisms of interaction between
viruses and plants can help to defend against viruses effectively. Molecular and biological
features of potato viruses will be characterised on the basis of:
•
the known and complete nucleotide and amino acid sequences of selected isolates,
•
the prediction of a 3-D structure,
•
the phylogenetic analysis and the studies of protein-protein, protein-RNA interactions,
•
the preparation and use of antibodies to nonstructural viral proteins.
The results obtained will lead to a detailed description of the viral life cycle and introduction
of simple and reliable methods of detection and classification. (Čeřovská, Moravec)
Other types of research will concentrate on protein and genome analysis of the fungus
Venturia inaequalis and apple varieties resistant to this pathogen in order to gain knowledge
of the molecular mechanisms underlying plant-pathogen interactions, and to identify
molecular markers of qualitative and quantitative resistance to the plant pathogen. These
markers will be used for further breeding. (Juříček, Juříčková, Tupý)
2.3
Contribution to organisation on international scientific activity concerning
research on Arabidopsis thaliana
Following the 2000 completion of the Arabidopsis genome sequence by the Arabidopsis
Genome Initiative, the international community of scientists has developed a long-range
plan for the „Multinational Coordinated Arabidopsis thaliana Functional Genomics Project“.
The mission of this project is to achieve complete understanding of the biology of the
flowering plant A. thaliana, which is used as a model plant. Arabidopsis researchers
representing 14 countries from all around the world involved in the project have already
established the Multinational Arabidopsis Steering Committee (MASC). Eastern European
Arabidopsis research groups and laboratories are not yet involved in the Project and are not
represented on MASC. After correspondence with Dr. Rebecca Joy, coordinator of the
MASC, Dr. Fellner has initiated a project called Eastern Europe Arabidopsis Community
(EEAC). The goal of this project is to involve Eastern European countries in international
Arabidopsis research and in the Arabidopsis Functional Genomics Project. (Fellner)
2.4
Potential practical applications
Some of the approaches leading to practical use of our research have already been
mentioned above. They include:
•
production of pharmaceutically useful proteins (scFv antibodies, viral proteins for
vaccination, other edible vaccines),
•
expression of verified sequences in edible tissues of transgenic plants (besides
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•
•
production tomato plants, other feasible plant species will be tested),
development of pseudovirion system of human papillomavirus for oral administration
of DNA/vaccines,
establishment and further development of the methods of production of
pharmaceutically important proteins in liquid culture of the moss Physcomitrella
patens (this will be achieved by extending the possibilities of targetted modification of
the genome of the moss due to homologous recombination, which will be studied in
connection with DNA repair in plants).
The possible use of modified plant vectors for the production of therapeutically effective
substances will be looked into (use of suppressors of post-transcriptional gene silencing for
optimal expression of recombinant proteins in plants). (Angelis)
The appearance of heavy metals in the environment and their harmful effects on organisms
are one of the serious ecological problems. The genotoxic effect of heavy metals,
contaminating the soil in the area of North Bohemia, will be explored, as well as the
genotoxic effect of by-products of water disinfection by chlorine and chloramin. The Comet
Assay, hellow DNA test for the estimate of DNA damage and apoptosis, somatic mutation
frequency test and homologous recombination test on tobacco (Nicotiana tabacum var.
Xanthi) will be used to study the above-mentioned effects. Potato plants will be used for in
situ experiments in contaminated soil. (Stavreva, Gichner)
Transgenosis is potentially very useful tool how to introduce required properties (as e.g.
resistance to insects or herbicides, some qualitative or quantitative features) into plants.
Transgenic potato plants with improved agricultural characteristics will be produced:
•
selected potato cultivars carrying a modified gene for phosphofructokinase from
Lactobacillus bifidus (gene LbPFK), adapted for translational apparatus of a plant;
these plants will be transformed with a vector carrying no selection marker of antibiotic
resistance,
•
potato plants carrying silk proteinase inhibitor gene from Galleria mellonella
(protection against pathogenic microorganisms). (Navrátil)
The transgenosis technique of microprojectile bombardment of transgenes will be further
optimised. The expression and penetration of introduced genes will be monitored in the
progeny of transgenic plants. Both molecular and phenotypical analyses will be performed
in haploid, dihaploid, polyhaploid and diploid populations. (Ohnoutková)
The process of breeding needs quality and homogeneous starting material. For this purpose
the material obtained by androgenesis in vitro is very suitable. The key factors inducing and
affecting the sporophyte development of male gametes – by androgenesis in vitro – will be
explored in seedy plants. The ability of dioecious plants with heterogametous male sex to
androgenously develop microspores with different sex chromosomes will also be studied.
The in vitro method for obtaining haploid, dihaploid and polyhaploid Solanum plants will be
introduced. This method will enable hybridisation between wild-type diploid species carrying
the resistance genes and tetraploid cultivars of potato. Isogenisation of some Triticale with
desired gene (chromosomal) translocations will be achieved by induced androgenesis.
Together with other applied research teams and breeding stations, dihaploid lines of
promising barley and wheat will be prepared. (Ohnoutková)
The long-term project on breeding, advanced testing, legal protection and commercialisation
through licensing of apple varieties resistant against fungal diseases and with low
requirements for plant protection chemical use and, therefore, suitable for organic farming
will continue. The project also includes breeding of varieties with a compact, columnar
growth habit derived from McIntosh Wijcik mutant. The aim of the programme is to improve
growing characteristics of the trees, marketing quality of fruit and more durable resistance to
scab by combining the Vf resistance derived from Malus floribunda with polygenic tolerance
of genetic sources we have selected previously. The genetic background of resistance of
selected hybrids will be characterised with the use of molecular markers. (Tupý, Juříček)
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C6.
Strategies and methods to be applied to carry out IRP
Due to the complexity of the research proposal, only the more general methods and
approaches are described in detail. Specific methods and approaches are, when necessary,
described directly in the research proposal (part C5). The research will run in parallel in all
studied areas and it will be our aim both to integrate all the directions as much as possible
and to use all available methods, apparatuses and approaches most effectively. It will be an
obvious part of most research directions to improve the existing methods and to develop
new ones.
1. Determination of the concentrations of phytohormones and other growth regulators
Determination of the levels of phytohormones (namely auxin, cytokinins,ABA and ethylene)
and other regulators (polyamines, melatonin) is an integral part of the research on regulation
of plant growth and development. The methods for determination of these substances are
well established in the IEB and on a very good level. Recently, a new method of extract
purification on two-parameter columns was elaborated and we can now determine
cytokinins, IAA and ABA in one extract. Liquid chromatography is used for the
determination, with mass spectrometric detection for cytokinins and fluorimetric detection for
auxin. Gas chromatography is used for ABA and ethylene determinations. At present, a new
determination method is being developed for ABA and IAA using GC/MS. A method was
developed for immunohistochemical localisation of cytokinins (free bases) in vivo and a
similar method will be developed for IAA as well. There will be new antibodies produced
against further types of cytokinins, IAA, ABA jasmonic acid and brassinosteroids. These
antibodies will be used for preparation of immunoaffinity columns for extract purification, for
ELISA tests and for imunohistochemical determination in situ.
2. Studies of transport, metabolism and mechanism of action of growth regulators
For the studies of transport and metabolism of hormones, radioactively labelled substances
will often be used. These are, in many cases, synthesised in the Izotope laboratory of IEB.
(it mostly concerns substances labelled with tritium and with very high specific radioactivity).
For detection of these substances, HPLC will be used with a radiometric detector. The
Isotope laboratory will synthesise not only labelled substances, but also substances needed
for various research directions, especially new groups of CDK inhibitors.
The mechanism of action of phytohormones will be studied at the level of binding sites
(receptors). For these studies, radioactively labelled ligands will be used and also transgenic
tobacco cell cultures with changed expression of the gene for the receptor. A similar
strategy will be used in the studies of auxin and cytokinin influx and efflux carriers. In this
case, inhibitors of auxin transport (TIBA, NPA, brefeldin) will be also used. Other inhibitors
will be used for studies of cytokinin biosynthesis in chloroplasts, inhibitors of both pathways
of isoprenoid biosynthesis, lovastatin and fosfidomycin, respectively. Also, in studies of
polyamine synthesis inhibitors, difluoromethyl ornithine and cyclohexyl amine - will be used.
3. Use of mutants
In several research directions, mutants will be used. In most cases it will be mutants of
Arabidopsis thaliana, namely T-DNA mutants abp15 and abm19 with changed reaction to
light and changed sensitivity to auxin, mutants with altered auxin transport and mutants
defective in reparation genes: AtSpo3-11, AtArf1, Att3B, AtTop6B, AtRad17, AtBrcA1 and
AtFandC2 and mutants in non-homologous recombination of double-strand DNA breaks
AtKu70 and AtKu80. Spontaneous tomato mutant 7b-1, with altered sensitivity to ABA and
increased resistance to osmotic stress, will also be used.
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4. Transgenic plants
Besides mutants, transgenic plants are also an important tool in modern plant biology.. In
the phytohormone research, transgenic plants of A. thaliana and tobacco carrying the genes
for isopentenyl transferase (IPT), cytokinin oxidase (CKX), and proteins involved in auxin
transport will be used under the control of various promoters – constitutive (CaMV35S
promoter) and inducible (promoters Palc and PSAG12 as well as dexamethason-inducible one).
We will also use transgenic tobacco plants with the genes for zeatin-O-glucosyl transferase
or specific glucosidase (Zm-P60.1) and transgenic tobacco and Arabidopsis plants with
increased expression of genes for proteins of signalling pathways. Plants carrying
constructs of various genes with green or yellow fluorescent protein (GFP, YFP) or rsRed
will also be prepared and these will be used for studies of the expression of studied genes
and their localisation using fluorescence microscopy. Plants with fusions of promoters of
some genes and GUS (β-glucuronidase) will also be constructed for this purpose. In
addition, cereals will be prepared carrying genes PHYT (for phytase, increasing the
phosphate availability) and DAP A (for dihydropicolinate synthase A, increasing lysine
content) and also potato plants carrying the gene for phosphofructokinase, which lowers
level of reducing sugars. Last, but not least, transgenic plants will be used for elaborating a
system of production of edible (oral) vaccines (namely tomato). Transformation will be
carried out using both Agrobacterium tumefaciens and gene gun.
5. Microscopic techniques
In the area of phytohormone research, immunohistochemical determination of cytokinin free
bases and of auxins (see above) will be performed. In signalling studies the activity of
phospholipases in vivo will be measured using fluorescently labelled substrates from the
BODIPY Comp. The study of expression of the constructs with fluorescent proteins (GFP,
YFP, dsRed) will also be performed at microscopic level. Not just a fluorescent microscope
will be used, but also a two-photon confocal microscope and techniques such as FRET
(fluorescence resonance energy transfer) and FRAP (fluorescence recovery after
photobleaching).
Microscopic analysis will be also used in studies of cytoskeleton (staining of actin and
tubulins), especially in the course of cell cycle and in signalling. Image analysis techniques
are elaborated for qualitative and quantitative evaluation of microscopic images.
6. Methods of plant genomics
Besides those approaches already described, the method of DNA chips (microarrays),
hybridisation techniques (Western, Northern blots), PCR and RT-PCR will be used for
studies of gene expression. Functional analysis of some genes will be performed using
small interfering RNA (RNAi). For some genes recombinant proteins will be prepared in
yeasts and used for further studies. Antibodies will be prepared against some proteins and
these will be used in localisation studies in situ. Functional interaction of proteins will be
studied in the two-hybrid system. Flow cytometry will be used to sort chromosomes and
sorted chromosomes from various plants will be used for the preparation of pure,
chromosome specific, high molecular DNA and for construction of chromosome specific
cDNA BAC libraries. Sorted chromosomes will also be used for localisation studies of some
genes using FISH (fluorescence in situ hybridisation).
42
AV0Z50380511
D4.
List of major implemented R&D results related to the subject of IRP achieved by
the members of research team, within the period of 1999-2003
PUBLICATION LIST 1999-2003:
1999:
IMPACTED JOURNALS:
no.
title
1 Angelis K.J., Dušínská M., Collins A.R.:
2
3
4
5
6
7
8
9
10
11
Single cell gel electrophoresis: Detection of DNA damage at different levels of
sensitivity.
Electrophoresis 20: 2133-2138, 1999.
Angelis K.J., McGuffie M., Menke M., Schubert I.:
Studies of DNA repair in various plants using the comet assay.
Neoplasma 46: 72–73, 1999.
Auer C. A., Motyka V., Březinová A., Kamínek M.:
Endogenous cytokinin accumulation and cytokinin oxidase activity during
shoot organogenesis of Petunia hybrida.
Physiol. Plant. 105: 141-147, 1999.
Bavrina T.V., Lozhnikova V.N., Macháčková I., Gryanko T.I.:
Tobacco transformants to study the role of phytohormones in flowering and
seed formation.
Russ. J. Plant Physiol. 46: 189-193, 1999.
Benková E., Witters E., Van Dongen W., Kolář J., Motyka V., Brzobohatý
B., Van Onckelen H.A., Macháčková I.:
Cytokinins in tobacco and wheat chloroplasts: occurrence and changes due to
light/dark treatment.
Plant Physiol. 121: 245-251, 1999.
Bílková J., Albrechtová J., Opatrná J.:
Histochemical detection and image analysis of nonspecific esterase activity
and the amount of polyphenols during annual buds development in Norway
spruce.
J. Exp. Bot. 336: 1129-1138, 1999.
Blažková J., Krekule J., Macháčková I., Procházka S.:
Auxin and cytokinins in the release of apical dominance in pea – a differential
response due to bud position.
J. Plant Physiol. 154: 691-696, 1999.
Bögre L., Calderini O., Binarová P., Mattauch M., Till S., Kiegerl S., Jonak
C., Pollaschek C., Barker P., Huskisson N.S., Hirt H., Heberle-Bors E.:
A MAP kinase is activated late in mitosis and becomes localized to the plane
of cell division.
Plant Cell 11: 101-113, 1999.
Burketová L., Šindelářová M., Ryšánek P., Šindelář L.:
Changes in ribonuclease and glucose-6-phosphate dehydrogenase activities
induced by beet necrotic yellow vein virus in sugar beet.
Biol. Plant. 42: 423 – 430, 1999.
Burketová L., Šindelářová M., Šindelář L.:
Benzothiadiazole as an inducer of β-1,3-glucanase and chitinase isozymes in
sugar beet.
Biol. Plant. 42: 279-287, 1999.
Cvikrová M., Binarová P., Cenklová V., Eder J., Macháčková I.:
Reinitiation of cell division and polyamine and aromatic monoamine levels in
alfalfa explants during the induction of somatic embryogenesis.
Physiol. Plant. 105: 330-337, 1999.
43
IF 1999
3.447
0.448
2.460
0.094
4.434
2.482
1.143
10.463
0.414
0.414
2.460
AV0Z50380511
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Cvikrová M., Binarová P., Eder J., Vágner M., Hrubcová M., Zoń J.,
Macháčková I.:
Effect of inhibition of phenylalanine ammonia lyase activity on growth of alfalfa
cell suspension culture: Alterations in mitotic index, ethylene production, and
contents of phenolics, cytokinins, and polyamines.
Physiol. Plant. 107: 329-337, 1999.
Cvrčková F., Žárský V.:
Ntrop1, a tobacco (Nicotianna tabacum) cDNA encoding a Rho subfamily
GTPase expressed in pollen (accession No AJ222545) (PGR 99-079).
Plant Physiol. 120: 633, 1999.
Čeřovská N., Moravec T., Filigarová M., Ryšlavá H., Grosclaude J.:
Partial antigen characterization of different potato virus Y-NTN isolates with
monoclonal antibodies by means of competitive binding tests and
immunoblotanalysis.
Acta Virologica 43: 391-393, 1999.
Dewitte W., Chiappetta A., Azmi A., Witters E., Strnad M., Rembur J.,
Noin M., Chriqui D., Van Onckelen H.:
Dynamics of cytokinins in apical shoot meristems of a day neutral tobacco
during floral transition and flower formation.
Plant Physiol. 119: 111-122, 1999.
Dršata J., Netopilová M., Tolman V.:
Stereoisomers of 4-fluoroglutamic acid: influence on Escherichia coli
glutamate decarboxylase.
Pharmazie 54: 713-714, 1999.
Ehrenbergová L., Vaculová K., Zimolka J., Müllerová E.:
Výnosové znaky a jejich vztahy k jakostním ukazatelům zrna bezpluchého
ječmene jarního.
Rostl. Výr. 45: 53-59, 1999.
Ephritikhine G., Fellner M., Vannini C., Lapous D., Barbier-Brygoo H.:
The sax1 dwarf mutant of Arabidopsis thaliana shows altered sensitivity of
growth responses to abscisic acid, auxin, gibberellins and ethylene and its
partially rescued by exogenous brassinosteroid.
Plant J. 18: 303-314, 1999.
Fujikura Y., Doležel J., Číhalíková J., Bögre L., Binarová P.:
Vicia faba germination: Synchronized cell growth and localization of nucleolin
and alpha-tubulin.
Seed Sci. Res. 9: 297-305, 1999.
Gichner T., Ptáček O., Stavreva D.A., Plewa M.J.:
Comparison of DNA damage in plants as measured by single cell gell
electrophoresis and somatic leaf mutation induced by monofunctional
alkylating agents.
Environ. Mol. Mutagen. 33: 279-286, 1999.
Gichner T., Velemínský J.:
Monitoring the genotoxicity of soil extracts from two heavily polluted sites in
Prague using the Tradescantia stamen hair and micronucleus (MNC) assays.
Mut. Res. 426: 163-166, 1999.
Haisel D., Pospíšilová J., Synková H., Čatský J., Wilhelmová N., Plzáková
Š.:
Photosynthetic pigments and gas exchange of in vitro grown tobacco plants
as affected by CO2 supply.
Biol. Plant. 42: 463-468, 1999.
Hrubá P., Tupý J.:
N-glycoproteins specific for different stages of microspore and pollen
development in tobacco.
Plant Sci. 141: 29-40, 1999.
Jansa J., Gryndler M., Matucha M.:
Comparison of the lipid profiles of arbuscular mycorrhizal (AM) fungi and soil
saprophytic fungi.
Symbiosis 26: 247-264, 1999.
44
2.460
4.434
0.476
4.434
0.446
0.192
5.098
0.942
1.998
2.107
0.414
1.015
0.766
AV0Z50380511
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26
27
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Lebeda A., Křístková E., Doležal K.:
Peroxidase isozyme polymorphysm in Cucurbita pepo cultivars with various
morphotypes and different level of field resistance to powdery mildew.
Scientia Horticulturae 81: 103-112, 1999.
Lysák M.A., Doleželová M., Horry J.P., Swennen R., Doležel J.:
Flow cytometric analysis of nuclear DNA content in Musa.
Theor. Appl. Genet. 98: 1344-1350, 1999.
Lysák M.A., Číhalíková J., Kubaláková M., Šimková H., Künzel G.,
Doležel J.:
Flow karyotyping and sorting of mitotic chromosomes of barley (Hordeum
vulgare L.).
Chrom. Res. 7: 431-444, 1999.
Nouzová M., Kubaláková M., Doleželová M., Koblížková A., Doležel J.,
Macas J.:
Cloning and characterisation of new repetitive sequences in field bean (Vicia
faba L.).
Ann. Bot. 83: 535-541, 1999.
Palomino G., Doležel J., Cid R., Brunner I., Mendez I., Rubluo A.:
Nuclear genome stability of Mammillaria san-angelensis (Cactaceae)
regenerants induced by auxins in long-term in vitro culture.
Plant Sci. 141: 191-200, 1999.
Pospíšilová J., Čatský J.:
Development of water stress under increased atmospheric CO2 concentration.
Biol. Plant. 42: 1-24, 1999.
Pospíšilová J., Synková H., Haisel D., Čatský J., Wilhelmová N., Šrámek
F.:
Effect of elevated CO2 concentration on acclimation of tobacco plantlets to ex
vitro conditions.
J. Exp. Bot. 50: 330: 119-126, 1999.
Pospíšilová J., Tichá I., Kadleček P., Haisel D., Plzáková Š.:
Acclimatization of micropropagated plants to ex vitro conditions.
Biol. Plant. 42: 481-497, 1999.
Rupp H.-M., Frank M., Werner T., Strnad M., Schmülling T.:
Increased steady state mRNA levels of the STM and KNAT1 homeobox
genes in cytokinin overproducing Arabidopsis thaliana indicate a role for
cytokinins in the shoot apical meristem.
Plant J. 18: 557-563, 1999.
Říhová L., Tupý J.:
Manipulation of division symmetry and developmental fate in cultures of
potato microspores.
Plant Cell Tiss. Org. Cult. 59: 135-145, 1999.
Stavreva D.A., Strnad M., Havlíček L., Gichner T.:
SCGE analysis of genomic damage induced in cultured tobacco suspension
cells by three purine analogues – olomoucine, bohemine and roscovitine and
by hydrogen peroxide, and ethyl methanesulfonate.
Neoplasma 46: 92-93, 1999.
Svobodová H., Albrechtová J., Kumstýřová L., Lipavská H., Vágner M.,
Vondráková Z.:
Somatic embryogenesis in Norway spruce: Anatomical study of embryo
development and influence of polyethylene glycol on maturation process.
Plant Physiol. Biochem. 37(3): 209-221, 1999.
Synková H., Van Loven K., Pospíšilová J., Valcke R.:
Photosynthesis of transgenic Pssu-ipt tobacco.
J. Plant Physiol. 155: 173-182, 1999.
Šesták Z., Čatský J.:
Bibliography of reviews and methods of photosynthesis. 81.
Photosynthetica 36: 291-319, 1999.
Bibliography of reviews and methods of photosynthesis. 82.
45
0.347
2.082
1.576
1.326
1.015
0.414
2.482
0.414
5.098
0.498
0.448
1.347
1.143
0.734
0.734
AV0Z50380511
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Šindelář L., Šindelářová M., Burketová L.:
Changes in activity of glucose-6-phosphate and 6-phosphogluconate
dehydrogenase isozymes upon potato virus Y infection in tobacco leaf tissues
and palisade parenchyma protoplasts.
Plant Physiol. Biochem. 37: 195-201, 1999.
Šindelářová M., Šindelář L., Burketová L.:
Changes in glucose, fructose and saccharose metabolism in tobacco plants
infected with potato virus Y.
Biol. Plant. 42: 431-439, 1999.
Tolman V., Hanuš J., Sedmera P.:
A hetero diels-alder access to (Z)-zeatin and (Z)-isozeatin.
Collect. Czech. Chem. Commun. 64: 696-702, 1999.
Vaněk T., Valterová I., Vaňková R., Vaisar T.:
Biotransformation of (-)limonene using Solanum aviculare and Dioscorea
deltoidea immobilized plant cells.
Biotechnol. Lett. 21: 625-628, 1999.
Vondráková Z., Krekule J:
The role of hypocotyl and roots in photoperiodic floral induction of
Chenopodium rubrum.
Plant Biol. 1: 96-98, 1999.
Witters E., Vanhoutte K., Dewitte W., Macháčková I., Benková E., Van
Dongen W., Esmans E.L., Van Onckelen H.A.:
Analysis of cyclic nucleotides and cytokinins in minute plant samples using
phase system switching capillary ion spray-LC-MSMS.
Phytochem. Anal. 10: 143-148, 1999.
Zonia L., Tupý J., Staiger C.J.:
Unique actin and microtubule arrays co-ordinate the differentiation of
microspores to mature pollen of Nicotiana tabacum.
J. Exp. Bot. 50: 581-594, 1999.
OTHERS:
47 Bögre L., Calderini O., Meskiene I., Binarová P:
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52
Regulation of cell division and cytoskeleton by mitogen-activated protein
kinases in higher plants. In : Results and Problems in Cell Differentiation, vol.
27, Hirt H. (ed.): MAP Kinases in plant signal transduction Springer-Verlag
Berlin Heidelberg, pp. 95-117, 1999.
Bögre L., Ligterink W., Meskiene I., Binarová P.:
Mitogen-activated protein kinases in plant signal transduction: are they
mitogenic? In: Strnad M., Peč P., Beck E., (eds.): Advances in Regulation of
Plant Development, Peres Publications, Prague, pp. 169-181, 1999.
Cvikrová M., Hrubcová M.:
The role of phenolic substances in the processes of differentiation and
morphogenesis. In: Strnad M., Peč P., Beck E., (eds.): Advances in
Regulation of Plant Development, Peres Publications, Prague, pp. 213-220,
1999.
Doležel J., Číhalíková J., Weiserová J., Lucretti S.:
Cell cycle synchronization in plant root meristems.
Methods Cell Sci. 21: 95-107, 1999.
Doležel J., Macas J., Lucretti S.:
Flow analysis and sorting of plant chromosomes In: Robinson J.P.,
Darzynkiewicz Z., Dean P.N., Dressler L.G., Rabinovitch P.S., Stewart C.C.,
Tanke H.J., Wheeless L.L. (eds.): Current Protocols in Cytometry, pp. 5.3.1 –
5.3.33, John Willey and Sons, Inc., New York, 1999.
Fellner M.:
Research of mechanisms of auxin action: isolation and characterization. In:
Advances in Regulation of Plant Growth and Development. Strnad M., Peč P.,
Beck E. (eds.), Peres Publ., Prague, pp. 139-156, 1999.
46
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0.916
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0.798
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Fowke L.C., Attree S., Binarová P.:
Light and electron microscopic studies of somatic embryogenesis in spruce.
In: Morphogenesis in Plant Tissue Cultures, Soh W.-Y., Bhojwani S.S., (eds.),
Kluwer Acad Publ., pp. 95-114, 1999.
Galuszka P., Frébort I., Šebela M., Strnad M., Peč P.:
Cytokinin oxidase: the key enzyme in the biodegradation of cytokinins. In:
Advances in Regulation of Plant Growth and Development. Strnad M., Peč P.,
Beck E. (eds.), Peres Publ., Prague, 1999.
Gaudinová A.:
Effects of phytohormones on nitrate reductase. In: Strnad M., Peč P., Beck E.,
(eds.): Advances in Regulation of Plant Development, Peres Publications,
Prague, pp. 59-66, 1999.
Hajdúch M., Havlíček L., Vesley J., Novotný R., Mihál V., Strnad M.:
Synthetic cyclin dependent kinase inhibitors – New generation of potent anticancer drugs. Drug resistance in leukemia and lymphoma III, vol. 457, pp.
341-353, 1999.
Hrubá P.:
Exprese genů během mikrosporogenese a vývoje pylu.
Biol. listy 64: 201-214, 1999.
Kolář J., Johnson C.H., Macháčková I.:
Presence and possible role of melatonin in short-day flowering plant,
Chenopodium rubrum.
Adv. Exp. Med. Biol. 460: 391-393, 1999.
Kolářová H., Kotala L., Strnad M., Bancířová M., Lasovský J.:
Fluorescenční metody studia buněčného poškození.
Sborník lékařský 99: 437-442, 1999.
Kolářová H., Kubínek R., Lenobel R., Bancířová M., Strnad M., Jírová D.,
Lasovský J.:
In vitro photodynamic therapy with phthalocyanines on the MCF7 cancer cells.
Internet J. Photochem. Photobiol.
http://www.photobiology.com/photobiology99/contrib/kolarova/index.htm.
1999.
Kong L., Attree S.M., Eveans D.E., Binarová P., Yeung E.C. Fowke L.C.:
Somatic embryogenesis in white spruce: Studies of embryo development and
cell biology. In: Jain S.M., Gupta P.K., Newton R.J. (eds.) Somatic
embryogenesis of Woody plants, vol. 4, Kluwer Acad Publ., pp. 1-28, 1999.
Lebeda A., Doležalová I., Křístková E., Vinter V., Vránová O., Doležal K.,
Tarkowski P., Petrželová I., Trávníček B., Novotný R., Janeček J.:
Complex research of taxonomy and ecobiology of wild Lactuca spp. genetic
resources. In: Lebeda A., Křístková E. (eds.): Eucarpia Leafy Vegetables ´99,
UP Olomouc, pp. 117-131, 1999.
Lebeda A., Kubaláková M., Křístková E., Navrátilová B., Doležal K.,
Doležel J., Lysák M.:
Morphological and physiological characteristics of plants issued from an
interspecific hybridization of Cucumis sativus x Cucumis melo. In: Abak K.,
Büyükalaca S. (eds.) Proc. First Int. Symp. on Cucurbits, Acta Hort. 492: 149155, 1999.
Lucretti S., Gualberti G., Nardi L., Trionfetti Nisini P., Doležel J.:
Ploidy analysis and chromosome sorting in plant species.
In: Starace G. (ed.): Manuale GIC „Citometria a Flusso“. pp. 445-473, GIC
Gruppo Italiano di Citometria, Roma, 1999.
Lucretti S., Nardi L., Trionfetti P., Moretti F., Gualberti G., Doležel J.:
Bivariate flow cytometry DNA/BrdU analysis of plant cell cycle.
Methods Cell Sci. 21: 155-166, 1999.
Macháčková I., Kolář J.:
Photoperiodism, rhythmicity and melatonin – comparison of animals and
plants. In: Frontiers in Plant Physiology, Strnad M., Péč P., Beck E. (eds.),
Advances in Regulation of Plant Development, Peres Publications, Praha,
pp.131-138, 1999.
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Macháčková I.:
Exogenous and endogenous hormones in the regulation of morphogenesis in
vitro.
3rd Symposium „Recent Advances in Plant Biotechnology“, Stará Lesná,
Slovak Rep., p. 9-12, 1999.
Malá J., Cvrčková H., Březinová A., Hrubcová M., Eder J., Vágner M.,
Cvikrová M.:
Biochemical characteristics of oak somatic embryos. In: Applications of
Biotechnology to Forest Genetics, Vitoria –Gasteiz 22-25 September 1999,
Spain, pp 56-58, 1999.
Matucha M., Uhlířová H.:
Sekundární atmosférické polutanty a zdravotní stav lesa: cesta, příjem a osud
kyseliny trichloroctové ve smrku ztepilém (Picea abies /L./ Karst). Ovzduší
‘99, Brno, 7.-10.2.1999, Sborník Konf. Recetox Brno, pp.86-90, 1999.
Novotný J., Vagera J., Ohnoutková L.:
Iron as a key factor in andogenesis in cereals. Proc. Nové poznatky
z genetiky a šlachtěnia poĺnohospodárských rostlín. VÚRV Piešťany, pp. 3336, 1999.
Pospíšilová J., Čatský J.:
Effect of increased atmospheric CO2 concentration on water use efficiency of
plants. In: Pessarakli M. (ed.): Handbook of Plant and Crop Stress: Second
Edition. Marcel Dekker, New York - Basel - Hong Kong, pp. 1163-1184, 1999.
Schoofs H., Panis B., Strosse H., Mayo Mosqueda A., Lopez Torres J.,
Roux N., Doležel J., Swennen R.:
Bottlenecks in the generation and maintenance of morphogenic banana cell
suspensions and plant regeneration via somatic embryogenesis therefrom.
Infomusa 8: 3-7, 1999.
Strnad M., Kryštof V., Havlíček L.:
Control of Tumour Development in Plants and Animals: A Comparative
Treatise. In: Advances in Regulation of Plant Growth and Development.
Strnad M., Peč P., Beck E (eds.), Peres Publ., Prague, 1999.
Šesták Z.:
Chlorophyll fluorescence kinetics depends on age of leaves and plants. In:
Argyroudi-Akoyunoglou J., Senger H., (eds.): The Chloroplast: From
Molecular Biology to Biotechnology. Kluwer Acad. Publ., Dordrecht, pp. 291296, 1999.
Šesták Z.:
Vědecká komunikace. Jak psát a přednášet o vědě. Academia, Praha, 205 p.,
1999.
Vágner M., Vondráková Z., Špačková J., Cvikrová M., Eder J., Lipavská
H., Albrechtová J., Svobodová H., Macháčková I.:
Norway spruce somatic embryogenesis: endogenous levels of
phytohormones during somatic embryo development. Current Plant Science
and Biotechnology in Agriculture, vol. 36: Plant Biotechnology and In Vitro
Biology in the 21st Century – Proc. IXth Int. Congress Int. Assoc. Plant Tissue
Cult. Biotechnol., Jerusalem, Israel, 14-19 June 1998, Altman A., Ziv M., Izhar
S. (eds.), Kluwer Acad. Publ., Dordrecht, Boston, London, pp. 93-96, 1999.
Vaňková R.:
Cytokinin glycoconjugates – distribution, metabolism and function. In: Strnad
M., Peč P., Beck E., (eds.): Advances in Regulation of Plant Development,
Peres Publications, Prague, pp. 67-78, 1999.
Zažímalová E., Březinová A., Motyka V., Kamínek M.:
Control of cytokinin biosynthesis and metabolism. In Hooykaas P., Hall M.A.,
Libbenga K.R., (eds.): Biochemistry and Molecular Biology of Plant
Hormones, Ser. New comprehensive biochemistry, vol. 33, Elsevier, pp. 141160, 1999.
Žárský V., Cvrčková F.:
Small GTPases in plant cell morphogenesis. In: Strnad M., Peč P., Beck E.,
(eds.): Advances in Regulation of Plant Development, Peres Publications,
Prague, 1999.
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IMPACTED JOURNALS:
no.
title
80 Angelis K.J., McGuffie M., Menke M., Schubert I.:
81
82
83
84
85
86
87
88
89
90
91
Adaptation to alkylation damage in DNA measured by the comet assay.
Environ. Mol. Mutag. 36(2): 146-150, 2000.
Astot C., Doležal K., Moritz T., Sandberg G.:
Deuterium in vivo labelling of cytokinins in Arabidopsis thaliana analysed by
capillary liquid chromatography/frit-fast atom bombardment mass
spectrometry.
J. Mass Spectrom. 35(1): 13-22, 2000.
Astot C., Doležal K., Nordström A., Wang Q., Kunkel T., Moritz T., Chua
N., Sandberg G.:
An alternative cytokinin biosynthesis pathway.
Proc Natl. Acad. Sci. USA 97: 14778-14783, 2000.
Bálint-Kurti P.J., Clendennen S.K., Doleželová M., Valárik M., Doležel J.,
Beetham P.R., May G.D.:
Identification and chromosomal localization of the monkey retrotransposon in
Musa sp.
Mol. Gen. Genet. 263(6): 908-915, 2000.
Barták P., Pěchová D., Tarkowski P., Bednář P., Kotouček M., Stránský
Z., Vespalec R.:
Determination of the first dissociation constant of 6-benzylaminopurine - A
comparison of methods
Anal. Chim. Acta 421(2), 221-229, 2000.
Binarová P., Cenklová V., Hause B., Kubátová E., Lysák M., Doležel J.,
Bögre L., Dráber P.:
Nuclear gamma-tubulin during acentriolar plant mitosis.
Plant Cell 12(3): 433-442, 2000.
Blažková A., Vondráková Z., Krekule J.:
The shoot apex as a marker of the responsivity to photoperiodic treatment
inducing flowering of Chenopodium rubrum L.
Biol. Plant. 43(1): 31-34, 2000.
Dršata J., Netopilová M., Tolman V.:
Influence of stereoisomers of 4-fluoroglutamate on rat brain glutamate
decarboxylase.
J. Enzyme Inhib. 15(3): 273-282, 2000.
Franěk F., Hohenwarter O., Katinger H.:
Plant protein hydrolysates: Preparation of defined peptide fractions promoting
growth and production in animal cells cultures.
Biotechnol. Progress 16(5): 688-692, 2000.
Frank M., Rupp H.M., Prinsen E., Motyka V., Van Onckelen H.,
Schmulling T.:
Hormone autotrophic growth and differentiation identifies mutant lines of
Arabidopsis with altered cytokinin and auxin content or signaling.
Plant Physiol. 122(3): 721-729, 2000.
Gichner T., Menke M., Stavreva D.A., Schubert I.:
Maleic hydrazide induces genotoxic effects but no DNA damage detectable by
the Comet assay in tobacco and field beans.
Mutagenesis 15(5): 385-389, 2000.
Gichner T., Ptáček O., Stavreva D.A., Wagner E.D., Plewa M.J.:
A comparison of DNA repair using the comet assay in tobacco seedlings after
exposure to alkylating agents or ionizing radiation.
Mut. Res. 470(1): 1-9, 2000.
49
IF 2000
2.278
2.638
10.789
2.462
1.849
11.093
0.424
1.733
1.897
4.831
2.226
1.506
AV0Z50380511
92
93
94
95
96
97
98
99
100
101
102
103
104
105
Grospietsch M., Lipavská H., Opatrná J.:
Effect of paclobutrazol on soluble sugars and starch content of de novo
regenerating pototo stem explants.
Biol. Plant. 43(1): 137-139, 2000.
Hanuš J., Siglerová V., Matucha M.:
N-6-alkyladenosines and adenines labelled with tritium
J. Label. Comp. Radiopharm. 43(5): 523-531, 2000.
Havlíčková H., Cvikrová M., Eder J:
Polyamine accumulation in ears of winter wheat cultivars infested by Sitobion
avenae (F.).
Z. PflKrankh. PflSchutz. 107: 505-513, 2000.
Honys D., Combe J.P., Twell D., Čapková V.:
The translationally repressed pollen-specific ntp303 mRNA is stored in nonpolysomal mRNPs during pollen maturation.
Sex. Plant Reprod. 13(3): 135-144, 2000.
Honys D., Čapková V.:
Temporal changes in the RNA distribution between polysomes and
postpolysomal ribonucleoprotein particles in tobacco male gametophyte.
Biol. Plant. 43(4): 517-522, 2000.
Hrubcová M., Cvikrová M., Eder J., Zon J., Macháčková I.:
Effect of inhibition of phenylpropanoid biosynthesis on peroxidase and IAAoxidase activities and auxin content in alfalfa suspension cultures.
Hušková R., Pěchová D., Kotouček M., Lemr K., Doležal K.:
Voltametrické chování a stanovení některých cytokininů na rtuťové elektrodě.
Chem. Listy 94, 1004-1009 (2000).
Janas K.M., Cvikrová M., Palagiewicz A., Eder J.:
Alterations in phenylpropanoid content in soybean roots during low
temperature acclimation.
Plant Physiol. Biochem. 38(7-8): 587-593, 2000.
Kamínek M., Březinová A., Gaudinová A., Motyka V., Vaňková R.,
Zažímalová E.:
Purine cytokinins: a proposal of abbreviations.
Plant Growth Regul. 32: 253-256, 2000.
Klemš M., Balla J., Macháčková I., Eder J., Procházka S.:
The uptake and metabolism of A-3-benzylaminopurine in tobacco (Nicotiana
tabacum L.) and cucumber (Cucumis sativus L.) explants.
Plant Growth Regul. 31(3): 135-142, 2000.
Kohout L., Chodounská H., Macek T., Strnad M.:
On steroids part CDX - Synthesis of (20S)-2 alpha,3 alpha-dihydroxy-6-oxo-7oxa-7a-homo-5 alpha-pregnane-20-carboxylic acid as a brassinosteroid part
of ligand for binding to affinity chromatography carriers.
Coll. Czech. Chem. Comm. 65(11): 1754-1761, 2000.
Kovářová H., Hajdúch M., Kořínková G., Halada P., Krupičková S.,
Gouldsworthy A., Zhelev A., Strnad M.:
Proteomics approach in classifying the biochemical basis of the anticancer
activity of the new olomoucine-derived synthetic cyclin-dependent kinase
inhibitor, bohemine.
Electrophoresis 21: 3757-3764, 2000.
Kubaláková M., Lysák M.A., Vrána J., Šimková H., Čihalíková J., Doležel
J.:
Rapid identification and determination of purity of flow-sorted plant
chromosomes using C-PRINS.
Cytometry 41(2): 102-108, 2000.
Kubišta J., V., Havlíček L., Hanuš J., Halada P., Kuzma M.:
Rearrangement and loss of bromine radical and CO from some bromobenzyl
alcohols following electron ionisation.
Eur. J. Mass Spectrom. 6(2): 135-141, 2000.
50
0.424
0.756
0.526
2.260
0.424
1.262
0.278
1.262
0.837
0.837
0.960
3.385
2.557
0.629
AV0Z50380511
106 Lipavská H., Svobodová H., Albrechtová J., Kumstýřová L., Vágner M.,
0.750
107
1.274
108
109
110
111
112
113
114
115
116
117
118
119
Vondráková Z.:
Somatic embryogenesis in Norway spruce: Carbohydrate status during
embryo maturation and the effect of polyethyleneglycol treatment.
In Vitro Cell. Dev. Biol. 36(4): 260-267, 2000.
Lysák, M.A., Rostková, A., Dixon, J.M., Rossi, G., Doležel, J.:
Limited genome size variation in Sesleria albicans.
Ann. Bot. 86: 399-403, 2000.
Martinec J., Feltl T., Scanlon C.H., Lumsden P.J., Macháčková I.:
Subcellular localization of a high affinity binding site for D-myo-inositol 1,4,5trisphosphate from Chenopodium rubrum.
Plant Physiol. 124(1): 475-483, 2000.
Mašek T., Smýkal P., Janotová I., Honys D., Čapková V., Pechan P.M.:
Isolation of a Brassica napus L. cDNA encoding a putative high-mobilitygroup HMG I/Y protein.
Plant Sci. 159(2): 197-204, 2000.
Menke M., Angelis K.J., Schubert I.:
Detection of specific DNA lesions by a combination of comet assay and FISH
in plants.
Environ. Molec. Mutag. 35(2): 132-138, 2000.
Mes T.H.M., Kuperus P., Kirschner J., Štěpánek J., Oosterveld P.,
Štorchová H; den Nijs J.C.M.:
Hairpins involving both inverted and direct repeats are associated with
homoplasious indels in non-coding chloroplast DNA of Taraxacum
(Lactuceae: Asteraceae).
Genome 43(4): 634-641, 2000.
Novotná Z., Valentová O., Martinec J., Feltl T., Nokhrina K.:
Study of phospholipases D and C in maturing and germinating seeds of
Brassica napus.
Biochem.Soc.Trans. 28: 817-818, 2000.
Novotný J., Vagera J., Ohnoutková L.:
Effects of free and chelated iron on in vitro androgenesis in barley and wheat.
Plant Cell Tissue Org. Cult. 63(1): 35-40, 2000.
Otyepka M., Kryštof V., Havlíček L., Siglerová V., Strnad M., Koča J.:
Docking-based development of purine-like inhibitors of cyclin-dependent
kinase-2
J. Med. Chem. 43(13): 2506-2513, 2000.
Pospíšilová J., Haisel D., Synková H., Čatský J., Wilhelmová N., Plzáková
Š., Procházková D., Šrámek F.:
Photosynthetic pigments and gas exchange during ex vitro acclimation of
tobacco plants as affected by CO2 supply and abscisic acid.
Pospíšilová J., Synková H., Rulcová J.:
Cytokinins and water stress.
Biol. Plant. 43(3): 321-328, 2000.
Sergeeva L.I., de Bruijn S.M., Koot-Gronsveld E.A.M., Navrátil O.,
Vreugdenhil D.:
Tuber morphology and starch accumulation are independent phenomena:
Evidence from ipt-transgenic potato lines.
Physiol. Plant. 108(4): 435-443, 2000.
Smýkal P., Mašín J., Hrdý I., Konopásek I., Žárský V.:
Chaperone activity of tobacco HSP18, a small heat-shock protein, is inhibited
by ATP.
Plant J. 23(6): 703-713, 2000.
Soukupová J., Cvikrová M., Albrechtová J., Rock B.N., Eder J.:
Histochemical and biochemical approaches to study of phenolic compounds
and peroxidases in needles of Norway spruce (Picea abies (L.) Karst).
New Phytol. 146: 403-414, 2000.
51
4.831
1.259
2.278
0.945
4.134
0.444
2.149
0.444
0.424
1.476
5.629
2.149
AV0Z50380511
120 Šesták Z., Čatský J.:
0.402
121
0.424
122
123
124
125
126
127
128
129
130
Bibliography of reviews and methods of photosynthesis - 83.
Influence of auxin-like herbicides on tobacco mosaic virus multiplication.
Biol. Plant. 43(3): 467-470, 2000.
Correlation between activity of ribonucleases and potato virus Y biosynthesis
in tobacco plants.
Physiol. Mol. Plant Pathol. 57(5): 191-199, 2000.
Štorchová H., Hrdličková R., Chrtek J., Tetera M., Fitze D., Fehrer J.:
An improved method of DNA isolation from plants collected in the field and
conserved in saturated NaCl/CTAB solution.
Taxon 49(1): 79-84, 2000.
ten Hoopen R., Manteuffel R., Doležel J., Malysheva L., Schubert I.:
Evolutionary conservation of kinetochore protein sequences in plants.
Chromosoma 109(7): 482-489, 2000.
Tolman V., Sedmera P.:
Chemistry of 4-fluoroglutamic acid. Part 3. Preparation of the diastereomers of
4-fluoroglutamine and 4-fluoroisoglutamine. An enzymatic access to the
antipodes of 4-amino-2-fluorobutyric acid.
J. Fluorine Chem. 101(1): 5-10, 2000.
Tolman V., Šimek P.:
Chemistry of 4-fluoroglutamic acid. Part 4. Resolution of the racemic erythro
and threo forms through their diastereomeric salts.
J. Fluorine Chem. 101(1): 11-14, 2000.
Trávníček Z., Maloň M., Biler M., Hajdúch M., Brož P., Doležal K., Holub
J., Kryštof V., Strnad M.:
Synthesis, characterization and biological activity of two nickel (II) complexes
with 6-(2-chlorobenzylamino)purine.
Transit. Metal Chem. 25: 265-269, 2000.
Ulman P., Čatský J., Pospíšilová J.:
Photosynthetic traits in wheat grown under decreased and increased CO2
concentration, and after transfer to natural CO2 concentration.
Biol. Plant. 43(2): 227-237, 2000.
Vrána J., Kubaláková M; Šimková H., Čihalíková J., Lysák M.A., Doležel
J.:
Flow sorting of mitotic chromosomes in common wheat (Triticum aestivum L.).
Genetics 156(4): 2033-2041, 2000.
Wittink F.R.A., Knuiman B., Derksen J., Čapková V., Twell D., Schrauwen
J.A.M., Wullems G.J.:
The pollen-specific gene Ntp303 encodes a 69-kDa glycoprotein associated
with the vegetative membranes and the cell wall.
Sex. Plant Reprod. 12(5): 276-284, 2000.
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131 Angelis K. J.:
132
133
The use of single cell gel electrophoresis for the study of DNA repair in plants.
Plant Mol. Biol. Reporter 18(2): S03-6, 2000.
Burketová L., Feltlová M., Štillerová K.:
Detekce viru žluté nekrotické žilkovitosti řepy in situ.
Biol. Listy 65: 263-265, 2000.
Dobrev P., Kamínek M.:
Cytokinin purification: new approaches on old grounds.
Biol. Listy 65: 230-233, 2000.
52
1.970
0.863
3.157
0.851
0.851
0.561
0.424
4.687
2.260
-
AV0Z50380511
134 Doležel J., Lysák M.A., Vrána J., Kubaláková M., Šimková H., Číhalíková,
-
135
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136
137
138
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140
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142
143
144
145
146
147
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Flow cytogenetics of agricultural crops. – In: Proceedings of the Mendel
Centenary Congress.
Vortr. Pflanzenzüchg. 48: 247-256, 2000.
Hanuš J., Kryštof V., Hajdúch M., Veselý J., Strnad M.:
Substituted nitrogen heterocyclic derivatives, way of their preparation, these
derivatives for uses as drugs, pharmaceutical composition and combined
pharmaceutical preparation containing these derivatives and uses of these
derivatives for drug production. PCT/CZ00/00002, Czech Patent Application,
January 25th, 2000.
Havlíček L., Kryštof V., Siglerová V., Lenobel R., Van Onckelen H.,
Slegers H., Esmans E., Strnad M.:
New heterocyclic compounds with anticancer, immunosupressive, antiviral
and antiinflammatory properties, processes for their preparation and methods
for therapy. BE-1307-00, Patent Application, January 14th 2000
Hoyerová K., Kamínek M., Březinová A., Dobrev P., Gaudinová A.:
Comparison of different methods of extraction, purification and determination
of cytokinins.
Biol. Listy 65: 237-230, 2000.
Kamínek M., Gaudinová A., Dobrev P.:
Cytokinin-binding proteins in cereal grains: Specific physiological functions
and methodological approaches.
Biol. Listy 65: 240-242, 2000.
Malá J., Cvikrová M., Kálal J., Cvrčková H., Eder J.:
The influence of PVP on rooting of oak microcuttings.
Comm. Inst. Forest. 19: 5-14, 2000.
Cvikrová M.:
Endogenous contents of phytohormones and phenylpropanoids in sessile oak
somatic embryos in relation to their conversion potential.
J. Forestry Sci. 46(5): 197-204, 2000.
Malá J., Kálal J., Cvrčková H., Cvikrová M., Eder J:
The effect of reduction of exuded phenolic substances level on rooting of oak
microcuttings. Proc. Int. Symp. Methods and Markers for Quality Assurance in
Micropropagation. (Eds. Cassels A.C., Doyle B.M., Curry P.F.),
Acta Hort. 530: 353-360, 2000.
Martinec J., Feltl T., Nokhrina E., Zažímalová E., Macháčková I.:
Plant inositol signaling – biochemical study of phospholipase C and D-myoinositol-1,4,5-trisphosphate receptor.
Korean J. Plant Tissue Cult. 27(5): 375-377, 2000.
Motyka V., Kamínek M.:
Extraction and determinatrion of cytokinin oxidase activity in wheat grains and
tobacco callus culture.
Biol. Listy 65: 248-250, 2000.
Motyka V.:
Methods for determination of cytokinin oxidase and zeatin reductase activity in
plant tissues - and overview.
Biol. Listy 65: 245-247, 2000.
Sedlářová M., Binarová P., Lebeda A.:
Imunochemical methods in plant cytopathology.
Biol. Listy 65(3-4): 262-263, 2000.
Soukupová J., Cvikrová M., Albrechtová J., Rock B.N., Eder J.:
Influence of air polution on phenolic compound composition in needles of
Norway spruce. In: Polyphenols Communications 2000, FreisingWeihenstephan (Germany), September 11-15, pp. 599-601, 2000.
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Regulation of cell division by growth substances.
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53
-
-
-
-
-
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-
-
-
-
AV0Z50380511
148 Svirshchevskaya A.M., Doležel J.:
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149
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150
151
152
153
Production and performance of gynogenetic sugarbeet lines.
J. Sugarbeet Res. 37: 117-133, 2000.
Trčková, M., Kamínek, M.:
Nitrate uptake and nitrogen allocation in wheat as affected by exogenous
cytokinins. In: Nitrogen and Sustaineble Ecosystems. M.A. Martins-Loucao &
S.H. Lips, pp. 261-268, 2000.
Vaňková R., Kuncová G.:
Two dimensional fluorescence spectroscopy – non-traditional method for
determination of cell viability.
Biol. Listy 65: 299 - 301, 2000.
Zažímalová E., Petrášek J.:
Estimation of activity of auxin uptake and efflux carriers in the cells of VBI-0
tobacco strain.
Biol. Listy 65: 253-257, 2000.
Zonia L.:
The actin cytoskeleton during differentiation of microspores to mature pollen.
In: Actin: A Dynamic Framework for Multiple Plant Cell Functions. Eds.
Staiger, C.J., Baluška, F., Volkmann, D., Barlow, P.W. Kluwer Academic
Publishers, Dordrecht. Pp. 361-371, 2000.
Žárský V., Soukupová H.:
Stress as a trigger of pollen embryogenesis.
Korean J. Plant Tissue Cult. 27: 411-413, 2000.
-
-
-
-
2001:
IMPACTED JOURNALS:
no.
title
Blažková
A.,
Macháčková
I.,
Eder
J.,
Krekule J.:
154
Benzyladenine-induced inhibition of flowering in Chenopodium rubrum in vitro
is not related to the level of isoprenoid cytokinins.
Plant Growth Regul. 34: 159-166, 2001.
155 Bubner M., Fuksová K., Matucha M., Heise K. H., Bernhard G.:
Synthesis of [1,2-C-14]trichloroacetic acid.
J. Labelled Comp. & Radiopharm. 44(11): 811-814, 2001.
156 Čeřovská N., Moravec T., Filigarová M., Petrzik, K.:
Nucleotide sequences of 5´ terminal parts of coat protein genes of various
isolates of NTN strain of Potato virus Y.
Acta Virologica 45: 55-59, 2001.
Čeřovská
N., Moravec T., Velemínský J.:
157
Expression of Potato virus A (PVA) coat protein in Escherichia coli and
possibility of its use as foreign antigen carrier.
Eur. J. Biochem. 268(1): 214, 2001.
158 Eliáš M., Cvrčková F., Obermeyer G., Žárský V.
Microinjection of guanine nucleotide analogues into lily pollen tubes results in
isodiametric tip expansion.
Plant Biol. 3(5): 489-492, 2001.
159 Engstová, H., Žáčková M., Růžička M., Meinhardt A., Hanuš J., Kramer
R., Ježek P.:
Natural and azido fatty acids inhibit phosphate transport and activate fatty
acid anion uniport mediated by the mitochondrial phosphate carrier.
J. Biol. Chem. 276(7): 4683-4691, 2001.
Fellner
M., Sawhney V.K.:
160*
Seed germination in a tomato male-sterile mutant is resistant to osmotic, salt
and low-temperature stresses.
Theor. Appl. Genet. 102: 215-221, 2001.
54
IF 2001
0.761
0.839
0.644
2.849
1.828
7.258
2.438
AV0Z50380511
161* Fellner M., Zhang R., Pharis R.P., Sawhney V.K.:
2.433
162
1.018
163
164
165
166
167
168
169
170
171
172
173
174
Reduced de-etiolation of hypocotyl growth in a tomato mutant is associated
with hypersensitivity to, and high endogenous levels of, abscisic acid.
J. Exp. Bot. 52: 725-38, 2001.
Fidlerová A., Smýkal P., Tupý J., Čapková V.:
Glycoproteins 66 and 69 kDa of pollen tube wall: properties and distribution in
angiosperms.
J. Plant Physiol. 158 : 1367-1374, 2001.
Forczek S.T., Matucha M., Uhlířová H., Albrechtová J., Fuksová K.,
Schröder H.P.:
Biodegradation of trichloroacetic acid in Norway spruce/soil system.
Biol. Plant. 44(2): 317-320, 2001.
Franěk F., Strnad M., Havlíček L., Siglerová V., Eckschlager T.:
Concentration- and time-dependent activities of bohemine, a novel cytostatic
agent.
Cytotechnology 36: 115-122 (2001).
Gichner T., Stavreva D.A., Van Breusegem F.:
o-Phenylenediamine-induced DNA damage and mutagenicity in tobacco
seedlings is light-dependent.
Mutat. Res. 495(1-2): 117-125, 2001.
Haisel D., Hofman P., Vágner M., Lipavská H., Tichá I., Schäfer C.,
Čapková V.:
Ex vitro phenotype stability is affected by in vitro cultivation.
Biol. Plant. 44(3): 321-324, 2001.
Henselová M., Vizárová, G., Macháčková I.:
The effect of growth regulator Rastim 30DKV on the level of endogenous
phytohormones in tomato (Solanum lycopersicum L.).
Rostlinná Výroba 47: 411-417, 2001.
Honys D.:
Isolation of proteins comprising native gene-specific messenger
ribonucleoprotein particles using paramagnetic beads.
Plant Sci.: 161(3) 605-611, 2001.
Horký M., Wurzer G., Kotala V., Anton M., Vojtěšek B., Vácha J.,
Wesierska-Gadek J.:
Segregation of nucleolar components coincides with caspase-3 activation in
cisplatin-treated HeLa cells.
J. Cell Sci. 114: 663-670, 2001.
Chmela Z., Veselý J., Lemr K., Rypka M., Hanuš J., Havlíček L., Kryštof
V., Michnová L., Fuksová K., Lukeš J.:
In vivo metabolism of 2,6,9-trisubstituted purine-derived cyclin-dependent
kinase inhibitor bohemine in mice: Glucosidation as the principal metabolic
route.
Drug Metabol. Dispos. 29: 326-334, 2001.
Kadleček P., Tichá I., Haisel D., Čapková V., Schäfer Ch.:
Importance of in vitro pretreatment for ex vitro acclimation and growth.
Plant Sci. 161: 695-701, 2001.
Kejnovský E., Vrána J., Matsunaga S., Souček P., Široký J., Doležel J.,
Vyskot B.:
Localization of male-specifically expressed MROS genes of Silene latifolia by
PCR on flow-sorted sex chromosomes and autosomes.
Genetics 158: (3) 1269-1277, 2001.
Kotala V., Uldrijan S., Horký M., Trbušek M., Strnad M., Vojtěšek B.:
Potent induction of wild-type p53-dependent transcription in tumour cells by a
synthetic inhibitor of cyclin-dependent kinases.
Cell Mol. Life Sci. 58: 1333-1339, 2001.
Kryštof V., Strnad M.:
Inhibitors of cyclin-dependent kinases.
Chem. Listy 95: 295-300, 2001.
55
0.426
0.703
1.624
0.426
0.237
1.384
6.213
2.989
1.384
4.803
4.539
0.317
AV0Z50380511
175 Maloň M., Trávníček Z., Maryško M., Zbořil R., Mašláň M., Marek J.,
1.394
176
1.181
177
178
179
180
181
182
183
184
185
186
187
188
189
Doležal K., Rolčík J., Kryštof V., Strnad M.:
Metal complexes as anticancer agent 2. Iron(III) and copper(II) bio-active
complexes with 6-benzylaminopurine derivatives.
Inorg. Chim. Acta 323(1-2): 119-129, 2001.
Matucha M., Uhlířová H., Bubner M.:
Investigation of uptake, translocation and fate of trichloroacetic acid in Norway
spruce (Picea abies /L./ Karst.) Using 14C-Labelling.
Chemosphere 44(2): 217-222, 2001.
Menke M., Cheng I-P., Angelis K.J., Schubert I.:
DNA damage and repair in Arabidopsis thaliana as measured by the Comet
Assay after treatment with different classes of genotoxins.
Mutat. Res. 493(1–2): 87-93, 2001.
Mužáková V., Kandar R., Vojtíšek P., Skalický J., Vaňková R., Cegan A.,
Červinková Z.:
Antioxidant vitamin levels and glutathione peroxidase activity during
ischemia/reperfusion in myocardial infarction.
Physiol. Res. 50(4): 389-396, 2001.
Pospíšilová J., Rulcová J., Vomáčka L. :
Effect of benzyladenine and hydroxybenzyladenosine on gas exchange of
bean and sugar beet leaves.
Biol. Plant. 44(4): 523-528, 2001.
Procházková D., Sairam R., Srivastava G.C., Singh D.V.:
Oxidative stress and antioxidant activity as the basis of senescence in maize
leaves.
Plant Sci. 161: 765-771, 2001.
Ptáček O., Stavreva D. A., Jin Kyu Kim, Gichner T.:
Induction and repair of DNA damage as measured by the Comet assay and
the yield of somatic mutations in gamma-irradiated tobacco seedlings.
Mut. Res. 491: 17-23, 2001.
Roux N., Doležel J., Swennen R., Zapata-Arias F.J.:
Effectiveness of three micropropagation techniques to dissociate
cytochimeras in Musa spp.
Rulcová J., Pospíšilová J.:
Effect of benzylaminopurine on rehydration of bean plants after water stress.
Biol. Plant. 44: 75-81, 2001.
Sedlářová M., Binarová P., Lebeda A.:
Changes in microtubular alignment in Lactuca spp. (Asteraceae) epidermal
cells during early stages of infection by Bremia lactucae (Perenosporaceae).
Phyton - Ann Rei Bot. 41(1): 21-34, 2001.
Suda J., Lysák M.A.
A taxonomic study of the Vaccinium sect. Oxycoccus (Hill) W.D.J. Koch
(Ericaceae) in the Czech Republic and adjacent territories.
Folia Geobot. 36(3): 303-320, 2001.
Synková H., Valcke R.:
Response to mild water stress in transgenic Pssu-ipt tobacco.
Physiol. Plant. 112: 513-523, 2001.
Bibliography of reviews and methods of photosynthesis – 84.
Photosynthetica 39 (1): 131-160, 2001.
Bibliography of reviews and methods of photosynthesis – 85.
Photosynthetica 39 (4): 615-640, 2001.
Šindelářová M., Šindelář L.:
Changes in composition of soluble intercellular proteins isolated from healthy
and TMV-infected Nicotiana tabacum L. cv. Xanthi-nc.
Biol. Plant. 44, 567- 572, 2001.
56
1.624
1.027
0.426
1.384
1.624
0.631
0.426
0.275
0.467
1.760
0.807
0.807
0.426
AV0Z50380511
190 Široký J., Lysák M.A., Doležel J., Kejnovský E., Vyskot B.:
1.835
191
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192
193
194
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196
197
Heterogeneity of rDNA distribution and genome size in Silene spp.
Chromosome Res. 9: 387-393, 2001.
Temsch E.M., Obermayer R., Doležel J., Greilhuber J.:
Application of an optical immersion-gel in a flow cytometer with horizontally
oriented objective.
Biotechnic & Histochemistry 76: 11-14, 2001.
Trávníček Z., Maloň M., Šindelář Z., Doležal K., Rolčík J., Kryštof V.,
Strnad M., Marek J.:
Preparation, physicochemical properties and biological activity of copper(II)
complexes with 6-(2-chlorobenzylamino)purine (HL1) or 6-(3chlorobenzylamino)purine (HL2). The single-crystal X-ray structure of
[Cu(H+L2)(2)Cl-3]Cl-2H(2)O.
J. Inorg. Biochem. 84 (1-2): 23-32, 2001.
Vagera J., Nesvadba Z., Martinek P., Ohnoutková L.:
In vitro haploid zygotic embryogenesis due to pollination with maize pollen
and induced in vitro androgenesis in Czech wheat breeding genotypes.
Rostlinná Výroba 47(5): 193-200, 2001.
Vaňková R., Kuncová G., Opatrná J., Süssenbeková H., Gaudinová A.,
Vaněk T.:
Two-dimensional fluorescence spectroscopy - a new tool for the determination
of plant cell viability.
Plant Cell Rep. 20(1): 41-47, 2001.
Werner T., Motyka V., Strnad M., Schmülling T.:
Regulation of plant growth by cytokinin.
Proc. Natl. Acad. Sci. USA 98: 10487-10492, 2001.
Wolf K., Kolář J., Witters E., Van Dongen W., Van Onckelen H.,
Macháčková I.:
Daily profile of melatonin levels in Chenopodium rubrum L. depends on
photoperiod.
J. Plant Physiol. 158: 1491-1493, 2001.
Zonia L., Cordeiro S., Feijo J. A.:
Ion dynamics and hydrodynamics in the regulation of pollen tube growth.
Sex. Plant Reprod. 14(1-2): 111-116, 2001.
OTHERS:
198 Bubner M., Meyer M., Heise K.H., Jander R., Matucha M., Vlasáková V.,
199
200
201
Fuksová K., Nitsche H.:
Radiolabelling of Polychlorinated Biphenyls: Synthesis of [14C]PCB
Congeners 11 and 77. Proc. Seventh Int. Symp. on the Synthesis and
Applications of Isotopes and Isotopically Labelled Compounds, June 18 - 22,
2000, Dresden, Germany, (R. Voges and U. Pleiss, Eds.), J. Wiley, pp.260263, 2001.
Cvrčková F., Eliáš M., Hála M., Obermeyer G. Žárský V.:
Small GTPases and Conserved Signalling Pathways in Plant Cell
Morphogenesis: From Exocytosis to the Exocyst.
In ”Cell Biology of Plant and Fungal Tip Growth” (A. Geitmann, M. Cresti, B.
Heath eds.). IOS Press Amsterdam, pp. 105-122, 2001.
Doležel J., Lysák M., Doleželová M., Valárik M., Šimková H., Vrána J.:
Analysis of Musa genome using flow cytometry and molecular cytogenetics. In: Cellular Biology and Biotechnology Including Mutation Techniques for
Creation of New Useful Banana Genotypes. Report of the Third FAO/IAEA
Research Co-ordination Meeting. Pp. 85-93. IAEA, Vienna, 2001.
Doležel J., Lysák M.A., Kubaláková M., Šimková H., Macas J., Lucretti S.:
Sorting of Plant Chromosomes. – In: Darzynkiewicz, Z., Crissman, H.A.,
Robinson, J.P. (eds): Methods in Cell Biology. Vol. 64. Third Edition, Part B.
Pp. 3 – 31. Academic Press, San Diego, 2001.
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1.729
0.237
1.375
10.896
1.018
1.753
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202 Forczek S.T., Matucha M., Uhlířová H., Albrechtová J., Fuksová K.,
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207
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211
212
213
Schröder P.:
Distribution and Fate of Trichloroacetic Acid in Norway Spruce (Picea Abies
(L.) Karst.) Using 14C-Labelling.
In: Proc. Seventh Int. Symp. on the Synthesis and Applications of Isotopes
and Isotopically Labelled Compounds (U. Pleiss, R. Voges, Eds.) J. Wiley, p.
610-613, 2001.
Franěk F., Katinger H.:
Specific effects of synthetic oligopeptides in animal cell culture. Independent
modulation of production and growth. Animal Cell Technology: From Target to
Market. Proc. 17th ESACT Meeting Tylosand, Sweden, June 10-14, 2001,
Kluwer Acad. Publ., The Netherlands, pp.164-167, 2001.
Kolář J., Macháčková I.:
Occurrence and possible function of melatonin in plants. A review.
Endocytobiosis & Cell Res. 14: 75-84, 2001.
Kolářová H., Kubínek R., Navrátil L., Strnad M., Réblová K., Škopek J.:
Laser-induced photodynamic effect.
In: Laser Florence 2000: A window on the laser Medicine World,, Lonog, L.,
Hofstetter, A.G., Pascu, M.L., Waidelich, W.R.A. (eds.), Porceedings of SPIE,
vol. 4606: 144-147, 2001.
Krekule J., Macháčková I.:
The localization of endogenous rhythm of flowering in Chenopodium rubrum.
Evidence gained in experiments with direct electric current.
Endocytobiosis & Cell Res. 14: 51-56, 2001.
Kubaláková M., Vrána J., Číhalíková J., Lysák M.A., Doležel J.:
Localisation of DNA sequences on plant chromosomes using PRINS and CPRINS.
Methods Cell Sci. 23: 71-82, 2001.
Lebeda A., Doležalová I., Křístková E., Janeček J., Vinter V., Vránová O.,
Doležal K., Tarkowski P., Petrželová I., Trávníček B., Novotný R.:
Biodiversity of genetic resources of wild Lactuca spp. In: Swiecicki, W.,
Naganowska, B., Wolko, B. (Eds.): Broad Variation and Precise
Characterization – Limitation for the Future. Eucarpia, Section Genetic
Resources, Poznan (Poland), 53-56, 2001.
Macháčková I., Krekule J.:
Úloha světla v regulaci růstu rostlin.
Zprávy Čes. Bot. Společ. Praha 36: 31-47, 2001
Martinek P., Ohnoutková L., Bednář J., Vyhnánek T.:
Increasing grain quality parameters by intra- and interspecific hybridisation in
wheat.
Proceeding of International Conference of Genetic Collections, Isogenic and
Alloplasmic Lines, Novosibirsk, Russia, July 30 - August 03, 195-197, 2001.
Martinek P., Ohnoutková L., Bednář J., Vyhnánek T.:
Vlastnosti Tritordea v podmínkách České republiky a možnosti jeho využití.
(Characteristic of tritordeum and its potential use in the Czech Republic).
Nové poznatky z genetiky a šľachtenia poľnohospodárskych rastlín.
Zborník zo 6. odborného seminára VÚRV Piešťany 10. Máj 2001, (6):49-53,
2001 (ISBN 80-88790-19-0)
Matucha M., Uhlířová H., Fuksová K., Bubner M.:
Application of 14C-Labelling In Investigation Of Forest Decline: Uptake,
Translocation And Fate Of Trichloroacetic Acid In Norway Spruce (Picea Abies
(L.) KARST.) Proc. Seventh Int. Symp. on The Synthesis and Applications of
Isotopes and Isotopically Labelled Compounds, June 18 - 22, 2000, Dresden,
Germany, (U. Pleiss, J. Voges, Eds.) J. Wiley 2001, pp. 604-609. 2001.
Matucha M., Uhlířová H., Lomský B., Fuksová K., Forczek S.T.,
Albrechtová J.:
Těkavé chlorované uhlovodíky a zdravotní stav lesa: účinek kyseliny
trichloroctové na smrk ztepilý (Picea abies /L./ Karst),
Konf. Ovzduší 2001, Brno, 14.-16.5.2001, Sborník konf. Recetox Brno 2001,
str. 182-188, 2001.
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214 Moravec T., Filigarová M., Dědič P., Čeřovská N.:
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216
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218
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220
221
222
Possibility of the use of recombinant coat protein of Potato Mop-top virus
(PMTV) for virus characterization. In: 11-th EAPR Virology section meeting,
Book of abstracts 80-82, Třešť, Czech Republic, 7-13-th October 2001.
Müllerová E., Novotný J., Vagera J., Harwood W.A., Ohnoutková L.:
Induction of androgenesis in transgenic barley plants.
In: Bohanec, B., Ed. Biotechnological Approaches for Utilization of Gametic
Cells. Office for Official Publications of the European Communities,
Luxembourg. 29-32, 2001.
Němcová L., Faragó J., Ohnoutková L., Müllerová E.:
High-frequency regeneration through somatic embryogenesis from immature
scutellum-derived calli in Slovak and Czech cultivars of barley (Hordeum
vulgare L.).
In: Book of Abstracts of 4th International Symposium in the Series “Recent
Advances in Plant Biotechnology”, Třeboň, Czech Republic, September 1721, 130, 2001.
Ohnoutková L., Müllerová E., Faragó J.:
Transgenoze ječmene – současny stav a perspektivy. (Genetic
Transformation of barley – situation and perspektive. Nové poznatky z
genetiky a šľachtenia poľnohospodárskych rastlín.
Zborník zo 6. odborného seminára VÚRV Piešťany 10. Máj 2001, pp. 25-28,
2001.
Ohnoutková L., Novotný J., Vagera J., Kučera L.:
Is a could pre-treatment needed for induction of in vitro androgenesis in barley
and wheat?
In: Bohanec, B., Ed. Biotechnological Approaches for Utilization of Gametic
Cells. Office for Official Publications of the European Communities,
Luxembourg. 33-37, 2001.
Svirshchevskaya A.M., Doležel J.:
Karyological characterization of sugar beet gynogenetic lines cultured in vitro.
J. Appl. Genet. 42: 21-32, 2001.
Štorchová H., Procházka F., Horn K., Pavlíčko A.
Diphasiastrum oellgardii - nový druh moravské květeny. (Diphasiastrum
oellgardii - a new species of Moravian flora).
Zprávy Čes. Bot. Spol. 36: 77 - 80, 2001.
Uldrijan S., Nenutil R., Fait V., Kotala V., Rejthar A., Vojtěšek B.:
Melanomové fragmenty a jejich krátkodobá kultivace jako nástroj pro
sledování odezvy na cytotoxické látky s rozdílným mechanismem účinku.
Klin. Onkologie 1, 25-30, 2001.
Vrána J., Šimková H., Kubaláková M., Číhalíková J., Lysák M., Doležel J.:
Sorting of mitotic chromosomes in common wheat (Triticum aestivum L.)
using flow cytometry.
In: Bedö, Z., Láng, L. (eds.): Wheat in a Global Environment. Pp. 531-536.
Kluwer Academic Publishers, Dordrecht, 2001.
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2002:
IMPACTED JOURNALS:
č.
citace
Ayaz
F.A.,
Glew
R.H.,
Huang
H.S.,
Chuang L.T., VanderJagt D.J., Strnad
223
224
M.:
Evolution of fatty acids in mediar (Mespilus germanica L.) mesocarp at
different stages of ripening.
Grasas Aceites 53: 352-356, 2002.
Baroja-Fernandez E., Aguirreolea J., Martínková H., Hanuš J., Strnad M.:
Aromatic cytokinins in micropropagated potato plants.
59
IF 2002
0.286
1.582
AV0Z50380511
225 Bavrina T.V., Milyaeva E.L., Macháčková I., Pustovoitova T.N., Gurko
N.A., Kasumova I.V., Zhdanova N.E.:
Effect of phytohormone biosynthesis genes (ipt and iaaM plus iaaH) on the
sexual reproduction of transgenic tobacco plants.
Russ. J. Plant Physiol. 49(4): 484-491, 2002.
Čeřovská
N., Filigarová M., Hadámková R.:
226
Optimum conditions for the storage of potato virus YNTN strain.
Biol. Plant. 45: 315-316, 2002.
227 Čeřovská N., Moravec T., Velemínský J.:
Polyclonal antibodies to a recombinant coat protein of Potato virus A.
Acta Virologica 46: 31; 2002.
228 Dobrev P., Kamínek M.:
Fast and efficient separation of cytokinins from auxin and abscisic acid and
their purification using mixed-mode solid-phase extraction.
J. Chromatography A 950(1-2): 21-29, 2002.
229 Dobrev P., Motyka V., Gaudinová A., Malbeck J., Trávníčková A.,
Kamínek M., Vaňková R.:
Transient accumulation of cis and trans-zeatin type cytokinins and its relation
to cytokinin oxidase activity during cell cycle of synchronized tobacco BY-2
cells.
230 Doležal K., Astot C., Hanuš J., Holub J., Peters W., Beck E., Strnad M.,
Sandberg G.:
Identification of aromatic cytokinins in suspension cultured photoautotrophic
cells of Chenopodium rubrum by capillary liquid chromatography/frit - fast
atom bombardment mass spectrometry.
231 Doležalová I., Lebeda., Janeček J., Čihalíková J., Křístková E., Vránová
O.:
Variation in chromosome numbers and nuclear DNA contents in genetic
resources of Lactuca L. species (Asteraceae).
Genet. Resources Crop Evol. 49(4): 383-395, 2002.
232* Fellner M., Sawhney V.K:
The 7B-1 mutant in tomato shows blue-light-specific resistance to osmotic
stress and abscisic acid.
Planta 214: 675-682, 2002.
Filek
M., Biesaga-Koscielniak J., Marcinska I., Krekule J., Macháčková I.:
233
Direct electric current partly replaces the chilling effect in vernalisation of
winter wheat.
J. Plant Physiol. 159(7): 795-797, 2002.
234 Franěk F., Fismolová I., Eckschlager T.:
Antiapoptotic and proapoptotic action of various amino acids and analogs in
starving MOLT-4 cells.
Arch. Biochem. Biophys. 398(1): 141-146, 2002.
Franěk
F., Katinger H.:
235
Specific effects of synthetic oligopeptides on cultured animal cells.
Biotechnol. Progress 18(1): 155-158, 2002.
Franěk
F., Siglerová V., Havlíček L., Strnad M., Eckschlager T., Weigl E.:
236
Effect of the purine derivative myoseverin and of its analogues on cultured
hybridoma cells.
Coll. Czech. Chem. Comm. 67(2): 257-266, 2002.
237 Gichner T., Mühlfeldová Z.:
Induced DNA damage measured by the Comet assay in 10 weed species.
Biol. Plant. 45 (4): 509-516, 2002.
238 Govindjee, Šesták Z., Peters W.R.:
The early history of "Photosynthetica", "Photosynthesis Research", and their
publishers.
Photosynthetica 40(1), 1-11, 2002.
60
0.102
0.583
0.660
3.098
1.582
0.850
0.579
2.960
0.941
2.606
1.734
0.848
0.583
0.773
AV0Z50380511
239 Hartung F., Angelis K.J., Meister A., Schubert I., Melzer M., Puchta H.:
7.007
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An archaebacterial topoisomerase homolog not present in other eucaryotes is
indispesable for cell proliferation of plants.
Curr. Biology 12: 1787-1791, 2002.
Hofman P., Haisel D., Komenda J., Vágner M., Tichá, I., Schaffer C.,
Čapková V.:
Impact of in vitro cultivation conditions on stress responses and on changes in
thylakoid membrane proteins and pigments of tobacco during ex vitro
acclimation.
Biol. Plant. 45: 189-195, 2002.
Janas K.M., Cvikrová M., Palagiewicz A., Szafranska K., Posmyk M.M.:
Constitutive elevated accumulation of phenylpropanoids in soybean roots at
low temperature.
Plant Sci. 163(2): 369-373, 2002.
Kolář Z., Murray P.G., Maďarová J., Lukešová M., Hlobilková A.,
Řiháková P., Flavell J.R., Strnad M., Student V., Vojtěšek B.: Nuclear
receptors in early hormone refractory prostate cancer and their relationship to
apoptosis-related proteins.
Neoplasma 49: 172-177, 2002.
Kryštof V., Lenobel R., Havlíček L., Kuzma M., Strnad M.:
Synthesis and biological activity of olomoucine II.
Bioorg. Med. Chem. Lett. 12(22): 3283-3286, 2002.
Kubaláková M., Vrána J., Číhalíková J., Šimková H., Doležel J.:
Flow karyotyping and chromosome sorting in bread wheat (Triticum aestivum
L.).
Theor. Appl. Genet. 104(8): 1362-1372, 2002.
Link V.L., Hofmann M.G., Sinha A.K., Ehness R., Strnad M., Roitsch T.:
Biochemical evidence for the activation of distinct subsets of mitogenactivated protein kinases by voltage and defense-related stimuli.
Plant Physiol. 128(1): 271-281, 2002.
Maďarová J., Lukešová M., Hlobilková A., Strnad M., Vojtěšek B.,
Lenobel R., Hajdúch M., Murray P.G., Perera S., Kolář Z.:
Synthetic inhibitors of CDKs induce different responses in androgen sensitive
and androgen insensitive prostatic cancer cell lines.
J. Clin. Pathol. 55: 227-234, 2002.
Macháčková I., Krekule J.:
Sixty-five years of searching for the signals that trigger flowering.
Maloň M., Trávníček Z., Marysko M., Marek J., Doležal K., Rolčík J.,
Strnad M.:
Synthesis, characterization and antitumour activity of copper(II) 6-(4chlorobenzylamino)purine complexes. X-ray structure of 6-(4-chlorobenzylamino)purinium perchlorate.
Transit. Metal Chem. 27(6), 580-586, 2002.
Matschke J., Macháčková I.:
Changes in the content of indole-3-acetic acid and cytokinins in spruce, fir and
oak trees after herbicide treatment.
Biol Plant. 45(3): 375-382, 2002.
Mes T.H.M., Kuperus P., Kirschner J., Štěpánek J., Oosterveld P.,
Štorchová H., Nijs den J.C.M.:
Detection of genetically divergent clone mates in apomictic dandelions.
Mol. Ecol. 11(2): 253-265, 2002.
Neumann P., Požárková D., Vrána J., Doležel J., Macas J.:
Chromosome sorting and PCR-based physical mapping in pea (Pisum
sativum L.).
Chrom. Res. 10(1): 63-71, 2002.
Petrášek J., Březinová A., Holík J., Zažímalová E.:
Excretion of cytokinins into the cultivation medium by suspension-cultured
tobacco cells.
Plant Cell Rep. 21(1): 97-104, 2002.
61
1.556
0.679
2.051
2.249
5.800
2.549
0.102
0.949
0.583
3.014
1.828
1.340
AV0Z50380511
253 Petrášek J., Elčkner M., Morris D.A., Zažímalová E.:
2.960
254
0.102
255
256
257
258
259
260
261
262
263
264
265
266
Auxin eflux carrier activity and auxin accumulation regulate cell division and
polarity in tobacco cells.
Planta 216(2): 302-308, 2002.
Piruzian E.S., Goldenkova I.V., Lenets A.A., Cvikrová M., Macháčková I.,
Kobets N.S. Mett V.L. Musiichuk K.A.:
Physiological and biochemical characteristics of tobacco transgenic plants
expressing bacterial dioxygenase.
Požárková D., Koblížková A., Roman B., Torres A.M., Lucretti S., Lysák
M., Doležel J., Macas J.:
Development and characterization of microsatellite markers from
chromosome 1-specific DNA libraries of Vicia faba.
Biol. Plant. 45(3): 337-345, 2002.
Ptáček O., Mühlfeldová Z., Dostálek J., Čechák T., Gichner T.:
Monitoring DNA damage in wood small-reed (Calamagrostis epigejos) plants
growing in a sediment reservoir with substrates from uranium mining.
J. Environ. Monitoring 4(4): 592-595, 2002.
Rolčík J., Lenobel R., Siglerová V., Strnad M.:
Isolation of melatonin by immunoaffinity chromatography.
J. Chromatography B 775(1): 9-15, 2002.
Rypka M., Veselý J., Chmela Z., Riegrová D., Červenková K., Havlíček I.,
Lemer K., Hanuš J., Černý B., Lukeš J., Michalíková K.:
In vitro biotransformations of 2,6,9-trisubstituted purine derived cyclindependent kinase inhibitor bohemine by mouse liver microsomes.
Xenobiotica 32: 1017-1031, 2002.
Semorádová Š., Synková H., Pospíšilová J.:
Responses of tobacco plantlets to change of irradiance during transfer from in
vitro to ex vitro conditions.
Photosynthetica 40(4): 605-614, 2002.
Scherer G.F.E., Paul R.U., Holk A., Martinec J.:
Down-regulation by elicitors of phosphatidylcholine-hydrolyzing phospholipase
C and up-regulation of phospholipase A in plant cells.
Biochem Biophys. Res. Comm. 293(2): 766-770, 2002.
Soukupová J., Albrechtová J., Svobodová H., Opatrná J.:
Anatomical and histochemical changes of Norway spruce buds induced by
simulated acid rain
Biol. Plant. 45: 77-84, 2002.
Stavreva D.A., Gichner T.:
DNA damage induced by hydrogen peroxide in cultured tobacco cells is
dependent on the cell growth stage.
Mutation Res. 514: 147-152, 2002.
Synková H., Pospíšilová J.:
In vitro precultivation of tobacco affects the response of antioxidative enzymes
during acclimation to ex vitro.
J. Plant Physiol. 159(7): 781-789, 2002.
Šesták, Z., Čatský, J.:
Bibliography of reviews and methods of photosynthesis - 86.
Šindelář L., Šindelářová M.:
Correlation of viral RNA biosynthesis with glucose-6-phosphate
dehydrogenase activity and host resistance.
Planta 215: 862-869, 2002.
Changes in glucose-6-phosphate dehydrogenase/6-phosphogluconate
dehydrogenase ratio and the glucose-6-phosphate, 6-phosphogluconate and
fructose-6-phosphate contents in tobacco plants infected with potato virus Y.
Biol. Plant. 45(4): 575-580, 2002.
62
0.583
1.348
1.913
1.919
0.773
2.935
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267 Šindelářová M., Šindelář L., Burketová L.:
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268
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277
Glucose-6-phosphate dehydrogenase, ribonucleases and esterases upon
tobacco mosaic virus infection and benzothiodiazole treatment in tobacco.
Biol. Plant. 45(3): 423-432, 2002.
Štorchová H., Chrtek J., Bartish I.V., Tetera M.,Kirschner J., Štěpánek J.:
Genetic variation in agamospermous taxa of Hieracium sect. Alpina
(Compositae) in Tatry Mts. (Slovakia).
Plant Syst. Evol. 235(1-4): 1-17, 2002.
Trávníček Z., Maloň M., Popa I., Doležal K., Strnad M.:
Preparation and cytotoxic activity of nickel (II) complexes with 6benzylaminopurine derivatives.
Transit. Metal Chem. 27(8): 918-923, 2002.
Uldrijan S., Kotala V., Vojtěšek B.:
Regulation of the p53 tumour suppresor stability and activity.
Chem. Listy 96: 145-149, 2002.
Valárik M., Šimková H., Hřibová E., Šafář J., Doleželová M., Doležel J.:
Isolation, characterization and chromosome localization of repetitive DNA
sequences in bananas (Musa spp.).
Chrom. Res. 10: 89-100, 2002.
Vermeulen K., Strnad M., Havlíček L., van Onckelen H., Lenjou M., Nijs
G., Van Bockstaele D.R., Berneman Z.N.:
Plant cytokinin analogues with inhibitory activity on cyclin dependent kinases
exert their antiproliferative effect through induction of apoptosis initiated by the
mitochondrial pathway: determination by a multiparalalic flow cytometric
analysis.
Exp. Hematol. 30(10): 1107-1114, 2002.
Vermeulen K., Strnad M., Kryštof V., Havlíček L., Van der Aa A., Lenjou
M., Nijs G., Rodrigus I., Stockman B., van Onckelen H., Van Bockstaele
D.R., Berneman Z.N.:
Antiproliferative effect of plant cytokinin analogues with an inhibitory activity
on cyclin-dependent kinases.
Leukemia 16(3): 299-305, 2002.
Vítová L., Stodůlková E., Bartoníčková A., Lipavská H.:
Mannitol utilisation by celery (Apium graveolens) plants grown under different
conditions in vitro.
Plant Sci. 163(4): 907-916, 2002.
Vláčilová K, Ohri D., Vrána J., Čihalíková J., Kubaláková M., Kahl G.,
Doležel J.:
Development of flow cytogenetics and physical genome mapping in chickpea
(Cicer arietinum L.)
Chrom. Res. 10(8): 695-706, 2002.
Zonia L., Cordeiro S., Tupý J., Feijo J.A.:
Oscillatory chloride efflux at the pollen tube apex has a role in growth and cell
volume regulation and is targeted by inositol 3,4,5,6-tetrakisphosphate.
Plant Cell 14: 2233-2249, 2002.
Zubko E., Adams C.J., Macháčková I., Malbeck J., Scollan C., Meyer P.:
Activation tagging identifies a gene from Petunia hybrida responsible for the
production of active cytokinins in plants.
Plant J. 29(6): 797-808, 2002.
OTHERS:
278 Ajalin I., Kobza F., Doležel J.:
Ploidy identification of doubled chromosome number plants in Viola x
wittrockiana Gams. M1-generation. .
Hort. Sci. (Prague) 29: 35-40, 2002.
63
0.949
0.336
1.828
3.366
4.693
1.556
1.828
10.751
5.850
-
AV0Z50380511
279 Cvikrová M., Binarová P., Cenklová V., Eder J., Doležel J., Macháčková
-
280
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281
282
283
284
285
286
287
288
289
290
I.:
Effect of 2-aminoindan-2-phosphonic acid on cell cycle progression in
synchronous meristematic cells of Vicia faba roots. Polyphenols
Communications, pp. 55-57, Marrakech-Morocco, 2002.
Čeřovská N., Moravec T., Synková H., Velemínský J., Pokorná D.,
Šmahel M.:
Engineering of the recombinant coat protein of potato virus A (PVA) as a
carrier for foreing antigen.
Zborník XVIII. Biochemický Zjazd. pp. 7-10, Stará Lesná 2002.
Doležel J., Alkhimova O., Janda J., Valárik M., Hřibová E., Bartoš J.,
Doleželová M., Šafář J., Šimková H.:
Physical mapping and comparative karyotype analysis in Musa.
In: Abstracts of the 3rd International Symposium on the Molecular and Cellular
Biology of Banana. Pp. 16-17. Catholic University Leuven, Leuven, 2002.
Doležel J., Valárik M., Vrána J., Šafář J., Hřibová E., Gasmanová N., Van
den Hoew I., Doleželová M., Swennen R., Šimková H.:
Analysis of Musa genome using flow cytometry and molecular cytogenetics.
In: Abstracts of the FAO/IAEA 4th Research Co-ordination Meeting on Cellular
Biology and Biotechnology Including Mutation Techniques for Creation of New
Useful Banana Genotypes.
Infomusa 11 (Promusa 9): 16 – 17, 2002.
Eliáš M., Potocký M., Cvrčková F., Žárský V.:
Molecular diversity of phospholipase D in angiosperms.
BMC Genomics 3: art. no.2, 1-15, 2002.
Franěk F., Strnad M., Havlíček L., Siglerová V.:
Antiproliferative and growth-stimulating activities of synthetic cytokinin
analogs. In: Shirata, S., et al. (eds.), Animal Cell Technology: Basic and
Applied Aspects. Vol. 12. Kluwer Acad. Publ., The Netherlands, pp. 315-319,
2002.
Haisel D., Tichá I.:
The effect of irradiance on growth parameters and photosynthetic pigments
content during acclimatization of micropropagated plants to ex vitro.
In: Zima, M., Černá, K. (ed.): Ecophysiology on Plant Stress, pp. 88-89. SPU,
Nitra 2002.
Krekule J., Ćulafić L.:
Control of flowering : past and present with special consideration of
photoperiodism and phytohormones. In : Quarrie, A., Krstić,B. and V. Janjić
(eds): Plant Physiology in the New Millenium. Pp. 77-84, Vizartis, Belgrade,
2002.
Cvikrová M.:
Endogenous contents of phytohormones and phenylpropanoids in sessile oak
somatic embryos in relation to their conversion potential.
Polyphenols Communications, pp. 47-49, Marrakech-Morocco, 2002.
Matucha M., Uhlířová H.:
Volatile chlorinated carbohydrogens and forest decline.
Biol. Listy 67: 161-176, 2002.
Ohnoutková L., Mullerová E, Vagera J., Martinek P.:
Anther culture response of Tritordeum (Tritordeum Ascherson et Graebner)
and its comparison with wheat (Triticum aestivum L.) and barley (Hordeum
vulgare L.) Proceedings of “Triticeae 4th International Symposium 2001,
Cordoba, Spain, September 10-12., pp. 269 – 271, 2002.
Ondřej V., Navrátilová B., Tarkowski P., Doležal K., Lebeda A.:
In vitro pollination as a tool of overcoming crossing barriers between Cucumis
cativus L. and Cucumis melo L.
Acta Fac. Rerum Nat. Univ. Comenianae Bot. 41: 81-88, 2002.
64
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AV0Z50380511
291 Pospíšilová J., Vágner M.:
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292
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293
294
295
296
297
298
Vliv kyseliny abscisové a benzyladeninu na rychlost fotosyntézy a transpirace
a na vodivost průduchů během vodního stresu. (Effect of abscisic acid and
benzyladenine on photosynthetic and transpiration rates and stomatal
conductance during water stress.. In: Hnilička F. (ed.): Vliv abiotických a
biotických stresů na vlastnosti rostlin, pp. 106-110. ČZU, Praha 2002.
Pospíšilová J.:
Can cytokinins interact with abscisic acid during regulation of stomatal
opening? In: Zima, M., Černá, K. (ed.): Ecophysiology of Plant Stress. pp. 3436, SPU, Nitra 2002.
Procházková D., Wilhelmová N.:
Comparison of resistance to water stress of two wheat cultivars.
Zeszyty Problemowe Postepow Nauk Rolniczych 2002: 217-221, 2002.
Roux N., Toloza A., Doležel J., Swennen R., Lepoivre P., Zapata-Arias
F.J.:
Usefulness of embryogenic cell suspension for the induction and selection of
mutants in Musa spp.
In: Abstracts of the FAO/IAEA 4th Research Co-ordination Meeting on Cellular
Biology and Biotechnology Including Mutation Techniques for Creation of New
Useful Banana Genotypes.
Infomusa 11 ( Promusa 9): 17 – 18, 2002.
Šafář J., Piffanelli P., Glaszmann J.C., Doležel J.:
Construction of BAC library for the B genome of banana (Musa balbisiana).
Biology of Banana, pp. 18-19, Catholic University Leuven, Leuven, 2002.
Šafář J.:
Construction of BAC DNA libraries and their characterization using DNA
markers.
In: Proceedings of the workshop “The Use of Molecular Markers in Plant
Biology, Breeding and Germplasm Conservation. Pp. 197-204. Agritec, Ltd.,
Šumperk, 2002 (in Czech).
Uhlířová H., Matucha M., Forczek S.T.:
Účinky chloroctových kyselin na smrk ztepilý (Picea abies /L./ Karst.)
Zprávy lesnického výzkumu 47, 16-20, 2002.
Valárik, M., Hřibová, E., Alkhimova, O., Bartoš, J., Doleželová, M., Šafář,
J., Janda, J., Šimková, H., Doležel, J.:
Repetitive DNA sequences and karyotype evolution in the genus Musa.
Biology of Banana, pp. 20-21. Catholic University Leuven, Leuven, 2002.
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2003 + papers accepted or in press:
IMPACTED JOURNALS:
no.
title
299 Binarová P., Cenklová V., Sulimenko V., Dryková D., Volc J., Dráber P.:
300
301
Distribution of gamma-tubulin in cellular compartmens of higher plant cells.
Cell Biol. Int. 27(3): 167-169, 2003.
Blagoeva E., Malbeck J., Gaudinová A., Vaněk T., Vaňková R.:
Cyclin-dependent kinase inhibitor, roscovitine, in combination with exogenous
cytokinin, N-6-benzyladenine, causes increase of cis-cytokinins in immobilized
tobacco cells.
Biotechnol. Lett. 25(6): 469-472, 2003.
Burketová L., Štillerová K., Feltlová M.:
Immunohistological localization of chitinase and β-1,3-glucanase in
rhizomania-diseased and benzothiadiazole treated sugar beet roots.
Physiol. Molec. Plant Pathol. (in press)
65
IF 2002
1.017
0.802
1.634
AV0Z50380511
302 Burketová L., Štillerová K., Feltlová M., Šindelářová M.:
Immunohistological analysis of chemically induced proteins in sugar beet.
Biol. Plant. 47: 243-251, 2003/2004.
303 Cenklová V., Binarová P., Havel L.:
Actin distribution in mitotic apparatus of Norway spruce (Picea abies (L.)
Karst.) somatic embryos.
Biol. Plant. 46 (2): 167-174, 2003.
304 Cvikrová M., Binarová P., Cenklová V., Eder J., Doležel J., Macháčková
I.:
Effect of 2-aminoindan-2-phosphonic acid on cell cycle progression in
synchronous meristematic cells of Vicia faba roots.
Plant Sci. 164(5): 823-832, 2003.
305 Cvikrová M., Malá J., Hrubcová M., Eder J., Zoń J, Macháčková I.:
Effect of inhibition of biosynthesis of phenylpropanoids on sessile oak somatic
embryos.
Červenková
K., Belejová M., Chmela Z., Rypka M., Riegrová D., Michnová
306
K., Michalíková K., Surová I., Brejcha A., Hanuš J., Černý B., Fuksová K.,
Havlíček L., Veselý J.:
In vitro glycosidation potential towards olomoucine-type cyclin-dependent
kinase inhibitors in rodent and primate microsomes.
Physiol. Res. 52 (4): 467-474, 2003.
307 Čeřovská N., Moravec T., Rosecká P., Dědič P., Filigarová M.:
Production of polyclonal antibodies to a recombinant coat protein of potato
mop-top virus (PMTV).
J. Phytopathol. 151(4): 195-200, 2003.
Čeřovská
N., Moravec T., Rosecká P., Filigarová M., Pečenková T.:
308
The nucleotide sequences of the coat protein coding regions of six Potato
mop-top virus (PMTV) isolates.
Acta Virologica 47(1): 37-40, 2003.
309 Doležel J., Bartoš J., Voglmayer H., Greilhuber J.:
Nuclear DNA content and genome size of trout and human.
Cytometry 51: 127-128, 2003.
310 Doležel J., Carter N.P., Ferguson-Smith M.:
Chromosomes go with the flow.
Chrom. Res. (in press).
Doležel
J., Kubaláková M., Bartoš J., Macas J.:
311
Flow cytogenetics and plant genome mapping.
Chrom. Res. (in press)
312 Dryková D., Cenklová V., Sulimenko V., Volc J., Dráber P., Binarová P.:
Plant γ-tubulin interacts with α-,β-tubulin dimers and forms membrane
associated complexes.
Plant Cell 15(2): 465-480, 2003.
313 Eliáš M., Drdová E., Žiak D., Bavlnka B., Hála M., Cvrčková F.,
Soukupová H., Žárský V.:
The exocyst complex in plants.
Cell Biol. Int. 27(3): 199-201, 2003.
Fellner
M., Horton L.A., Cocke A.E., Stephens N.R., Ford E.D., Van
314*
Volkenburgh E.:
Light interacts with auxin during leaf elongation and leaf angle development in
young corn seedlings.
Planta 216: 366-376, 2003.
315 Filek M., Biesaga-Koscielniak J., Marcinska I., Krekule J., Macháčková I.,
Dubert F.:
The effects of electric current on flowering of grafted of non-vernalized winter
rape.
Biol. Plant. 46(4): 625-628, 2003.
66
0.583
0.583
1.556
1.582
0.984
0.567
0.660
1.933
1.828
1.828
10.751
1.017
2.960
0.583
AV0Z50380511
316 Franěk F., Eckschlager T., Katinger H.:
1.734
317
0.848
318
319
320
321
322
323
324
325
326
327
328
329
Enhancement of monoclonal antibody production by lysine-containing
peptides
Biotechnol. Progr. 19(1): 169-174, 2003.
Franěk F., Eckschlager T., Kohout L.:
24-Epibrassinolide at subnanomolar concentrations modulates growth and
production characteristics of a mouse hybridoma.
Collect. Czech Chem. Commun. 68(11): 2190-2200, 2003.
Gichner T. :
DNA damage induced by indirect and direct acting mutagens in catalasedeficient transgenic tobacco. Cellular and acellular Comet assays.
Mutation Res. 535(2): 187-193, 2003.
Gichner T., Patková Z., Kim J.K.:
DNA damage measured by the Comet assay in eight agronomic plants.
Biol. Plant. 47(2): 185-188, 2003/2004.
Gichner T.:
Differential genotoxicity of ethyl methanesulphonate, N-ethyl-N-nitrosourea
and maleic hydrazide in tobacco seedlings based on data of the Comet assay
and two recombination assays.
Mutat. Res. – Genet. Toxicol. Env. Mutag. 538(1-2): 171-179, 2003.
Honys D., Twell D.:
Comparative analysis of Arabidopsis pollen transcriptome.
Plant Physiol. 132(2): 640-652, 2003.
Kamínek M., Trčková M., Fox J.E., Gaudinová A.:
Comparison of cytokinin-binding proteins from wheat and oat grains.
Physiol. Plant. 117(4): 453-458, 2003.
Kirschner J., Štěpánek J., Mes T.H.M., den Nijs J.C.M., Oosterveld P.,
Štorchová H., Kuperus P.:
Principal features of the cpDNA evolution in Taraxacum (Asteraceae,
Lactuceae): a conflict with taxonomy.
Plant Syst. Evol. 239(3-4): 231-255, 2003.
Kolář J., Johnson C.H., Macháčková I.:
Exogenously applied melatonin (N-acetyl-5-methoxytryptamine) affects
flowering of the short-day plant Chenopodium rubrum L..
Physiol. Plant. 118(4): 605-612, 2003.
Kubaláková M., Valárik M., Bartoš J., Vrána J., Číhalíková J., MolnárLáng M., Doležel J.:
Analysis and sorting of rye (Secale cereale L.) chromosomes using flow
cytometry.
Genome 46: 893-905, 2003.
Lalanne E., Honys D., Johnson A., Borner G., Dupree P., Grossniklaus
U., Twell D.:
AtPIGC and AtPIGA, two components of the glycosylphosphatidylinositol
anchor biosynthetic pathway, are required for pollen germination and tube
growth in Arabidopsis.
Plant Cell 2003, (accepted).
Laukens K., Lenobel R., Strnad M., Van Onckelen H., Witters E.:
Cytokinin affinity purification and identification of a tobacco BY-2 adenosine
kinase.
FEBS Lett. 533(1-3): 63-66, 2003.
Lexa M., Genkov T., Malbeck J., Macháčková I., Brzobohatý B.:
Dynamics of endogenous cytokinin pools in tobacco seedlings: a modeling
approach.
Ann. Bot. 91: 585-597, 2003.
Kuthanová A., Gemperlová L., Zelenková S., Eder J., Macháčková I.,
Opatrný Z., Cvikrová M.:
Cytological changes and alterations in polyamine contents induced by
cadmium in tobacco BY-2 cells.
Plant Physiol. Biochem. (accepted)
67
1.636
0.583
1.636
5.800
1.565
1.114
1.565
1.815
10.751
3.912
1.476
1.582
AV0Z50380511
330 Mandák B., Pyšek P., Lysák M., Suda J., Krahulcová A., Bímová K.:
1.476
331
5.271
332
333
334
335
336
337
338
339
340
341
342
343
344
Variation in DNA-ploidy levels of Reynoutria taxa in the Czech Republic.
Ann. Bot. 92(2): 265-272, 2003.
Matsunaga S., Isono E., Kejnovský E., Vyskot B., Doležel J., Kawano S.,
Charlesworth D.:
Duplicative transfer of a MADS box gene to a plant Y chromosome.
Mol. Biol. Evol. 20(7): 1062-1069, 2003.
Matucha M., Forczek S.T., Gryndler M., Uhlířová H., Fuksová K.,
Schroder P.: Trichloroacetic acid in Norway spruce/soil-system I.
Biodegradation in soil.
Chemosphere 50(3): 303-309, 2003.
Moravcová D., Kryštof V., Havlíček L., Moravec J., Lenobel R., Otyepka
M., Strnad M.: Pyrazolo[4,3-d]pyrimidines as new generation of cyclindependent kinase inhibitors.
Moravec J., Kryštof V., Hanuš J., Havlíček L., Moravcová D., Fuksová K.,
Kuzma M., Lenobel R., Otyepka M., Strnad M.:
2,6,8,9-Tetrasubstituted purines as new CDK1 inhibitors.
Moravec T., Čeřovská N., Boonham N.:
The detection of recombinant, tuber necrosing isolates of Potato virus Y
(PVYNTN) using a three-primer PCR based in the coat protein gene.
J. Virol. Meth. 109: 63-68; 2003.
Motyka V., Vaňková R., Čapková V., Petrášek J., Kamínek M., Schmulling
T.:
Cytokinin-induced upregulation of cytokinin oxidase activity in tobacco
includes changes in enzyme glycosylation and secretion.
Physiol. Plant. 117(1): 11-21, 2003.
Naganowska B., Doležel J., Świecicki W.K.:
Development of molecular cytogenetics and physical mapping of ribosomal
RNA genes in Lupinus.
Biol Plant. 46(2): 211-215, 2003.
Novák J., Vlasáková V., Tykva R., Ruml T.:
Degradation of juvenile hormone analog by soil microbial isolate.
Chemosphere 52: 151-159, 2003.
Novák O., Tarkowski P., Tarkowská D., Doležal K., Lenobel R., Strnad M.:
Quantitative analysis of cytokinins in plants by liquid chromatography/single
qadrupole mass spectrometry.
Anal. Chimica Acta 480(2): 207-218, 2003.
Novotná Z., Hynek R., Martinec J., Potocký M., Valentová O.:
Plant PIP2-dependent phospholipase D activity is regulated by
phosphorylation.
FEBS Lett. 554(1-2): 50-54, 2003.
Novotná Z., Martinec J., Profotová B., Žďárová Š., Kader J.-C., Valentová
O.:
In vitro distribution and characterization of membrane associated PLD and PIPLC in Brassica napus.
J. Exp. Bot. 54: 691-698, 2003.
Palomino G., Doležel J., Méndez I., Rubluo A.:
Nuclear genome analysis in Agave tequilana Weber through flow cytometry.
Caryologia 56(1): 37-46, 2003.
Pechová R., Kutík J., Holá D., Kocová M., Haisel D., Vičánková A.:
The ultrastructure of chloroplasts, content of photosynthetic pigments, and
photochemical activity of maize (Zea mays L) as influenced by different
concentrations of the herbicide amitrole.
Photosynthetica 41(1): 127-136, 2003.
Petrášek J., Černá A., Schwarzerová K., Elčkner M., Morris D.A.,
Zažímalová E.:
Do Phytotropins Inhibit Auxin Efflux by Impairing Vesicle Traffic?
Plant Physiol. 131: 254-263, 2003.
68
1.461
2.051
2.051
1.938
1.565
0.583
1.461
2.114
3.912
2.852
0.267
0.773
5.800
AV0Z50380511
345 Pokorná J., Schwarzerová K., Zelenková S., Petrášek J., Janotová I.,
3.015
346
0.583
347
348
349
350
351
352
353
354
355
356
402
357
Čapková V., Opatrný Z.:
Sites of actin filament initiation and re-organization in cold-treated tobacco
cells.
Plant Cell Environ., in press 2004.
Pospíšilová J.:
Participation of phytohormones in the stomatal regulation of gas exchange
during water stress.
Biol. Plant. 46(4): 491-506, 2003.
Pospíšilová J.:
Interaction of cytokinins and abscisic acid during regulation of stomatal
opening in bean leaves.
Potocký M., Eliáš M., Novotná Z., Profotová B., Valentová O., Žárský V.:
Phosphatidic acid produced by PLD is necessary for pollen tube growth.
Planta 217(1): 122-130, 2003.
Procházková, D., Wilhelmová, N.:
Changes in antioxidative protection in bean cotyledons during natural and
continuous irradiation-accelerated senescence.
Biol. Plant. 48: accepted, 2004
Roman B., Satovic Z., Požárková D., Macas J., Doležel J., Cubero J.I.,
Torres A.M.:
Development of a composite map in Vicia faba, breeding applications and
future prospects.
Theor. Appl. Genet. (in press).
Roux N., Toloza A., Radecki Z., Zapata-Arias F.J., Doležel J.:
Rapid detection of aneuploidy in Musa using flow cytometry.
Plant Cell Rep. 21: 483-490, 2003.
Ryšlavá H., Muller K., Semorádová Š., Synková H., Čeřovská N.:
Photosynthesis and activity of phosphoenolpyruvate carboxylase in Nicotiana
tabacum L. leaves infected by Potato virus A and Potato virus Y.
Photosynthetica (in press)
Sáenz L., Jones L.H., Oropeza C., Vláčil D., Strnad M:
Endogenous isoprenoid and aromatic cytokinins in different plant parts of
Cocos nucifera (L.).
San Martin A.P.M., Adamec L., Suda J., Mes T.H.M., Štorchová H.:
Genetic variation within the endangered species Aldrovanda vesiculosa
(Droseraceae) as revealed by RAPD analysis.
Aquatic Bot. 75(2): 159-172, 2003.
Schröder P., Matucha M., Forczek S.T., Uhlířová H., Fuksová K.,
Albrechtová J.:
Uptake, translocation and fate of trichloracetic acid in Norway spruce/soil
system.
Chemosphere 52(2): 437-442, 2003.
Schwarzerová K., Pokorná J., Petrášek J., Zelenková S., Čapková V.,
Janotová I., Opatrný Z.:
The structure of cortical cytoplasm in cold-treated tobacco cells: the role of
the cytoskeleton and the endomembrane system.
Cell Biol. Int. 27(3): 263-265, 2003.
Stirk W.A., Novák O., Strnad M., van Staden J.:
Cytokinins in makroalgae.
Plant Growth Regul. 41(1):13-24, 2003.
Strnad M., Kohout L.:
Simple brassinolide analogue 2α,3α-dihydroxy-17β-(3methylbutyryloxy)-7oxa-B-homo-5α-androstan-6-one inducing the bean second internode
splitting.
69
0.773
2.960
0.583
2.264
1.340
0.773
0.850
1.014
1.461
1.017
0.850
0.850
AV0Z50380511
358 Sun J., Niu Q.W., Tarkowski P., Zheng B., Tarkowská D., Sandberg G.,
5.800
359
0.773
360
361
362
363
364
365
366
367
368
369
370
Chua N.-H., Zuo J.:
The Arabidopsis AtIPT8/PGA22 gene encodes an isopentenyltransferase that
is involved in de novo cytokinin biosynthesis.
Plant Physiol. 131: 167-176, 2003.
Synková H., Pechová R., Valcke R.:
Changes in th echloroplast structure in PSSU-ipt tobacco during plant
ontogeny.
Šimková H., Čihalíková J., Vrána J., Lysák M., Doležel J.:
Preparation of high molecular weight DNA from plant nuclei and
chromosomes isolated from root tips.
Biol. Plant. 46(3): 369-373, 2003.
Hexokinases of tobacco leaves: changes in the cytosolic and non-cytosolic
isozyme complexes induced by tobacco mosaic virus infection.
Biol. Plant. 47: 413-419, 2003/2004.
Changes in glucose-6-phosphate dehydrogenase, ribonucleases and
esterases and content of viruses in potato virus Y infected tobacco
superinfected with tobacco mosaic virus.
Biol. Plant. 47: 99-104, 2003/2004.
Influence of antiviral factor on tobacco mosaic virus RNA biosynthesis in
tobacco.
Biol. Plant. 46(1): 95-100, 2003.
Tarkowská D., Doležal K., Tarkowski P., Åstot C., Schmülling T., Holub
J., Fuksová K., Sandberg G., Strnad M.:
Identification of new aromatic cytokinins in Arabidopsis thaliana, poplar leaves
and Agrobacterium tumefaciens strains by LC-(+)ESI-MS and capillary liquid
chromatography/frit - fast atom bombardment mass spectrometry.
Physiol. Plant. 117(4): 579-590, 2003.
Tarkowská D., Kotouček M., Doležal K.:
Elecrochemical reduction of 6-benzylaminopurine at mercury electrodes and
its analytical application.
Coll. Czechoslovak Chem. Comm. 68(6): 1076-1093, 2003.
Trávníček B., Lysák M.A., Číhalíková J., Doležel J.:
Karyo-taxonomic study of the genus Pseudolysimachion (Scrophulariaceae)
in the Czech Republic and Slovakia.
Folia Geobot. (in press).
Trávníček Z., Maloň M., Zatloukal M., Doležal K., Strnad M., Marek J.:
Mixed ligand complexes of platinum(II) and palladium(II) with cytokininderived compounds Bohemine and Olomoucine: X-ray structure of
[Pt(BohH(+)-N7)Cl-3]center dot 9/5H(2)O {Boh=6-(benzylamino)-2-[(3(hydroxypropyl)-amino]-9-isopropylpurine, Bohemine}
J. Inorg Biochem. 94: (4) 307-316, 2003.
Tykva R., Šimek P., Benetová B., Holík J., Hlaváček J., Havlíček L.:
Comparison for following the metabolism of oostatic peptides in Neobellieria
bullata by mass spectrometry and radiolabelling.
Coll. Czechoslovak Chem. Comm. (in press)
Tykva R., Vlasáková V., Novák J., Havlíček L.:
RadioHPLC for ecotoxicity assessment of insect growth regulators.
J. Chromatography A (in press)
Überal I., Vrána J., Bartoš J., Šmerda J., Doležel J., Havel L.:
Isolation of chromosomes from Picea abies L. and their analysis by flow
cytometry.
Biol. Plant. (in press).
70
0.583
0.583
0.583
0.583
1.565
0.848
0.564
2.204
0.848
3.098
0.564
AV0Z50380511
371 Vagera J., Novotný J., Ohnoutková L.:
0.632
372
5.850
373
374
375
376
377
378
Induced androgenesis in vitro in mutated populations of barley, Hordeum
vulgare L.
Plant Cell, Tissue Org. Cult. (in press) 2003
Valárik M., Bartoš J., Kovářová P., Kubaláková M., de Jong J.H., Doležel
J.:
High-resolution FISH of super-stretched flow sorted plant chromosomes.
Plant J. (in press)
Veach Y., Martin R.C., Mok D.W.S., Malbeck J., Vaňková R., Mok M.C.:
O-glukosylation of cis-zeatin in maize: characterization of genes, enzymes,
and endogenous cytokinins.
Plant Physiol. 131: 1374-1380, 2003.
Vomáčka L., Pospíšilová J.:
Rehydration of sugar beet plants after water stress: effect of cytokinins.
Biol. Plant. 46(1): 57-62, 2003.
Werner T., Hanuš J., Holub J., Schmülling T., van Onckelen H., Strnad
M.:
New cytokinin metabolites in ipt transgenic Arabidopsis thaliana plants.
Physiol. Plant. 118(1): 127-137, 2003.
Werner T., Motyka V., Laucou V., Smets R., van Onckelen H., Schmülling
T.:
Cytokinin-deficient transgenic Arabidopsis plants show multiple
developmental alterations indicating opposite functions of cytokinins in
regulating shoot and root meristem activity.
Plant Cell 15: 2532-2550, 2003.
Zažímalová E., Napier R.:
Points of regulation for auxin action.
Plant Cell Rep. 21(7): 625-634, 2003.
Zonia L., Feijó J.A.:
State and spectral properties of chloride oscillations in pollen.
Biophys. J. 84(2): 1387-1398, 2003.
OTHERS:
379 Blagoeva E., Dobrev P. I., Vaňková R.:
380
381
382
383
384
Inhibition of cytokinin-glucose conjugation in derooted radish, tobacco and
bean seedlings.
Bulg. J. Plant Physiol. 29: (1-2), in press.
Coufal D., Matucha P., Uhlířová H., Lomský B., Forczek S.T., Matucha M.:
GUHA analysis of coniferous forest damage: Effects of trichloroacetic acid,
sulphur, fluorine and chlorine on needle loss of Norway spruce.
Neural Network World 13: 89-102, 2003.
Dobrev P.I., Motyka V.:
Stanovení aktivity cytokininoxidasy/dehydrogenasy v rostlinách pomocí HPLC
spojené s průtokovým radioaktivním detektorem. (In Czech)
Biol. listy 68: 163-166, 2003.
Doležel J., Kubaláková M., Vrána J., Bartoš J.:
Flow cytogenetics. In: Goodman R.M. (ed.): Encyclopedia of Plant and Crop
Science. Marcel Dekker Inc., New York (in press)
Doležel J., Macas J., Lysák M., Neumann P., Kubaláková M., Nouzová M.,
Šimková H., Koblížková A., Požárková D., Číhalíková J., Lucretti S.:
Sorting of Plant Chromosomes and Construction of Chromosome-Specific
DNA Libraries. In: Speel, E.J.M. and Hopman, A.H.N. (eds.): Chromosome
Analysis Protocols, Humana Press, Totowa, USA (in press).
Doležel J., Šafář J., Janda J., Bartoš J., Kubaláková M., Číhalíková J.,
Šimková H., Sourdille P., Bernard M., Chalhoub B.:
Development of flow cytogenetics for wheat genome mapping. In:
Proceedings of the Tenth International Wheat Genertics Symposium. Pp. 6568. Istituto Sperimentale per la Cerealicoltura, Rome, 2003.
71
5.800
0.583
1.565
10.751
1.340
4.643
-
-
-
-
-
AV0Z50380511
385 Galbraith D.W., Bartoš J., Doležel J.:
-
386
-
387
388
389
390
391
392
393
394
395
396
397
Flow cytometry and cell sorting in plant biotechnology. In: Sklar L.A. (ed.):
Flow Cytometry in Biotechnology. Oxford Univ. Press, Inc., New York (in
press)
Gaudinová A., Vaňková R., Dobrev P.I., Motuyka V.:
Extrakce a stanovení aktivity cytokinin N- a O-glukosyltransferas v rostlinných
pletivech. (In Czech)
Biol. Listy 68: 176-179, 2003.
Genkov T., Vágner M., Dubová J., Malbeck J., Moore I., Brzobohatý B.:
Increased ethylene production can account for some of the phenotype
alterations accompanying activation of a cytokinin biosynthesuis gene, during
germination and early seedluing development in tobacco. In: Biology and
Biotechnology of the Plant Hormone Ethylene III, Eds.: Vendrell M., Klee H.,
Pech J.C., Romojaro F., Proc. of the NATO Advanced Workshop on Biology
and Biotechnology of the Plant Hormone Ethylene, 23-27 April 2002, IOS
Press, Amsterdam, 2003.
Honys D., Twell D.:
Male gametophyte. In Goodman R.M. (ed.): Encyclopedia of Plant and Crop
Science, Marcell Dekker Inc., New York, USA (in press)
Chalhoub B., Allouis S., Šafář J., Janda J., Bellec A., Sarda X., Arar C.,
Lefévre A., Rouault P., Pateyron S., Dupin A., Burgio G., Georget C.,
Sourdille P., Faivre-Rampant, Caboche M., Moore G., Bernard M., Doležel
J.:
Towards precise analysis of the wheat genome: preparation of genomic
resources for structural and functional characterization. In: Proceedings of the
Tenth International Wheat Genertics Symposium. Pp. 229-233. Istituto
Sperimentale per la Cerealicoltura, Rome, 2003.
Kolář J., Macháčková I:
Diurnal rhythms in plants and melatonin. In: Macháčková I., Romanov G.A.
(eds.), Phytohormones in Plant Biotechnology and Agriculture. Proc. of the
NATO - Russia Workshop, May 2002, Moscow, Kluwer, Dordrecht, The
Netherlands, pp. 79-87
(in press)
Krinke O., Novotná Z., Valentová O., Martinec J.:
Fluorometrická metoda pro in vitro měření ligandem otevíraných iontových
kanálů pro Ca2+ v rostlinách.
Biol. listy 68(3): 195-199, 2003.
Lebeda, A., Doležalová, I., Dziechciarková, M., Doležal, K., Frček, J.:
Morphological variability and isozyme polymorphisms in maca and yacon.
Czech J. Genet. Plant Breed. 39: 1-8, 2003.
Matucha M., Gryndler M., Forczek S.T., Uhlířová H., Fuksová K.,
Schröder P.: Chloroacetic acids in environmental processes.
Environ. Chem. Letters 1,127-130,2003.
Morris D.A., Friml J., Zažímalová E.:
The transport of auxins. In Davies P.J. (ed): Plant Hormones: Biosynthesis,
Signal Transduction, Action! New edition. Kluwer Acad. Publ., NY. (in press).
Morris D.A., Zažímalová E.:
Physiological and molecular genetic aspects of auxin transport: recent
developments. In: Macháčková I., Romanov G.A. (eds): Phytohormones in
Plant Biotechnology and Agriculture. Kluwer Acad. Publ. In press 2003.
Pejchar P., Valentová O., Martinec J.:
Stanovení aktivit rostlinných fosolipas in situ s využitím fluorescenčně
značených substrátů.
Biol. listy 68(3): 227-234, 2003.
Pospíšilová J., Dodd I.C.:
Role of plant growth regulators in stomatal limitation of photosynthesis during
water stress. In: Pessarakli, M. (ed.): Handbook of Photosynthesis, Second
Edition, Revised and Expanded. Marcel Dekker, New York , in press.
72
-
-
-
-
-
-
-
AV0Z50380511
398 Uhlířová H., Novotný R., Matucha M.:
-
399
-
400
401
Projevy poškození lesních dřevin pod vlivem abiotických stresů. Seminář „Vliv
abiotických a biotických stresů na vlastnosti rostlin“, Výzk. úst. rostlinné
výroby a ČZU Praha, 8.10.2003, Sborník přednášek, str. 76-89, 2003.
Vágner M., Fischerová L., Špačková J., Vondráková Z.:
Somatic embryogenesis of Norway spruce. In: Protocols of Somatic
Embryogenesis – Woody Trees, Eds. Jain S.M., Gupta P.K., Kluwer Acad.
Publ. (in press)
Vaňková R., Gaudinová A.:
Využití dvojrozměrné fluorescenční spektroskopie pro stanovení množství a
viability buněk tabáku.
Biol. listy 68(3): 250-253, 2003.
Zažímalová E., Petrášek J., Morris D.A.:
The dynamics of auxin transport in tobacco cells.
Bulg. J. Plant Physiol., Spec. Iss., 207-224, 2003.
-
-
* - indicates the papers of M. Fellner published during his study stay abroad without adress of IEB (they are listed
above as his research became the part of research plan for years 2005-2010)
PATENTS AND LICENCES 1999-2003:
patents:
FUKSOVÁ, K., HAVLÍČEK, L., KRYŠTOF, V., LENOBEL, R., STRNAD, M.: Azapurine
derivatives. PO12166GB NJN.
HANUŠ, J., KRYŠTOF, V., HAJDÚCH, M., VESELÝ, J., STRNAD, M.: Substituted nitrogen
heterocyclic derivatives and pharmaceutical use thereof. WO 00/43394.
DOLEŽAL, K., POPA, I., HOLUB, J., LENOBEL, R., WERBROUCK, S., STRNAD, M.:
Heterocycklické sloučeniny na bázi N6-substituovaného adeninu, způsoby jejich přípravy, tyto
deriváty pro použití jako léčiva, kosmetické přípravky a růstové regulátory. PV 2001-2818.
HAVLÍČEK, L., KRYŠTOF, V., SIGLEROVÁ, V., LENOBEL, R., VAN ONCKELEN, H.,
RERNEMAN, Z., SLEGERS, H., ESMANS, E., STRNAD, M., WERMEULEN, K.: Purine
derivatives, process for their preparation and use thereof. WO 01/49688.
MORAVCOVÁ. D., HAVLÍČEK, L., LENOBEL, R., KRYŠTOF, V., STRNAD, M.: Novel
pyrazolo-pyrimidine derivatives with antiinflamatory, anticancer, immunosuprresive and
neurogenerative properties and their use thereof. EP 24128-099.
MORAVCOVÁ. D., HAVLÍČEK, L., LENOBEL, R., KRYŠTOF, V., STRNAD, M.: Disubstituted
pyrazolo-pyrimidine derivatives with CDK inhibitory activity and their use thereof. EP 37456099.
DOLEŽAL, K., POPA, I., ZATLOUKAL, M., LENOBEL, R., HRADECKÁ, D., VOJTĚŠEK, B.,
ULDRIJAN, S., MLEJNEK, P., WERBROUCK, S., STRNAD, M.: "Substituční deriváty N6adenosinu, způsob jejich přípravy, jejich použití pro přípravu léčiv, kosmetických přípravků a
růstových regulátorů, kosmetické přípravky a růstoví regulátory tytzo přípravky obsahující.
PV 2002-4273
DOLEŽAL, K., POPA, I., HOLUB, J., LENOBEL, R., WERBROUCK, S., STRNAD, M.:
"Heterocyclic compounds based on N6-substituted adenine. PCT/CZ02/00045
73
AV0Z50380511
Apple varieties - patents and licences:
Patents:
Maďarsko – 3 plant patents: TOPAZ P 9800450; RAJKA P 9800585;
RUBINOLA P 9800586
Plant Breeder’s Rights (analogy of plant patents):
EU – 5 certificates ”Community Plant Variety Rights“: RUBINOLA EU 5824; GOLDSTAR
EU 7380; OTAVA EU 7381; RAJKA EU 8880; LENA EU 10713
Švýcarsko – 4 Plant Breeder’s Rights: RUBINOLA 00.20.1293; OTAVA 01.20.1393; RAJKA
02.20.1474; GOLDSTAR 01.20.1394
Polsko – 4 Plant Breeders Rights: TOPAZ OS 00001; RUBINOLA OS 00002; RAJKA
OS 00003; GOLDSTAR OS 00004
Slovakia – 4 Plant Breeders Rights: 4528; SVATAVA 4755; GOLDSTAR 5161; RAJKA 5169
Czech Republic – 2 Plant Breeders Rights: BIOGOLDEN 647; LENA 54/2002
Licences - foreign:
country
Belgium
Netherlands
Germany
Poland
U.S.A.
Great Britain
total
no of licenses
1
2
3
2
1
1
10
no of varieties
1
7
3
3
1
1
16
year
1999
2002,2003
2001,2003
2000,2001
1999
2000
Licences in CR :
country
CR
no of licenses
41
no of varieties
4
74
year
1999-2003

Following text represents the selection of materials compiled

Transcription

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