December 25 - Indian Society of Pulses Research and Development
Transcription
December 25 - Indian Society of Pulses Research and Development
Volume 25 Number 4 ISSN 0970-6380 Online ISSN 0976-2434 Journal of Food Legumes Journal of Food Legumes Volume 25 Number 4 December 2012 December 2012 Indian Society of Pulses Research and Development I SPR D 1987 Indian Institute of Pulses Research Kanpur, India www.isprd.in INDIAN SOCIETY OF PULSES RESEARCH AND DEVELOPMENT (Regn. No.877) The Indian Society of Pulses Research and Development (ISPRD) was founded in April 1987 with the following objectives: To advance the cause of pulses research To promote research and development, teaching and extension activities in pulses To facilitate close association among pulse workers in India and abroad To publish “Journal of Food Legumes” which is the official publication of the Society, published four times a year. Membership : Any person in India and abroad interested in pulses research and development shall be eligible for membership of the Society by becoming ordinary, life or corporate member by paying respective membership fee. Membership Fee Indian (Rs.) Foreign (US $) Ordinary (Annual) 350 25 Life Member 3500 200 Admission Fee 20 10 Library/ Institution 3000 100 Corporate Member 5000 - The contribution to the Journal, except in case of invited articles, is open to the members of the Society only. Any non-member submitting a manuscript will be required to become annual member. Members will be entitled to receive the Journal and other communications issued by the Society. Renewal of subscription should be done in January each year. If the subscription is not received by February 15, the membership would stand cancelled. The membership can be revived by paying readmission fee of Rs. 10/-. Membership fee drawn in favour of Treasurer, Indian Society of Pulses Research and Development, through M.O./D.D. may be sent to the Treasurer, Indian Society of Pulses Research and Development, Indian Institute of Pulses Research, Kanpur 208 024, India. In case of outstation cheques, an extra amount of Rs. 40/- may be paid as clearance charges. EXECUTIVE COUNCIL : 2010-2012 Chief Patron Dr S Ayyappan Co-patron Dr N Nadarajan President Dr JS Sandhu (Acting) Joint Secretary Mr Brahm Prakash Patron Dr SK Datta Vice President Dr JS Sandhu Secretary Dr AK Choudhary Treasurer Dr KK Singh Councillors Zone I : Zone II : Zone III Zone IV : : Dr (Mrs) Livinder Kaur PAU, Ludhiana Dr HK Dixit IARI, New Delhi Vacant Dr Vijay Prakash ARS, Sriganganagar Zone V : Zone VI : Zone VII Zone VIII : : Dr KK Nema RAK College, Sehore Dr Ch Srinivasa Rao CRIDA, Hyderabad Vacant Dr Anoop Singh Sachan IIPR, Kanpur Editor-in-Chief : Dr. NP Singh Editors Dr Dr Dr Dr Dr Dr Dr Dr Dr A Amarendra Reddy, ICRISAT, Hyderabad AB Rai, IIVR, Varanasi AK Tripathi, CSAUAT, Kanpur CS Praharaj, IIPR, Kanpur IP Singh, IIPR, Kanpur Jagdish Singh, IIPR, Kanpur KB Saxena, ICRISAT, Hyderabad Li Zhenghong, RIRI, PRC China MK Singh, IIPR, Kanpur Dr Dr Dr Dr Dr Dr Dr Dr Dr MA Iquebal, IASRI, New Delhi Mohd Akram, IIPR, Kanpur P Duraimurugan, DRR, Hyderabad Rajindar Peshin, SKUAT, Srinagar RK Varshney, ICRISAT, Hyderabad RS Raje, IARI, New Delhi Sarvjeet Singh, PAU, Ludhiana SC Gupta, ARS, Durgapura VK Shahi, RAU, Pusa Journal of Food Legumes (Formerly Indian Journal of Pulses Research) Vol. 25 (4) December 2012 CONTENTS RESEARCH PAPERS 1 Status, scope and strategies of arid legumes research in India- A review 255 D. Kumar and A.B. Rodge 2. Transferability of cowpea and azuki bean derived SSR markers to other Vigna species 273 Ravindra Bansal, Sudhir Kumar Gupta and T. Gopalakrishna 3. Genetic diversity studies in blackgram (Vigna mungo L. Hepper) 279 M. Srimathy, M. Sathya and P. Jayamani 4. Stability analysis for seed yield and its component traits in rice bean (Vigna umbellata) 282 Wungsem Rungsung and S.A.P.U. Changkija 5. Sequence comparison of coat protein gene of Mungbean yellow mosaic India virus isolates infecting mungbean and urdbean crops 286 Naimuddin and M. Akram 6. Bio-efficacy of some insecticides against H. armigera (Hubner) on chickpea (Cicer arietinum L.) 291 P.S. Singh, R.K. Shukla and N.K. Yadav 7. Comparison of nodC and 16S rDNA gene analysis of rhizobia associated with legumes of arid and semi-arid regions of Haryana 294 Rajesh Gera, Ranjana Bhatia and Varun Kumar 8. Phenology, dry matter distribution and yield attributes under normal and drought stress conditions in Lentil (Lens culinaris Medik.) 300 Vijay Laxmi 9. Efficacy of post emergence herbicides on weed control and seed yield of rajmash (Phaseolus vulgaris L.) 306 Baldev Ram, S.S. Punia, D.S. Meena and J.P. Tetarwal 10. Enhancing water use efficiency and production potential of chickpea and fieldpea through seed bed configurations and irrigation regimes in North Indian Plains 310 J.P. Mishra, C.S. Praharaj and K.K. Singh 11. Variability in the nutrients, antinutrients and other bioactive compounds in soybean (Glycine max (L.) Merrill) genotypes 314 Reeti Goyal, Sucheta Sharma and B.S. Gill 12. Effect of presoak treatment on cooking characteristics and nutritional functionality of rice bean V.D. Pawar , M.K. Akkena, P.M. Kotecha, S.S. Thorat and V.V. Bansode 321 13. Factors associated with economic motivation of legume growers in desert area of Rajasthan 326 Subhash Chandra, P.Singh and J.P. Lakhera 14. Farmers participatory approach in seed multiplication of pulses in Bundelkhand region - A case study 330 Purushottam, S.K. Singh, C.S. Praharaj and Lakhan Singh 15. Tropical Legumes 2 pigeonpea seed system in India: An analysis 334 M.E. Holmesheoran, M.G. Mula, C.V.S. Kumar, R.P. Mula and K.B. Saxena 16. Dissemination of pulse production technologies in Uttar Pradesh : A micro-level analysis 340 Rajesh Kumar, S.K. Singh, Purushottam and Uma Sah 17. Pigeonpea (Cajanus cajan L.) price movement across major markets of India 344 D.J. Chaudhari and A.S. Tingre SHORT COMMUNICATIONS 18. Genetic variability, character association and path coefficient analysis in faba bean 348 B.K. Chaubey, C.B. Yadav, K. Kumar and R.K. Srivastava 19. A comparative study of hybrid and inbred cultivars for germination and other related traits of pigeonpea 351 M. Bharathi and K.B. Saxena 20. Screening of chickpea (Cicer arientinum L.) genotypes for identification of source of resistance to Botrytis grey mould 355 Lajja Vati, K.P.S. Kushwaha and Abhijeet Ghata 21. Management of Fusarium wilt of lentil using antagonistic microorganisms in Tarai region of Uttarakhand 358 Ankita Garkoti and H.S. Tripathi 22. Effect of phosphorus and zinc on yield and economics of mothbean under semiarid conditions 361 L.R. Yadav, Poonam Choudhary, Santosh, O.P. Sharma and Meenu Choudhary List of Referees for Vol. 25 (4) i Reviewer Index (2012) ii Author Index (2012) iv Subject Index (2012) vi ISPRD Fellowship awards, 2012 Obituary viii x Journal of Food Legumes 25(4): 255-272, 2012 Status, scope and strategies of arid legumes research in India- A review D. KUMAR and A.B. RODGE1 Central Arid Zone Research Institute, Jodhpur-342 003, India; 1Department of Food Chemistry and Nutrition, MAU, Pabhani-431 402, India; E-mail: [email protected] (Received : May 25, 2012 ; Accepted : November 15, 2012) ABSTRACT Arid legumes are adjudged for sustaining growth and production, especially in driere co-systems encountered with harsh and hostile growing environments. Sustained breeding efforts have been made to improve these crops so as to make them more productive. In guar [Cyamopsis tetragonoloba (L.) Taub.], early maturing (85-90 days) varieties like, ‘RGC-936’, ‘HG-365’ and ‘HG-563’ suited to 300-400 mm rainfall, medium maturing (90-105 days) varieties like, ‘RG-1002’, ‘RGC-1003’, ‘RGC-1038’, ‘HG-870’ and ‘HG-884’ suited to 400-450 mm rainfall and long duration (110-120 days) varieties like, ‘H-220’ and ‘RGC-986’ suited to 500-550 mm rainfall have been developed. Guar cultivar ‘RGC-1066’ is also suitable for mechanical harvesting. In cowpea [Vigna unguiculata (L.) Walp.], varieties with improved plant types and early maturity (60-62 days) like, ‘RC-101’, medium maturity like, ‘Co (CP-7)’ (67-73 days) and ‘GC-3’ (90-95 days and adapted to whole country) and late maturity (>95 days) like, ‘V-585’, ‘V-240’ and ‘KBC-2’ with dual purpose have also been developed. In moth bean [Vigna aconitifolia (Jacq.) Marechal], varieties with erect upright growth (‘FMO-96’ matured in 58-60 days), erect growth (‘RMO40’, ‘RMO-225’ and ‘CAZRI-Moth-3’ in 60-63 days), semi-erect growth (‘CAZRI Moth-2’ in 66-68 days) and semi-spreading growth (‘CAZRI Moth-1’ in 72-74 days) suiting to 150-400 mm rainfall have been developed. In horsegram [Macrotyloma uniflorum (Lam.) Verd], promising varieties maturing in 85-90 days (‘AK-21’ and ‘AK-42’), 90-100 days (‘PHG-9’, ‘CRIDA-118R’ and ‘BJPL-1’) and 105-111 days (‘VLG-15’ and ‘VLG-19’) have also been released. Besides this, protocols for rapid callus induction and regeneration systems have also been developed in arid legumes. QTLs for resistance to thrips damage have been identified in cowpea. Efficient cropping sequences like, guar-mustard and guar-wheat are widely followed in northern India. In southern states, growing short duration horsegram with rice by replacing cowpea is beneficial. In Orissa and parts of Andhra Pradesh, horsegram is successfully cultivated in rice/ maize/sorghum sequences. Beneficial effects of intercropping guar with pearl millet/sorghum/maize/castor have also been observed. Moth bean+pearl millet (3:1) intercropping system is effective in arid situations. However, cowpea+castor (6:1) is found to have highest monetary return over other intercropping systems. Foliar application of ZnSO4 @ 0.5% at 25 or 45 days of sowing (DAS) has been useful with 15-20% higher seed yield in guar, cowpea and moth bean. Basalin @ 1.0 kg a.i./ha in moth bean and guar; and pendamethalin @ 0.75 kg a.i./ha + one hand weeding in cowpea are useful for effective weed control with leas t weed index. Orga nic, inorganic, biocontrol management and IPM strategies have also been developed against major pests and diseases. Guar gum (galactomannan) - Dr. D. Kumar, Emeritus Scientist at Central Arid Zone Research Institute (CAZRI), Jodhpur, obtained M. Sc. (Ag) and Ph. D. Degrees from Banaras Hindu University. He served R.B.S. College Bichpuri, Agra as Jr Plant Breeder during September 1976 to 24 November, 1981 and Haryana Agricultural University Reg. Res. Sta., Bawal as Assistant Oilseeds Botanist during 1981-1984. He joined CAZRI as Scientist S-2 on 26, November 1984. He served CAZRI in different capacities including Principal Scientist and Project Coordinator, National Network Research Project on Arid Legumes upto superannuation (31 July, 2010). He founded the “Indian Arid Legumes Society” in February, 2000 and remained Secretary upto superannuation. In capacity of Organizing Secretary, he organized 4 National Symposia on Arid Legumes. He has authored/edited 19 technical books, brought out 12 production technology bulletins, authored more than 50 chapters in books and proceedings and has more than 150 full length research papers in refereed journals. He has guided 7 Ph. D students. He was a member of Institute Management Committee of NRC on Rapeseed-Mustard, Bharatpur for 6 years and NRC for Groundnuts, Junagadh for 3 years. In individual capacity, he has developed 3 promising varieties of moth bean, CAZRI Moth-1, CAZRI Moth-2 and CAZRI Moth3 and rigistred two unique germplasm of moth bean. He has visited Israel, Zambia, Kenya and Ethiopia and Tanzania and successfully introduced moong bean (K-851) in summer season of 2011 in Gambella region of Ethiopia, and guar in Tanzania, during 2012. Ch. Devi Lal Award for outstanding performance of AICRP was bestowed upon him on 16 July, 2009 for excellent performance of National Network Project on Arid Legumes.He is recipient of Haldhar Times Ratan Award, 2012. He is involved in introduction of guar in non-traditional regions of Anantapur, Karnool, Mahboob Nagar, RangaReddi (A.P.), Yavatmal (M.S.), Raipur (CS) and Madurai (T.N). Dr. A. B. Rodge obtained M.Sc. degree (Food Science) from University of Saskatchewan, Saskatoon, Canada and Ph. D. (Food Science) from Marathwada Agril. University (MAU), Parbhani. Presently he is working as Head, Dept. of Food Chemistry and Nutrition, College of Food Technology, MAU Parbhani. He has about three decades of experience in research, teaching and extension activities. As a Principal Scientist, he has been monitoring 2 National Projects, 1 National Network Project on Arid Legumes and 2 National Network Project on Harvesting, Processing and Value Addition of Natural Resins and Gums. He has published more than 60 research papers in reputed journals, and 30 other technical publications, one book and written six book chapters. He is recipient of travel grant from American Association of Cereal Chemists, Nashville, USA, President Honor roll from the American Oil Chemist Society, Annual meeting USA; and recipient of common Wealth Scholarship for higher studies in Canada. He is recipient of prestigious George F. Stewart International award as a finalist in Institute of Food Technologist Annual Meeting Chicago USA. He has been acting as Vice President of Indian Arid Legume Society, Jodhpur and as Research Advisory Committee member at Indian Institute of Natural Resins and Gums. 256 Journal of Food Legumes 25(4), 2012 a polysaccharide organic compound - is used in a number of industrial products where water is an important factor. However, varieties having more than 32.0% gum content and higher viscosity of guar gum (4000-5000 cP) are more preferred for export. Guar meal (a by-product of guar gum industries) can also replace edible oil cakes due to its higher crude protein (40-45%). Future strategies for these arid legumes are also discussed. Key words : Arid legumes, Cowpea, Guar, Horsegram, Moth bean Deep rooted, summer annual legumes grown under resource constraint situations are generally referred to as arid legumes. Commonly known as the crops of dry habitats, these are characterized with low cost source of livelihood of financially ridden arid farmers. For convenience, four legumes viz., guar or clusterbean, moth bean or dew bean, cowpea or lobia and horsegram or kulthi form a group of crops generally referred to as arid legumes in India. These crops adapted to specific set of environments are basically known for taming drought, sustaining soil productivity, stabilizing agricultural system and providing nutritional security. The crops provide nutritious green fodder/vegetables and are being used in many secondary and tertiary products. These virtues have made these crops from being locally and regionally important to front running dryland crops of great economic significance in India. In spite of a series of merits attached, these crops also suffer from certain biological bottlenecks which need to be addressed. Main obstacle is poor productivity of arid legumes resulting from poor source-sink relationship. The plant type is generally suiting for survival values but little for higher productivity. Second most important biological weakness is their long maturity subjecting these to terminal stress leading to poor adaptation and production. There are few but important plant diseases which may also cause heavy yield losses during congenial conditions. Thus, to bridge the gaps between potential and realized yields of these crops, immediate biological and management remedies may be taken up requiring insight to research information available for assessing existing status and necessary impetus in genetic improvement, crop husbandry, plant protection and biochemistry. In view of stagnated growth of these crops in reference to area, production, productivity and quality aspects, it appears imminent for a strategically move in required direction which would help in increasing productivity of arid regions contributing to self sufficiency in production of these pulses, increasing export potential of industrial products and income generation. This paper provides an insight to relook on its existing strategic research and development in arid legumes to provide a more sensible road map to bridge the research gaps in productivity at different levels. Genetic Resource Guar: More than 5000 accessions have been collected by National Bureau of Plant Genetics Resources (NBPGR) New Delhi, mainly from dry habitats of northern India. Two wild species viz., C. serrata and C. senegalensis were also introduced from USA. A total of 3714 accessions have been put for ex-situ conservation. Additionally, 4878 accessions with indigenous origin have also been conserved in mediumterm storages (Mishra et al. 2009). Evaluation of more than 730 indigenous and 20 exotic lines led to selection of lines maturing in less than 90 days. Effective evaluation of more than 375 accessions at S.K.Nagar, Gujarat, India against important diseases have resulted in promising resistance lines against bacterial leaf blight (‘GAU-9406’, ‘GG-1’, ‘RGC-1027’), alternaria leaf blight (‘GAUG-9406’, ‘GAUG-9005’, ‘GG-1’, ‘GAUG-9003’) and RootRot (‘GG-1’, ‘HGS-844’, ‘GAUG-9406’) were identified (Kumar 2008). Certain guar lines viz., ‘Sona’, ‘Suvidha’, ‘IC-09229/P3’, ‘Naveen’, ‘PLG-85’ and ‘RGC-471’ for seed type, and others like, ‘Pusa Mausmi’, ‘Pusa Sadabahar’, ‘Pusa Navbahar’, ‘IC-11388’, ‘PLG-850’ and ‘Sharad Bahar’ were released as promising varieties for vegetable purposes. Cowpea: A total of 2139 accessions have been evaluated for 24 descriptors and 3422 lines have been conserved ex situ. More than 67% varieties developed in cowpea owe their origin from evaluated germplasm lines. Some of promising varieties released following evaluation of exotic germplasm includes ‘Aseem’, ‘Bundel Lobia-1’, ‘C-152’, ‘Shweta’, ‘Co-2’, ‘Co-4’, ‘Pusa Phalguni’, ‘Pusa Sawni’, ‘Rituraj’, ‘S-288’ and ‘S-488’, ‘Charodi’, ‘Co-1’, ‘Co-5’, ‘cowpea-78’, ‘cowpea-88’, ‘FS-68’, ‘GC-1’, ‘Gomti’, ‘JC-2’, ‘JC-10’, ‘BBC-1’, ‘Bundel Lobia-1’ and ‘Paiyur-1’. Moth bean: Evaluation of more than 2011 accessions has exhibited wide diversity in growth, yield, quality traits and disease resistance (Gautam et al. 2000). However, 43 accessions introduced from Ceylon (Agarwal 1964), Mexico (Agarwal 1964), USA (Mishra et al. 2009), former USSR (Agarwal 1964) and Taiwan (Agarwal et al. 1987) are maintained and more than 1100 accession of moth bean have been evaluated at NBPGR. A total of 1540 moth bean accessions have also been conserved ex-situ in the national repository. Almost 2143 accessions have been maintained as active collections in medium-terms-storage facilities available at different centres of NBPGR. The varieties like, ‘Type-3’, ‘T-9’, ‘Baleswar-12’, ‘MG-1’, ‘Jadia’, ‘Jawala’, ‘IPCMO-880’ and ‘IPCMO-912’ have been developed directly or through selections made from local germplasm (Kumar 2005). Horsegram: More than 1500 accessions collected by NBPGR are being maintained at New Delhi (> 500 accessions), Akola (650 accessions) and Thrissur (500 accessions). TNAU, Coimbatore is also maintaining almost 320 accessions at RARS, Piyaur. Similarly, GKVK Bangalore is maintaining almost 120 accessions. Majority of these improved varieties released at Kumar & Rodge : Status, scope and strategies of arid legumes research in India- A review National and/or State level owe their origin from local germplasm directly or selection thereafter. These varieties are for Bihar (‘BR-5’, ‘BR-10’ and ‘Madhu’), Himachal Pradesh (‘HPK-2’, ‘HPK-4’, ‘HPK-5’ and ‘HPK-6’), Andhra Pradesh (‘PDM-1’and ‘VZM-1’), Jharkhand (‘K-82’ and ‘Birsa Kulthi’), Orissa (‘S-27’, ‘S-28’, ‘S-39’ and ‘S-1264’), Rajasthan (‘Maru Kulthi’, ‘KS-2’, ‘AK-21’and ‘AK-42’), Uttranchal (‘VL Ghat1’), Karnatka (‘Hebbal Hurali-2’, ‘PHG-9’ and ‘KBH-1’) and Tamil Nadu (‘Co-1’, ‘35-5-122’ and ‘35-5-123’). Genetic Improvement Interspecific hybridization: Earlyness is required to be transferred from wild species (S. serrata and S. senegalenesis) to cultivated species (C. tetragonolaba) of guar. Hence, Sandhu (1988) while studying the details of hybridization between 2 species (C. tetragonoloba and C. serrata) reported failure of hydridization through conventional methods. Other approaches like, bud pollination, amputation of stigma and style, use of organic solvents also failed to overcome the stigmatic incompatibility barriers. Scope for interspecific hybridization is, therefore, limited in guar unless specific tools are used. This prompted application of nonconventional methods of hybridization including ovary rescue, protoplast fusion etc. for genetic improvement. Interspecific hybridization in cowpea is required for transfer of resistance against pod-borer and sucking bugs from wild to cultivated species. Ng (1990) reported fertile hybrids by crossing cultivated cowpea [Vigna unguiculata sp. unguiculata (L.) Walp.] with wild relatives viz.,V. unguiculata spp dekind tiana (Harms) Verdc., Vig na unguiculata spp stenophylla (Harvey) M.M. & S., and Vigna unguiculata spp tenuis (E. Mey.) M.M. & S. At IITA, Nigeria, crosses between wild species (Vigna unguiculata ssp dekindtiana) and cultivated cowpea were made to transfer resistance against pod-borer and pod-sucking bugs. The introgressive backcrossing (IBC) resulted in the traits of recurrent parent being fixed as early as in IBC2 . Fertility increased with each IBC generation. A new method was termed congruity backcrossing (CBC) which involved crossing with each parent species in alternate generations (Kumar and Dixit 2004). Increase in fertility of the species used as female in the original crosses were observed in subsequent generations. Mahudeswaran et al. (1973) derived a ‘Co 2’ variety from 3way crosses involving V. catjang (Burm. f.) Walp., V. unguiculata and V. sesquipedalis (L.) Fruw. Improved plant types and earliness: Arid legumes known for high biomass production suffer from poor partitioning towards economic parts and are therefore, branded as poor yielders. Alteration in their plant types in concomitant with early maturity in reference to the agro-ecoregion and rainfall distribution is desired. Research carried out in moth bean has resulted in the development of new genotypes characterized with erect growth habit and early maturity (58-62 days) 257 compared with traditional genotypes with spreading and late maturing habits (90-100 days). These altered genotypes, in spite of curtailment in growth period by about 30-35 days, have great yield advantages. For instance, ‘CZM 32’ and ‘CZM 18’ mutant lines flower in 28 days, take 63 days to maturity, have 12.5 g/day productivity and may yield almost 780 kg/ha compared to late maturing spreading types (with 51 days, 94 days, 3.72 g/day and 380 kg/ha, respectively) (Kumar 2002). Thus, ‘CZM-32’ has been registered as a new potential genotype (IGNR no. 01024) at the NBPGR (Kumar 2003). Similarly, ‘RMM 12’ an early and synchronous-maturing type having only main stem has also been registered as new plant material (IGNR No. 04095) with maximum daily uptake of Na (0.08 × 10-9 mole) as compared to others with 0.04-0.09 × 10-9 mol (Tarafdar and Kumar 2003). ‘CAZRI Moth-1’- an improved semi-erect type - is also known for (Kumar 2001) in built resist ance against yel low mosaic viru s (YMV). A comprehensive list of promising varieties of guar, moth bean, cowpea and horsegram is also given in Table 1. Drought tolerance: Efforts have been made to make arid legumes more productive by inducing earliness so as to escape terminal drought by manipulating sowing dates and bringing in uniformity in flowering for matching these with irrigated crops. In moth bean for instance, dry-matter distribution towards economic parts, nutrients movement from soil to root (Tarafdar and Kumar 2003) and retention of flowering period (30-32 days) are of practical significance and are exploited. In cowpea, higher total plant dry matter in season (Agarwal 1987), larger tap root system under wilt (Itani 1992), lower value of 13 C in concomitant with higher levels of water-use efficiency, leaf area and xylem ABA (Nagugi et al. 1999), detached leaf test (Shekhawat et al. 2002) and screening during summer season at 3-4% available soil moisture have been successful. However, morphological and physiological characters contributing for drought tolerance have varied inheritance pattern (monogenic and polygenic) and gene action. Stomatal sensitivity and osmotic adjustment (OA) are dominant and drought tolerance is controlled by single dominant gene (Rds1 and Rds2) in cowpea (Mai-Kodomi et al. 1999). Breeding for high yielding varieties with drought tolerance is complex due to difficulties in combining drought tolerance and seed yield traits as both are governed by polygenes. There is also a need to identify high yielding genotype giving high yield under stress conditions. Hence, increasing yield potential under nonstress situation is being explored as a simple method with other approaches viz., collection of germplasm from dro ught habit ats representing bot h dro ught and heat environments, screening genotypes under summer season representing drought complex, predicting seed yield of drought-tolerant lines during rainfed rainy season situation, hybridization between high-yielding and drought-tolerant lines, and hybridization between traditional lines representing avoidance phenomenon and early-maturing lines representing escape mechanisms. Since expression of yield and yield traits 258 Journal of Food Legumes 25(4), 2012 Table 1. Improved varieties of arid legumes suitable for different cropping regions Sl. Average No. Rainfall* (mm) Rajasthan 1 170-200 2 3 200-250 250-300 Region/ district Cropped Produ- Varieties area* ctivity* (Year of (000 ha) (kg/ha) release) Churu 315.00 235 Jaisalmer 190.00 100 Bikaner Barmer 411.00 325.00 215 135 Ganganagar Hanumangarh 180.00 319.00 RGC-936 (1992) HG-365 (1998) RGC-936 RGC-365 RGC-563 (2005) 5 6 300-350 375-400 400-450 Remarks 85-90 Suitable for arid Rajasthan 80-85 85-90 80-85 85-90 675 RGC-1031 (2005) 105-108 Seed whitish in color, leaves wide, field tolerant to BLB and ALB Suitable for summer cultivation and wide spacing Suitable for irrigated conditions RGC-1002 HG-20-2 110-112 Suitable to high rainfall, high gum content, moderately resistant to BLB, RR and ALB with seed yield 1300-1600 kg/ha 115-120 Tall growing, requiring better management, seed yield 1100-1200 kg/ha Gum content 31.41% suited for wider spacing and irrigated conditions Dual purpose and suitable for canal command areas, HG-365 RGM-112 (2005) Nagaur 155.00 420 Jodhpur 183.00 180 Sikar 78.00 311 Jhunjnu 62.00 280 Pali 67.45 708 Jalore 69-50 600-650 Jaipur Bhilwara 55.14 37.00 780 600 8 700-800 Alwar 34.66 1000 RGC-986 Bhiwani Mohindergarh Sarsa Hisar 90.00 30.00 101.00 70.00 900 985 1400 1200 HG-365 HG-563 HG-884 HG-2-20 80-85 80-85 95-100 108-112 Banaskantha 61.60 604 GG-2 95-100 Kutch 58.10 610 HG-563 HG-365 RGC-936 Churu 293.00 470 Jaisalmer 170.00 121 Bikaner 283.00 215 Barmer 208.00 194 Rajasthan 1 170-200 2 200-250 High viscosity (3500 cP) and ruling variety of Haryana - RGC-1038 (2006) HG-884 (2007) RGC-1002 (1999) RGC-1017 (2002) RGC-1038 807 870 7 Haryana 9 200-250 10 200-225 11 300-350 12 250-300 Gujarat 13 Most drought hardy, flowers light pink in color and seed yield 900-1200 kg/ha Medium height, spreading type and seed yield 1000-1200 kg/ha - Suited to low rainfall zones, heavy podding behavior, improved in gum content and its quality 80-85 85-90 Moderately susceptibility to BLB disease, leaves light green and seed yield 1100-1300 kg/ha 100-105 Suitable for Rajasthan state, single stem type, tall growing, brisk podding and seed yield of 1200-1500 kg/ha 95-100 Somewhat photo insensitive and seed yield of 1200-1500 kg/ha 95-100 Improved in gum and quality and seed yield potential 1400-1500 kg/ha 100-105 Moderately resistant to BLB, improved in gum content and seed yield 800-1300 kg/ha 95-100 Moderately resistant to BLB, PM and seed yield 1200-1400 kg/ha 95-100 - RGC-1066 (2007) 4 Guar Maturity Important traits (days) FMO-96 (1996) CAZRI Moth-3 (2003) RMO-40 (1994) RMO-225 (1999) - Branched type, determinate growth and Moderately resistance to BLB disease 80-85 80-85 85-90 Mothbean 58-59 60-62 61-62 64-65 Suitable for Haryana, high gum and viscosity profile (4050 cP). - Suitable to mechanical harvesting in canal command areas with close planting Suitable to summer season and wide spacing (40-50 cm) High gum content (30-31%) and viscosity (3000-3500 cP) - - - Erect upright and synchronized growth Released for whole Rajasthan. Suitable for intercropping Erect and synchronized growth, escapes Suitable for extreme drought YMV and seed yield 700-800 kg/ha situations Less biomass, erect growth and seed yield 600-750 kg/ha Field tolerance to YMV, synchronized growth and seed yield 650-700 kg/ha Suitable for extreme drought situation Suitable for low rainfall Kumar & Rodge : Status, scope and strategies of arid legumes research in India- A review Sl. Average Region/ No. Rainfall* district (mm) 3 250-300 Ganganagar 4 300-350 5 350-450 Sl. No. Region/ district Cropped area* (000 ha) 0.23 Productivity* (kg/ha) 446 Hanumangarh 39.00 417 Jodhpur 159.00 251 Nagaur 215.00 218 Sikar Pali Jalore 0.93 0.32 0.32 289 239 470 Rajasthan 1 Andhra Pradesh 2 3 Tamil Nadu Karnatka Varieties (Year of release) Maturity (days) Co(CP-7) (2006) 67-72 Co(CP-7) Vamban-1 (1999) GC-3 (1997) 67-72 90-100 90-95 KBC-2 (1998) 95-105 GC-3 Vamban-1 90-95 90-100 4 Kerala Subhra 70-90 5 Madha Pradesh, Chattisgarh, Orissa GC-3 V-130 (1994) 90-95 95-100 6 Rajasthan (Sikar, RC-101 (2001) 60-62 Jodhpur, Jaipur region) V-240 (1994) 90-95 7 GC-5 (2003) Gujarat (North Gujarat, GC-5 Mehsana, Banaskantha) GC-3 Sl. No. Region/ district Cropped Produarea* (000 ctivity* ha) (kg/ha) Rajasthan 1 Andhra Pradesh 39.31 444 2 206.41 460 3 Karnatka Tamil Nadu 75-80 75-80 90-95 44-79 500 Varieties (Year of release) CAZRI Moth-3 RMO-435 (2002) FMO-96 RMO-40 CAZRIMoth-2 (2002) RMO-435 RMO-257 (2005) CAZRIMoth-1 (1999) Maturity Important traits (days) 60-62 259 Remarks - - 64-65 Leaves dark green and seed yield 600- Suitable for Rajasthan 650 kg/ha. - 66-68 First variety from hybridization, dark M ost high yielding and green color, seed yield 800-1200 kg/ha suitable for high inputs and better soils Good for seed and fodder, seed yield Suitable for intercropping and 600-650 kg/ha dual purpose Inputs responsive, natural source of High protein 25.0%, suitable YMV, seed yield 500-550 kg/ha to medium June planting and good source of fodder 64-65 63-65 73-75 Cowpea Important traits Remarks Moderately resistance to LCV, erect, synchronized growth habit and seed yield 9501200 kg/ha Seed contain 28.0% crude protein with minimum tannin content (0.75 mg/g) Long pods and seed yield 950-1000 kg/ha Release for whole Rajasthan, dual purpose High digestibility (90.0%), with less cooking time 44.0% Dual purpose Adapted to whole country, short stature, YMV resistant and seed yield 750-1150 kg/ha Asynchronous growth, long pods, tolerant to rust and seed yield 650-1100 kg/ha Very promising for Kerala, medium tall height, MR to anthracenose and seed yield 800-1000 kg/ha Adapted to low rain fall situations, seed red colour and seed yield 900-1100 kg/ha Synchronized growth, non-viny, escapes CYMV and seed yield 750-800 kg/ha, white seed color Red seed, dark green leaves and seed yield 10001200 kg/ha Bold seeds and seed yield 1200-1400 kg/ha Horsegram Varieties Maturity Important traits (Year of period release) (days) CRIDA 1-18R (2007) PHG-9 (1997) BGM-1 (1990) CRIDA 1-18R Paiyur-2 (1994) CRIDA 1-18R BJPL-1 (2009) 95-100 95-105 100-110 Released for whole Karnataka, dual purpose. Seed cP 25-26% Released for whole Kerala, dual purpose Dual purpose Released for whole Rajasthan and for mechanical harvesting. Dual purpose Suitable to kharif and summer Remarks Synchronized, 20% more yield over Most promising for southern states PHG-9 and seed yield 750-1150 kg/ha. Winy type, photo sensitive, suitable to southern states Bears tendrils, tolerant to YMV and Dual purpose seed yield 600-700 kg/ha 95-100 105-110 More suited to rabi season 95-100 95-98 27% more yield over PHG-9 and seed Dry fodder yield of 1200-1500 yield 800-1050 kg/ha kg/ha with 1.70 mg/g tannin content 260 Journal of Food Legumes 25(4), 2012 Sl. No. Region/ district 4 Madhya Pradesh Chattishgarh and Maharashtra Cropped Produarea* ctivity* (000 ha) (kg/ha) 62.00 310 Varieties (Year of release) AK-42 (2005) Maturity period (days) 90-95 AK-21 (1999) 85-90 5 6 Orissa Hilly regions 58.79 11.76 300 580 AK-21 85-90 VLG-10 (2006) 115-130 VLG-15 (2008) 105-115 VLG-19 (2010) 100-105 Important traits Remarks Suited for moderate rainfalls, seed color brisk red and seed yield 600-850 kg/ha Suitable to northern, western and central India and seed yield 650-800 kg/ha Field tolerance to anthracnose and stem rot and seed yield 1000-1200 kg/ha 30% higher yield over AK-21, field tolerance to anthracnose, seed yield 800-1400 kg/ha Moderately resistance to anthracnose and RR, seed yield 1000-1300 kg/ha CP 30.0%, Fat 3.65, grain type Most drought hardy and suitable to low rains Suitable for June sowing, dual purpose Suitable for June sowing, dual purpose and 86.2% digestibility Seed protein 26.6%, 83.4%digestibility *Long range statistics on rainfall, area and productivity are usually poorly expressed under stress situations, selection of drought related traits may be carried out under drought conditions while their yield potential is assessed under non stressed situations (Kumar 2008). Improved seed yield: Arid legumes show increased seed yield under existing situations with little or no fertility and poor agronomic inputs. Improved guar varieties like, ‘RGC 1002’, ‘RGC 1071’, ‘HGS 365’, ‘RGC 1038’ and ‘HG 884’ have yield potential of 1000-1400 kg/ha compared to traditional varieties (800-900 kg/ha). Earlyand high yielding varieties of guar like, ‘HG-365’, ‘HG-563’ and ‘HG 884’ have higher average productivity (1200 kg/ha) in Haryana state in comparison to their counterparts (370 kg/ha) in Rajasthan state of India. Similarly, varieties like, ‘Co(CP-7), ‘V585’, ‘GC 3’and ‘KBC 2’ in cowpea, ‘CAZRI Moth 3’, ‘CAZRI Moth 2’, ‘RMO 435’, ‘RMO 40’ and ‘RMO 225’ in moth bean, and ‘AK 42’, ‘AK 21’, ‘CRIDA-1-18’ and ‘VLG-19’ in horsegram have higher yield potential (Table 1). Improved quality: Guar needs quality improvement in respect of industrial guar gum production having potential to export. Similarly, guar meal is also an important animal feed rich in crude protein (40-50%) and green pods are largely used as a rich source of Fe and Zn in the form of delicious green vegetable. However, significant strides in enhancement of gum and protein content in guar seeds has not been made due to certain inverse relationships. There is a positive correlation between seed yield and per cent gum content (Mittal et al. 1971), while there is a negative association between seed weight and gum content hampering breeding efforts for large seeds. Seed color is also not associated with seed gum content. Both additive and non-additive gene effects are involved in determination of gum content. Viscosity is maximum in ‘HG365’ (3500-4000 cP) while genotype ‘HG-884’ has maximum gum content (31.41%). In moth bean, cowpea and horsegram, systematic evaluation have resulted in isolation of some promising lines viz., cowpea strain ‘PGCP-1’ and ‘Co (CP-7)’ with maximum seed protein (28%) and lower tannin content (0.75 mg/g) in the latter. In case of horsegram, maximum crude protein (31.12%) in ‘CRHG-7’ and minimum tannin content (1.44 mg/g) in ‘AK-21’ are observed (Table 2) (Chinnaswamy et al. 2011and Rodge 2009). In general, more tannin content is related to more cooking time in case of horsegram. Maximum seed protein (25%) is also observed in moth bean ‘CAZRI Moth-1’ (Kumar 2001). Table 2. Arid Legume varieties with improved quality traits Seed crude Seed cooking Seed tannin protein (%) time (min) contents (mg/g) Cowpea Co(CP-7) 28.2 28.2 0.75 PGCP-1 28.0 28.0 0.70 Moth bean CAZRI Moth-1 25.0 13.0 0.33 RMO-225 24.5 15.0 0.24 Horsegram CRHG-7 31.1 15.0 2.27 AK-21 30.2 13.0 1.44 Crop Varieties Biotechnology 1. Callus induction and regeneration protocols: In case of guar, MS media containing 6-benzylaminopurine (13.3 µM or 3 mg/L) in combination with indole-3-acetic acid (11.4 µM or 2 mg/L) with cotyledon node explants gives the highest frequency of multiple shoot regeneration (Deepak et al. 2003). More efficient regeneration is reported following culturing callus on MS medium containing 1-napthlenacetic acid (13.0 µM) in combination with 6-benzylaminopurine (5.0 mM) with a range of 82.1-88.4% of callus clumps producing 20-25 shoots (Deepak et al. 2005). The medium containing 2, 4-D (10.0 mM) in combination with 6-benzylaminopurime (5.0 mM) with embryo or cotyledon explants is the most suitable for induction of callus in guar. In moth bean, embryogenic callus cultures are established from the cotyledonarynode as explant on semi-solid MS medium supplemented with 0.75 mg/L 2,4-D and 1.5 mg/L 6-benzylaminopurine (BA) and with various additives (50 mg/L ascorbic acid and 25 mg/L each of adenine sulphate, citric acid and L - arginine). Numerous somatic embryos are differentiated on basal nutrient medium supplemented with 0.25 mg/L 2,4 – D and 0.5 mg/L of kinetic Kumar & Rodge : Status, scope and strategies of arid legumes research in India- A review (Kin). Transfer of these embryos onto fresh MS basal medium containing 0.2 mg/L BA and 2.0 mg/L gibberellic acid enable these to achieve complete maturation (Choudhary et al. 2009). In vitro regeneration of plants via somatic embryogenesis through cell suspensions culture is achieved in horsegram. Embryogenic calluses are induced on leaf segments on solid MS medium with 9.0 mM 2, 4-D. Differentiation of somatic embryos occur on transfer of embryogenic calli to liquid MS medium containing 2,4-D. Conclusive results indicates that a medium supplemented with 7.9 mM 2,4-D, 3.0% sucrose, 40 mg 121 L-glutamine and 1.0 m Mabscisic acid is effective to achieve high frequency of somatic embryo induction, maturation and further development (Mohamedi et al. 2004). Similarly, cowpea plants are regenerated from in vitro cultured explants of primary leaves. Primary leaves, including the intact petiole, is excised from three-day-old seedlings and cultured on Gamborg’s B5 basal medium containing 8×10-7M 2,4,5trichlorophenoxyacetic acid, 1×10-2 M L-glutamine and 1 × 104 M adenine sulfate. Callus is formed at petiole end and prolific shoot regeneration occurs when this callus is transferred to B5 basal medium containing 5×10-6 M 6-benzyl-aminopurine (BAP). Regenerated shoots rooted in growth-regulator-free B5 basal medium are also established in soil (Muthukumar et al. 1995). 261 for isolation of drought tolerant gene P5 Cs encoding 1pyrroline 5-carboxylate synthesis protein through proline biosynthesis pathway for transfer of drought tolerance in Nicotiana tabacum (Hong et al. 2000). The same gene is also utilized for development of transgenic rice for salt tolerance (Karthikeyan et al. 2011). Hence, P5Cs is an important source of drought and salt tolerance capable of being effectively transferred through Agrobacterium tumefaciens strain LBA 4404 harbouring the binary vector pCAMBIA 1301/P5Cs. Guar: A 1.6 kb guar mannam synthase (MS) promoter region has been isolated. This MS promoter sequence is over expressed in alfalfa (Medicago sativa L.). The quantitative GUS assay reveales that the MS promoter directs GUS expression especially in the endosperm in transgenic alfa, hence guar MS promoter could prove useful in directing endosperm-specific expression of transgenes in legumes (Naoumkina and Dixon 2011). The procedure of transformation of large seeded endospermous guar is also reported. Using Agrobacterium tumefaciens with T-DNA construct harbouring a-glucuronidase gene (uid A) and a neomycin phosphotrans ferase gene, maximum transformation frequency with cotyledomary explants are obtained using 145 mg/l kanamycin sulfate as selective agent. Carbenicillin and cefotaxine used for elimination of Agrobacterium after co-culture increases transformation frequency up to 10-folds in total. The presence of transgenes in the primary transformations is demonstrated by genomic DNA analysis of GUS-positive shoots. The high influx of energy into storage protein synthesis in guar seed has also been reflected by a high representation of genes annonated as involved in signal transduction carbohydrate metabolism, translation and ribosome structure. Thus, guar unigenes involved in galactomannan metabolism are identified. It is reported that among storage proteins, the most abundant protein is a conglutin accounting for 3.7% of the total ESTs (Naoumkina et al. 2007). Cowpea: QTL for economic traits have been identified to make the selection procedures more effective through MAS loci (Citadin et al. 2011). For instance, three quantitative trait loci (QTL) for resistance to Thrips tabaci and Frankliniella schultzei are identified using a cowpea recombination inbred population. The AFLP genetic linkage map and foliar feeding damage ratings are used to identify genomi c regions contributing towards resistance to thrips damage. Three QTLs (Thr-1, Thr-2 and Thr-3) are identified on linkage groups 5 and 7 accounting for between 9.1 and 32.1% of the phenotypic variance. AFLP markers ACC-CAT7, ACG-CTC-5, and AGGCAT1 are co-located with QTL peaks for Thr-1, Thr-2 and Thr-3, respectively. These results will provide a resource for mo lecu lar marker devel opment and t he genet ic characterization of foliar thrips resistance in cowpea (Muchero et al. 2010). In another case, a genetic linkage map is constructed using SSR markers and a recombinant inbred (RI) population derived from a cross between the breeding line 524B, and 219-01. Polymorphic SSRs obtained are used to construct a genetic map consisting of 11 linkage groups (LGs) spanning 677 cM with an average distance between markers of 3 cM. Six QTL for seed size reveal phenotypic variation ranging from 8.9 to 19.1%. Four QTL for pod shattering are also identified with the phenotypic variation ranging from 6.4 to 17.2%. The QTLs for seed size and pod shattering have been identified in two areas LGs-1 and 10 facilitating the use of MAS to eliminate undesirable wild phenotypes in breeding activities involving utilization of traits from wild germplasm (Andargie et al. 2011). A highly efficient Agrobacteriummediated cowpea transformation method for introduction of the cowpea á-amylase inhibitor-1 (áAI-1) gene into a commercially important cowpea cultivar, Pusa Komal generates fertile transgenic plants. The use of constitutive expression of addi tional vir genes in resi dent pSBI vector in Agrobacterium strain LBA4404, thiol compounds during cocultivation and a geneticin based selection system also results in two-fold increase in stable transformation frequency. Expression of áAI-1 gene under bean phytohemagglutinin promoter results in accumulation of áAI-1 in transgenic seeds. The transgenic protein is active as an inhibitor of porcine áamylase in vitro. Transgenic cowpeas expressing áAI-1 strongly inhibit the development of C. maculates and C. chinensis in insect bioassays (Solleti et al. 2008). Moth bean: Being extremely drought hardy crop, it is utilized Horsegram: Based on rDNA, IGS, RFLP by means of three 2. Identification and transfer of genes and QTL 262 Journal of Food Legumes 25(4), 2012 restriction enzymes, 69 isolates of rhizobia in horsegram are grouped in five clusters. By sequence analysis of 16S-23S, rDNA, IGS identifies the genotypes of rhizobia distributed into five different lineages of Brady rhizobium genus. Nearly 87% of indigenous horsegram isolates (IGS types I, II, III & V) could not be related to any other species within the genus Bradyrhizobium. Phylogeny based on house keeping ginll and recA genes confirms those results found by the analysis of the IGS sequence. All these isolated rhizobia nodulate Macrotyloma and Vigna spp. The isolates within each IGS type varied in their ability to fix nitrogen. Selection for high symbiotic effective strains could reward horsegram production (Chinnaswamy et al. 2011). Crop Production Guar and moth bean require well drained light textured sandy to loamy plain lands having even sloppy profiles (of 510% area) in Rajasthan, Haryana and Gujarat. Cowpea and horsegram are adapted to large range of climates from arid to semi-arid regions of western dry tracts of Gujarat to humid region in eastern parts of Orissa and West Bengal, and from north plains of Jammu and hill region of Himachal to deep south in Tamil Nadu and Kerala. The details of agrotechnologies are discussed here. Sowing windows: Early planting of guar and moth bean may cause profuse growth resulting in poor harvest but late planting may not attain usual growth and result in severe yield reductions (Kumar 2009). However, results reported byYadav et al. (2003) and Yadav et al. (2002) on early maturing guar varieties (‘HG 365’, ‘HG 563’ and ‘RGC 936’) revealed that planting after 25th June resulted in higher seed yield. In Southern India, winter cowpea is planted in OctoberNovember following N-E monsoon. Horsegram in rainy and winter seasons is sown from August to October and sometimes during November particularly in southern states (Table 3). Planting in row spacing as paired rows (Singh et al. 2003) or solid rows is better over broadcasting method. Thus, optimum plant stand is desired (Yadav et al. 1989a) for higher yield and both inter and intra-row spacing are different for different plant types (branched and unbranched). Under late sowing condition, closer spacing may be recommended. Early maturing varieties (‘HG-563’ and ‘RGC-936’) perform better under 60 cm inter row spacing than that of 40 cm (Kumar et al. 2003a). Usually 15 and 10 cm intrarow spacing are optimum for branched and unbranced guar varieties. Location specific trial on cowpea at Bangalore and Pattambi also indicates that 30 cm wider spacing is better over 45 cm (Table 3). However, in case of horsegram, seed yield is less influenced by row spacing as usually a seed rate of 40 kg/ha is adequate for higher seed yield. In plateau region of Bihar, 20 cm interrow spacing (50 kg seed/ha) proved optimum with seed yield of 2.06 t/ha (Rafey and Srivastva 1989). Cropping system: Cultivation of legumes improves N status of the soil thereby, reducing its requirement for the succeeding crops (Faroda and Singh 2003). Guar fixes around 30-70 kg N/ ha through biological N fixation and leaves a residual effect equivalent to about 15-20 kg N/ha (Rao 1995). In rainfed conditions, most prominent rotations viz., pearl millet + guar fallow, guar- fallow/mustard, sesame + guar- taramira, guartaramira, guar-mustard, guar + grass- fallow etc are dominant (Table 3). A long term study on cropping system reveals that pearl millet-guar cropping system gives 11% higher seed yield over monocropping of pearl millet only. Contents of organic carbon and available soil phosphorus may considerably improve (Saxena et al. 1997) through proper cropping systems. Location specific studies on intercrops at Bawal, Haryana indicates that intercropping one row of pearl millet and paired rows of guar is better for realizing guar yield by 27.2% (with 8.7 q/ha additional yield of pearl millet) compared to sole stand in paired row (Singh et al. 2003). At Bikaner, Rajasthan, strip-cropping of pearl millet and guar in 4 : 4 row ratio gives highest pearl millet yield over other combinations (4 : 6 or 4 : 8 ratio) (Yadav 1992), whereas alternate row planting (1:1) of pearl millet : guar is superior compared to other planting systems (Singh and Joshi 1994). Although guar yield was highest in 4:4 row strip-cropping of pearl millet: guar, but the net returns were highest in alternate-row intercropping system (Singh and Joshi, 1994a). At Bikaner, growing of guar and pearl millet in 2:2 planting system has the highest yield advantage (Yadav and Beniwal 2003). Similarly, uniform planting of castor+guar in 1:1 ratio and application of 10 kg N and 30 kg P2O5/ha to guar component has significantly yield advantage in terms of yield and additional gross income (‘1564/ ha) over the sole crop of guar (Kumar 2009). Cowpea grown with 1 or 2 rows between 2 rows of pigeonpea/maize/sorghum/ pearl millet may give 0.5-0.7 t seed yield/ha of cowpea without affecting the yield of companion crop. However, seed yield of pulses can be increased considerably (by 25%) by cultivating 1 or 2 rows of cowpea between paired rows of main crop (Yadav 1992). Studies also reveals that highest land equivalent ratio (1.532) was recorded with cowpea + amaranth (4:1) and is at par with other intercropping systems viz., cowpea + okra (4:1), cowpea + amaranth (5:1 and cowpea + okra (5.1) in Kerala (Table 3). Trials at Bangalore during 2007-08 and 2008-09 also reveals that cowpea + castor (6:1) has higher gross monetary returns per ha (‘ 18020/- and ‘18090/- during both years) compared to sole cowpea (‘15760/- and ‘15080/-) and other cowpea based intercropping systems (Kumar 2009). A study at Fatehpur-Shekhawati (Rajasthan) reveals that moth bean mixed with pearl millet (moth bean at 2/3 seed rate + pearl millet at 1/3 seed rate) gives the highest moth bean-equivalent seed yield compared to other crops when mixed with pearl millet (Shekhawat et al. 2002). Input management: Based on two years’ data at CAZRI, Jodhpur, application of 20 kg N and 40 kg P2O5 /ha has Kumar & Rodge : Status, scope and strategies of arid legumes research in India- A review 263 Table 3: Region wise high input and low input technologies and common cropping systems for arid legumes Guar Sl. Regions/ No. districts Technology Optimum LITs/ HITs sowing window Protection Improved Planting priority variety inputs Fertility inputs Cropping sequences/rotation Inter/Mixed cropping 3 4 5 6 7 8 9 10 1 Churu Jaisalmer Bikaner Badmer LITs June end to 25 July (within 2-3 days of 30-40 mm rainfall) (ST-RR) ‘RGC-936’ ‘RGM112’ ‘ HG-365’ ‘HG-563’ Line sowings, 60 × 10 cm, hand weeding, deep interculture up to 30-40 dos Monocropping, Guar – Guar, Guar – Guar – Bajra, Guar - Bajra Guar + Bajra (3:1) If delayed rains up to 1st week of August Guar + Moth, Bean + Bajra + Til + Cowpea (25% seed of each crop) 2 Ganganagar Hanumangah HITs June end to midJuly (within 2-3 days of 30-40 mm rainfall). Beginning to mid of March for summer crop ST-RR ST-BLB Full protection package ‘RGC1066’ ‘RGC1031’ Line sowing, Full fertility 25 × 5 cm, package deep interculture up to 30-40 days full production package Sole cropping Guar – Mustard Guar – Wheat Crop substitutions: Groundnuts Cotton, Bajra Limited mix or intercropping 1 2 Rajasthan 3 Nagaur HITs Jodhpur Sikar Jhaunjnu June end to mid ST-RR July ST-BLB (within 2-3 days of 30-40 mm rainfall). Beginning to mid of March for summer crop ‘RGC1002’ ‘RGC1003’ ‘RGC1017’ ‘RGC-936’ Line sowing, 45 × 10 cm, deep interculture up to 40 days 4 Pali Jalore Jaipur Bheelwara Alwar HTs End of June to mid of July (with rains or preirrigation). Beginning to mid of March for summer crop ST-RR ST-BLB Full protection package ‘RGC1038’ ‘HG-884’ ‘HG-1031’ ‘RGC-986’ Lime sowing, Fertility 40 × 10 cm, package Deep interculture up to 40 days, Production package Guar-Wheat Guar-Mustard HIT End of June to mid July, Beginning to mid of March for summer crop ST-RR ST-BLB Full protection package ‘HG-365’ ‘HG-563’ ‘HG-884’ Line sowing, Full 35-40×10 cm Fertility Deep package interculture up to 40 days Production package Guar + Bajra (3:1) Substitution: Cotton, Bajra, Til, Groundnuts Guar + Sorghum (3:1) Crop sequence: Guar (‘HG-884’, ‘HG2-20’) – Wheat, Guar (‘HG-365’, ‘HG563’) – Mustard, for saline water Guar - Wheat LTTs End of July to ST-RR mid of August. ST-BLB Beginning to mid of March for summer crop ‘HG-365’ ‘RGC-936’ ‘GG-2’ ‘GG-1’ Line sowing, 60 × 10 cm, Deep intercullure upto 40 dos Haryana 5 Sarsa Hisar Bhiwani Rewari Gujarat 6 Katch Banaskantha Urea spray Guar- Mustard @1-2% at Guar – Guar 50-60 Guar – Bajra DOS Guar – Guar – Bajra/Sorghum Guar-B. tournifortii Spray of Guar-Bajra , Guar urea @1- (GG-1-Mustard 2% at 50- (irrigated) 60 DAS North Gujrat : Guar-Potato, Agrisilviculture system Guar with P. cineraria Guar : Bajra (3:1) Guar + Bajra / Sorghum (3:1) Guar + Bajra (3:1) Guar + Sorghum (2:1) Mixed cropping for North Gujrat Bajra + Moth bean + Cowpea + Guar (0.40, 3.0, 5.0 & 3.75 kg/ha respectively) 264 Journal of Food Legumes 25(4), 2012 Mothbean 1 2 Rajasthan 1 Jaisalmer Bikaner Barmer Jodhpur Pali Sikar 3 4 5 6 7 8 9 LITs 5 July to 25 July (2-3 days of 30-35 mm rain falls) ST- ‘RMO-40’ RR ‘RMO-225’ ‘RMO-435’ ‘FMO-96’ ‘CAZRI’ ‘Moth-3’ Line sowing, 60×5 cm 1-2% urea spray spacing at 35-45 DAS Deep interculture up to 30 DAS and hand weeding 2 Churu Nagaur Ganganagar Hanumangarh Jalore HITs End of June to mid July (2-3 days of 30-40 mm rainfall) ST- ‘CAZRI’ RR ‘Moth-2’ ‘CAZRI’ ‘Moth-1’ Line sowing, Full fertility package 45 × 5 cm Deep interculture, Production full planting package 3 Gujarat Haryana LTTs Mid July to end of July ST- RMO-40 RR CAZRI Moth-3 Line sowing, 40 × 10 cm deep interculture 10 Sole cropping Moth beanGuar Moth bean Mustard Mono cropping Strip cropping Moth bean + C. ciliaris Mixed cropping Bajra (? ) + Moth bean (? ) seed Sole cropping Moth bean + Bajra Moth bean - and Castor (1:3 and 1:1) Mustard Moth bean – Moth : Bajra Wheat (3:1 or 2:2) Moth bean Sunflower Urea spray @1.0% at flowering Horsegram 1 Southern states Andhra LIT/ Pradesh HIT 2 Karnatka 3 Beginning to mid ST against July anthracmose (ST-Anth) ST-RR ‘CRIDA-118R’ ‘Palm-1’ ‘Palm-2’ HIT First fortnight of ST-Anth August ST-RR ‘PHG-9’ ‘CRIDA-118R’ Tamil Nadu LIT Second fortnight ST-Anth of Oct. to second ST-RR fortnight of Nov. ‘Paiyur-2’ 4 Madhya Pradesh and Chattisgarh LIT Second fortnight ST- Anth of June to first ST-RR fortnight of July ‘AK-42’ ‘AK-21’ 5 Orissa & Jharkhand LIT 3rd week of August to Ist week of Sept. (late kharif) ‘AK-21’ ‘AK-42’ 6 Hill region HIT First fortnight of ST-Amilha June with complete package ST-Anth ST-RR VLG-15 VLG-19 Line sowing, 35 × 5 cm late planting 30 × 5 cm with production package Line sowing, 35 × 5 cm Deep interculture up to 40 days with production package Line sowing, 37.5 × 10 cm spacing Preimergence herbi cide (Basalin/ @ 1.5 kg ai/ha) Line sowing, 40 × 10 cm spacing Deep interculture up to 40 days and hand weeding Protection package Kharif Maize – winter Horsegram Kharif Sorghum – winter Horsegram Protection package Horsegram – Finger millet Ragi - Horsegram - Horsegram – Finger millet Ragi - Horsegram - Kharif Maize – winter Horsegram Kharif Sorghum – winter Horsegram Kharif Finger millet – winter Horsegram Upland RiceHorsegram Maize-Horsegram Ragi-Horsegram Line sowing, 40 × 10 cm spacing Deep interculture up to 40 days and hand weeding Line sowing, Full 30 × 5 cm spacing fertility package with full planting package Mono crop Hybrid Sorghum + Groundnut (2:2) + 1 row of Horsegram when grounds at flowering Maize + Horsegram (1:1) Horsegram + Ragi (6:1) Horsegram + Castor (6:1) Maize + Horsegram (1:1) Horsegram + Ragi (6:1) Horsegram + Castor (6:1) Niger + Horsegram (2:1) Castor (VI-1) + Horsegram (1:1) Marvel grass + Horsegram (1:1) Amaranthus+ Horsegram Horsegram + Finger millet + Horsegram + Groundnuts Cowpea 1 Southern States Andhra LIT Pradesh HIT HIT Kharif- June end to July end RabiOct-Nov Summer Whole March ST-RR ‘Co(CP-7)’ ST-Anth Full package plant ‘Co(CP-7)’ protection ‘Co (CP-7)’ Line sowing, 40 × 15 cm Fertility Deep interculture and package hand weeding Line sowing: 30 × 10 cm whole package Rice-Cowpea Rice-fellow-summer Cowpea Rice-Rice-Cowpea Pigeon pea (PP) + Maize + Sorghum (2 rows) + Cowpea (2 rows) Kumar & Rodge : Status, scope and strategies of arid legumes research in India- A review 1 2 2 Tamil Nadu 3 LIT 4 5 Kharif: June end ST-RR to July end ST-Anth Rabi: Oct-Nov to Summer Whole March 6 ‘Co (CP-7)’ 7 Line sowing, 40 × 15 cm Deep interculture and hand weeding Line sowing: 30 × 10 cm whole package 8 - 9 Rice-Rice-Cowpea 3 Karnatka LITs June end to July ST-RR end coconut floor ST-Anth – May – Sept. ‘KBC-2’ Line sowing, 30 × 15 cm spacing deep interculture and hand weeding - 4 Kerala LITs Homestead garden whole year ‘Subhra’ Line sowing, 45 × 10 cm spacing deep interculture and hand weeding - Kharif- Sole Cowpea Rabi: Sole Cowpea Summer- Sole Cowpea Maize + Cowpea – Maize Cowpea – Maize + Cowpea ‘V-585’ ‘V-240’ Line sowing, 30×10 cm deep interculture Production package ‘RC-101’ ‘GC-31’ Line sowing, 30x5 cm hand weeding ‘RC-101’ ‘GC-3’ Line sowing, 60 × 10 cm hand weeding 5 Northern and Eastern Hills, UP, HITs Kharif: Punjab and Beginning to Bihar mid-July LITs Summer18-31 March ST-RR ST-Anth with protection package ST-ALS 6 Rajasthan LIT KharifFirst week of July to end of July Summer- Mid Feb to end of March ST-RR 7 Gujarat HIT Mid July to mid August Summer Mid Feb ST-RR ST-Anth GC-3 GC-5 Fertility package Maize – Wheat – Cowpea Cowpea – Mustard/ Wheat – Moong bean 1-2% urea Berseem + Japanirye – Jowar + spray at flowering Cowpea Maize + Cowpea Jowar + Cowpea Bajra + Cowpea 1-2% urea Cowpea – Mustard – spray summer fellow Cowpea – Mustard summer Moong bean Line sowing, Fertility package 40 × 10 cm Deep interculture Production package Cowpea – Wheat – Moong bean Maize – Wheat Cowpea 265 10 Paired rows of PP + Maize + Sorghum + ½ rows of Cowpea Bhindi + Cowpea (1:1) Amaranthus + Cowpea (1:4) Cowpea + Ragi (5:1) Cowpea + Castor (6:1) Cowpea + Amaranth (4:1) Maize paired rows + Cowpea Cowpea + Sorghum (1:1) Cowpea + Pigeon pea (2:1) Bajra + Moth bean + Cowpea + Til + Guar (0.90 + 3.0 + 5.0 + 3.75 + 0.10 kg/ha resp.) Paired row’s of Maize + 1 row of Cowpea Intercropping Pigeon pea + Cowpea (2:1 or 2:2) ST- RR: Seed treatment against root-rot: 2-3 g Thiram/Bavistin per kg seed, ST-Anth: Seed treatment against Anthracenose fungal disease of horsegram and cowpea: Mancozeb/Benomyl @ 2-3 g/kg seed; ST-BLB: Seed treatment against Bacterial leaf blight disease of guar: 100 ppm streptocycline for one hr; Ful l package of plant protecti on: Alternaria Leaf Spot: Spray of Mencozeb @0.2%. BLB: Spray of streptocycline @ 150 ppm + 0.2% vitavax. Cercospora Leaf Spot (CLS): Spray of Dithane M-45 or Dithane Z-78 (ai) @ 0.2%. White flies: Spray rogor @0.02%;Full package of fertility: 20 kg N + 40 kg P 2O5 + 20 kg ZnSO4 per ha, + PSB, 1-2% urea spray at flowering; Ful l package of planting: Chemical weedicide: basalin @ 1 to 1.5 kg/ha ai pre-plant application by dissolving in 700 liter water/ha + 1 handweeding, one light suppl. irrigation at pod formation stage increased seed yield of guar substantially (25.2%) while water use efficiency was much higher (21.7%) over that of control (Singh and Khan 2003). Work at Bikaner reveals that 40 kg P2O5/ha increases seed yield of guar (by 38.4 and 22.7% compared to control and 20 kg P2O5/ha, respectively). Similarly, three years (1992-94) study at S. K. Nagar reveals that guar fertilized with 40 kg P2O5/ha records the highest seed yield (by 28% over control). Similarly, at Agra, application of 60 kg P2O5/ha increases seed yield of guar (by 38.1 and 13.6% over control and 30 kg P2O5/ha respectively). At Gwalior, guar yield increases with each successive increase of 20 kg P2O2 up to a level of 60 kg P2O5/ha and increase in yield at this level is much higher (44.9%) than control. Besides this, gum and protein contents are also increased (Bhadoria et al. 1997). It is also reported that application of 60 kg P2O5/ha is optimum for higher seed yield by 27.3 and 11.9% over control and 30 kg P2O5/ha, respectively (Kumawat and Khangarot 2001). Guar grown on S-deficient soil of Gwalior, responds up to a level of 40 kg S/ha in terms of growth, yield attributes and seed yield. The yield increase at 40 kg S/ha is shown to be optimum, respectively. Based on three years’ (1992-1994) results at Bawal, application of 20 kg S/ha in guar crop resulted in significantly higher seed yield (1.212 t/ha) over control (10.82 q/ha) (1 q=100 kg). 266 Journal of Food Legumes 25(4), 2012 Micronutrients studies conducted at Hisar, Bawal, Durgapura and Gwalior on guar reveals that at all the locations, one spray of 0.5% ZnSO4 either at 25 or 45 DAS gives higher seed yield over control but is statistically equivalent to soil application of ZnSO4 @ 25 kg/ha. A result from S. K. Nagar also shows that cowpea may be applied with 25 kg ZnSO4/ha as basal dose to obtain higher net returns and BCR. If Zn is not applied at sowing, it can be alternatively applied in 2 sprays at 25 and 45 DAS. At Bangalore, soil application of 25 kg ZnSO4/ha, gives higher yield, followed bycombined application of ZnSO4 and FeSO4 and 2 sprays of 0.5% ZnSO4 at 25 and 45 DAS. Application of Fe and Zn in moth bean also brings significant increase in yield at Hisar. Optimum doses of Fe and Zn also help in less infection of guar against Alternaria leaf spot (Gupta and Gupta 1999). Maximum seed yield of guar at Gwalior is obtained when the crop receives 3 irrigations each at vegetative, flowering and seed development stage. However, 2 irrigations at flowering and seed development stages are optimum at Durgapura, whereas one irrigation at seed development at S. K. Nagar while that at flowering stage at Bawal is sufficient to obtain higher seed yield. Weed control: Season-long competition of weeds in guar may cause reduction in yield by 30 to 50% and the losses may also go up to 70-90%. Critical crop-weed competition lies between 15 to 30 DAS in loamy sand and sandy loam soils. Thus, removing weeds at 20 or 30 DAS is helpful in raising pods/ plant, water use efficiency and seed yield (Yadav 1998). Studies conducted in different agro-climatic zones/soil types have established the superiority of mechanical/cultural means over chemicals (Kumar et al. 1996). The work on chemical weed control also indicates that pre-plant incorporation of fluchloralin 1.0 kg a.i./ha (Basalin) is an effective herbicide to control weeds in guar (Yadav et al. 1998) under rainfed condition at Bawal, Haryana and is at par with hand weeding at 30 DAS (Kumar et al.1996). Increase in seed yield due to one hand weeding and fluchloralin is 49 and 45% respectively, over weedy check. Two hoeings and fluchloralin at 1.5 kg/ha are also useful at Hisar, Haryana with higher seed yields of 1.89 and 1.84 t/ha and net income of ‘2900 and 2845/ha, respectively over control (Yadav et al. 1997). However, the weed control efficiency of the two hoeings (80.7%) and fluchloralin at 1.5 kg/ha (79.9%) are better over those of than one hoeing (64.2%). Pre-plant incorporation of fluchloralin at 1.0 kg a.i./ha is beneficial for 84% reduction in weed population but seed yield was at par with one or two hand weedings, whereas pendimethalin causes phytotoxic effect in guar (Yadav et al. 1998). Seed inoculation: Inoculation of guar seed with VAM fungi has improved dry matter production and seed yield (Rao and Tarafdar 1993). Similarly, inoculation of seed with arbuscular mycorrhizal fungus (AMF) and Rhizobium culture also significantly improve nodules, dry weight and seed yield of guar over no inoculation, and further application of FYM @ 4 t/ha along with inoculation of AMF and Rhizobium culture has beneficial effect on seed yield over NP application (20 kg N and 40 kg P2O5/ha) (Tarafdar and Rao 2001). Increase in yield through Rhizobium ranges from 8 to 15% depending upon the intensity and distribution of rainfall (Rao 1995). In a field study conducted at Jaipur, Kumawat and Khangarot (2001) gets higher guar seed yield by Rhizobium inoculation along with increased N, P, S, protein and gum content (in guar seed) and higher total uptake of N, P and S (Kumawat and Khangarot 2002). Three years study reveals that sole FYM or sole seed treatment with PSB may not influence seed yield of cowpea, bu t their combined applicatio n increases seed yield significantly over control. Results of multilocation trials conducted on integrated nutrient management at Hisar, Gwalior, Durgapura, S. K. Nagar and Bawal to know the response of Rhizobium inoculation and PSB in guar indicates that inoculation with both Rhizobium and PSB is helpful in increasing seed yield (by 22.35%). Plant Protection Arid legume diseases: Powdery mildew [Erysiphe polygoni DC, Sphaerotheca fuliginea (Schlect) Pollacci, Leveillula taurica (Lev) Arnaud] is an important disease of cowpea and horsegram, particularly, in southern India. Sometimes guar may also show infection particularly, in late sown conditions. Appearance of dirty-white, circular, floury patches at the time of pod formation and maturity are common symptoms (Gupta and Rohilla, 2008). Similarly, root rot complex [Macrophomina phaseolina (Tass) Goid, R. solani, Fusarium caeruleum, Sclerotium rolfsii, Neocosmospora vasinfecta] is also common. Among root diseases, root rot and wilt are major diseases. In case of the former, root tissues are decomposed, preventing normal growth of host plants. In case of latter, leaves and green parts lose their turgidity, become flaccid and ultimately fall down. R. solani is highly pathogenic in guar and may kill 90% seedlings within three weeks. F. caeruleum may cause wilting to almost 78% plants. Dry root rot is also attributed to high soil temperature (35-40°C) and low soil moisture content (0.5-1%). Yield losses due to anthracnose [Colletotrichum lindemuthianum Sacc & Mogn. (Bri & Cav.)] may range from 35 to 50% (Gupta, 2009). Initially water soaked lesions appear on the pods, later-on, become brown and enlarging to form circular spots of varying size. In more severe conditions, the infected leaf petiole and stem parts may wither off. Seedlings very often are blighted due to infection soon after the seeds germinate (Gupta and Rohilla, 2008). Similarly, in Cercospora leaf spot [Cercospora canescens Ell. & Martin, C. Dolichi Ell & EVr], two pathogens viz., C. canescens and C. cruenta have been reported to cause leaf spot in cowpea growing areas of Fiji, Kenya, Brazil, Nigeria, Zimbabwe, India, Bangla Desh, Iran, Malaysia, Thialand (Saxena et al. 1998). The disease is Kumar & Rodge : Status, scope and strategies of arid legumes research in India- A review favoured by humid weather and the spots is spread by wind and water splash. Fusarium wilt [Fusarium oxysporum f sp trachciphilum (Smith) Syn. & Han.] in another important disease where more than 6 species of Fusarium is reportedly associated with cowpea seeds, seedlings and adult plants that cause wilt at different stages of plant growth. Cation exchange-capacity (CEC) may play an important role in management of this disease in soil. It is more severe in nutrient-deficient soils with CEC 4.9. The higher CEC nullifies the adverse effect of this fungus when used as seed treatment (Monga and Grover 1993). Similarly, in alternaria leaf spot [Alternaria cyamopsidis Rangaswami & Rao. A. cyamopsidis (Ell. & Ev.) Elliot], heavy yield losses to the tune of 50-55% are reported. It appears every year in mild to severe form and the pathogen is seed born in nature. Disease infection is observed more in late sown conditions, high humidity (70-90%) and moderate temperature range of 25-31°C. Early sown crops particularly, during last June may escape from this disease. Bacterial disease like, Xanthomonas axonopodis pv. vignicola; X. campestris pv. cyamopsidis Dye and X. phaseoli has the favourable temperature range of 25-30% and relative humidity of 70-90%. As the bacterial pathogen is seed born, the disease appears both as leaf spot and blight simultaneously. Disease spots appear intraveinal and are round, water soaked or oily in appearance and well defined on the dorsal surface of the leaves. Black streaks which develop later on, may lead to cracking of stem and these black streaks may also develop on pods. With regards to viral diseases, cowpea mosaic virus (CMV) is very important as yield reductions to the extent of 40-100% is reported Two strains of this virus i.e., CPMV-Sb and CPMV-Vu are reported (Agarwal 1964). Under natural condition, two distinct types of necrotic local lesions or chlorotic local lesions are observed. The virus is readily sap transmissible. The diagnostic sp includes cowpea cv. Black Eye and Early Ramshon and Chenopodium quinoa. Similarly, mungbean yellow mosaic (MYM) disease may appear at any stage of plant growth. If it appears in initial stage, plants may not flower and yield losses may reach as high as 90%. High incidence of yellow mosaic in early sown and late sown moth bean is probably due to higher population of white fly in the field. Guar is found most effective trap crop for checking the white fly. Moth bean variety CAZRI Moth-1 is also reported possessing resistance to yellow mosaic. Arid legume pests: Colonies of aphids (Aphis craccivora Kock) are found on leaves, stems and pods of guar and cowpea. The pest is most effective in early growth of plants (July-September), having several generations during the cropping season. Nymphs and adults suck cell sap from lower surface of the leaves, top shoots; consequently plants become discolored and weak. Similarly, in case of Pod borer [Helicoverpa armigera or H. Vitrata (Hübner)], young larvae 267 feed on the foliage and later on damage flowers buds, pods and feed on developing seeds inside pods and may reduce seed yield to the tune of 60%. A single larva may destroy 3040 pods before reaching maturity. There may be as many as 8 generations in one year. Other pests include white flys [Bemisia tabaci (Gennadius)] where, both nymphs and adults feed on cell sap from under surface of the leaves. White flies also act as vectors for spread of viral diseases. These flies also excrete honeydew on which black moulds grow interfering with photosynthesis. Leaf perforator [Dichomeris ianthest (Mery)] is also an important pest of guar as the newly emerged larvae crawl and ultimately settle down near mid rib or near thick leaf vein, spinning a web there. There is more damage due to this pest in early stage of crop growth. White grub (Holotrichia consanguinea Blanch.) is destructive and a polyphagous pest for which killing of adults through trapping in light lamp is easiest way. The fungus Metarrhizum anisopliae (Mestch) has been fo und pathogenic o n the adults. Bruchi ds [Collosobruchus maculates Fab., C. chinensis (L.), C. analis Fab.] are also seen in cowpea, moth bean and horsegram seeds. Remedial measures against the above major diseases and insect pests is given in Table 4. Quality of arid legumes Cooking is known to improve the palatability and nutritional quality of food legumes. However, prolonged cooking may result in decreased protein quality and loss of vitamins and minerals. Dehusking and splitting of pulses into cotyledons reduce the cooking time considerably. In certain legumes like horsegram, this practice is also not effective. Pressure cooking, addition of chemicals to cooking water or soaking of common legumes in solution of chemicals have been recommended for reducing the cooking time or producing instant cooking of beans. Soaking and germination of horsegram and moth bean reduced the levels of tannin and phytase. Native and germinated legumes can be incorporated in bakery products to increase the protein level (Deshpande et al. 2005, Ghatage et al. 2005, Rodge and Wankhede 2003). Resistant Starch (RS): Legumes contain substantially higher levels of resistant starch (RS) than do cereal seeds, flours and seed-based food products. They (RS) have drawn broad interest worldwide for both their functional properties and potential health benefits. Initial clinical studies have demonstrated that the same have properties similar to dietary fiber and promising physiological benefits in human beings which may result in prevention of various diseases. There are four types of resistant starch RS1, RS2, RS3, and RS4. Legumes come under RS1 which resists digestion, protects from digestive enzymes by other components normally present in a matri x of typical starch sources. Among the bestcharacterized forms of RS are those which are derived from legumes. The RS provide many health benefits such as improving bowel health, blood lipid profile, and increased 268 Journal of Food Legumes 25(4), 2012 Table 4: Important diseases and insect pests in arid legumes and their remedial measures Sl. No. Diseases 1 Root rot complex Dry root rot Wilt Crops Guar Cowpea 2 Cowpea Horsegram 3 4 5 6 7 Anlhraconose Remedial measures Seed treatment in Bavistin (0.2%), Vitavax (0.2%) Fungicide spray of Brassicol (0.2%) Foltaf (0.2%) Tiride (0.2%) Seed treatment for Carbendazim @ 2 g/kg seed Mancozeb /Thiram/ Benomyl @ 2-3 g/kg seed Foliar spray of Benomyl (0.2%) Mancozeb (0.2%) Cercospora leaf spot Cowpea Seed treatment of Horsegram Moth Captan/thiram @ 2.5 bean g/kg seed Foliar spray of Benomyl (0.2% ai) Dithane M-4.5 (0.2% ai) Dithane Z-78 (0.2% ai) Powdery mildew Cowpea Foliar spray of Horsegram (in Tridemorph (0.05%) south India, in Kerthane (0.5%) late sown Calixin (0.1%) condition) Benomyl (0.2%) Alternaria leaf spot Guar Seed treatment of Cowpea Thiram/Dithane Z-78/ Horsegram illex @ 0.3% Foliar application of Iprodione (0.2%) soil application and foliar application of or Zn in combination Seed treatment of Bacterial leaf blight Guar (BLB), Cowpea Ceresin wet (0.2%) Bacterial Leaf Spot Thiram (0.2%) (BLS) Streptocycline (0.1%) Foliar spray: Two spray of ZnSO4 at25 & 45 DOS (0.5%) Streptocycline spray (250 ppm) B. subtilis + white sterile fungs Mung bean Cowpea Moth Foliar spray of yellow mosaic virus bean Rogor (0.2%) (MYV) Methaldemeton (0.1%) micronutrient absorption (magnesium and calcium) in the colon. These factors may affect the risk of developing diseases su ch as co lorectal cancer, cardio vascular diseases, osteoporosis and inflammatory bowel diseases. Increase in fecal bulk RS are important in preventing constipation and hemorrhoids and in diluting potentially toxic compounds that might promote the formation of cancer cells. RS may be of benefit to healthy individuals who are trying to achieve and maintain a healthy body weight. RS containing foods have low glycogenic index thus release of glucose is slow, resulting References Lodha and Sharma (2000) Insect pests Aphids Crops Guar Cowpea Remedial measures Malathion spray (0.05%) Rogor spray (0.02%) Use of entomogenous fungi Entomophthora, Cephalosporium Use of predators: Coccinellids, syrphids lace wings. Neem based insecticide like nimbecidine Release of egg parasitoid Trichodermona chilonii @ 0.3 million adults/ha Release of larval parasitoid Campolestis chloridaeuchida Gupta and Rohilla (2008) Pod borer Cowpea Moth bean Gupta et al. (1998) Leaf perforator Guar Spray malathion @0.05% Endosulfan @ 0.07% Gupta and Rohilla (2008) Whitfly Cowpea Moth bean Horsegram Foliage spray of Dimethoate 30 EC @ 250 ml or oxydemeton 25 EC @ 300 ml/0.4 ha Gupta et al. (1999) White grub Guar Dusting with methyl parathion @ 2% or Spraying quinolphos @ 0.05% for killing beetles Use of pathogenic fungus Metarrhizum anisopliae (Meslch) Gupta et al. (2007) Lodha (2001) Bruchids Cowpea Moth bean Horsegram Store seed at moisture content (10.0%) Seed treatment with neem leaves Seed treatment with edible oil content @5-6 g/kg seed in lowered insulin response and greater access and use of stored fat. This helps in management of diabetes and impaired glucose tolerance. Hence, it is used in the treatment of obesity and weight management (Rodge 2009). Guar Gum: Guar gum is a white to yellowish-white powder and is nearly odorless. Fine finished guar gum powder is available in different viscosities and granulometries depending on the desired viscosity development and applications. Guar gum is a natural high molecular weight hydrocolloidal polysaccharide composed of galactan and mannan units Kumar & Rodge : Status, scope and strategies of arid legumes research in India- A review combined through glycosidic linkages, which may be chemically described as galactomannan. Guar gum is a cold water soluble polysaccharide. This ability to hydrate without heating makes it very useful in many industrial uses. Dissolved guar gum forms a line of high viscosity and viscosity is a function of temperature, time and concentration. Solution with different gum concentrations can be used as emulsifiers and stabilizers because they prevent oil droplets from coalescing. Guar gum is also used as suspension stabilizer and is an economical thickener and stabilizer. Different guar gum powders are manufactured as per their industrial uses such as thickening, texturing, stabilizing and enhancing suspension. There are two types of guar powders viz., food grade guar gum powder which is used in industries like food, cosmetics, pharma etc. Here the particle size of food grade gum powders are 200 mesh having 2000-5000 viscosity (cP) and with 300 mesh having 3500-5000 cP values. The other one is industrial grade gum powder which is used mainly in industries like paper, mining, explosive, fire fighting, oil drilling etc and are available with particle size of 100 mesh with viscosity values of 3000-7000 cP. Minimum standard for good quality guar gum as defined by US FCC and by European Union Specification includes moisture 14.0% maximum (max.) ash 1.5% max., acid insoluble residue 47% max., galactomannan 75.0% min, protein 7.0% max., arsenic 3 ppm max., lead 10 ppm max., zinc 25 ppm max., copper and zinc 50 ppm max (Rodge 2008). Guar gum is used in almost all systems where water is an important factor. Food industries share almost 30-40%, petroleum and mining industries share around 20-25% and textile industries share almost 18-20% of total guar gum consumption (Table 5). Export of guar gum: India exports almost 75-85% of its guar gum including other derivatives annually. India exported 0.258 million tonnes of guar gum worth ‘ 139 million during 2008-09. The same dramaticallyincreased to almost 0.35 m t worth ‘28000 Table 5. Use of hydrocolloids in food products Dairy industry Beverages Bakery products Confectionery Meat and fish products Ice-cream stabilizer, milk shake, Ice milk (Soft icecream), ice pops and water ice chocolate, milk drink, flavoured milk drinks, instant desert puddings, cooked desert puddings, cottage cheese, cheese spread, whipped cream yoghurt Soft drinks with fruit pulp, soft drinks, fruit juices and nectars, foam stabilizer, beer clarification, wines, juices and vinegar, aging of spirits Bread doughs, cake batters, fruit cakes, yeast raised dough nuts, pipe fillings, fruit fillings, bakery jellies, flat icings, cookies, frozen pies fillings Candy jells and jellies, Frozen confection, candy glaze, chewing gums, cough drops, gum drops, candy mints Fish preservation, canned fish, meat and poultry Coated jellied meat, preservative coating for meat and poultry, synthetic meat fibers and products Source: Halis and Feramuz (2007) 269 million during 2010-11 (Fig 1). It is due to increased demand of guar gum from USA to 0.45 m t annually against 0.075 m t earlier due to increased demands from drilling of petroleum fields in middle east countries. During 2006-07, top importing countries of guar products were USA followed by China, Germany, Italy and Netherlands. 3000 2500 2000 1500 1000 500 0 Fig 1. Exports of guar gum from India (Source APEDA) Guar meal: The germ and outer seed-coat of guar seed together constitute guar meal. Removal of gum from guar seeds increases the protein content of the residual byproduct, i.e. guar meal. It is light, grayish material with beany flavour. The guar seeds result in 62-68% of guar meal having a rich source of protein content by about 35-46%. It contains about 1.5 times more protein than guar seed and is compared well with other vegetable protein sources like oilseed cake used in poultry diets. The proximate composition and nutritive value of defatted guar meal, protein isolates and protein concentrate are best for monogastric animals. It is observed that the guar oil contained 55.1% linoleic acid compared to 51.8% in sunflower oil. The total unsaturated fatty acids are 78.7 and 92.0% in gaur and sunflower oil, respectively. Guar oil contained 3.36% linolenic acid. The iodine value and refractive index of the guar oil is well comparable with that of sunflower oil. Arid legumes in non-traditional areas: Frequent climate changes, need for more production and enhanced international demands for industrial products have prompted to explore the possibility of introducing these crops in newer regions and seasons. Development of early maturing varieties (60-65 days) of cowpea (RC 101, PGCP-3) has helped in introduction of this legume as summer cowpea in northern western plain zones and foot-hills with 3-4 irrigations only. Seed yield obtained to the tune of 700-1000 kg/ha support their successful acceptance in these regions. Similarly, field trials of guar during rainfed rainy season of 2011 and summer season (5 irrigations including pre-sowing) of 2012 at CAZRI, Jodhpur have given new concept of guar cultivation in summer season (Table 6). Seed yield of guar can be increased by 2.5-3 times, gum content by 1.7%, guar gum yield by 320 kg/ha and viscosity of guar gum by 339 cP in summer over rainy season. It has been due to favorable climatic conditions and controlled availability of soil moisture, proved useful in reducing diseases and insect 270 Journal of Food Legumes 25(4), 2012 Table 6. Seed yield, gum content and viscosity of guar genotypes during rainfed conditions and summer irrigated conditions at CAZRI, Jodhpur during two seasons Genotypes RGC-1066 RGC-986 RGC-936-1-5-1 HG-884 HGS-563 RGC-936 (Ch.) Mean C.D.(0.05) * Seed yield (kg/ha) Kharif 2011 Summer* 2012 487.19 1021.0 483.75 1275.0 495.94 1916.67 534.38 1520.83 482.81 1950.00 546.25 1439.58 505.0 1520.5 36.11 584.41 Kharif Gum content (%) 2011 Summer* 2012 29.41 30.57 28.70 30.05 27.90 30.98 29.09 31.29 29.14 29.70 29.24 31.17 28.9 30.60 NS 1.36 Viscosity of guar gum (cP) Kharif 2011 Summer* 2012 3522 3639 3402 3440 3169 3231 3130 3474 3015 3226 3336 4600 3262.3 3601.6 NS 902.34 5 irrigations including pre-sowing irrigation, no fertility and plant protection measures were adopted pests infection and leading to better source sink relationship. Guar is successfully supplementing rainfed groundnut in Annantpur and other parts of Rayalsema region, Government of Andhra Pradesh is supporting guar in traditional rainfed groundnut regions. It has been successfully adopted during summer 2012 in Yavatmal and nearbydistricts of Maharashtra. There are still other examples like Vadodra (Gujarat), Raipur (Chattisgarh) where guar is being cultivated. Future strategies Increasing productivity of arid legumes by about 3-5 folds over existing levels for each drop of water, unit of time, area and inputs, to face the ever increasing demands of these crops. Development of guar varieties maturing in 70-75 days so as to prevent from losses caused due to terminal stress and varieties must have inbuilt resistance potential against root rot and bacterial leaf blight diseases. Development of cowpea varieties wit h mo re synchronous maturity and compact plant types having 60-70 days maturity, resistance against CYMV, with colorless, bold seeds for table purpose. Curtailing maturity period of horsegram to 75-80 days with improved conversion having resistance against anthracnose and powdery mildew diseases in addition to showing thermo-in sensitivity. The value added products/derivatives like hydroxyl propyl used in oil drilling field may be taken up. Similarly in food applications hydrocolloids, stabilizer food additives may be developed and exported. Developing sustained production technology for subsistence farming community and corporate world as well. Finding out remedial measures against anti nutritional factors in horsegram seed, making it alternative source of edible food pulses particularly for tribal areas and popularization of its medicinal and food values. Exploiting Bt technology against pod borer (Armiigera vitrata) and developing Bt cowpea lines. Developing non-conventional breeding strategies for transferring desired traits from wild sp. C. serreta L. into cultivated sp. C. tetragonoloba L. for early maturity, more pods and resistance against BLB. 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Journal of Crop Sciences 181: 209-214. Journal of Food Legumes 25(4): 273-278, 2012 Transferability of cowpea and azuki bean derived SSR markers to other Vigna species RAVINDRA BANSAL, SUDHIR KUMAR GUPTA and T. GOPALAKRISHNA Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre, Mumbai – 400 085, India; E-mail: [email protected] (Received: July 10, 2012; Accepted: December 18, 2012) ABSTRACT The genus Vigna contains many important grain legumes in the tropical and sub-tropical regions. Among the tropical grain legumes, the most studied species of Vigna are the cowpea and azuki bean. Also genomic resources of only these two Vigna species are available that can be further used for genomic analysis of other Vigna species. This study represents the transferability study of cowpea and azuki bean SSR markers to other Vigna species. Fifty SSR markers developed in cowpea using sequences available in cowpea CGKB database and 95 azuki bean SSR markers from other literature were checked for transferability to nine other Vigna species. It was found that 17 (34%) cowpea SSR and 61 (64.21%) azuki SSR markers were transferable to other Vigna species. These SSR markers derived from cowpea and azuki bean would serve as a valuable tool for genetic analysis and marker assisted selection of agronomically important traits in other Vigna species like blackgram, mungbean, rice bean etc. for which there is either little or no sequence information is available. Key words: Azuki bean, Cowpea, SSR markers, Transferability, Vigna species The genus Vigna contains the most important legumes in the sub-tropical and tropical regions. The legume genus Vigna comprises about 75-80 species originating from regions of Africa and Asia. The genus has been sub-divided into 7 subgenera based on their centers of origin (Marechal et al. 1981). Many Vigna species are cultivated for food. Many of them are valued as forage, cover and green manure crops in many parts of the world. Annual worldwide production of the various Vigna species is likely to approach 20 million hectares and virtually all of this production is in developing countries (Richard 2002). Azuki bean (Vigna angularis) is an annual food legume and is considered to have been domesticated in China, Korea or Japan from its wild ancestral form, V. angularis var. nipponensis (Ohwi) Ohwi & Ohashi. A number of SSRs have been developed in adzuki bean. Cowpea [Vigna unguiculata (L.) Walp] is also one of the most important crops and predominantly a hot weather crop. It is more tolerant than the other legume to drought, water logging, infertile soils, and soil acidity stress. They are widely grown in eastern Africa and south-east Asia primarily as a leafy vegetable (Richard 2002). Simple sequence repeats or microsatellite repeats are defined as regions within DNA sequences where short sequences (1-6 bp; monomers to hexamers) are repeated in tandem array. These markers are also termed simple sequence length polymorphism (SSLP), short tandem repeats (STRs), and simple sequence repeats (SSRs) or sequence-tagged microsatellite site (STMS). SSR markers are widely used in gene mapping (Tanksley et al 1995), analysis of genetic diversity (Wang et al 2006), and marker-assisted selection in breeding programs (Sun et al 2006). Uniform abundance in the genome, codominant in nature, locus specificity and high reproducibility of these markers make them very suitable to check the transferability. Genomic resources are very limited in Vigna species (Gupta and Gopalakrishna 2008; Datta and Gupta 2009) and are main hurdle in their improvement. However, because of high rate of transferability, SSR markers developed in one species can be effectively used in other related species (Souframanien and Gopalakrishna 2 009, Gupta and Gopalakrishna 2010). Therefore, aim of the present study was to develop the SSR markers in cowpea, and evaluate the transferability of these cowpea SSRs and already reported azuki bean SSR markers to other Vigna species. MATERIALS AND METHODS Sequences retrieval and primer designing: Cowpea gene space sequences were downloaded from Cowpea Genespace/ Genomi cs Knowl edge Base (http: // cowpeagenomics.med.virginia.edu/CGKB/). SSR locator programme were used to find tandem nucleotide repeat in the sequencese. Primer3 software (http://frodo.wi.mit.edu/primer3/) was used to design the PCR primer. For designing primers, user defined parameters was used viz. optimum primer length was 20 mer (range was 18-25 mer), optimum annealing temperature was 60°C (range was 55-62°C), optimum GC content was 50% (range was 30-80%) and rest of the parameters had the default value. The azuki bean SSR primers used in the study were taken from Han et al. (2005). Plant material and DNA extraction: Ten Vigna spp. genotypes were used in the study are listed in Table 1. Total genomic DNA was extracted from young seedlings using the modified CTAB method (Doyle and Doyle 1987). The quality 274 Journal of Food Legumes 25(4), 2012 Table 1. List of Vigna species used in the study S. N. Vigna species used in the study* Source 1 Vigna unguiculata (V-240) NBPGR, India 2 Vigna vexillata NBPGR, India 3 Vigna aconitifolia TNAU, India 4 Vigna trilobata NBPGR, India 5 Vigna glabrescence NBPGR, India 6 Vigna umbellata (EC-634639) NBPGR, India 7 Vigna angularis (EC-634633) AVRDC, Taiwan 8 Vigna mungo (TU 94-2) BARC, India 9 Vigna radiata var. radiata (TMB-37) BARC, India 10 Vigna radiata var. setulosa NBPGR, India * Number in the paranthesis indicate accession number/cultivar of DNA was checked on 1% agarose gel and the quantity was determined using UV spectrophotometer (Unicam UV 300, UK). SSR marker analysis: PCR reactions were performed in 20 µl volume containing 10 mM Tris-HCl (pH 9.0), 50 mM KCl, 1.5 mM MgCl 2, 0.2 mM of each dNTP, 0.5 unit Taq DNA polymerase (Bangalore Genei, Bangalore, India), 50 ng template DNA, 20 ng each of forward and reverse primer. PCR amplifications were performed in an Eppendorf Mastercycler Gradient (Eppendorf, Hamburg, Germany) using the following thermal profile: 1 cycle of 95°C for 2 min, followed by 39 cycles of 94°C for 30 s, 50-65°C for 30 s (depending on primer), 72°C Table 2. List of cowpea SSR markers developed and used in the study Primer name VuM-01F VuM-01R VuM-02F VuM-02R VuM-03F VuM-03R VuM-04F VuM-04R VuM-05F VuM-05R VuM-06F VuM-06R VuM-07F VuM-07R VuM-08F VuM-08R VuM-09F VuM-09R VuM-10F VuM-10R VuM-11F VuM-11R VuM-12F VuM-12R VuM-13F VuM-13R VuM-14F VuM-14R VuM-15F VuM-15R VuM-16F VuM-16R VuM-17F VuM-17R VuM-18F VuM-18R VuM-19F VuM-19R VuM-20F VuM-20R VuM-21F VuM-21R VuM-22F VuM-22R VuM-23F VuM-23R VuM-24F VuM-24R VuM-25F VuM-25R Primer sequence (5’….3’) AACAAGATGTGGCATGCTGA TGAAAACGGAAAAGGGATCA GAAACTAGCACCAAATCCAACA GAGCAAAAGCCTCCATCACT GCACCCAATCAAACACACAC GAAGCGGATTTGAGAGTTGG GCAGGGGCAACAATACATTA GTTGGACTACCCCAAATGCT GCGGGATTCTATTCCAGTGA TCCATTGGGTTTCTCAACCT TGAAAGTTGAGAAGGGGACAA CATTCAGGTTCAGCTCACGA TGTTTCCAACAGGATTAGCC AAGGCCAATAATTGCACAAG TCAAAAACACAGGTCCTCCA CATCCCGTGAAATTCAACAA TTGAGCACAAGTTCATCGAG TGATTGCCTAAACGACACAC TCAAAACTTCAACCCAGACA AAAAAGGAAGTCCATTGCTC GGGCAGGAGCTGCATATAAC CCTGCAACAACAAAAATGGA CATGGCAATTTGCAACAAAG CTAAAGTGCCGTGACGATGA ACTCAACGTGTGTGAATAGGC CCCTCACAAGAAGAAACAGAA CGGGCAAGATAACCAATTAGAC AGTTGTCAGACCAACCTGCAT ATGTTGCTGGACAAATCTCTGA TGTGCCAACTGATTCTCTGC GGACATTTCCGGATGTCAAC CTTTGCCATTCACTTTCACG GGATATCATAGCAAGTCGAA AAGGAGTGCATCCTAAACTC GGCACCCCAGTTCAGGAT TTGCGAACTTGTTCATGTGG AGAACCCAGCATACCTGCAT CCTCGCCAATGATTCTGAG CCAAGAGGAAAAGGTATCAGACA GCATTCTTGCACAAGGAGTCT AAACCAGATGTCTCTGTTTCTTCTC GCGTAACACAGGCGTTATCA CAATCACCATTCACCAAACA TATTGGGACTCAGGTCTTGG CGTACCTAATGTGAAGGTTCGTT AAGGCAAAAAGCTCTTGCAG GGTTTATCACCACCTCAACA CGATGAATTTTAGCCATCAG AGGGATGAGTTCCTTCAACG AAGAAGTGGTGAGGGCACAG Primer name VuM-26F VuM-26R VuM-27F VuM-27R VuM-28F VuM-28R VuM-29F VuM-29R VuM-30F VuM-30R VuM-31F VuM-31R VuM-32F VuM-32R VuM-33F VuM-33R VuM-34F VuM-34R VuM-35F VuM-35R VuM-36F VuM-36R VuM-37F VuM-37R VuM-38F VuM-38R VuM-39F VuM-39R VuM-40F VuM-40R VuM-41F VuM-41R VuM-42F VuM-42R VuM-43F VuM-43R VuM-44F VuM-44R VuM-45F VuM-45R VuM-46F VuM-46R VuM-47F VuM-47R VuM-48F VuM-48R VuM-49F VuM-49R VuM-50F VuM-50R Primer sequence (5’….3’) TTTTAAGCATTGCCACCAGA AACAACAACCGCATATCCCTA TCCATCCACCATTTTCCATC ATGGGAATGCCCGAGAGT TAGAACACTCTTGGGGGTTA CGGAGAAAGAGGAAGTACAA TTTTTCTCGACACACGGTGA TTTCCCCCTCTCTCACACAC TGCAACATCCACTAATAGACCA TTGCTCAACATAAAGGACGAC TGGTTCACTTCCCATATTGTCA AGGCAGAGACGAAGGAGTGA AATGAAATCAGCCCAAGGAA ATGGCTTTTGTCTTGCCTTC AAAGGTGGGGGATTATGAGG TGTCCAATCCTGATGGATGA CCTGATGGATTTACAGACATGC GGTGAGGGCAATACCTGTGT AAGACTTTCGTGGTGCAGGT AAGTGGCATGGAAGATGGAG TGTGCCAAAAGGAAAAGACA GGGATGGTATGTTCCTCACG TCATTGGTACGTTCAAAGCAA TGGATCCCTACTCAATTTCTCC TGCTTAAAGGAGAAATACTCGACTT CTGTCCTCATGTTGAAAACCTCT CGAAAAAGCATGATCAACCA CCCCTTTCGCTAAAATTTCC TTCTACATGGTTTTGGGGTCA GAGCTTGCCCTCAAGAATTG TGAGGTGTGCACTTTTAACTCC TTCTCACACATACACACGCAAT ATGTTATACGCCGGCAAAGT TCTGGGTGCTTTGGAAAATC AGCTTTGCACTAATCCATCTTAGTC CAAGATCATTTTTCGCGACTC ACCAAACCATCCGTGAAGTG TGGTTGTCCACGAATATGTGTC TGGTTGGAAGTCTCACATCAA GCATATGCATCTCGTATGTAGGTC TTTGGTTTCACATGTTGAGG TTCTTGGGAATATGTTCAGG TGTTTTTCGCTATGCCTCAA GGCAGCTAGATTCGTCCTTG ACCTACTCACGAATATCCACAG CACCGATAATCTCCAAAACA TGAGATTGGTGTTGAATGCT CAATGAACTAAACCCCTTCTTC TTAGGGACCAAAAGGAATGA TAATCGCACACATCAGCCTA Bansal et al.: Transferability of cowpea and azuki bean derived SSR markers to other Vigna species 275 Table 3. Cross-species transferability of cowpea SSR markers to other Vigna species S.N. Primer V. vexillata name 1. VuM5 + 2. VuM7 3. VuM8 4. VuM11 + 5. VuM15 + + 6. VuM18 7. VuM19 + + 8. VuM25 9. VuM37 10. VuM40 + 11. VuM44 + + 12. VuM51 13. VuM57 + 14. VuM58 + 15. VuM59 + 16. VuM60 17. VuM61 + V. aconitifolia + + + + + + + + + + + + V. trilobata + + + + + + + + + + + + V. glabrescence + + + + + + + + + + + + + + V. umbellata + + + + + + + + + + + + + V. angularis + + + + + + + + + + + + V. mungo + + + + + + + + + + + + + + V. radiata var. radiata + + + + + + + + + + + V. radiata var. setulosaa + + + + + + + + + + + Table 4. List of azuki bean SSR markers used in the study S. N. 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17. 18. 19. 20. 21. 22. 23. 24. 25. 26. 27. 28. 29. 30. 31. 32. Primer No. CEDCAA1 CEDG2 CEDAAT2 CEDAAG4 CEDG5 CEDGAT8 CEDC8 CEDG11 CEDC11 CEDC14 CEDG15 CEDG16 CEDC16 CEDG18 CEDG21 CEDG22 CEDG23 CEDG24 CEDC28 CEDG30 CEDG33 CEDC33 CEDG35 CEDC35 CEDG36 CEDG37 CEDG40 CEDG41 CEDG42 CEDC55 CEDG59 CEDG63 Serial No. 33. 34. 35. 36. 37. 38. 39. 40. 41. 42. 43. 44. 45. 46. 47. 48. 49. 50. 51. 52. 53. 54. 55. 56. 57. 58. 59. 60. 61. 62. 63. 64. Primer No. CEDG66 CEDG71 CEDG73 CEDG80 CEDG81 CEDG98 CEDG99 CEDG100 CEDG106 CEDG107 CEDG112 CEDG114 CEDG117 CEDG121 CEDG124 CEDG125 CEDG131 CEDG132 CEDG134 CEDG146 CEDG147 CEDG150 CEDG158 CEDG165 CEDG171 CEDG172 CEDG174 CEDG184 CEDG186 CEDG187 CEDG191 CEDG193 for 1 min and a final extension of 72°C for 7 min. PCR products were separated on 2% agarose gel using 1X Tris-borate-EDTA (TBE) buffer, stained with ethidium bromide and photographed in a gel documentation system (Syngene, UK). Serial No. 65. 66. 67. 68. 69. 70. 71. 72. 73. 74. 75. 76. 77. 78. 79. 80. 81. 82. 83. 84. 85. 86. 87. 88. 89. 90. 91. 92. 93. 94. 95. Primer No. CEDG195 CEDG196 CEDG197 CEDG198 CEDG201 CEDG202 CEDG203 CEDG205 CEDG212 CEDG215 CEDG232 CEDG238 CEDG243 CEDG244 CEDG245 CEDG247 CEDG251 CEDG253 CEDG257 CEDG259 CEDG261 CEDG262 CEDG265 CEDG269 CEDG270 CEDG280 CEDG285 CEDG286 CEDG294 CEDG298 CEDG305 RESULTS AND DISCUSSION Transferability of cowpea SSRs within genus Vigna: A total of 50 cowpea SSR markers were developed in this study (Table 2) and were screened on nine other Vigna species to determine 276 Journal of Food Legumes 25(4), 2012 Table 5. Cross-species transferability of azuki bean SSR markers to other Vigna species S. N. Primer name V. unguiculata V. vexillata V. aconitifolia V. trilobata V. glabrescence V. umbellata V. mungo 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17. 18. 19. 20. 21. 22. 23. 24. 25. 26. 27. 28. 29. 30. 31. 32. 33. 34. 35. 36. 37. 38. 39. 40. 41. 42. 43. 44. 45. 46. 47. 48. 49. 50. 51. 52. 53. 54. 55. 56. 57. 58. 59. 60. CEDG2 CEDAAG4 CEDGAT8 CEDC8 CEDC11 CEDG11 CEDG15 CEDG21 CEDG24 CEDG35 CEDG36 CEDG37 CEDG40 CEDG42 CEDC55 CEDG59 CEDG66 CEDG71 CEDG73 CEDG81 CEDG98 CEDG99 CEDG100 CEDG106 CEDG107 CEDG114 CEDG117 CEDG121 CEDG124 CEDG132 CEDG146 CEDG147 CEDG165 CEDG171 CEDG172 CEDG174 CEDG186 CEDG196 CEDG197 CEDG198 CEDG201 CEDG202 CEDG203 CEDG205 CEDG212 CEDG215 CEDG243 CEDG245 CEDG247 CEDG251 CEDG257 CEDG259 CEDG261 CEDG262 CEDG265 CEDG269 CEDG270 CEDG285 CEDG298 CEDG305 + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + V. radiata var. radiata + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + V. radiata var. setulosa + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + Bansal et al.: Transferability of cowpea and azuki bean derived SSR markers to other Vigna species Fig 1. Azuki bean markers SSR markers CEDGAT8 & CEDC550 showing trans ferability to other Vig na species (The species number is according to Table 2). their transferability. Out of 50 primer pairs examined, only 17 primer pairs were found to give clear amplification. Out of 17 cowpea SSR markers tested, 4 gave amplification in all Vigna species. Around 9 gave amplification in 8 species and 5 gave amplification in 7 species. Whereas, only 2 sho wed amplification in 6-5 species and 4 were found to be transferable in less than 5 species. The cross species transferability pattern of cowpea SSR markers is given in Table 3 and PCR amplification of cowpea SSR markers on Vigna species is shown in Fig 1. Transferabilty of azuki bean SSRs within genus Vigna: To determine azuki SSR marker transferability, 95 SSR markers (Table 4) were screened on nine other Vigna species. Out of 95 primer pairs examined, 61 primer pairs were found to give clear amplification. Out of 61 azuki bean SSR markers, 28 amplified in all Vigna species, thirteen gave amplification in 8 species and nine showed amplification in 7 species, and 8 primer pairs gave amplification in 6-5 species. Only 3 primers showed amplification in less than 5 species. The cross species transferability pattern of azuki bean SSR markers is given in Table 5 and PCR amplification of azuki bean SSR markers on Vigna species is shown in Figure 2. In many genetic studies, one of the major rate limiting steps is the development of markers for use in a new study system. The study gets more complicated when there is no or very little information about the genome sequence is available for the development of markers. Moreover many marker systems are more or less species specific, limiting their use in other related species. Because of high transferability of SSR markers, these problems can be minimized. The ability to use the same microsatellite primers in different plant species depends on the extent of sequence conservation in the primer binding sites flanking the microsatellite loci and the stability of these sequences during evolution (Choumane et al. 2000, Decroocq et al. 2003). Microsatellite primer pairs used in the current study originated from cowpea and azuki bean, and were found to be transferable to other Vigna species. This indicates the conservation of microsatellite sequences between the Vigna species during evolution. The conservation of microsatellite sequences also has been observed across other legumes (Choumane et al. 2000, Ford et al. 2002, Phansak et al. 2005). However, the transferability of azuki derived SSR Fig 2. 277 Cowp ea SSR markers VuM7 & VuM18 sho wing transferability to other Vigna species (The species number is according to Table 2). markers was high (63%) compared to cowpea SSR markers (34%). This may be because cowpea belongs to the sub genus Vigna, and azuki bean and other species like mungbean, blackgram, rice bean, moth bean belongs to the subgenus Ceratotropis of the genus Vigna,. Thus, compared to cowpea, phylogenetically azuki bean is closer to other Vigna species and therefore, markers developed from azuki bean showed more transferability. These results indicate that these microsatellite markers will be helpful for genetic analysis of other Vigna species like mungbean, black gram, rice bean etc. for which sequence information is not available for SSR marker development. The transferability of microsatellite markers across species may increase their utility and potentially decrease the development cost. The microsatellite markers conserved between the species also serve as a valuable tool for comparative mapping studies (Dirlewanger et al. 2004, Yu et al. 2004, Gupta et al. 2008). REFERENCES Choumane W, Winter P, Baum M and Kahl G. 2000. 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RFLP and microsatellite mapping of a gene for soybean mosaic virus resistance. Phytopathology 84: 60-64. Journal of Food Legumes 25(4): 279-281, 2012 Genetic diversity studies in blackgram (Vigna mungo L. Hepper) M. SRIMATHY, M. SATHYA and P. JAYAMANI* Department of Pulses, Centre for Plant Breeding and Genetics, Tamil Nadu Agricultural University, Coimbatore 641003, India; E-mail: [email protected] (Received: February 18, 2012 ; Accepted : January 05, 2013) ABSTRACT Divergence analysis of 46 genotypes including 20 genotypes of blackgram and 26 accessions of V. mungo var. silvestris, a wild progenitor species for eleven biometrical traits was carried out using Mahalanobis D2 statistics. The genotypes were grouped into twelve clusters. The cluster I was the largest with 25 accessions of V. mungo var. silvestris while, other clusters consisted of two cultivated genotypes. Cluster XII had only one accession viz., V. mungo. var. silvestris acc 10. This study showed clear grouping of V. mungo var. silvestris accessions from the cultivated blackgram (V. mungo) genotypes. Cluster XI recorded the maximum intra cluster distance of 12.58 followed by cluster X with a distance of 11.39. The highest inter cluster distance was found between cluster IX and cluster XII (28.71) followed by cluster XI and XII (23.79) and cluster V and XII (23.30). Based on cluster mean and divergence, it was concluded that the hybridization between accessions of V. mungo var. silvestris in clusters I and XII and cultivated genotypes in the other clusters could produce desirable recombinants for plant type, important economic traits and grain yield. Key words: Blackgram, Cluster analysis, D2 analysis, Genetic divergence, V.mungo var. silvestris Blackgram (Vigna mungo (L.) Hepper) popularly known as urdbean or mash, is a grain legume domesticated from V. mungo var silvestris (Chandel, 1984). This wild progenitor is resistant to bruchid infestation and also tolerant against abiotic stresses. Blackgram is a rich source of protein (20.8 to 30.5 per cent) with total carbohydrates ranging from 56.5 to 63.7 per cent. It is also a good source of phosphoric acid and calcium. India is the largest producer and consumer of blackgram in the world. It is the fourth important pulse crop in India, cultivated as a sole crop and intercrop covering an area of about 3.24 million hectares and producing 1.46 million tons. However, the productivity is very low with 526 kg/ha (Anonymous, 2010). Many breeding efforts have been carried out to improve the yield level of this crop and to break the yield plateau, but it could not be done because of narrow genetic base of parents used in hybridization. Genetic diversity is an important factor and also a prerequisite in any hybridization programme. Inclusion of diverse parents in hybridization programme serves the purpose of producing desirable recombinants. Multivariate analysis by means of Mahalanobis D2 statistic is a powerful tool in quantifying the degree of divergence at genotypic level. Therefore, an attempt has been made in the present investigation with a view to estimate genetic divergence among a set of 46 genotypes including cultivated genotypes of blackgram and its wild progenitor accessions for eleven biometrical traits. MATERIALS AND METHODS Forty six genotypes of cultivated blackgram and its wild progenitor, collected two decades ago, were evaluated at Department of Pulses, Tamil Nadu Agricultural University, Coimbatore in a randomized block design (RBD) with two replications. Each genotype was sown in paired rows of four meter length with a spacing of 30 x 10 cm. Recommended package of practices were followed to raise a healthy crop. Five randomly taken plants were considered to record data for days to 50 per cent flowering, days to maturity, plant height (cm), number of branches per plant, number of clusters per plant, number of pods per cluster, number of pods per plant, pod length (cm), number of seeds per pod, 100 seed weight (g) and yield per plant (g). The mean values of five plants were taken for the analysis of genetic divergence following Mahalanobis (1936). The genotypes were grouped into different clusters following Tocher’s method as described by Rao (1952). RESULTS AND DISCUSSION Genetic diversity is the basic requirement for successful breeding programme. Collection and evaluation of germplasm lines and genotypes of any crop is a pre-requisite for any programme, which provides a greater scope for exploiting genetic diversity. The multivariate analysis (D2) is a powerful tool to measure the genetic divergence within a set of genotypes (Murthy and Arunachalam, 1966). The present study was planned to examine the trend of genetic divergence in 20 genotypes of cultivated blackgram and 26 accessions of V.mungo var. silvestris, a wild progenitor species. The genotypes were grouped into twelve clusters indicating large amount of genetic diversity among the genotypes (Table 1). Elangaimannan et al. (2008) also reported grouping of 55 blackgram genotypes into seven clusters, where cluster I was the largest (34 genotypes) followed by clusters IV (eight genotypes), II (six genotypes), V (four genotypes), while rest of the clusters had one genotype each. Grouping of accessions by multivariate method in the present study is of practical 280 Journal of Food Legumes 25(4), 2012 value to the breeders. Representative accessions may be chosen from particular cluster for hybridization programme. In the present study, cluster I was the largest with 25 accessions of V. mungo var silvestris while, other clusters consisted of two genotypes each except cluster XII which had only one accession viz., V. mungo var silvestris acc. 10. High level of variability was observed for several morphological and biometrical traits among the accessions of V. mungo var silvestris in cluster I. Even though all the accessions of V. mungo var silvestris formed a single cluster, a good level of variability was also observed in the mean values of different traits. Similar kind of separate grouping of cultivated genotypes and V. mungo var silvestris accessions was observed in a dendrogram based on SSR analysis (data not shown). Contribution of various biometrical characters towards genetic divergence is presented in Table 3. Among the characters studied, yield per plant contributed maximum towards divergence, followed by number of pods per cluster, 100 seed weight and days to 50 per cent flowering. Similar results were also reported earlier by Ghafoor and Ahmed (2005), Konda et al. (2007), Shanthi et al. (2006). Plant height and number of branches per plant contributed minimum to the genetic divergence leading to the inference that in general, the variability for these characters are low in blackgram genotypes used in this study. The intra and inter cluster D2 values among the clusters are presented in the Table 2. Cluster XI recorded the maximum intra cluster distance of 12.58 followed by cluster X with a distance of 11.39. There was one solitary cluster (cluster XII) possessing no intra cluster value. This accession had smaller seed size and found to be resistant to bruchid infestation. The highest inter cluster distance was found between cluster IX and cluster XII (28.71) followed by cluster XI and XII (23.79), cluster VII and XII (23.35) and cluster V and XII (23.30). The inter cluster distance of all other clusters with cluster XII showed higher values when compared to the inter cluster distance between other clusters. The least inter cluster distance was found between cluster II and IV (6.19). The mean values of 11 characters for twelve clusters are presented in the Table 3. Cluster IV recorded the highest mean value (38.00) for days to 50 % flowering followed by cluster XII (37.50) and clusters VII and X (37.00), whereas cluster XI recorded the lowest mean value for days to 50 % flowering (31.50). Cluster IV recorded the highest mean value for days to maturity (68.00) followed by cluster XII (67.50) and clusters VII and X (67.00), while Cluster XI recorded the minimum days to maturity (61). Cluster IV recorded the maximum plant height of 30.88 cm followed bycluster XII (29.77 cm), while cluster XI recorded the minimum plant height of 20.22 cm. The highest mean value for number of branches per plant was recorded by cluster VI and XII (1.84) and the lowest was recorded by cluster XI (1.42). The highest mean for number of clusters per plant was recorded by cluster V (13.00) followed by cluster VII (12.00) Table 1. Constitution of D2 clusters of 46 genotypes of blackgram Cluster number Number of genotypes Name of the genotype I 25 Vigna mungo var. silvestris acc 1, acc 2, acc 3, acc 4, acc 5, acc 6, acc 7, acc 8, acc 9, acc 11, acc 12, acc 13, acc 14, acc 15, acc 16, acc 17, acc 18, acc 19, acc 20, acc 21, acc 22, acc 23, acc 24, acc 25, acc 26, II 2 AC-305, PLS-44 III 2 Cotton leaf – 32, K 951 IV 2 P-169, Co 5 V 2 P-226, PLS 364/92 VI 2 P-123, T9 VII 2 P-153, VBN (Bg) 4 VIII 2 P-307/1-1/, VBN 3 IX 2 P-202, Co-02/103 X 2 AC-43, VBN (Bg) 5 XI 2 P 133/13, CoBG 653 XII 1 V. mungo var. silvestris acc 10 Table 2. Average intra (in bold) and inter cluster D2 distances Cluster I II III IV V VI VII VIII IX X XI XII I 8.95 II 12.86 3.21 III 11.18 6.31 4.24 IV 12.18 6.19 6.98 4.35 V 14.86 9.85 8.42 9.49 4.82 VI 10.24 9.49 7.97 9.31 10.55 5.73 VII 15.45 6.87 9.37 6.63 8.27 12.04 5.81 VIII 11.56 6.23 6.99 8.63 9.83 8.59 9.38 8.16 IX 17.53 9.21 11.55 10.79 10.45 11.60 9.19 10.86 9.94 X 12.71 9.44 7.75 8.42 11.68 10.04 11.73 10.17 13.80 11.39 XI 11.90 12.33 11.69 13.58 12.67 9.36 14.62 10.67 14.31 14.60 12.58 XII 17.71 23.16 20.63 20.16 23.30 22.77 23.35 22.88 28.71 21.42 23.79 0.00 Srimathy et al.: Genetic diversity studies in blackgram (Vigna mungo L. Hepper) 281 Table 3. Cluster mean values for 11 biometrical characters in blackgram Cluster I II III IV V VI VII VIII IX X XI XII Contribution of traits toward divergence Days to 50% flowering 34.16 36.25 35.50 38.00 34.25 34.50 37.00 34.00 35.75 37.00 31.50 37.50 4.54 63.96 66.25 65.50 68.00 64.25 64.50 67.00 64.00 65.75 67.00 61.50 67.50 Plant height (cm) 27.47 23.33 23.00 30.88 22.43 29.20 29.67 26.96 23.58 27.71 20.22 29.77 Number of branches/ plant 1.62 1.42 1.67 1.58 1.67 1.84 1.67 1.57 1.83 1.59 1.42 1.84 0.39 0.10 0.19 Days to maturity Number of Number of Number of clusters/ pods/ pods/ plant cluster plant 7.72 4.51 34.99 8.50 3.46 29.50 6.42 2.84 18.77 9.50 3.59 34.52 13.00 2.59 28.67 8.50 4.22 36.50 12.00 2.90 34.50 8.84 3.70 26.00 11.50 3.35 39.50 8.09 4.21 23.25 10.67 4.14 41.80 5.73 2.50 26.00 1.06 17.10 5.80 Pod length (cm) 4.04 4.86 4.64 4.44 4.58 4.54 4.54 4.46 4.72 4.55 4.59 3.15 0.48 Single Number of 100 seed plant yield seeds/pod weight (g) (g) 5.66 3.96 6.88 6.00 5.13 5.16 5.78 4.68 5.92 5.98 4.80 6.71 5.88 4.90 8.53 6.10 4.88 8.91 6.08 5.18 6.09 5.78 4.98 5.51 5.80 5.80 8.14 5.85 4.75 7.24 5.63 4.58 8.43 5.30 2.00 4.32 0.68 17.29 52.37 and maximum mean of number of pods per cluster was recorded by cluster I (4.51) while, the lowest mean for number of pods per cluster was recorded by cluster XII (2.50). The highest mean value for number of pods per plant was recorded by cluster XI (41.80) and cluster III recorded the lowest mean (18.77). mungo var silvestris can be used in inter sub-specific hybridization program to transfer genes for resistance to biotic and tolerance to abiotic stresses, improved plant type and also to broaden the genetic base in blackgram. Cluster II recorded the maximum mean value for pod length (4.86 cm) followed by cluster IX (4.72 cm) and the lowest pod length was recorded by cluster XII (3.15 cm). Cluster VI recorded the maximum value for number of seeds per pod (6.10) followed bycluster VII (6.08) while, cluster XII recorded the lowest mean value for number of seeds per pod (5.30). Cluster IX recorded the maximum hundred seed weight (5.80 g) and cluster VI recorded maximum single plant yield (8.91 g) while, Cluster XII recorded lowest mean value for hundred seed weight (2.0 g) and single plant yield (4.32 g). The accession V. mungo var silvestris acc 10 recorded lowest mean value for most of the traits viz., number of clusters/plant, number of pods/cluster, pod length, 100 seed weight and yield/plant. The uniqueness of the accession could be the reason for the formation of separate cluster XII, when compared to all other accessions of V. mungo var silvestris in the cluster I. Anonymous. 2010. Project Coordinator ’s Report. AICRIP on MULLaRP. IIPR, Kanpur. Pp-20. From the present investigation, it was concluded that blackgram displayed a wide range of diversity for few traits and there were few accessions with unique characters. Vigna mungo var silvestris accessions were distinctly separated from the other blackgram genotypes. Hence, the accessions of V. REFERENCES Chandel KPS. 1984. Role of wild Vigna species in the evolution and improvement of mung (Vigna radiata (L.) Wilczek) and urdbean (V. mungo (L.) Hepper). Annals of Agricultural Research 5: 98-111. Elangaimannan R, Anbuselvan Y and Karthikeyan P. 2008. Genetic diversity in blackgram (Vigna mungo (L.) Hepper). Legume Research 31 (1): 57-59. Ghafoor A, Sharif A, Ahmed Z, Zahid MA and Rabbani MA. 2001. Genetic diversity in blackgram (Vigna mungo (L.) Hepper). Field Crops Research 69: 183-190. Konda CR, Salimath PM and Mishra MN. 2007. Genetic diversity in blackgram (Vigna mungo (L.) Hepper). Legume Research 30 (3): 212-214. Mahalanobis PC. 1936. On the generalized distance in statistics. Proc. Natl. Acad. Ins India. 12: 49-55. Murthy BR and Arunachalam V.1966. The nature of genetic divergence in relation to breeding system in crop plants. Indian Journal of Genetics 26: 188-189. Rao CR. 1952. Advanced statistical methods in Biometrical Research. John Wiley and sons Inc., New York. Shanthi P, Jebaraj S and Manivannan N. 2006. Genetic diversity in urdbean (Vigna mungo (L.) Hepper). Legume Research 29: 181185. Journal of Food Legumes 25(4): 282-285, 2012 Stability analysis for seed yield and its component traits in rice bean (Vigna umbellata) WUNGSEM RUNGSUNG and S.A.P.U. CHANGKIJA Department of Genetics and Plant Breeding, SASRD, Medziphema-797106, Nagaland, India; E-mail: [email protected] (Received: June 25, 2012; Accepted: October 25, 2012) ABSTRACT Fifty germplasm lines of rice bean were evaluated in a randomized complete block design (RCBD) at three locations during kharif season for two consecutive years. Stability analysis was carried out as per Eberhart and Russell (1966) model. The pooled analysis of variance due to genotypes was found highly significant for all the characters indicating the presence of considerable genetic variability among the genotypes. Highly significant pooled deviation for all the characters except days to flowering, pods/cluster, pod length, seeds/pod and seed weight was observed suggesting that the performance of the various genotypes under study fluctuated significantly from their respective linear path of response to environments. The genotype × environment interactions for most of the characters were also highly significant revealing differential interactions of genotypes with changes in the environments. Among all the genotypes studied, ‘NRB-34’ and ‘NRB-35’ were found most stable for yield/plant. Key words: G × E interaction, Rice bean, Seed yield, Stability analysis Rice bean, a leguminous crop and also known as climbing mountain bean, mambi bean and oriental bean, is native to south-east Asia (Bolivar and Luis 2010). The crop possesses excellent food and fodder values and is grown for fodder, green manure and cover crop. The dry seeds are eaten boiled as dhal (soup) and young immature pods are consumed as vegetables (Gupta et al. 2009). Rice bean, thus, occupies an important place in the Indian food system, and studies conducted so far on the bean revealed the existence of high genetic variability. An investigation on its stability analysis will, therefore, help in sorting out the most promising and stable genotypes from the genetically variable populations. Therefore, the present investigation was taken up for stability analysis of seed yield and its component traits in some of the rice bean germplasm lines. MATERIALS AND METHODS The experimental materials consisted of a set of 50 germplasm lines of rice bean. The seeds of these lines were sown and raised as kharif crop on dry terraces under rain-fed and normal sown conditions in the randomized complete block design (RCBD) with three replications under the same set of agronomic and cultivation practices at three locations, namelyMedziphema, Chumukedima and Kohima (Nagaland) during 2009 and 2010. Six competitive plants (two from each replication) from each germplasm line were randomly selected. Data were recorded on seed yield (g) and its component traits, viz., days to flowering, days to maturity, plant height, clusters/ plant, pods/plant, pods/cluster, pod length (cm), seeds/pod, biomass/plant (g) and 100-seed weight (g), were observed and recorded at different phenological events. Stability analysis was carried out as per Eberhart and Russell (1966) using SPAR-2 (Statistical Package for Agricultural Research) developed at the Indian Agricultural Statistics Research Institute, New Delhi by Ahuja et al. (2005). The mean and deviation from regression of each genotype were considered for stability, and linear regression was used for testing the varietal response: (i) genotypes with high mean, bi =1 and non-significant S2d (not significantly deviating from zero) were considered ‘average responsive’ (adaptable or suitable over all environmental conditions), (ii) genotypes with high mean, regression coefficient greater than unity (bi>1) and nonsignificant S2d were rated ‘highly responsive’ (suitable for favourable environments but yielding poor in unfavourable environments), (iii) genotypes with high mean, bi<1 with nonsignificant S2 d were ‘low responsive’ (not favourably responsive to improved environmental conditions and hence could be regarded as specifically adapted to poor/unfavourable environments) and (iv) genotypes with any bi value with significant S2d were unstable. RESULTS AND DISCUSSION The development of varieties/genotypes, which can be adapted to a wide range of diversified environments, is the ultimate goal of plant breeders in any crop improvement programme (Danyali et al. 2012) and it is imperative to evaluate varieties/genotypes over different environments to ascertain their consistency and stability of performance before some potential genotypes are released for commercial cultivation (Patel and Acharya 2011). The pooled variance due to genotypes was found highly significant for all the characters (Table 1). Highly significant pooled deviation for all the characters except days to flowering, pods/cluster, pod length, seeds/pod and seed weight was o bserved. These observations were in close agreement with the findings of Yan et al. (1995) who observed highly significant pooled deviation for days to maturity, clusters/plant, pods/cluster, pods/plant and yield in French bean genotypes evaluated under contrasting soils. This highly significant pooled Rungsung and Changkija: Stability analysis for seed yield and its component traits in rice bean (Vigna umbellata) 283 Table 1. Pooled analysis of variance (mean squares) for stability of various characters in rice bean genotypes Sources of Variation Df Genotype (G) 49 5 Environment(E) G x E (linear) 245 Pooled deviation 200 Pooled error 600 Days to Days to Plant height Pods/ (cm) flowering maturity Cluster 425.00** 53.04** 1.11 1.00 2.32 888.37** 12137.83** 77.01** 165.91** 1.89** 18.57** 2.38** 14.77** 2.90 18.58 2.29** 0.76 0.01 0.01 0.02 Mean squares (MS) Clusters/ Pods/ Pod plant length plant (cm) 419.88** 8450.27** 13.76** 34.77** 2276.67** 0.67 1.96** 37.48** 0.01 1.69** 24.79** 0.02 1.84 42.41 0.02 Seeds/ Biomass/ 100-seed Seed yield/ plant weight Pod Plant (gm) (gm) (gm) 6.55** 252.25** 365.12** 40016.45** 0.39 119.39** 0.71 4333.92** 0.03 4.35** 0.06 169.50** 0.02 5.31** 0.10 75.77** 0.03 5.58 0.11 86.92 *, **: Significant at P = 0.05 & 0.01, respectively Table 2A. Estimates of stability parameters of rice bean genotypes for various characters Genotype NRB-1 NRB-2 NRB-3 NRB-4 NRB-5 NRB-6 NRB-7 NRB-8 NRB-9 NRB-10 NRB-11 NRB-12 NRB-13 NRB-14 NRB-15 NRB-16 NRB-17 NRB-18 NRB-19 NRB-20 NRB-21 NRB-22 NRB-23 NRB-24 NRB-25 NRB-26 NRB-27 NRB-28 NRB-29 NRB-30 NRB-31 NRB-32 NRB-33 NRB-34 NRB-35 NRB-36 NRB-37 NRB-38 NRB-39 NRB-40 NRB-41 NRB-42 NRB-43 NRB-44 NRB-45 NRB-46 NRB-47 NRB-48 NRB-49 NRB-50 Pop. mean Days to flowering Mean bi S2d 122.06 1.13** -1.70 115.83 0.72 -1.08 97.72 0.23 2.54 111.95 0.41 -1.24 125.06 0.90* -1.53 124.11 0.76 -0.89 109.28 0.13 -0.93 123.95 1.17** -2.04 125.39 1.04* -0.62 115.94 0.88* -1.89 108.89 1.34** -1.43 126.00 0.48 -1.68 125.72 0.33 -1.72 102.61 1.09* -1.60 107.28 0.46 -1.30 132.56 0.84 -1.54 114.89 0.79 -1.72 124.89 0.40 -1.09 118.78 0.37 -0.46 122.67 0.65 -1.60 105.11 0.77 -1.80 123.89 0.56 -1.97 123.28 0.94* -1.92 113.83 0.28 -2.16 120.72 1.22** -2.00 122.17 0.97* -2.13 121.89 0.89* -1.18 121.28 1.14** -2.02 121.28 1.21** -1.98 122.39 0.78 -1.32 121.06 1.23** -0.63 103.45 1.07* -1.48 128.39 1.47** -1.05 114.44 1.35** -1.82 126.61 1.89** -1.10 105.50 0.61 -1.56 106.06 0.61 -2.06 113.00 0.95* -1.56 124.50 1.99** -1.97 125.17 0.69 0.31 122.72 1.20** -1.65 115.50 1.70** -2.09 123.50 1.13** -2.10 121.00 0.95* -1.46 122.56 1.78** -2.09 122.56 1.29** -1.87 126.61 1.54** 0.27 95.11 1.19** -1.39 115.72 1.28** -1.92 121.06 1.51** -1.73 118.12 0.54 -1.42 Days to maturity Mean bi S2d 158.61 1.07 -1.64 142.33 1.30* -1.05 121.78 0.97 -2.52 147.33 0.89 -1.85 163.83 0.45 -1.74 165.67 1.11* -2.31 136.22 0.81 -2.79 165.06 0.50 -1.31 165.17 0.82 -2.17 154.89 0.75 -1.50 145.22 0.95 -1.94 162.17 1.21* 0.09 159.44 1.41* -1.69 132.22 1.47** -2.64 141.67 1.53** -2.30 168.11 1.73** -2.43 145.72 0.83 -1.63 164.11 0.54 -2.62 161.11 1.83** -0.31 163.94 0.60 -1.00 141.78 1.38* -2.43 162.56 1.17* -2.82 164.72 0.96 -1.91 139.22 1.20* -1.78 152.50 0.44 -2.23 158.28 1.32* -0.88 148.44 1.31* -0.24 163.33 0.57 -2.52 153.72 0.48 -1.88 161.94 1.49** -2.06 159.89 1.48** 1.36 130.11 -0.33 -0.75 165.06 0.46 -1.51 145.94 0.72 -1.94 151.33 2.22** -0.77 136.94 0.85 -1.28 139.06 0.19 3.77 135.72 1.10* 0.84 161.56 0.50 -1.01 162.22 0.43 -0.67 144.72 1.03 -2.37 157.22 1.16* 4.37* 147.06 1.72** -1.10 161.28 1.18* 0.00 145.61 1.46** -2.52 145.78 0.21 1.77 150.56 1.15* 1.66 120.83 1.82** -0.78 141.50 1.09 1.81 163.67 0.84 -1.48 151.54 0.65 -1.21 Plant height (cm) Mean bi S2 d 117.20 0.61 -16.08 129.33 0.59 -16.63 178.11 0.94 -15.59 68.14 0.75 -7.56 118.81 0.60 -17.17 192.64 1.32 -9.20 76.36 0.70 -6.63 272.72 0.56 -9.47 211.14 1.02 -14.49 254.70 -0.54 25.22 145.25 0.63 -10.59 137.78 0.99 -17.30 179.17 1.56 -17.86 167.72 0.30 -5.88 185.67 1.62 -13.52 211.47 1.54 -7.64 154.97 -0.41 0.97 172.67 0.11 -9.17 164.36 0.41 2.85 5.39 196.06 1.42 98.70 0.40 -14.31 143.83 2.22* -6.86 192.81 -0.28 4.13 181.70 1.03 -9.88 204.67 1.71 -8.49 185.97 2.04* -17.88 193.61 0.71 0.48 177.97 2.16* 4.08 244.22 2.78** -8.20 223.92 1.77 -4.69 107.06 1.32 -15.05 184.72 0.67 -1.21 165.67 1.53 -13.17 199.67 0.42 -10.53 187.80 -0.15 -13.18 106.16 -0.23 -7.58 107.64 1.28 -15.10 99.61 0.14 -15.15 124.00 0.30 -3.30 173.89 0.99 -3.39 183.22 1.67 -2.66 191.92 1.35 -13.98 154.33 -1.10 2.42 252.22 0.70 -2.74 203.30 2.65** 2.21 185.47 0.16 -15.95 176.97 1.34 6.45 117.44 0.16 -18.05 150.11 1.32 -14.44 161.50 1.27 -4.13 168.29 0.74 -7.61 Pods/Cluster Mean bi S2 d 2.54 1.15** -0.01 2.47 0.75* -0.01 3.45 0.78* 0.00 2.50 1.26** -0.05 1.79 1.05** -0.01 3.46 1.24** -0.01 2.42 1.19** -0.01 3.55 1.13** -0.01 3.56 0.94* -0.01 2.56 1.18** -0.02 2.98 0.96** 0.00 3.38 0.59 0.00 3.35 1.25** -0.01 3.53 0.86* -0.01 3.37 0.70 -0.02 3.52 0.93* -0.01 3.09 2.05** 0.00 3.32 0.78* -0.01 3.44 0.88* -0.02 3.61 0.90* -0.01 1.64 0.83* -0.01 2.61 1.06** -0.01 3.23 0.75* -0.01 3.56 1.05** 0.00 3.35 0.85* -0.02 1.63 0.92* -0.02 2.48 1.13** -0.01 3.51 1.10** -0.02 2.69 0.84* -0.01 2.98 0.23 -0.01 1.97 1.38** -0.01 2.94 1.31** 0.00 3.49 1.04** -0.01 3.51 0.96** 0.00 3.56 0.75* -0.01 2.80 0.87* 0.00 2.65 0.76* -0.01 2.60 0.64 -0.02 2.50 0.99** -0.01 2.79 1.58** 0.00 2.52 1.57** -0.01 2.33 0.69 -0.01 3.45 1.19** -0.01 4.16 1.18** 0.00 3.53 0.69 -0.01 3.03 0.75* 0.00 2.46 0.84* -0.01 3.27 1.03** -0.01 4.20 0.95* -0.02 2.61 1.16** -0.01 3.00 0.59 -0.01 Clusters/plant Mean bi S2d 36.72 1.33 -1.43 34.81 2.80** 0.56 40.94 2.00** -1.76 35.94 0.87 2.08 30.50 1.99** -0.61 30.08 3.04** 2.03 36.69 0.75 -1.42 43.83 2.37** -0.12 37.67 1.44* -1.68 36.33 2.04** 0.74 44.22 1.06 -1.68 35.44 1.15 0.42 33.11 1.69* -1.03 25.42 1.75* -1.50 23.06 1.08 -1.34 38.53 0.96 -0.72 32.61 -0.26 -1.15 42.39 0.83 2.04 31.86 0.53 2.34 34.14 0.67 -0.49 34.11 0.39 -0.97 24.56 0.96 -1.74 35.17 -0.55 -1.24 34.92 1.82** 0.58 51.25 0.66 -1.47 31.56 0.68 -0.16 27.06 0.76 0.24 35.14 0.69 -0.62 49.19 1.05 3.86* 43.97 0.10 1.31 27.14 0.77 -0.71 28.86 0.95 -0.21 34.50 0.46 -0.19 45.53 0.14 -0.92 41.72 0.53 -1.32 44.06 1.73* -0.15 26.97 1.73* 0.65 22.17 0.59 2.49 17.61 0.63 -1.17 37.94 1.25 -1.28 18.75 0.45 -1.08 35.47 1.02 -0.43 26.53 0.55 -0.51 34.36 0.35 -0.42 25.89 2.07** 0.29 12.81 0.15 -0.93 26.33 0.96 -0.35 48.78 0.27 -1.31 41.86 -0.04 -0.40 25.86 0.72 5.18** 33.89 0.63 -0.35 Pods/plant Mean bi S 2d 87.91 1.17** -28.09 80.58 1.17** -12.69 135.83 1.24** -8.81 84.52 1.18** 6.98 49.39 0.96** -36.13 98.87 2.02** -17.60 83.26 1.07** -40.06 150.22 2.05** -12.54 128.47 1.30** -35.46 87.77 1.41** -27.01 126.33 1.19** 7.10 114.02 0.89** -29.74 105.42 1.47** -25.98 84.49 1.20** -35.32 72.23 0.88** -36.09 130.24 1.19** -19.59 95.32 1.31** -34.79 135.29 1.22** -11.36 103.94 0.73* 8.30 118.01 1.05** -15.20 50.43 0.73* -35.98 58.84 0.81* -37.74 108.24 0.37 -27.40 119.05 1.64** 20.77 165.97 1.05** -39.52 45.75 0.71* -36.09 61.57 0.72* -22.91 117.91 0.98** -25.26 126.88 0.98** -11.36 125.26 -0.04 -0.35 48.02 0.83* -41.10 79.41 1.02** -24.09 115.07 0.88** -4.63 154.11 0.74* -7.57 143.33 0.92** -24.39 118.18 1.28** 51.85 65.84 0.93** -13.49 52.19 0.58 -13.90 0.56 -34.83 38.65 100.69 1.71** -29.95 41.81 0.73* -37.41 77.01 0.58 -34.25 86.00 0.85** -7.41 137.51 1.09** -18.64 85.88 1.03** -10.52 33.17 0.16 -40.20 59.52 0.74* -26.64 154.00 0.98** -22.51 168.95 0.53 -8.57 63.63 0.73* 11.93 97.50 0.39 -19.12 284 Journal of Food Legumes 25(4), 2012 Table 2B. Estimates of stability parameters of rice bean genotypes for various characters Genotype Pod length (cm) Mean bi S2 d NRB-1 -0.02 9.61 0.75 NRB-2 -0.02 9.29 0.98 NRB-3 -0.01 9.30 0.61 NRB-4 -0.02 8.28 1.16* NRB-5 -0.02 8.23 0.79 NRB-6 -0.02 8.62 0.57 NRB-7 -0.01 8.43 0.87 NRB-8 -0.01 9.51 1.17* NRB-9 -0.02 10.24 0.72 NRB-10 -0.02 11.24 0.76 NRB-11 -0.02 9.26 0.83 NRB-12 -0.02 7.36 1.10 NRB-13 8.05 1.73** -0.01 NRB-14 -0.02 9.25 1.19* NRB-15 -0.01 8.45 0.30 NRB-16 -0.02 7.41 1.08 NRB-17 -0.02 7.39 0.99 NRB-18 11.25 1.19* -0.01 NRB-19 8.47 -0.02 1.02 NRB-20 8.81 2.01** 0.07** NRB-21 7.35 -0.02 1.22* NRB-22 8.39 -0.02 0.93 NRB-23 12.69 -0.02 0.84 NRB-24 9.28 -0.02 0.80 NRB-25 10.49 1.25* -0.02 NRB-26 10.60 -0.02 0.72 NRB-27 8.91 0.01 1.01 NRB-28 11.08 2.02** -0.01 NRB-29 12.57 -0.02 0.83 NRB-30 13.12 1.31* -0.01 NRB-31 11.03 1.45* -0.01 NRB-32 6.44 -0.02 0.56 NRB-33 -0.02 8.50 0.97 NRB-34 -0.01 10.08 1.12* NRB-35 -0.02 8.41 1.07 NRB-36 -0.01 8.23 0.91 NRB-37 -0.02 6.51 0.92 NRB-38 -0.02 7.54 0.91 NRB-39 -0.01 7.55 1.08 NRB-40 -0.02 8.56 0.87 NRB-41 -0.02 8.29 0.70 NRB-42 -0.01 10.52 0.72 NRB-43 -0.01 10.45 0.94 NRB-44 0.00 10.15 1.25* NRB-45 0.00 9.52 0.66 NRB-46 0.00 9.09 0.45 NRB-47 -0.01 10.04 1.65** NRB-48 -0.01 10.15 0.79 NRB-49 -0.01 10.42 0.81 1.14* NRB-50 -0.02 7.46 Pop. mean 9.24 0.82 -0.01 Mean 8.32 7.87 8.01 6.52 5.94 4.55 6.51 6.39 6.50 6.83 7.51 6.50 6.24 8.13 7.01 6.23 6.32 7.42 7.22 7.40 6.30 4.47 8.92 7.79 7.24 7.36 6.33 6.71 8.01 8.77 6.68 5.55 7.57 6.43 5.54 7.31 5.22 6.26 6.03 7.69 6.75 6.90 7.17 6.36 5.61 5.50 6.07 8.35 9.03 5.65 6.82 Seeds/pod bi 0.99 2.01** -0.51 1.37 1.23 1.96** 1.07 0.88 1.64* 1.46 0.25 3.77** 0.79 0.30 2.46** 1.38 0.59 -0.10 -0.19 0.49 1.81* 1.17 0.30 2.24** 0.04 0.60 0.91 2.67** 0.30 1.66* -0.28 -0.34 0.48 1.48 0.30 1.31 0.41 0.65 2.13** 1.59* 0.18 2.82** 0.23 -0.05 0.98 1.18 0.34 0.51 1.03 1.15 0.60 S2d -0.03 0.00 -0.02 -0.03 -0.01 -0.03 -0.01 -0.02 -0.03 -0.02 0.01 0.08** -0.02 -0.01 -0.01 -0.02 -0.02 -0.01 -0.03 -0.03 -0.03 -0.02 -0.01 0.01 -0.02 -0.02 0.00 0.11** -0.01 -0.03 0.00 -0.02 -0.03 -0.03 0.02 -0.03 -0.01 -0.02 -0.02 -0.03 0.01 -0.02 0.00 0.00 -0.01 -0.01 -0.01 -0.02 -0.02 -0.01 -0.02 Biomass/plant (g) Mean bi S2d 24.17 0.86 -4.66 27.22 0.85 -3.32 31.70 0.16 -0.83 18.42 1.42 -3.43 24.39 0.68 -3.09 31.80 0.77 14.25** 26.97 -0.64 47.86** 43.14 2.08** 29.06** 37.64 0.15 -1.14 42.61 0.80 -4.99 26.03 0.03 -3.27 28.92 0.71 -3.64 30.48 1.33 20.24** 25.42 0.93 7.35 34.39 0.50 0.98 39.75 1.33 -3.12 26.92 0.73 -5.17 35.42 1.25 -3.79 32.56 0.53 -1.88 34.56 1.54* -5.39 19.81 0.63 -3.89 27.06 0.27 -0.83 34.28 1.03 -3.05 32.67 1.70* -4.43 37.03 1.07 -5.26 32.72 1.41 -3.91 36.39 0.44 -0.07 33.64 1.30 -3.37 38.97 0.47 -2.22 42.61 1.38 -3.82 23.86 1.26 -5.52 35.05 0.41 -1.99 33.00 1.61* -4.16 34.22 1.08 -4.36 35.42 1.13 5.32 22.67 0.71 -2.46 26.50 0.84 -5.20 18.06 1.31 -3.94 22.61 1.52* -3.61 35.19 1.85* -3.38 33.20 1.46 -2.56 34.72 2.19** 0.13 29.17 1.33 -4.33 48.89 1.80* -3.49 35.45 1.52* -3.36 31.86 0.13 6.04 33.00 1.70* -3.79 25.11 1.39 -2.51 27.97 -0.02 -1.41 31.86 1.92* -0.37 31.51 0.81 -2.50 deviation also indicates that considerable genetic diversity is locked up in the bean. Such non-linear deviation might also be of practical value to construct and test the utility of the multiple regression models to know more critically the complex mechanism of adaptation. The genotype × environment interactions for most of the characters were also highly significant which further substantiated differences among genotypes and their inconsistent response to different environments (Table 2A,B). However, the magnitude of 100-seed weight (g) Mean bi S 2d 7.37 0.70 -0.11 8.02 -0.45 -0.08 7.62 0.76 -0.09 10.35 0.19 -0.16 14.65 0.87 -0.06 26.98 0.61 -0.03 12.23 1.91 -0.06 23.23 0.90 -0.07 25.58 1.68 -0.06 29.04 1.65 -0.07 8.37 1.06 -0.11 7.42 0.04 -0.07 15.40 1.17 -0.09 11.87 0.00 -0.08 14.11 1.15 -0.05 6.34 0.28 -0.10 7.79 0.56 -0.05 21.22 0.90 -0.06 14.36 0.24 -0.10 11.34 0.88 -0.10 7.36 0.24 -0.10 25.79 1.36 -0.08 22.38 1.09 -0.09 18.32 1.11 -0.10 26.23 0.67 -0.08 20.09 1.13 -0.08 23.19 1.26 0.00 27.24 0.98 -0.07 28.36 1.24 -0.10 29.35 0.95 -0.10 22.18 -0.07 0.32 -0.10 7.31 1.17 7.33 -0.10 0.70 18.51 -0.03 0.34 20.39 -0.10 1.11 7.42 1.47 -0.10 11.34 -0.10 1.43 13.79 -0.08 1.89 11.24 -0.08 0.80 7.39 1.10 -0.09 11.95 -0.04 0.72 -0.02 25.75 1.89 -0.08 24.23 0.71 25.41 -0.09 0.74 -0.04 28.18 0.97 -0.08 27.71 2.06* -0.09 23.01 -0.49 -0.04 8.21 1.13 -0.10 11.37 1.29 -0.10 12.34 0.65 16.73 0.88 -0.08 Seed yield/plant (g) Mean bi S2d 50.01 0.62 -79.28 46.98 0.55 -66.80 79.11 0.65 -58.18 53.27 0.84 -75.98 39.39 0.69 -79.24 117.41 2.12** -29.05 62.60 0.87* -80.76 219.41 2.74** -73.10 209.53 2.22** -68.99 170.85 2.50** -16.03 75.77 0.68 -74.62 51.06 0.63 -80.28 97.54 1.24** -56.74 77.71 0.75 -71.86 67.80 0.96* -75.77 47.68 0.49 -80.84 43.09 0.59 -82.24 209.06 0.84 -68.71 103.95 0.62 -41.17 95.11 0.82 -64.65 19.68 0.34 -86.10 64.15 0.89* -73.43 212.10 0.79 -5.66 166.50 2.39** 87.09 311.27 1.65** 302.31** 63.72 -73.14 0.87* 86.64 -1.23 0.77 212.10 2.13** 190.55* 284.08 1.77** 195.62* 318.69 0.63 731.22** 67.13 -71.57 0.83* 28.34 -81.71 0.34 59.91 -78.32 0.44 179.84 0.98* -11.16 158.10 1.01* -5.23 60.31 -51.93 0.70 35.17 -75.24 0.46 -61.40 41.14 0.48 22.48 -83.04 0.39 53.44 -83.11 0.95* 30.00 -83.03 0.41 -62.21 133.33 1.34** 145.60 1.29** 5.34 218.12 1.48** -2.28 131.95 1.35** 71.51 46.63 -77.68 0.26 78.55 -75.78 0.72 101.48 0.85* -74.79 169.35 0.80 -67.46 40.72 -64.62 0.40 109.16 0.60 -54.58 genotype × environment interaction for flowering, pods/ cluster, pod length, seeds/pod and seed weight was low and non-significant indicating the consistent performance of genotypes over variable environments for these characters. As the stability of the genotypes for pod length was concerned, ‘NRB-2’, ‘NRB-34’ and ‘NRB-43’ were rated the most stable genotypes since they possessed good pod length and showed non-significant mean square deviations from zero along with regression coefficient values very close to unity Rungsung and Changkija: Stability analysis for seed yield and its component traits in rice bean (Vigna umbellata) (+0.98, +1.12 and +0.94, respectively). Similarly, ‘NRB-24’ and ‘NRB-34’ were found most stable for pods/cluster; ‘NRB-1’ and ‘NRB-49’ for seeds/pod; ‘NRB-12’, ‘NRB-16’ and ‘NRB42’ for clusters/plant; ‘NRB-14’, ‘NRB-32’ and ‘NRB-38’ for flowering; ‘NRB-3’, ‘NRB-11’, ‘NRB-38’, ‘NRB-41’ and ‘NRB49’ for maturity; ‘NRB-23’, ‘NRB-25’, ‘NRB-34’ and ‘NRB-35’ for biomass; ‘NRB-20’, ‘NRB-25’, ‘NRB-28’, ‘NRB-29’, ‘NRB35’, ‘NRB-44’ and ‘NRB-48’ for pods/plant and ‘NRB-8’, ‘NRB18’, ‘NRB-23’, ‘NRB-24’, ‘NRB-26’, ‘NRB-27’, ‘NRB-28’, ‘NRB30’, ‘NRB-35’ and ‘NRB-45’ for seed weight. A perusal of stability parameters for grain yield/plant shows that ‘NRB18’, ‘NRB-23’ and ‘NRB-49’ registered promising average grain yield and regression coefficients less than unity (+0.84, +0.79 and +0.80, respectively) with non-significant mean square deviations from regression, thus, revealing their less sensitivity to environmental changes and hence will be better adapted to poor environmental conditions. However, ‘NRB6’, ‘NRB-8’, ‘NRB-9’, ‘NRB-10’, ‘NRB-24’, ‘NRB-42’, ‘NRB43’, ‘NRB-44’ and ‘NRB-45’, which were also good performers for grain yield showed highly significant regression coefficient values (bi=+2.12, +2.74, +2.22, +2.50, +2.39, +1.34, +1.29, +1.48 and +1.35, respectively) and non-significant mean square deviations. This clearly implies that these genotypes are highly sensitive to environmental changes and therefore will be suitable for favourable environments. ‘NRB-25’ and ‘NRB30’, despite the fact of being the highest-yielding genotypes, had unpredictable performance across the environments as indicated by their highly significant deviations from regression and thus, were unstable for yield. The stability of yield is an important characteristic to be considered when judging the value of a cropping system relative to others (Piepho 2008). Among all the genotypes studied, ‘NRB-34’ and ‘NRB35’ were found most stable for yield/plant as they showed 285 high degree of stability irrespective of the environments. These genotypes may be suitable for cultivation in Nagaland during kharif season irrespective of locations, and may also be used directly for breeding stable genotypes of rice bean. In like manner, ‘NRB-3’, ‘NRB-11’, ‘NRB-38’, ‘NRB-41’and ‘NRB-49’ can be used in the breeding of rice bean for developing stable and early-maturing genotypes. REFERENCES Ahuja A, Malhotra PK, Bhatia VK and Parsad R. 2005. Statistical package for agricultural research (SPAR-2). Indian Agricultural Statistics Research Institute (IASRI), New Delhi. Bolivar A and Luis CZ. 2010. Impact of germination on phenolic content and antioxidant activity of 13 edible seed species. Food Chemistry 119: 1485–1490. Danyali SF, Razavi F, Segherloo AE, Dehghani H and Sabaghpour SH. 2012. Yield stability in chickpea (Cicer arietinum L.) and study of relationship among the univariate and multivariate stability parameters. Research in Plant Biology 2: 46-61. Eberhart SA and Russell WA. 1966. Stability parameters for comparing varieties. Crop Science 6: 36-40. Gupta S, Kozak M, Sahay G, Durrai AA, Mitra J, Verma MR, Pattnayak A, Thongbam PD and Das A. 2009. Genetic parameters of selection and stability and indication of divergent parents for hybridization in rice bean [Vigna umbellata (Thunb) Ohwi and Ohashi] in India. Journal of Agricultural Science 147: 581-588. Patel JB and Acharya S. 2011. Stability analysis for grain yield in fieldpea (Pisum sativum L.). Journal of Food Legumes 24: 150151. Piepho HP. 2008. Methods for comparing the yield stability of cropping systems. Journal of Agronomy and Crop Science 180: 193–213. Yan X, Bube BE and Lynch JP. 1995. Genetic variation for phosphorus efficiency of French bean in contrasting soil types II: yield response. Crop Science 35: 1074-1099. Journal of Food Legumes 25(4): 286-290, 2012 Sequence comparison of coat protein gene of Mungbean yellow mosaic India virus isolates infecting mungbean and urdbean crops NAIMUDDIN and M. AKRAM Division of Crop Protection, Indian Institute of Pulses Research, Kalyanpur, Kanpur 208024, India; E-mail: [email protected] (Received: July 02, 2012 ;Accepted: November 14, 2012) ABSTRACT The coat protein (CP) gene sequence of ten isolates of Mungbean yellow mosaic India virus (MYMIV), five from different genotypes of each mungbean and urdbean were used to study the intra-field variation in MYMIV during kharif 2009. The CP gene was successfully amplified using primer pair NM1 5’ GTA TTT GCA (GT)CA (AT)GT TCA 3’ / NM2 5’ AGG DGT CAT TAG CTT AGC 3’ designed using DNA sequence of MYMIV isolates. The complete nucleotide sequence of the CP gene of all the MYMIV isolates had single open reading frame (ORF) of 774 bp and 257 amino acids. Analysis of CP gene sequences of isolates from mungbean and urdbean genotypes revealed that all the isolates among themselves had 99-100% homology at amino acid level and 97-99% similarity at nucleotide level. Isolates differed in amino acid composition only at four locations. These isolates had 96-100% similarity at amino acid level and 95-99% similarity at nucleotide level with known MYMIV isolates. Results indicated that CP gene was highly conserved among the isolates of MYMIV infecting different genotypes in a field at Kanpur. Key words: Amino acids, Coat protein, MYMIV, Nucleotides, PCR, Variation Yellow mosaic disease occurs across the Indian subcontinent and is a major constraint to the production of most of the warm-season legumes, particularly mungbean, urdbean and soybean. Estimation of actual losses due to yellow mosaic disease (YMD) in farmers’ field is difficult as these losses vary from year to year and from variety to variety. However, based on the incidence of YMD in mungbean, urdbean and soybean, an annual loss of over US $ 300 million is estimated in these crops (Varma et al. 1992). Yellow mosaic disease occurs in a number of leguminous plants such as mungbean, urdbean, cowpea (Nariani 1960, Nene 1973), soybean (Suteri 1974), horsegram (Muniyappa et al. 1975), lablab bean (Capoor and Varma 1948) and French bean (Singh 1979). The YMD in pulse crops is caused by whitefly transmitted Begomoviruses such as Mungbean yellow mosaic virus (MYMV), Mungbean yellow mosaic India virus (MYMIV) and Horsegram yellow mosaic virus (HgYMV) across India (Malathi and John 2008). These viruses are distinct species of the genus Begomovirus under the family Geminiviridae and have bipartite genome that consists of two components, viz., DNA A and DNA B. In DNA A there are five to seven ORFs; two in virion sense (AV1-coat protein and AV2-pre-coat protein) and others in complementary sense (AC1-replication initiation protein, AC2transcription activator protein, AC3-replication enhancer protein, AC4 and AC5). Coat protein (CP) is a multifunctional protein and is involved besides encapsidation in three major functions of viral pathogenicity-viral DNA replication, intra-, inter-cellular movement and long distance transport of viral genome. Most importantly, the vector transmission specificity is controlled by CP gene. Differences in the disease reaction of genotypes of mungbean and urdbean in different locations particularly in southern and northern zones of India are commonly reported. This difference can be to some extent due to the different virus species operating in these areas, MYMIV in north (Usharani et al. 2004) and MYMV in south (Karthikayan et al. 2004). Difference in the severity of yellow mosaic in mungbean and urdbean in a given location is also observed. To address this we have tried to look whether there is a possibility of intra-field variability among MYMIV isolates at Kanpur. CP gene i s the mo st conserved sequence in the genus Begomovirus (Harrison et al. 2002) and is also known to control functions of virus pathogenicity. Also, this gene has long been considered important for establishing the identity of a distinct geminivirus (Rybicki 1998). CP gene was, therefore, selected to analyze the variability in the isolates of MYMIV infecting mungbean and urdbean in a field at Kanpur. MATERIALS AND METHODS MYMIV isolates: During kharif 2009, young leaves showing characteristic yellow mosaic symptoms were collected from the field-infected plants of five mungbean (T44, Meha, LGG 407, PDM 54, Kopergaon) and five urdbean (Shekhar, PDU 1, AKU 9904, Barabanki Local, TPU 4) genotypes and brought to the laboratory and used to isolate total DNA using E.Z.N.A.® Pl ant DNA Mini prep Kit (U.S.A.) according to t he manufacturer’s instructions. DNA from one healthy plant of each genotype was also isolated. Total DNA was used as template in PCR reactions. Primers: A set of degenerate primers (NM1 5’GTA TTT GCA (GT)CA (AT)GT TCA AGA 3’/NM2 5’AGG (AGT)GT CAT TAG CTT AGC 3’) was designed using DNA sequence of different MYMIV isolates. Primers were designed to get the complete coat protein gene of MYMIV without cloning by Naimuddin et al.: Sequence comparison of coat protein gene of MYMI virus isolates taking about 100 extra nucleotides on both sides of the gene. The complete CP gene sequence was extracted from the direct sequencing data of the PCR product. Thermal conditions: PCR was performed in T1 Thermalcycler, Biometra® programmed for 35 cycles with one step of initial denaturation for 2.5 min., and denaturation for 45 s at 94°C, 1 minute annealing at 54°C and 1 min for extension at 72°C followed by one step final extension for 10 min at 72°C. PCR assays were conducted with Easy-DoTM PCR PreMix (SBS Greentech Co., Ltd.) in total reaction mixture volume of 50 µl. Experimental control was a PCR master mix, in which the template DNA was 2 µl (50ng/µl) of healthy leaves of corresponding genotypes. Following inputs were added to make a total reaction mixture volume of 50 µl : DNA template (50ng/µl)-2µl, Primer NM1 (50 pmole/µl)-1µl, primer NM2 (50 pmole/µl)-1µl and dH2O - 46µl. Electrophoresis, sequencing and analysis: PCR amplicons were analyzed in 1% agarose gel in Tris-acetate EDTA (TAE) containing ethidium bromide @ 0.1%. The gel was observed under UV trans-illuminator and photographed. PCR product of CP gene was purified using E.Z.N.A®. Gel Extraction Kit (USA) and sequenced through M/S R.D. Applied Biosciences, Ltd. New Delhi. Sequence data were blasted using NCBI data base. Multiple sequence alignment and phylogram were generated using CLUSTAL W version 1.8 3 (http: // www.genome.jp/tools/clustalw). Cluster phylogram illustrating the phylo geneti c rel ationship was inferred using the Neighbor-Joining method (Saitou and Nei 1987) based on the multiple alignments of nucleotide sequences of CP genes of the MYMIV isolates under study with corresponding gene of known isolates of MYMIV and MYMV. The bootstrap consensus tree was inferred from 1000 replicates (Felsenstein 1985). The evolutionary distances were computed using the p-distance method (Nei and Kumar 2000) and are in the units of the number of transitional differences per site. The analysis involved 26 nucleotide sequences conducted in MEGA5 (Tamura et al. 2011). Sequences of CP gene of the isolates of MYMIV infecting five genotypes of mungbean and urdbean in an experimental field at Kanpur were compared. 287 RESULTS AND DISCUSSION Symptomatology and PCR amplification: The first symptom of YMD in mungbean and urdbean appeared as small interveinal yellow spots in young leaves. In subsequently emerging leaves symptoms became more conspicuous and appeared as irregular yellow and green patches. Inter-veinal area or even the whole lamina often turned completely yellow in highly susceptible genotypes. The puckering in lamina which is some times reported in YMD (Malathi and John 2008) was not observed in any of the genotypes used in the present study. There was however, observed some reduction in lamina size which was more in urdbean than in mungbean genotypes. Yellow patches on pods were observed more in the case of mungbean than in the urdbean genotypes. PCR products from all the YMD-affected samples of mungbean (5 nos.) and urdbean (5 nos.) when analyzed in gel yielded amplicon of expected size (~950bp), indicating involvement of MYMIV (Fig. 1). No amplicon was observed in PCR products from healthy plants indicating no infection by MYMIV in plants that were free from yellow mosaic symptoms. These results indicate that the primers designed worked well to amplify the fragment (containing CP gene) of DNA A of the virus genome. The primers used in this study have also been exploited successfully to detect the MYMIV infection in cowpea (Naimuddin and Akram 2009) and in accessions of wild Vigna (Naimuddin et al. 2011). These primers may be routinely used for the PCR-based detection of MYMIV in pulses. Fig 1. Gel electrophoresis of PCR amplified products of CP gene of MYMIV using NM1/NM2 primer pair. Lane M=1 kb DNA ladd er, lane 1-5= CP amp lified products with DNA of diseased mungbean plants, lane 6-10= amplified products with DNA of dis eased urdbean plants. Table 1. Per cent identity of coat protein gene of MYMIV isolates at amino acid (below) and nucleotide levels (above) MYMIV-Ub01 MYMIV-Ub02 MYMIV-Mb03 MYMIV-Mb01 MYMIV-Mb02 MYMIV-Ub05 MYMIV-Mb04 MYMIV-Ub04 MYMIV-Mb05 MYMIV-Ub03 MYMIVUb01 100 99 99 99 99 99 99 100 99 100 MYMIVUb02 97 100 99 99 99 99 99 99 99 99 MYMIVMb03 98 97 100 99 99 99 99 99 99 99 MYMIVMb01 97 98 97 100 99 100 100 99 100 99 MYMIVMb02 98 99 97 99 100 99 99 99 99 99 MYMIVUb05 98 98 98 98 98 100 100 99 100 99 MYMIVMb04 97 98 97 98 98 98 100 99 100 99 MYMIVUB04 99 98 99 98 98 98 99 100 99 100 MYMIVMb05 98 98 97 98 98 98 98 98 100 99 MYMIVUb03 97 98 97 98 98 97 97 97 97 100 288 Journal of Food Legumes 25(4), 2012 Comparison of CP gene sequences: The PCR products were purified and got directly sequenced. The CP gene sequences were easily extracted from the sequence data. The isolates from mungbean genotypes LGG 407, Meha, Kopergaon, PDM 54 and T 44 were designated as MYMIV-Mb01, -Mb02, -Mb03, -Mb04 and -Mb05 and isolates from urdbean genotypes AKU 9904, Barabanki Local, TPU 4, Shekhar and PDU 1 were designated as MYMIV-Ub01, -Ub02, -Ub03, -Ub04 and -Ub05 and their sequences were submitted to NCBI data base under the accession nos. GQ387501, GQ387502, GQ387503, GQ387504, GQ387505 andGQ387506, GQ387507, GQ387508, GQ387509, GQ387510, respectively. The complete nucleotide sequence of the CP gene of all the MYMIV isolates had single open reading frame (ORF) of 774 base pairs and 257 amino acids. This is in conformity with the earlier reports (Ilyas et al. 2010). The cluster phylogram based on multiple alignment of the nucleotide sequence of the CP gene of 10 isolates under study and 8 isolates of each MYMIV and MYMV indicated that all the 10 isolates belonged to MYMIV species as they formed cluster with other known isolates of MYMIV (Fig. 2). Fig 2. Cluster phylogram illustrating the phylogenetic relationship between MYMIV and MYMV isolates Blast search results showed that the isolates under study had 95-99% identity at nucleotide level and 96-100% identity at amino acid level with other isolates of MYMIV available at NCBI database. Comparison of CP gene of all the ten isolates under study revealed among themselves 97-99% similarity at nucleotide level and 99-100% similarity at amino acid level (Table 1). Isolate MYMIV-Ub01 had identical amino acids with MYMIV-Ub04 and MYMIV-Ub03. Amino acids sequence of isolate MYMIV-Mb01 was identical to MYMIVUb05, MYMIV-Mb04, and MYMIV-Mb05. Review of literature indicated non-availability of information on intra-field diversity in MYMIV based on CP gene sequence. However, recently Ilyas et al. (2010) studied diversity based on full sequence of DNA A of MYMIV isolates from different parts of Pakistan and indicated 93.8-99.5% nucleotide sequence identities with MYMIV sequences available in the database. CP gene of MYMIV isolates from different hosts was shown to have 97-99% nucleotide sequence similarity (Sachan et al. 2010). The amino acids of CP gene of the isolates under study differed among themselves at 4 positions (Fig. 3). All the isolates have threonine at position 5 of amino acids except MYMIV-[Mb02], which had proline at this position. Amino acid at position no. 6 was tyrosine in seven isolates, whereas in three isolates (Ub01, Ub03 and Ub04), this position had phenylalanine. The other differences in amino acids were observed at position no. 209 and 232. At position no. 209 all isolates had tyrosine except MYMIV-Mb02 in which this position had histidine. Similarly, at position no. 232, all isolates had methionine except MYMIV-Ub02 isolate, which had isoleucine. These results showed that there was little variation in coat protein. Harrison et al. (2002) also reported that the amino acid sequences of the coat protein of whitefly transmitted begomoviruses were more conserved than the remainder of the genome. In an earlier study, comparison of amino acids of CP gene of MYMIV isolates from four different hosts (cowpea, mungbean, urdbean and rajmash) revealed differences in amino acids at six positions (Sachan et al. 2010). Since CP gene is known to have multiple functions in viral pathogenesis and also in vector specificity, comparison of isolates operating in a field may yield some important information pertaining to the diversity in them. Deletion of two amino acids (75th and 150th) in N-terminal has been shown to affect the systemic spread and pathogenicity of MYMIV isolate in cowpea, mungbean and urdbean but not in rajmash (Haq et al. 2011). Coat protein gene offers an opportunity to decipher not only inter- and intra-field diversity in MYMIV isolates but also in the host preference (Haq et al. 2011). Further, it would be interesting to investigate the impact of natural change in amino acids at these positions on virus pathogenicity in different hosts. It may however, be concluded that the changes in amino acids in domains of coat protein that regulate viral pathogenesis may also be responsible for Naimuddin et al.: Sequence comparison of coat protein gene of MYMI virus isolates Fig 3. 289 Multiple alignment of amino acids sequences of MYMIV isolates. The total length of deduced amino acid in CP gene of MYMIV is 257. MYMIV isolates of mungbean (Mb01=LGG407, Mb02=Meha, Mb03=Kopergaon, Mb04=PDM54, Mb05=T44) and isolates of urdbean (Ub01=AKU9904, Ub02=Barabanki local, Ub03=TPU4, Ub04=Shakher, Ub05=PDU1). Dots indicate the similar amino acid. 290 Journal of Food Legumes 25(4), 2012 the temporal variation in disease expression in mungbean and urdbean in a given location. REFERENCES Capoor SP and Varma PM. 1948. A new virus disease of Dolichos lablab. Current Science 19:248-249. Felsenstein J. 1985. Confidence limits on phylogenies: An approach using the bootstrap. Evolution 39:783-791. Haque QMI, Jyothsna P, Ali A and Malathi VG. 2011. Coat protein deletion mutation of Mungbean yellow mosaic India virus (MYMIV). Journal of Plant Biochemistry and Biotechnology 20: 182-189. Harrison BD, Swanson MM and Fargette D. 2002. Begomovirus coat protein: serology, variation and functions. Physiological and Molecular Plant Pathology 60: 257-271. Ilyas M, Qazi J, Mansoor S and Briddon RW. 2010. Genetic diversity and phylogeography of begomoviruses infecting legumes in Pakistan. Journal of General Virology 91: 2091-2101. Karthikeyan AS, Vanitharani R, Balaji V, Anuradha S, Thillaichidambaram P, Shivaprasad PV, Parameswari C, Balamani V, Saminathan M and Veluthambi K. 2004. Analysis of an isolate of Mungbean yellow mosaic virus (MYMV) with a highly variable DNA B component. Archives of Virology 149: 1643-1652. Malathi VG and John P. 2008. Geminiviruses infecting legumes. In: Rao GP, Lava Kumar P, Holguin-Pena RJ eds. Characterization Diagnosis and Management of Plant Viruses-Vegetables and Pulses Crops, Studium Press LLC, USA, 97-123. Muniyappa V, Reddy HR and Shivshankar C. 1975. Yellow mosaic disease of Dolichos biflorus Linn. Current Research 4: 176. Naimuddin and Akram M. 2010. Detection of mixed infection of begomoviruses in cowpea and their molecular characterization based on CP gene sequences. Journal of Food Legumes 23: 191-195. Naimuddin, Akram M, Aditya Pratap, Chaubey BR and John JK. 2011. PCR based identification of the virus causing yellow mosaic disease in wild Vigna accessions. Journal of Food Legumes 24:14-17. Nariani TK. 1960. Yellow mosaic of mung (Phaseolus aureus L.). Indian Phytopathology 13: 24-29. Nene YL. 1973. Viral diseases of warm weather pulse crops in India. Plant Disease Reporter 57: 463-467. Nei M. and Kumar S. 2000. Molecular Evolution and Phylogenetics. Oxford University Press, New York. Rybicki EP. 1998. A proposal for naming geminiviruses: A reply by the Geminiviridae Study Group Chair. Archives of Virology 143:421424. Sachan M, Mishra M, Naimuddin and Akram M. 2010. Amino acid variability in coat protein gene of Mungbean yellow mosaic India virus infecting pulses crops. Trends in Biosciences 3:166-168. Saitou N and Nei M. 1987. The neighbor-joining method: A new method for reconstructing phylogenetic trees. Molecular Biology and Evolution 4:406-425. Singh RN. 1979. Natural infection of bean (Phaseolus vulgaris) by Mungbean yellow mosaic virus. Indian Journal of Mycology and Plant Pathology 9:124-126. Suteri BD. 1974. Occurrence of soybean yellow mosaic virus in Uttar Pradesh. Current Science 43:689-690. Tamura K, Peterson D, Peterson N, Stecher G, Nei M and Kumar S. 2011. MEGA5: Molecular Evolutionary Genetics Analysis using Maximum Likelihood, Evolutionary Distance, and Maximum Parsimony Methods. Molecular Biology and Evolution 28:27312739. Usharani KS, Surendranath B, Haq QMR, Malathi VG. 2004. Yellow mosaic virus infecting soybean in Northern India is distinct from the species infecting soybean in southern and western India. Current Science 86: 845-850. Varma A, Dhar AK, Mandal B. 1992. MYMV transmission and control in India. In: SK Green and D Kim (Eds), Mungbean Yellow Mosaic Disease, Taipei Taiwan: Asian Vegetable Research and Development Centre. Pp 8-27. Journal of Food Legumes 25(4): 291-293, 2012 Bio-efficacy of some insecticides against H. armigera (Hubner) on chickpea (Cicer arietinum L.) P.S. SINGH, R.K. SHUKLA and N.K. YADAV Department of Entomology and Agricultural Zoology, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi-221005, Uttar Pradesh, India; E-mail: [email protected] (Received: August 28, 2012; Accepted: November 13, 2012) ABSTRACT MATERIALS AND METHODS Chickpea ‘SAKI 9516’ was sown on Agricultural Research Farm of Banaras Hindu University during Rabi season 2010-11 and 2011-12 for the bio-efficac y of certain new molecules insecticides viz., HaNPV@ 250 LE/ha, spinosad 45 SC @ 100 g a.i./ha, fenvalerate 20 EC@300 g a.i./ha, quinalphos 25EC@450 g a.i./ha, emamectin benzoate 5SG @ 11 g a.i./ha, azadirachtin 1500 ppm@5ml/lit, cartap hydrochloride 50 SP @ 500 g a.i./ha, fipronil5 SC@50g a.i./ha and indoxacarb 14.5 EC @ 75 g were applied twice at 15 days interval against gram pod borer, H. armigera. Spinosad was found best among all the treatments with 81.2% reduction in larval population over control followed by indoxac arb, fipronil, e mamectin be nzoa te, cartap hydrochloride, fenvalerate, and quinalphos, azadirachtin and HaNPV. After 7 days of spraying of second application spinosad was found best again in reduceding 79.8% larval population followed by indoxacarb (with 78.3% reduction in larval populations). The highest yield was obtained in spinosad (1.79 t/ha) while low in azadirachtin (1.06 t/ha). The cost: benefit ratio was high in treatment fipronil (1: 8.2) while low in treatment indoxacarb (1: 5.3). The experiments were carried out under field conditions at the Agriculture Research Farm of Institute of Agricultural Sciences, BHU, Varanasi during the years 2010-12 on chickpea variety ‘SAKI 9516’ in randomized complete block design (RBD) having 10 treatments replicated thrice with plot size 7.2 m2 having 4 rows with 4 m long in each plot. The plant spacing between row to row and plants to plant were maintained at 40 cm and 15 cm, respectively. The crop received two sprays, the first being given at pod formation when the population crossed Economic Threshold Level while the second spray was imposed on the basis of insect population. The spray mixture of each treatment was prepared by mixing of required quantity of the insecticides formulations in water to make it equivalent to 500 liters. Insecticides were applied during early hours of the day where wind velocity was suitable for spraying. This helped in avoiding the drift of spray fluid to the adjacent plots. Due care was also taken to spray each plot uniformly and the sprayer was thoroughly washed after spraying of each insecticides. Key words: Bio-efficacy, Chickpea, H. armigera, Insecticides Insect pests are one of the major constraints which limit the production of chickpea. In India, it is attacked by 57 insect species and about half a dozen of them are considered to be of economic importance. Pod borer [Helicoverpa armigera (Hubner)] is the most prominent insect species that causes major economic damage to this crop. The yield loss in chickpea due to pod borer was reported as 10-60 per cent in normal weather conditions, while it was 50-100 per cent in favourable weather conditions, particularly in the states where frequent rains and cloudy weather prevailed during the crop season (Patel 1979). Reports showed that H. armigera has developed resistance to all the major insecticides classes and it has become increasingly difficult to control its population in India. H. armigera alone accounts for the consumption of half of the total pesticides used in India for the protection of different crops (Suryavanshi et al. 2008). For managing the insecticides resistance in better way, different group of insecticides should be preferred in lieu of a single group of insecticides. In this study, an attempt was made to study the bio-efficacy of certain new molecules against H. armigera in chickpea. The number of H. armigera larvae was counted on 5 randomly selected plants in each plot. Pretreatment larval count on 5 plants was made a day before spraying while post treatment counts were taken 7 days after applications. The per cent reduction in larval population was calculated on the basis of number of larvae recorded in treated and control plots. The dat a were stati stically analyzed after arc sin transformation. The significance was tested by referring to ‘F’ tables of Fisher and Yates (1963). To study the relative efficacy of various chemical treatments, the percentage pod damage and plot yield were recorded after the crop harvest in both cropping years. After harvesting, all the pod of 5 plants of individual plot were collected and pooled together. Finally, 100 pods were picked up randomly and per cent pod damage was recorded. For recording the yield, all the pods from individual treatment were threshed and grain weight obtained were converted into q/h. The cost: benefit ratio was calculated with the help of costs of inputs and yield obtained. The percentage reduction of the pod borer over untreated check in different treatments was calculated using 292 Journal of Food Legumes 25(4), 2012 Henderson and Tilton‘s (1955) formula as given below: T C Per cent efficacy = 1 a b 100 Tb C a Where, Ta = Population in the treated plot after spray Tb= Population in the treated plot before spray Ca = Population in the control plot after spray Cb= Population in the control plot before spray RESULTS AND DISCUSSION Per cent reduction in larval population: The pooled mean after 7 days of 1st application showed that all treatments were found significantly superior to control (Table 1). Spinosad 45 SC @ 100 g a.i./ha was the best treatment that reduced 81.2% larval population; however, it did not differ significantly with indoxacarb 14.5 EC @ 75 g a.i./ha, fipronil 5 SC@ 50 g/ha, emamectin benzoate 5 SG @ 11 g/ha1, cartap hydrochloride 50 SP @ 500 g a.i./ha, and fenvalerate 20 EC@300 g a.i./ha which reduced larval population by 79.7, 77.9, 76.9, 76.3 and 75.6 per cents, respectively. Quinalphos 25EC @450 g a.i./ha also reduced larval population by 74.4% while azadirachtin 1500 ppm and HaNPV @ 250 LE/ha reduced the population by 52.4 and 48.8%, respectively. The least effective and significantly inferior treatment in comparison to others was HaNPV @250 LE/ha (with 48.8% reduction in population). However, on the basis of pooled data, the reduction in larval population at 7 days after second spray showed that all the treatments were found significantly superior to control. Thus, spinosad 45 SC @ 100 g a.i/ha was the best treatment that reduced 79.8% larval population followed by indoxacarb 14.5 EC @ 75 g a.i./ha (with 78.3% reduction in larval population). The other treatments viz., fipronil 5 SC@ 50 g a.i./ha, emamaectin benzoate 5 SG @ 11 g a.i./ha, cartap hydrochloride 50 SP @ 500 g/ha, quinalphos 25EC@450 g a.i./ha, fenvalerate 20 EC@300 g a.i./ha, HaNPV @ 250 LE/ha and azadirachtin 1500ppm reduced larval population by 75.2, 74.5, 72.6, 70.1, 70.0, 58.1 and 54.1 per cent, respectively. The minimum reduction in larval population noted after second spray in azadirachtin was 54.1%. The present findings were in conformity with the findings of Kambrekar et al. (2012), Anandi et al. (2011), Deshmukh et al. (2010) and Singh et al. (2008). Pod damage: The bio-efficacy of different insecticidal treatment on per cent pod damage by H. armigera was evaluated under field condition. On the basis of pooled mean, it showed that all the treatments were found significantly superior to the control. The minimum pod damage was observed in spinosad 45 SC @ 100 g a.i. /ha (2.3%) while maximum damage was observed in azadirachtin 1500 ppm treatment (12.8%). The other treatments showing increased per cent pod damage included indoxacarb (3.6%), cartap hydrochloride (4.7%), fipronil (5.4%), emamectin benzoate (6.1%), fenvalerate (6.0%), quinalphos (8.3%) and HaNPV (10.7%). Present findings were in conformity with the findings of Singh et al. (2004) and Singh et al. (2008). Seed Yield: During both years when efficacy was tested in terms of grain yield, all the treatments proved significantly Table 1. Efficacy of insecticidal treatment on H.armigera larval population during 2010-12* Treatment details HaNPV@ 0.5ml/lit. [email protected] ml/lit. Fenvarlate @3ml/lit. [email protected] ml/lit. Emamactin [email protected] gm/lit. Azadirachtin@5 ml/lit. Cartap hydrochloride @ 2gm/lit. Fipronil@2ml/lit. [email protected]/lit. Untreated control SEm(±) CD (P=0.05) CV (%) Dosage g a.i./ha Per cent reduction in larval population over control Pod damage (%) Yield t/ha 7 days after first spraying 7 days after second spraying 2010-11 2011-12 Mean 2010-11 2011-12 Mean 2010-11 2011-12 Mean 2010-11 2011-12 Mean 250 LE 49.2 48.8 59.4 56.8 10.9 10.5 48.8 (44.3) 58.1 (49.7) 10.7 (19.3) 1.11 1.12 1.11 (44.5) (44.3) (50.4) (48.9) (19.3) (19.2) 100 81.7 81.2 80.3 79.3 2.3 2.2 81.2 (64.3) 79.8 (63.3) 2.3 (8.6) 1.74 1.84 1.79 (64.7) (64.3) (63.6) (62.9) (8.7) (8.5) 300 75.4 75.6 73.6 66.4 6.5 5.6 75.6 (60.4) 70.0 (56.8) 6.0 (14.2) 1.30 1.30 1.30 (60.3) (60.4) (59.1) (54.5) (14.8) (13.6) 450 73.9 74.4 72.1 67.9 8.8 7.7 74.4 (59.6) 70.1 (56.9) 8.3 (16.7) 1.42 1.41 1.41 (59.3) (59.6) (58.1) (55.5) (17.3) (16.1) 11 77.2 76.9 74.0 75.1 5.6 6.7 76.9 (61.3) 74.5 (59.7) 6.1 (14.3) 1.41 1.60 1.51 (61.5) (61.2) (53.3) (60.0) (13.6) (14.9) 1500 54.2 52.4 57.8 50.2 13.3 12.4 52.4 (46.4) 54.1 (47.4) 12.8 (21.0) 1.06 1.07 1.06 ppm (47.4) (46.4) (49.5) (45.1) (21.4) (20.7) 500 76.4 76.3 72.4 72.8 4.5 4.9 76.3 (60.9) 72.6 (58.4) 4.7 (12.5) 1.37 1.61 1.49 (61.0) (60.9) (58.3) (58.6) (12.2) (12.8) 50 78.5 77.9 76.2 74.3 5.4 5.4 77.9 (62.0) 75.2 (60.1) 5.4 (13.4) 1.55 1.62 1.58 (62.4) (62.0) (57.2) (59.5) (13.4) (13.4) 75 79.5 79.7 78.8 77.8 4.0 3.2 79.7 (63.2) 78.3 (62.2) 3.6 (10.9) 1.57 1.67 1.62 (63.1) (63.2) (62.6) (61.8) (11.5) (10.3) 28.4 28.4 28.4 (32.2) 0.82 0.83 0.83 (32.2) (32.2) 1.6 1.6 1.6 1.5 1.5 1.5 0.5 0.5 0.5 0.01 0.01 0.01 4.8 4.8 4.8 4.5 4.6 4.6 1.4 1.4 1.4 0.03 0.03 0.03 5.4 5.4 5.4 5.3 5.2 5.3 5.0 5.0 5.0 4.9 4.8 4.9 *Figures in parentheses are Arc sine transformation Singh et al.: Bio-efficacy of some insecticides against H. armigera (Hubner) on chickpea (Cicer arietinum L.) 293 Table 2. Benefit cost ratio of various insecticidal treatment on chickpea during 2010-12* Treatments detail HaNPV Spinosad Fenvarlate Quinolphos Emamactin Benzoate Azadirachtin Cartap hydrochloride Fipronil Indoxacarb Untreated control Increase in yield (t/ha) over control 10-11 11-12 Mean Cost of increased yield over control (`/ha)# 10-11 11-12 Mean Total cost of plant protection (`/ha)* 10-11 11-12 Mean Net Profit (`/ha) ICBR 1112 1:4.3 1:4.9 1:8.4 1:4.5 RANK 0.28 0.91 0.48 0.60 0.29 1.01 0.46 0.58 0.29 0.96 0.47 0.59 8520 27420 14370 17970 12138 42294 19446 24192 10329 34857 16908 21081 2100 6936 1860 4200 2300 7136 2060 4400 2200 7036 1960 4300 10-11 11-12 Mean 1011 6420 9838 8129 1:3.1 20484 35158 27821 1:3.0 12510 17386 14948 1:6.7 13770 19792 16781 1:3.3 Mean 1:3.7 1:3.9 1:7.6 1:3.9 IX VI II VIII 0.59 0.76 0.68 17610 32046 24828 4912 5112 5012 12698 26934 19816 1:2.6 1:5.3 1:3.9 VII 0.23 0.23 0.23 7020 10038 8529 1500 1700 1600 5520 8338 6929 1:3.7 1:4.9 1:4.3 V 0.54 0.78 0.66 16320 32634 24477 2800 3000 2900 13520 29634 21577 1:4.8 1:9.9 1:7.4 III 0.72 0.75 - 0.78 0.84 - 0.75 0.79 - 21720 22530 - 32928 35154 - 27324 28842 - 2860 4442 - 3060 4006 - 2960 4224 - 18860 29868 24364 1:6.6 1:9.8 1:8.2 18088 30512 24300 1:4.1 1:6.6 1:5.3 - I IV X *Includes cost of Sprayer, Labours for two applications. (Charges : Sprayer @`15/day/sprayer In 2010-11 and @`20/day/sprayer 2011-12, Labour charges @`125/day/labour In 2010-11 and @`200/day/labour in 2011-12), Quantity of water per ha: 500 lit., No. of labours per ha: 2/spray, # Price of chickpea @`30 /kg (2010-11) and @`42/kg (2011-12) superior to control (untreated). The maximum and minimum yield was obtained in both years from the plot treated with spinosad and azadirchtin, respectively. On the basis of mean data, the maximum yield obtained among the all treated plots was under spinosad 45 SC @ 100 g a.i./ ha (1.79 t /ha) while the minimum yield was obtained with azadirachtin 1500 ppm (1.06 t /ha). Present findings are in conformity with the findings of Kambrekar et al. (2012), Deshmukh et al. (2010) and Ladaji (2004) who reported that maximum yield could be obtained with spinosad in comparison to other treatments. Economics: The cost benefit ratio of various insecticidal treatments on chickpea during both years showed that use of insecticides against H. armigera larvae increased the yield. The pooled mean data showed maximum cost benefit ratio (1:8.2) in the plots treated with fipronil 5 SC@ 50 g a.i. /ha while the lowest cost benefit ratio (1: 3.7) was obtained in HaNPV @ 250 LE/ha treated plots. Present findings are in conformity with the findings of Singh et al. (2004) and Singh et al. (2008). Based on the two years study, it was inferred that spinosad 45 SC @ 100 g a.i. /ha was the best treatment so far to due maximum larval population reduction, minimum per cent of pod damage and maximum seed yield. However, the maximum cost: benefit ratio was under fipronil 5 SC@ 50 g a.i. /ha. REFERENCES Anandhi DMP, Elamathi S and Simon S. 2011. Evaluation of biorational insecticides for management of Helicoverpa armigera in chickpea: Annals of Plant Protection Sciences. 19: 207-209. Chaudhary RRP and Sachan RB. 1999. Comparative efficacy and economics of some insecticides against gram podborer Helicoverpa armigera (Hubner) in chickpea in western plain of Uttar Pradesh. Bhartiya Krishi Anusandhan Patrika 10: 159-164. Deshmukh SG, Sureja BV, Jethva DM and Chatar VP. 2010. Estimation of yield losses by pod borer Helicoverpa armigera (Hubner) on chickpea. Legume Research 33: 1, 67-69. Fisher RA and Yates F. 1963. Statistical tables for biological, agricultural and medical research. 6. Aufl. Oliver & Boyd, London. 146 S. Preis 30 s. Henderson CF and Tilton EW. 1955. Pests with acaricides against the brown wheat mite. J. Economic Entomology 48: 157-161. Kambrekar DN, Somanagouda G, Basavarajappa MP and Halagalimath SP. 2012. Effect of different dosages of Emamectin benzoate 5 SG and Indoxacarb 14.5 SC on pod borer (Helicoverpa armigera) infesting chickpea. Legume Research 35: 13-17. Ladaji RN. 2004. Management of chickpea pod borer, Helicoverpa armigera (Hubner) using indigenous materials and newer insecticides. M. Sc. (Ag) Thesis, UAS, Dharwad (India). Panse VG and Sukhatme PV. 1985, Statistical methods for Agricultural workers. ICAR, New Delhi. 381 pp. Patel RK.1979. Unusual outbreak of gram pod borer on gram in Madhya Pradesh. Science and Root cheek culture 45: 335-336. Singh H, Mahajan G and Singh I. 2004. Efficacy of different insecticides against the gram pod borer (Helicoverpa armigera) on chickpea (Cicer arietinum). Legume Research 27: 233-234. Singh S, Choudhary DP, Sharma C, Mahla RS and Mathur YS. 2008. Efficacy of different insecticides against Helicoverpa armigera (Hubner) on chickpea Cicer arietinum. Indian Journal of Entomology 70: 177-181. Suryavanshi DS, Bhede BV, Bosale SV and More DG. 2008. Insecticide resistance in field population of H. armigera (Hub.) (Lepidoptera : Noctuidae). Indian Journal of Entomology 70: 44-46. Journal of Food Legumes 25(4): 294-299, 2012 Comparison of nodC and 16S rDNA gene analysis of rhizobia associated with legumes of arid and semi-arid regions of Haryana RAJESH GERA, RANJANA BHATIA and VARUN KUMAR Department of Microbiology, CCS Haryana Agricultural University, Hisar-125004, India; E-mail: [email protected] (Received: February 29, 2012; Accepted: October 06, 2012) ABSTRACT In this study, nodC and 16 rDNA gene analysis of rhizobia isolated from legumes growing in arid and semi-arid regions of Haryana, India were compared. A total of 90 rhizobial isolates were obtained from nodules of different leguminous crops of arid and semi-arid zones of Haryana (India) and tested for nodulation efficiency in their respective crops. Out of the 90 isolates, 33 isolates forming better nodules were selected for the investigation of genetic diversity. The results of nodC restriction analysis of these isolates showed that Vigna unguiculata gro up lay s eparately from the Tri foli um alexandrinum and Cicer arietinum groups. However, restriction analysis of 16S rDNA revealed that the T. alexandrinum and the C. arietinum isolates formed separate phylotypes within the V. unguiculata group but with less similarity between them. From the comparison of nodC and 16S rDNA gene analysis of these isolates it has been concluded that nodC pattern shows some correlation with host plant range whereas 16S rDNA analysis results in wide diversity. Five of the isolates, one from each phylogenetic group, were identified on the basis of partial 16S rDNA gene sequencing. Key words: 16S rDNA, nodC, Phylogeny, RFLP, Rhizobia Haryana is a small state which falls under arid and semi arid zones of Northern India. Agriculture under arid zones is low input agriculture with minimum application of chemical fertilizers and pesticides. Role of microorganisms, especially in nitrogen fixation, plant health promotion and phosphate solubilization is very important under such conditions. Abiotic stresses like high temperature and drought during summer determine the microflora of soils. Cropping pattern and management practices such as fertilization, crop rotation and application of organic amendments may favor some microorganisms over the others. Crop production under rainfed conditions is solely dependent on microbial resources for nutrients. In arid and semi-arid zones of Haryana, the legumes are in rotation with non-leguminous crops under low input agriculture. Leguminous crops fix nitrogen and benefit the succeeding crops by providing residual nitrogen. Nonsymbiotic microorganisms have been known to benefit the cereals by increasing nutrient uptake. Symbiotic rhizobia have been classified as belonging to the á-subgroup of the Proteobacteria and to one of the six genera: Rhizobium, Sin orhi zobi um, Bradyrhizobiu m, Azorh izo biu m, Mesorhizobium and Allorhizobium (Garrity et al. 2003). However, recent studies have shown that the bacteria capable of nodulating legumes may also belong to other genera lying wi thin á- and â-Proteo bact eria (Chen et al. 200 3, Rasolomampianina et al. 2005, Sy et al. 2001, Trujillo et al. 2005 and Zakhia et al. 2004)). ã-Proteobacteria were found associated with legume nodules, although their presence and role is yet to be defined (Benhizia et al. 2004). Rhizobia are proteobacteria which establish symbiotic relationship with leguminous plants leading to the formation of root nodules where they fix atmospheric nitrogen (Dubey et al. 2010). Rhizobia that nodulate Trifolium alexandrinum, Cicer arietinum and Vigna unguiculata (Vigna radiata, Vigna aconitifolia, Cajanus cajan, Cyamopsis tetragonoloba and Vigna unguiculata) legumes are Rhizobium leguminosarum bv. trifolii, Rhi zobi um sp. and Brad yrhizob ium sp, respectively. Soil types, host and environmental conditions are the key factors to study the diversity of rhizobia. A better understanding of phylogenetic relationships among different rhizobia may be gained by using the 16S rRNA gene. Host specificity may not be related to 16S rRNA but phylogenies of nod genes have often been correlated with the host plant (Ueda et al. 1995, Zeze et al. 2001). To form an effective symbiosis, rhizobia require several classes of specific genes involved in the morphogenesis of nodules. The nod genes are unique to rhizobia and the phylogenies of nod A, nod B, nod C and nod D resemble each other. We used nod C gene for our studies. Nucleic acid based techniques such as nodC and 16S rDNA restriction analysis have been used to gain a better understanding of microbial comparative community structure and function in the edaphic component of soils and the rhizosphere. This study was carried out to identify rhizobia isolated from legumes grown in the semi-arid zones of Haryana on the basis of PCR-RFLP analysis of nodC and 16S rDNA genes. MATERIALS AND METHODS Nodule sampling and isolation of rhizobia: In order to obtain the nodule samples representing the spontaneous distribution of legumes in arid and semi-arid zones of Haryana, root nodules of Trifolium alexandrinum (berseem), Vigna radiata (mungbean), Cicer arietinum (chickpea), Vigna unguiculata (cowpea), Vigna aconitifolia (mothbean), Cajanus cajan (pigeonpea) and Cyamopsis tetragonoloba (guar) legumes Gera et al.: Comparison of nodC and 16S rDNA gene analysis of rhizobia associated with legumes of arid and semi-arid were non-selectively sampled. Segments of roots with attached nodules from the plants were excised and transported in plastic bags to the laboratory. Root nodules were collected and surface sterilized with 0.1% mercuric chloride (HgCl2), washed with sterile distilled water followed by surface treatment with 95% ethanol and further rinsing with sterile distilled water. Properly washed nodules were again washed 8-9 times with sterile distilled water to remove the traces of HgCl2. Bacteria were isolated from crushed nodules on YEMA media plates supplemented with 2.5 mg/100 ml Congo red (Vincent 1970). The isolates were purified by re-streaking on fresh plates and the purified isolates were maintained on YEMA slants at 4°C. Plant nodulation test: All the rhizobial isolates were tested for their nodulation efficiency in their respective crops. Rhizobial isolates were inoculated separately in YEM broth and incubated on rotary shaker at 28±2°C. Seeds of the legumes varietySC 5 (Cicer arietinum), HFG 156 (Cyamopsis tetragonoloba), HFB 600 (Trifolium alexandrinum), CS 88 (Vigna unguiculata), Manak (Cajanus cajan), RMO-225 (Vigna aconitifolia) and Basanti (Vigna radiata) were surface sterilized and kept for germination for 24 h. River sand was thoroughly washed with acid followed by 6-7 washings with water and sterilized in an oven at 180°C for one hour in trays. The sand was then filled in cups and nutrient solution was added to it. The cups were covered with paper and held in position with the help of a thread. These nodulation test assemblies were sterilized in an autoclave at 15 psi for 1 h. Germinated seedlings were transferred to the sterilized cups containing sand, along with 1-2 ml of broth of respective rhizobial isolates. These nodulation test assemblies were kept in pot house and watered daily with sterilized Sloger’s nitrogen free watering solution. After 50 days of growth, plants were uprooted and analyzed for nodulation. Isolation of Genomic DNA: Genomic DNA of rhizobial strains was isolated from log phase grown cells of rhizobia using CTAB method (Ausubel et al. 19 87). The DNA was resuspended in 50-100 µl of TE buffer and quantified. Amplification of nodC gene: Genomic DNA of all the rhizobia isolated from different crops was amplified for nodC gene using nodCF (5'-AYG THG TYG AYG ACG GTT C - 3') and nodCI (5'-CGY GAC AGC CAN TCK CTA TTG - 3') (Sarita et al. 2005) primers by Polymerase chain reaction (PCR). The reaction mixture (25 µl) for PCR contained 12.5 µl of Red Taq Ready Mix (Bangalore genei), 2.5 µl of each primer (10 pmol each), 1µl template DNA (50-70 ng/µl) and 6.5 µl sterile water. The conditions of thermal cycler for nodC gene amplification were: initial denaturation at 95°C for 3 min followed by 35 cycles of denaturation at 94°C for 1 min, annealing at 55oC for 1 min and extension at 72oC for 2 min with a final extension at 72oC for 3 min. The PCR was then set on hold at 4°C. The lid temperature was maintained at 105°C. Amplified gene was visualized in 2% agarose after electrophoresis using gel 295 documentation system (Minibis Pro). PCR-RFLP analysis of nodC gene: Restriction fragment length polymorphism (RFLP) is the identification of specific restriction patterns that reveal the difference between the DNA fragment sizes in individual organisms. The PCR amplified products of nodC gene were digested separately with two restriction enzymes MspI and RsaI. For the restriction digestion, 10 µl of amplified nodC product was treated with 1µl of both the enzymes separately and held on constant 37°C for 12 h in thermal cycler. The digested product was checked on 2% agarose gel and the different RFLP patterns were recorded using gel documentation system. Amplification of 16S rDNA: Universal primers BAC 27 F (5’AGA GTT TGA TCC TGG CTC AG - 3’) (Donachie et al. 2004) and 1488 R (5’- CGG TTA CCT TGT TAG GAC TTC ACC - 3’) (Herrera-Cervera et al. 1999) were used for the amplification of 16S rDNA gene in the thermal cycler. For PCR, 50 µl reaction mixture included 50-70ng/µl as template, 2 µl of each primer having concentration of 10 µM, 1 µl of 10 mM dNTPs, 5 µl of Taq buffer (10X) and 3 units of Taq DNA polymerase. The conditions of thermal cycler for 16S rDNA gene amplification were: initial denaturation at 94°C for 3min followed by 40 cycles of denaturation at 94°C for 30 s, annealing at 50°C for 30 s and extension at 72°C for 1 min with a final extension at 72°C for 10 min. This was followed with a final cooling at 4°C. Amplified gene was visualized in 1.5% agarose after electrophoresis using gel documentation system. PCR-RFLP analysis of 16S rDNA: PCR amplified products of 16S rDNA gene of various rhizobial isolates were digested separately with restriction enzymes MspI and RsaI. For the restriction digestion, 10 µl of amplified product was treated with 1µl of both the enzymes separately and held on constant 37°C for 12 h in thermal cycler. The digested product was checked on 2% agarose gel and the different RFLP patterns were recorded using gel documentation system. Banding patterns of different isolates were analyzed by Unweighted Pair Group Method with Arithmetic Mean (UPGMA) program cluster analysis using NTSYS-PC program. The results were applied to construct a dendrogram depicting the clustering of the various isolates on the basis of the similarity index of the nod C and 16S rRNA gene within them. Partial 16S rDNA gene seq uencing: Representative phylotypes from each phylogenetic group were selected from partial 16S rDNA gene sequencing using both forward BAC 27F and reverse 1488R primers. The blastn was used to compare the 16S rDNA gene nucleotide sequences of isolates. RESULTS AND DISCUSSION Isolation of symbiotic nitrogen fixing bacteria and Plant nodulation tests: In this study, ninety rhizobial isolates were obtained from the nodules collected from different host legumes grown in arid and semi-arid zones of Haryana, India. 296 Journal of Food Legumes 25(4), 2012 These isolates included 9 from T. alexandrinum, 38 from V. radiata, 24 from V. unguiculata, 7 from V. aconitifolia, 3 from C. arietinum, 3 from C. cajan and 6 from C. tetragonoloba. When tested for their nodulation efficiency in their respective crops, all the isolates were able to form nodules on their respective crops but with variable efficiency. Their nodulation potential indicated that these were rhizobia. Out of the 90 isolates, only 33 were able to form better nodules and these were selected for the phylogenetic analysis. Amplification of nodC and 16S rDNA gene in rhizobial isolates: DNA based methods are most suitable for the measurement of genetic relatedness (Sikora and Redzepovic 2003). In this investigation, 33 rhizobial isolates selected for phylogenetic analysis were tested for the presence of nodC and 16S rDNA genes using PCR. The amplification of nodC gene through PCR using specific primers revealed amplicons of 620bp in all the isolates. On the basis of the presence of nodC gene, the isolates were authenticated as rhizobia. The 16S rDNA gene of all the rhizobial isolates was amplified with universal primers and amplicons of 1460 bp were observed after comparison with low range ruler DNA (Bangalore Genei, India). Nodulation genes in rhizobia have been widely used to perform evolutionary analysis and to estimate their host ranges (Ueda et al. 1995, Laguerre et al. 2001, Chen et al. 2008). One of the nodulation genes, nodC, which encodes the enzyme involved in the first step of the Nod factor assembly, has often been chosen as a nodulation marker in different studies because it is essential for nodulation in most of the rhizobial species examined so far (Ueda et al. 1995, Moschetti et al. 2005, Kalita et al. 2006, Sarita et al. 2008). The amplification Table 1. Restriction patterns of rhizobial isolates revealed by RFLP analysis of PCR-amplified nodC genes Isolates Host plant M 1, M 2, M 3, M 5 M 7, M 12, M 13, M 15, M 20 M 8, M 14 M 16 M 17, M 10 C 2, C 4, C 5 C 11 C 12, C 13, C 14, C 15, C 17 B 5, B 13, B 14, B 23 Vigna radiata Vigna radiata A2 G1 G2 Mo 1, Mo 2 Rz 1 nodC genotype I II nodC RFLP Msp I Rsa I a a a b Vigna radiata Vigna radiata Vigna radiata Vigna unguiculata Vigna unguiculata Vigna unguiculata III IV V VI II VII b c d e a f b b c b b b Trifolium alexandrinum Cajanus cajan Cyamopsis tetragonoloba Cyamopsis tetragonoloba Vigna aconitifolia Cicer arietinum VIII g d IX V h d c c I a a II X a i b e Ten nod C genotypes were revealed of nodC gene using specific primers is a reliable and quick method for the identification of rhizobial strains (Harrison et al. 1992, Oliveira et al. 1999). Amplified ribosomal DNA restriction analysis (ARDRA) has been found to be a useful approach to study Rhizobium diversity more effectively and more authentically than the nodC gene (Sikora and Redzepovic 2003, Laguerre et al. 1993, Moschetti et al. 2005, Ltaief et al. 2007, Rashid et al. 2009, Nandwani and Dudeja 2009). We have used both nodC and 16S rDNA genes for diversity analysis of Rhizobia. PCR-RFLP analysis of nodC gene amplicons: Representative phylotypes have been identified for different crop rhizobia based upon RFLP analysis of the nodC gene. The PCR amplified products of nodC gene were digested with two restriction enzymes MspI and RsaI and different RFLP patterns were recorded (Table 1). Maximum number of isolates, which showed high similarity in the nodC gene, was obtained from hosts earlier classified as Vigna unguiculata miscellary group (Figure 1a). At the similarity value of 46%, the isolates were Table 2. Restriction patterns of rhizobial isolates revealed by RFLP analysis of PCR-amplified 16S rDNA genes Isolates Host plant 16S rDNA genotype M 1, M 2, M 3, Vigna radiata I M5 M7 Vigna radiata II M8 Vigna radiata III M 12 Vigna radiata IV M 13 Vigna radiata V M 14 Vigna radiata VI M 15 Vigna radiata VII M 16 Vigna radiata VIII M 17 Vigna radiata IX M 10 Vigna radiata X M 20 Vigna radiata XI C2 Vigna unguiculata XII C4 Vigna unguiculata XIII C5 Vigna unguiculata XIV C 11 Vigna unguiculata XV C 12 Vigna unguiculata XVI C 15 Vigna unguiculata XVII C 17 Vigna unguiculata XVIII C 13, C 14 Vigna unguiculata XIX B 5, B 23 Trifolium XX alexandrinum B 13 Trifolium XXI alexandrinum B 14 Trifolium XXII alexandrinum A2 Cajanus cajan XXIII G1 Cyamopsis XXIV tetragonoloba G2 Cyamopsis XXV tetragonoloba MO 1 Vigna aconitifolia XXVI Rz 1 Cicer arietinum XXVII Twenty seven 16S rDNA genotypes were revealed 16S rDNA RFLP Msp I Rsa I a a a b a a a a c a d e f f f f f g f f h b c d e f g h i j k l m n o p q r s t i t j u k l v w m x n o y e Gera et al.: Comparison of nodC and 16S rDNA gene analysis of rhizobia associated with legumes of arid and semi-arid 297 M1 M2 M3 M5 M7 M13 M14 M8 M12 M17 M15 M10 M20 C2 C12 C4 C17 C13 C14 C15 C5 C11 B5 B23 B13 B14 A2 G2 Mo1 Rz1 M16 0.56 Fig 1. 0.67 0.78 coefficient 0.89 G1 1.00 (a) (b) Dendrogram depicting grouping of various rhizobial isolates on the basis of (a) nodC-RFLP and (b) 16S rDNA-RFLP after restriction digestion with MspI and RsaI. The letters in the dendrogram represent B: Trifolium alexandrinum; Rz1: Cicer arietinum; A: Cajanus cajan; G: Cyamopsis tetragonoloba; Mo: Vigna aconitifolia; M: Vigna radiata and C: Vigna unguiculata. divided into two major clusters. The rhizobia from T. alexandrinum and C. arietinum legumes formed a separate cluster from the other legume crops indicating their different host range. Moreover, the T. alexandrinum isolates formed a single phylotype, the isolate Rz1 i.e. the Mesorhizobium strain from C. arietinu m sho wed 58 % simi larity to the T. alexandrinum isolates cluster. V. unguiculata isolates formed 2 major clusters having 3 and 5 isolates, respectively. The isolate C11 showed 100% similarity with V. aconitifolia isolates and few of the V. radiata isolates. On the other hand, V. radi ata iso lates showed 5 different phylo types with divergence started at about 90% similarity coefficient and formed 4 major clusters having 5, 3, 8 and 2 isolates, respectively. Isolates G1 and G2 from C. tetragonoloba exhibited 92% similarity among themselves but showed 100% similarity to V. radiata isolates M10, M17 and M1, M2, M3, M5, respectively. The isolate A2 from C. cajan showed a single phylotype with a 92% similarity coefficient. Isolate M1, M2, M3 and M5, isolates C13 & C14 and isolates B5 & B13 showed 100% similarityamong themselves, respectively, in both nodC and 16S rDNA restriction patterns indicating that they may represent the same strain. PCR-RFLP analysis of 16S rDNA gene amplicons: The PCR amplified products of 16S rDNA gene were digested with two restriction enzymes (MspI and RsaI) and different RFLP patterns were recorded (Table 2). Combined RFLP analysis of 16S rDNA gene of all the rhizobial isolates from different crops using MspI and RsaI revealed high polymorphism (Figure 1b). At the similarity level of 56%, two major groups were formed. V. radiata and V. unguiculata isolates formed separate clusters which were divided into various subgroups at about 90% similarity coefficient and showed 68% similarity among them. While all the T. alexandrinum isolates formed a single cluster with divergence at about 93% similarity coefficient, isolates from C. tetragonoloba, V. aconitifolia, C. cajan and Mesorhizobium strain from C. arietinum (Rz1), formed single phylotypes with 80% similarity coefficient. Isolates M16 from V. radiata and G1 from C. tetragonoloba formed separate group with 65% similarity among themselves. All the isolates from V. unguiculata and V. radiata grouped together, respectively. Partial nodC gene sequencing: Five isolates M2, M16, Mo1, C17 and B5 from V. radiata, V. aconitifolia, V. unguiculata and T. alexandrinum from each representative group were identified as Bradyrhizobium yuanmingense, Bradyrhizobium sp., Bradyrhizobium sp., Bradyrhizobium sp. and Rhizobium leguminosarum bv. trifolii on the basis of partial nodC gene sequencing using both forward and reverse nodC primers (Table 3). The PCR-RFLP analysis of nodC revealed that V. unguiculata group lay separately from the T. alexandrinum and C. arietinum, whereas that of 16S rDNA genes showed the T. alexandrinum and C. arietinum isolates as separate phylotypes within the V. unguiculata group but with less similarity between them. Rhizobial isolates from the different crops showed much more similarity in nodC gene with most of them exhibiting 80% similarity. On the other hand, in the 16S rDNA analysis, the rhizobial isolates from individual crops showed high similarity (up to 90%) among them but the diversity increased on comparison with isolates from other crops. Restriction patterns of nodC gene using both MspI and RsaI categorized all the tested isolates into ten groups (Table 1) while twenty seven groups were generated with 16S 298 Journal of Food Legumes 25(4), 2012 Table 3. Identity match of the sequenced rhizobial isolates S. No. 1 Isolate Crop M2 Vigna radiata 2 3 4 5 M16 Mo1 C17 B5 Vigna radiata Vigna aconitifolia Vigna unguiculata Trifolium alexandrinum % Identified Match to 16S Similarity rDNA 100% Bradyrhizobium yuanmingense 100% Bradyrhizobium sp. 99% Bradyrhizobium sp. 92% Bradyrhizobium sp. 97% R. leguminosarum rDNA using both MspI and RsaI restriction enzymes (Table 2). The wide diversity of rhizobial isolates on the basis of 16S rDNA analysis is supported by many other studies carried out in the same region or in different areas (Mutch et al. 2003, Moschetti et al. 2005, Ventorino et al. 2007, Shamseldin et al. 2009). Such comparative studies on nodC and 16S rDNA gene analysis of rhizobial from arid and semi-arid regions of Haryana state have not been reported earlier. Thus, thirty three rhizobial isolates from seven host legumes of arid and semi-arid regions of Haryana, India showed good nodulation potential. From the comparison of nodC and 16S rDNA gene analysis of these isolates, it has been concluded that nodC pattern shows some correlation with host plant range whereas 16S rDNA analysis results in wide diversity. ACKNOWLEDGEMENT The authors acknowledge NBAIM, Mau for providing financial support to carry out this work. REFERENCES Ausubel FM, Brent R, Kingston R. 1987. Current Protocols in Molecular Biology. Wiley, New York. 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Environmental Microbiology 3: 363-370. Journal of Food Legumes 25(4): 300-305, 2012 Phenology, dry matter distribution and yield attributes under normal and drought stress conditions in Lentil (Lens culinaris Medik.) VIJAY LAXMI Indian Institute of Pulses Research, Kanpur-208 024, Uttar Pradesh, India; E-mail: [email protected] (Received: November 14, 2011 ; Accepted: December 06, 2012) ABSTRACT Twenty lentil (Lens culinaris Medik.) genotypes were raised in field under normal (irrigated) and drought stress (non-irrigated condition) during rabi seasons of 2008-09 and 2009-10. The observations on phenological days, dry matter distribution in plant parts at different phenological stages, mean environmental temperature and total degree days were recorded. Seed yield were higher under normal as compared to drought condition. Multiple regressions between seed yield and different traits exhibited that all of them did not have significant contribution. Seed yield and seed yield determining factors were found to be different under normal and drought stress conditions. Under non-irrigated condition, lentil seed yield was determined by total dry matter production and its partitioning/day, mean temperature and total degree-days during podding to maturity. Under irrigated condition, lentil seed yield was determined by root dry matter at vegetative stage, total dry matter yield, per day dry matter production and its partitioning during seed filling period and total degree-days during flowering to podding. Multiple correlation (R) of traits were 0.9996 and 0.9984 and coefficient of determination (R2) were 0.9992 and 0.9964 under non-irrigated and irrigated conditions, respectively. Optimum values of the traits for seed yield maximization also varied amongst irrigated and non-irrigated conditions. Key words: Irrigation, Lentil, Yield component Lentil (Lens culinaris Medik.) is an important component of the rainfed farming system of West Asia and Africa and source of high-quality protein for humans (Hamdi et al.1992). Research indicated increase in lentil seed yield with increase in irrigation frequency and total water use (Salehi et al. 2006). There are strong linear relationships (r2 >0.90) between yield and moisture supply in lentil (Silim et al. 1993). Salehi et al. (2006) reported that the seedling and flowering stage were most sensitive to water availability and drought stress. However, a deficiency of water during any growth stages in legume species often result in loss of seed yield. They suggested that high seed yield is related to pods size and number and resistance to pests and disease. Since, the lentil is of in-determinate growth, supplying available water may result in higher vegetative and reproductive growth periods and drought stress during the flowering stage decreases this period (Kusmenglu and Muehlbauer, 1998). Water deficit highly influences seed yield components and causes reduced pods per plant, seeds per pod and 100-seed weight. Hudak and Patterson (1995) showed that irrigation during seed filling period, improves yield. Water deficit also results in the decline of number of flowers, pods, seeds per pod, and size of pods and seed weight (Desclaus et al. 2000). As the cell losses water, vacuole usually crumples more than cell wall which causes the silt in the protoplasm. It seems that such damage results in the death of cells (Lessani and Mojtahedi, 2003). Yield loss of the plants under water deficit is one of the most important events for the plant breeders to improve yield but difference in the yield potential mainly relates to the adaptation factors than merely to the stress itself in which drought resistance indices are used to determine resistant genotypes (Mitra, 2001). Stress appearance during the reproductive stage reduces seed weight (Katerji et al. 2000). The amount of the yield loss depends on the stress range and plant growth stage at which stress occurred. In fact, plant susceptibility to stress varies from germination to the maturity (Schmidtke et al. 2004). One of the main drought resistance factors in plants is the ability of cells to tolerate a large amount of lost water. In general, water requirement in grain legumes is less as compared to cereals but availability of soil moisture has been reported to be the pivotal point for realization of yield potential. There are reports for response of grain legumes to irrigation in terms of yield and this enhancement is due to an increase in growth attributes at different crop growth stages in chickpea (Bhattacharya and Singh 1997) and in lentil (Bhattacharya and Chandra 1997, Bhattacharya 1999). Variation in yielding ability was reported in chickpea and lentil (Bhattacharya 1998) under drought and irrigated conditions. It has been reported that various physiological processes differ with growth stages and their effects on seed yield of any crop plant are different (Sinha 1973). Since major area for lentil cultivation is largely confined to rainfed condition, and future prospect for increasing irrigation facility to this crop is remote possibility, therefore, increase in productivity of lentil for rainfed sustainable land use in agro ecological system is essential. Knowledge about yield determination factor(s) in lentil under normal and drought conditions is scanty and therefore, present study was undertaken to evaluate the relationship of lentil seed yield with dry matter distribution in plant parts, phenology, yield attributing traits, mean temperatures and total degree-days at different crop growth stages under normal and drought conditions. MATERIALS AND METHODS Field experiments were conducted during Rabi of 2008- Vijay Laxmi : Phenology, dry matter distribution and yield attributes under normal and drought stress 2009 and 2009-2010 at Indian Institute of Pulses Research, Kanpur Farm under two planting conditions viz., normal (irrigation) and drought (non-irrigation) involving twenty lentils [Lens culinaris L. (Medik).] genotypes viz. IPL 59, IPL 60, IPL121, IPL522, ILL7663, EC 208355, EC 208362, EC 520204, Bihar local, VL 4, B 77, Ranjan, WBL 58, IPL 203, IPL 403, IPL 404, IPL 517, 94/1468, P 2016, WBL 58. The soil of the experimental site was Inceptisol having low available nitrogen (150 kg N/ha), medium available phosphorus (22 kg P2O5/ha) and medium in available potassium (180 kg K2O/ha).Sowing was done under split-plot design, keeping irrigation as mainplot and genotypes as sub-plots, in three replications adopting standard recommended agronomical practices. Phenological observations viz., days to flowering, podding and seed filling period were taken during crop ontogeny. Dry matter (g/plant) in different plant parts viz., root, shoot, leaf and pod during vegetative (30 days after sowing), flowering, podding and at maturity were observed (mean of five plants at each stage) from each treatment after separating the plant parts and oven drying them at 800 C for constant dry weight. At maturity, yield attributing traits viz., number of pod/ plant, pod and pod wall weight (g/plant), were recorded (mean of ten plants). Per day dry matter production during seed to seed period (kg/ha/ day), per day dry matter partitioning (kg/ha/day), total dry matter yield (q/ha) and seed yield (kg/ha) were also recorded. Mean temperature and total degree-days during different crop growth stages viz., during vegetative to flowering, flowering to podding and during podding to maturity were recorded. Harvest index (%) for each treatment were also calculated (Jain 1975). Following mathematical relationships under nonirrigated and irrigated conditions are used. Non-irrigated condition: Y = -3106.51 – 3.352 X + 3.229X2 + 1.105 X3 + 0.195 X4 – 0.067 X5 (R = 0.9996, R2 = 0.9992) [Where X = Total dry matter yield, X2 = per day dry matter production, X3 = per day dry matter partitioning, X4 = Mean temperature during podding to maturity, X5 = Total degree-days during podding to maturity] Irrigated condition: Y = 166.40 – 0.023 X – 2.831X2 + 2.793 X3 + 1.031 X4 – 0.035 X5 (R = 0.9984, R2 = 0.9968) [Where X = Root dry matter at vegetative stage, X2 = Total dry matter yield, X3 = per day dry matter production, X4 = per day dry matter partitioning, X5 = Total degree-days during flowering to podding] Data of both the years were tested for their homogeneity, 301 were pooled over years, and were subjected for test of significance for various factors and their respective interactive effects, coefficient of correlation and regression under nonirrigated and irrigated conditions between seed yield and traits at various crop growth stages were estimated (Snedecor and Cochran 1978). Association percentage of the traits with seed yield was also calculated (Hays 1955). Multiple regressions of traits and estimation of optimum values of the traits for seed yield maximization under drought and irrigated conditions were worked through regression coefficients (Snedecor and Cochran 1978). RESULTS AND DISCUSSION Mean temperature as well as total degree-days during various phenological durations was higher under normal as compared to drought condition, excepting the total degreedays during podding to maturity where it was higher under non-irrigated as compared to irrigated condition. Mean temperature expressed as increase from vegetative to maturity, whereas total degree-days were minimum during flowering to podding. Phenological days, dry matter distribution amongst plant parts at different growth stages, yield attributes, mean temperature and to tal degree-days duri ng vario us phenological durations had significant differences for various treatments (Table 1a and 1b). Phenological days, except during seed filling stage, were observed to be earlier under drought as compared to irrigated condition. Dry matter in plant parts under irrigated as compared non-irrigated condition, did not differ significantly for irrigation, genotypes and their interactions. Yield attributing traits were higher under irrigated as compared to non-irrigated condition, except number of pods/plant, pod weight/plant, harvest index which were lower under irrigated as compared to non-irrigated condition. Biological yield was higher under irrigated as compared to non-irrigated, but harvest index was higher under nonirrigated as compared to irrigated condition. Mean values observed as well as predicted (based on mathematical relationship) for seed yield were numerically same and were more under normal as compared to drought condition and showed significant differences for irrigation, genotypes and their interaction. Percent coefficient of variation was higher for dry matter accumulation and lowest for mean temperature and to tal degree-days duri ng phenological durations. Multiple regression of seed yield with phenological days, dry matter of plant parts at different stages, yield attributing traits, mean temperature and total degreedays revealed that all traits do not express significant relationship with yield therefore, traits having significant relationship were considered under non-irrigated and irrigated conditions separately. Under non irrigated condition, total dry matter yield, per day dry mat ter producti on and partitioning, mean temperature and total degree-days during podding to maturity were the main traits determining lentil 302 Journal of Food Legumes 25(4), 2012 seed yield. Multiple correlation (R) and coefficient of determination (R2) of these traits with seed yield were 0.9996 and 0.9992, respectively. Under irrigated condition lentil seed yield was mainly determined by root dry matter at vegetative stage, total dry matter yield, per day dry matter production and partitioning and total degree-days during flowering to podding. Multipl e correlat ion (R) and coeffici ent of determination (R2) of these traits with seed yield were 0.9984 and 0.9968, respectively. Predicted seed yield, based on mathematical relationship were parallel to observed seed yield under different condition. Behaviour of these traits was different under different conditions. Under non-irrigated condition, except for total dry matter yield had positive, whereas total dry matter yield had negative relationship with lentil seed yield under irrigated condition, per day dry matter during seed to seed period and its partitioning during seed filling period had positive, but root dry matter at vegetative stage, total degree-days during flowering to podding had negative relationship with seed yield. Estimation of coefficient of correlation, regression between seed yield and traits having significant contribution (Table 2) revealed that under non-irrigated condition, correlation with total dry matter yield (0.740), per day dry matter production (0.741) and partitioning (0.961) were significant, whereas those of mean temperature during podding and maturity (0.160) and total degree-days (0.073) were not significant. Under irrigated condition, significant correlation Table 1a. Descriptive parameters of phenological days and dry matter distribution at different growth stages in lentil under nonirrigated (NI) and irrigated (I) conditions Range Minimum Maximum Mean Phenological days Flowering NI 88.0 96.0 91.5 I 90.0 97.0 96.0 Podding NI 104.0 112.0 106.22 I 110.0 118.6 116.5 Seed Filling NI 35.0 39.0 36.8 I 31.0 36.0 32.1 Root Dry Matter (g/plant) Vegetative NI 0.02 0.14 0.05 I 0.02 0.90 0.05 Flowering NI 0.08 0.590 0.22 I 0.10 0.37 0.21 Podding NI 0.05 0.40 0.14 I 0.07 0.24 0.14 Maturity NI 0.02 0.17 0.06 I 0.03 0.10 0.06 Shoot Dry Matter (g/plant) Vegetative NI 0.07 1.46 0.52 I 0.14 1.99 0.78 Flowering NI 0.21 4.06 1.441 I 0.40 5.53 2.17 Podding NI 0.16 3.25 1.15 I 0.30 4.44 1.77 Maturity NI 0.07 1.47 0.52 I 0.03 0.10 0.06 Leaf Dry Matter (g/plant) Vegetative NI 0.13 3.13 1.36 I 0.05 4.36 1.18 Flowering NI 0.17 4.07 1.77 I 0.07 5.67 1.53 Podding NI 0.08 1.90 0.83 I 0.03 2.65 0.71 Maturity NI 0.16 3.89 1.69 I 0.06 5.42 1.51 Pod Dry Matter (g/plant) Flowering NI 0.03 1.19 0.33 I 0.00 3.33 0.81 Podding NI 0.40 4.10 1.86 I 0.15 7.13 2.60 Maturity NI 2.33 7.90 5.46 I 5.00 13.73 8.33 Figures in parenthesis are the CD at 5% and NS are “non significant Irrigation Variances Genotypes CV (%) Interaction 445.90 (3.63) 935.51 (3.27) 504.10 (2.79) 9.85 (1.81) 20.12 (NS) 2.77 (1.26) 4.47 (NS) 4.68 (NS) 0.67 (NS) 0.00 (NS) 0.003 (NS) 0.001 (NS) 0.000 (NS) 0.00 (NS) 0.005 (NS) 0.002 (NS) 0.001 (NS) 0.000 (NS) 0.008 (NS) 0.004 (NS) 0.001 (NS) 1.531 (NS) 1.896 (NS) 7.598 (NS) 1.566 (NS) 0.090 (NS) 0.697 (NS) 0.444 (NS) 0.091 (NS) 0.084 (NS) 0.652 (NS) 0.415 (NS) 0.085 (NS) 0.764 (NS) 1.282 (NS) 0.284 (NS) 0.708 (NS) 0.487 (NS) 0.822 (NS) 0.180 (NS) 0 .773 (NS) 0.630 (NS) 1.064 (NS) 0.231 (NS) 1.063 (NS) 5.136 (NS) 12.336 (NS) 186.307 (0.67) 0.333 (NS) 1.877 (NS) 13.994 (NS) 0.476 (NS) 3.260 (NS) 14.952 (0.87) 1.70 2.58 3.22 15.45 13.42 13.15 11.24 15.15 12.54 12.23 13.14 13.15 14.22 15.14 13.50 15.03 14.91 7.93 Vijay Laxmi : Phenology, dry matter distribution and yield attributes under normal and drought stress with seed yield was expressed by total dry matter yield (0.575), per day dry matter production (0.572) and per day dry matter partitioning (0.985). Association percentages of these traits were also very high. Optimum values of lentil seed yield determining traits under non-irrigated and irrigated conditions for seed yield maximization revealed that the values were different from the observed mean under respective conditions. It was interesting to note that the optimum values under both non-irrigated and irrigated conditions were of higher magnitudes as compared to observed mean values of the traits. Except dry matter accumulation in different plant parts at di fferent phenological stages expressed signi ficant differences. Non-significant differences for dry matter distribution amongst plant parts at stages were not properly understood. Probably the genotypes under study had more or less similar interaction with prevailing environmental conditions. However, significant differences for mean temperature and to tal degree-days duri ng vario us phenological durations were in agreement to earlier reports (Bhattacharya and Chandra 1997, Bhattacharya 1997). In 303 mathematical relationship for drought and irrigated conditions, total dry matter yield had negative relationship and it confirmed the earlier findings (Bhattacharya and Chandra 1997, Bhattacharya 1997) that excess vegetative growth was detrimental for realization of yield potential. It was interesting to note that phenological days failed to express any significant contribution for lentil seed yield under both non irrigated and irrigated conditions. It was again interesting that under nonirrigated condition role of mean temperature and total degreedays during podding to maturity was almost negligible. Probably during this period, numbers of pods retained/plant were determined and/ or lower temperature during this period was detrimental on flower and pod shedding and thus, its effect on lentil seed yield was positive. However, a both maximum and minimum temperature during post flowering period had maximum effect on seed yield of normal sown chickpea (Bhattacharya and Pandey 1999). Under irrigated condition, mean temperature during any phenological duration was not responsible for lentil seed yield determination. However, importance of maximum and minimum temperature Table 1b. Descriptive parameters of yield attributing traits, mean temperature and total- degree-days at different growth stages, observed and predicted seed yield in lentil under non-irrigated (NI) and irrigated (I) conditions. Range Minimum Maximum Yield Attributed Traits Pods/plant NI 40.0 198.0 I 53.0 190.0 Pod weight (g/plant) NI 3.55 14.51 I 3.28 14.14 Pod wall (g/plant) NI 0.07 9.04 I 0.03 2.65 Biological yield NI 22.08 35.42 I 23.04 47.92 Per day DM Production NI 15.66 24.43 I 15.78 32.16 Per Day DM Partitioning NI 27.41 50.60 I 34.44 68.89 Harvest Index (%) NI 38.75 58.59 I 24.84 63.83 Observed Seed NI 1041.67 1770.67 Yield I 1102.08 2135.42 Predicted Seed NI 1042.84 1779.97 Yield I 1097.89 2139.89 Mean Temperature (oC) Vegetative to NI 14.37 14.55 flowering I 14.37 14.65 Flowering to NI 14.64 15.37 podding I 14.83 16.20 Podding to NI 18.98 20.88 maturity I 20.54 23.17 Total degree-day Vegetative to NI 776.0 873.3 flowering I 776.0 922.1 Flowering to NI 156.1 333.1 Podding I 234.8 491.7 Podding to NI 730.7 753.7 maturity I 711.2 850.6 Figures in parenthesis are the CD at 5% and NS are “non significant” Irrigation Variances Genotypes Interaction 114.9 105.4 8.30 7.80 2.6 0.71 28.59 38.11 20.06 25.5 37.14 52.73 47.80 45.10 1363.89 1688.75 1363.89 1688.75 2023.98 (5.44) 5.97 (0.48) 1.23 (NS) 20391.40 (7.30) 73.30 (0.52) 5464.528 (1.69) 159.53 (NS) 2374557.6 (61.02) 2374558.4 (104.37) 5921.04 (14.17) 14.82 (0.84) 4.85 (0.86) 14458.02 (1.49) 64.30 (1.05) 253.552 (3.67) 119.20 (3.88) 257971.96 (107.95) 256472.22 (108.45) 3735.71 (20.03) 26.19 (1.19) 14.80 (1.22) 3207.71 (2.11) 14.67 (1.48) 81.14 (5.48) 81.14 (5.48) 45864.12 (152.67) 46569.45 (153.37) 14.41 14.59 15.20 15.25 19.66 21.73 0.724 (0.011) 0.065 (NS) 96.825 (1.11) 0.013 (0.08) 0.041 (NS) 0.840 (0.67) 0.010 (0.11) 0.117 (0.39) 0.282 (NS) 812.37 888.11 233.92 339.17 739.80 722.53 29054.14 (55.27 249260.92 (91.28) 6711.54 (10.94) 2591.32 (32.66) 2520.44 (NS) 189.57 (NS) 1211.45 (NS) 2754.34 (NS) 217.43 (NS) Mean CV (%) 11.36 9.23 12.86 3.96 4.08 7.22 7.37 6.25 6.28 0.50 1.59 2.85 3.39 12.20 2.13 304 Journal of Food Legumes 25(4), 2012 during vegetative and 1st flowering and during 1st to 50% flowering in normal sown chickpea had been reported (Bhattacharya and Pandey 1999). Significant differences in total degree-days during different phenological durations were mainly due to changes in phenological days under nonirrigated and irrigated conditions as advancement of days increased per day temperature and also the total degree-days. It was interesting to note that total degree-days during flowering to podding were lower than that of during vegetative to flowering or during podding to maturity which probably indicated that in lentil flowering phase was under lower temperature regime and pod filling stage was under higher temperature regime. Per day dry matter production during seed to seed period and per day dry matter partitioning to developing seeds during seed filling period had expressed significant and positive contribution for lentil seed yield under both irrigated and non irrigated conditions. Phenological day (s) had also non-significant contribution for lentil seed yield under both non irrigated and irrigated conditions but they showed their importance in conjunction with per day dry matter production as well as their partitioning. This, phenological days had their function in controlling the amount and/or rate of dry matter production or it’s partitioning to developing seeds. It had been reported that levels of dry matter accumulation and be altered through changes in seeding dates and/or growing environment in chickpea (Singh and Bhattacharya 1995) as there was a strong genotype x environment interaction in chickpea and the same was also confirmed (Bahl 1984). Positive relationship of per day dry matter production was probably due to the fact that under non-irrigated condition, plant tend to complete its life cycle under lesser seed to seed period and it also accumulated less dry matter. Therefore, increasing per day dry matter production would lead to higher dry matter yield. Similarly, higher magnitude of dry matter partitioning from vegetative to reproductive plant parts would lead to higher seed yield and therefore, per day dry matter partitioning expressed positive relationship with lentil seed yield. Although the relationship of total dry matter yield with seed yield was observed to be negative, but the value of total dry matter yield for seed yield maximization was estimated to be higher than observed mean under both non-irrigated and irrigated conditions. Higher temperature and/or total degreedays during reproductive stage (flowering to podding) led to higher flower and pod abscission resulting in less number of pods/plant and seed yield; and the same was attributed to the negative relationship of total degree-days during flowering to podding with seed yield. Multiple correlations of traits under non-irrigated and irrigated conditions for seed yield were 0.9996 and 0.9984, respectively. Wide differences in calculated optimum values and observed mean values of traits under non-irrigated and irrigated conditions showed that there were scope for better lentil seed yield under respective conditions either through genetic improvement (pod number or weight/ plant) or through better crop management (days to flowering/ podding and seed filling period). Optimum value of pod wall weight/plant was less than observed mean values and similar observation was also reported for chickpea under different nitrogen levels (Bhattacharya 1983). It was concluded that physiological traits responsible for realization of potential lentil seed yield under non-irrigated and irrigated conditions were different. Under non-irrigated condition, it was mainly total dry matter yield, per day dry matter production and partitioning, mean temperature and total degree-days during podding to maturity. Under irrigated condition, lentil seed yield was determined by root dry matter at vegetative stage, total dry matter yield, per day dry matter Table 2.Optimum values of yield determining traits at different growth stages under non- irrigated and irrigated conditions in lentil for seed yield maximization Traits Components of regression Intercept Slope Observed Mean ‘r’ A (%) Maturity Maturity Maturity Podding to Maturity Podding to Maturity 0.740** 0.741** 0.961** 0.160 55.06 54.61 93.12 2.66 165.48 339.17 157.13 567.49 0.419 61.461 32.490 40.511 2859.1 20.0 27.14 19.66 3371.0 23.4 51.62 19.82 0.072 0.53 -1007.9 3.260 739.8 749.0 Vegetative Maturity Maturity -0.147 0.575** 0.572** 2.16 32.95 32.72 1870.83 104164.46 836.15 -3958.36 0.224 33.495 0.05 3811.1 25.5 0.07 5632.0 21.8 Maturity 0.985** 97.02 105.95 30.019 52.73 62.36 Flowering to -0.157 2.46 1952.08 podding r = Coefficient of correlation, A (%) = Association percentage, ** are significant at 1% -0.776 339.17 312.2 Non-irrigated condition Total dry matter yield Per day DM Production Per day DM partitioning Mean temperature Total degree-days Irrigated condition Root dry matter Total dry matter yield Per day DM Production Per day DM Partitioning Total degree-days Growth stage Optimum Value Vijay Laxmi : Phenology, dry matter distribution and yield attributes under normal and drought stress 305 production during seed to seed period and its partitioning during seed filling period and total degree-days during flowering to podding. 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Statistical Methods, 6th Edition, Oxford and IBH Publishing Co., Calcutta. Journal of Food Legumes 25(4): 306-309, 2012 Efficacy of post emergence herbicides on weed control and seed yield of rajmash (Phaseolus vulgaris L.) BALDEV RAM, S.S. PUNIA, D.S. MEENA and J.P. TETARWAL AICRP on MULLaRP, Agricultural Research Station (Maharana Pratap University of Agriculture & Technology) Ummedganj, Kota-324 001, Rajasthan, India; E-mail: [email protected] (Received: March 28, 2012; Accepted: December 04, 2012) ABSTRACT A field experiment was conducted during rabi of 2008-10 at Agricultural Research Station, Ummedganj, Kota to evaluate the efficacy of graded dose of imazethapyr and quizalofop ethyl against weed complex in rajmash. Application of imazethapyr 50 g/ha applied at 20 days after sowing (DAS) significantly reduced the weed density and their biomass recorded at 80 DAS as compared to both weedy check and graded dose of quizalofop ethyl applied at 20 and 30 DAS. The former also recorded significantly taller plants (43.0 cm), branches/plant (3.7), pods/plant (18.2), seeds/pod (4.2), seed index (28.5 g) and seed yield (710 kg/ha) over both weedy check and graded dose of quizalofop ethyl. In addition, imazethapyr 50 g/ha at 20 DAS had maximum net return (` 17360/ha) and B:C ratio (1.19) with least weed index (4.2%); and was superior to all treatments including hand weeding twice at 20 & 40 DAS. Key words: Imazethapyr, Quizalofop ethyl, Seed yield, Weed control efficiency, Weed index Rajmash (Phaseolus vulgaris L.) is grown throughout the year in India except during winter season in hills and rainy season in plains. Because of inefficient biological nitrogen fixation (BNF) through poor nodulations, it requires relatively higher dose of N fertilizer compared to other pulse crops along with sufficient quantity of P and K for its better growth and realization of yield potential. During its early growth stage, weed competes with it as it emerges simultaneously with the crop, leading to severe co mpet itio n between them (Kandasamy 2000). Since the initial growth of rajmash is very slow, the initial period of crop growth (30-45 DAS) is most crucial for crop-weed competition. In addition to slow initial crop growth, wider crop spacing also facilitates crop-weed competition which poses a serious limitation in rajmash production and thus, estimated seed yield loss may likely to go to the extent of 45-65 % under unweeded condition (Mishra 2006). During winter season, broad leaved weeds may become dominant in the early stages of crop growth because of their fast growth and deep root system. Usually farmers go for manual weeding under such a situation. However, availability of labour and cost involved make them to seek for other cheaper alternatives for weed control. The use of post emergence herbicides for season long weed control is thus, preferred over earlyuse of herbicides as pre-plant incorporation (fluchloralin & trifluralin) and preemergence (alachlor and pendimethalin) as the latter control weeds only during initial crop growth (up to 30 DAS). Hence, an integration of both pre-emergence herbicides along with one manual weeding is needed under a season long weed management strategy. There is also a possibility that use of single post emergence herbicide may replace the above and raise the income of both farmers and farms. Since work on post emergence herbicides especially in pulses is meager, an attempt is thus made to evaluate the efficacy of post emergence herbicides for effective control of weeds in rajmash. MATERIALS AND METHODS A field experiment was conducted at Agricultural Research Station, Ummedganj, Kota (250 18' N, 770 23' E and 271 m above mean sea level), Rajasthan, during rabi 2008-10 to evaluate the efficacy of new molecules of post emergence herbicide on rajmash. The clay loam (vertisols) neutral soil (pH 7.5) of the experimental field was low in organic carbon (4.1g/kg), medium in available P (20.5 kg/ha) and high in available K (292.5 kg/ha). The experiment was laid out in randomized complete block design (RCBD) comprising of 15 treatments viz., quizalofop ethyl at 40, 50 and 60 g/ha and imazethapyr at 25, 37.5 and 50 g/ha (both herbicides) applied at 20 and 30 DAS, pendimethalin 1.0 kg/ha as pre-emergence, hand weeding twice at 20 & 40 DAS and a weedy check with three replications. High yielding with semi dwarf erect type and tolerant to bean common mosaic virus ‘HUR 137’ Rajmash (115-125 days) was sown at a seed rate of 125 kg/ha with a row spacing of 30 cm in line during 3rd and 2nd week of November in 2008 and 2009, respectively. Half of recommended dose of N (60 kg/ha), full of P (60 kg P2O5/ha) and K (40 kg K2O/ha) through urea, diammonium phosphate and muriate of potash were drilled in the soil before sowing and the remaining N (60 kg/ha) was top dressed in two equal splits at first irrigation (25-30 DAS) and pod formation (65-70 DAS). The crop was raised under irrigated condition with recommended package of practices for the zone. Imazethapyr 10 % SL (at 25, 37.5 and 50 g a.i./ha) and quizalofop ethyl 5 % EC (at 40, 50 and 60 g a.i./ha) were sprayed 20 and 30 DAS, respectively by knapsack sprayer using a flat fan nozzle at 500 l/ha spray volume by diluting with water. Ram et al.: Efficacy of post emergence herbicides in rajmash Weeds were removed at 20 and 40 DAS under manual weed control (weed free). The total number of weeds from one square m area were counted species wise (monocot, dicot and sedges) separately with the help of iron quadrate at two places of the sample rows in each plot at 80 DAS and analyzed after subjecting the original data to square root transformation (vX + 0.5). For dry matter, weeds collected from one square m area were dried under the sun and then in an oven at 700 for 72 h, weighed (g/m2) and converted into kg/ha. Weed control efficiency (WCE) was calculated at 80 DAS and expressed as the percentage reduction in weed population due to weed management practices over control using formula: WCE (%)={Weed population in control plot (WPc)–weed population in treated plot (WPt)}/weed population in control plot × 100 Weed index (WI) was also calculated as per cent reduction in yield due to the presence of weeds in comparison with weed free plot. The economics of treatments was computed on the basis of prevailing market prices of inputs and outputs under each treatment. Analysis of variance was performed on all the collected data. Pooling was made over the years as similar trend was noticed during both the years. RESULTS AND DISCUSSION Weed control: The major broad leaved and grassy weeds observed in rajmash field included Chenopodium album (bathua), Chenopodium murale (kharthua), Cirsium arvense (kateli), Fumaria parviflora (gajri), Melilotus alba (senji), Lathyrus spp. (chatri-matri), Vicia sativa (ankari), Convolvulus arvensis (hirankhuri), Cyperus rotundus (motha) and Cynodon dactylon (doobgrass). Thus, broad leaved weeds (81.3%) were dominant compared to grassy(15.2%) and sedges 307 (3.5%) during both the years. All the weed control treatments significantly curtailed weed population and their dry weight compared to weedy check (Table 1). However, hand weeding twice at 20 & 40 DAS recorded lowest weed (monocot, dicot and sedges) population at 80 DAS. Amongst the herbicides, lowest dicot population was recorded with imazethapyr 50 g/ ha at 20 DAS; and was superior over rest of the herbicide treatments and weedy check as per cent reduction in total weed density was 90.9 over weedy check. Application of post emergence herbicide imazethapyr at 25-50 g/ha and quizalofop ethyl at 40-60 g/ha sprayed at 20 and 30 DAS also significantly reduced the total weed density to the tune of 36.0-57.9 and 18.3-30.5 % over weedy check, respectively whereas that of pre-emergence herbicide pendimethalin reduced the total weed density 45.7 per cent over weedy check. Imazethapyr being freely translocated in plants through roots and shoots could effectively controlled broad leaved as well as grassy weeds. Tiwari et al. (2007) observed that imazethapyr at 75 g/ha controlled broad leaved weeds in soybean whereas, Mishra and Chandra Banu (2006) reported efficient control of weeds by imazethapyr at 100 g/ha over other herbicides in summer irrigated urdbean. In the current investigation also, application of imazethapyr 50 g/ha effectively controlled the emerged grassy, sedges and broad leaved weeds. Thus, the results confirmed the findings of Ram et al. (2011) in field pea. With regards to weed biomass, imazethapyr 50 g/ha at 20 DAS recorded significantly lower weed biomass (36.8 g/ m2) and was closely followed by imazethapyr 50 g/ha at 30 DAS and imazethapyr 37.5 g/ha at 20 and 30 DAS over rest of the treatments and weedy check, respectively. Pendimethalin 1.0 kg/ha was also found significantly superior in reduction of total weed biomass (96.6 g/m 2 ) over weedy check. Table 1. Effect of post emergence herbicides on diverse weed flora, total weed density and weed control efficiency at 80 DAS in Rajmash (pooled) Treatments Quizalofop Ethyl 40 g/ha at 20 DAS Quizalofop Ethyl 50 g /ha at 20 DAS Quizalofop Ethyl 60 g/ha at 20 DAS Quizalofop Ethyl 40 g/ha at 30 DAS Quizalofop Ethyl 50 g/ha at 30 DAS Quizalofop Ethyl 60 g/ha at 30 DAS Imazethapyr 25 g a/ha at 20 DAS Imazethapyr 37.5 g/ha at 20 DAS Imazethapyr 50 g/ha at 20 DAS Imazethapyr 25 g/ha at 30 DAS Imazethapyr 37.5 g a/ha at 30 DAS Imazethapyr 50 g/ha at 30 DAS Pendimethalin 1.0 kg/ha PE Hand Weeding at 20 & 40 DAS Weedy check SEm (±) CD (P= 0.05) Monocot weed density (no/m2) 3.4*(8.4) 3.3 (7.8) 3.0 (6.2) 4.1 (12.6) 3.7 (10.3) 3.5 (9.1) 4.8 (18.6) 4.2 (13.5) 3.9 (11.3) 5.2 (21.9) 4.8 (18.7) 4.4 (15.3) 5.4 (23.7) 3.0 (6.2) 6.7 (38.3) 0.16 0.46 Dicot weed density (no/m2) 11.8 (126.5) 11.4 (119.0) 10.9 (108.8) 12.6 (146.1) 12.4 (141.7) 12.0 (132.8) 8.6 (64.7) 7.1 (43.2) 5.1 (20.8) 9.1 (74.5) 7.8 (53.4) 7.1 (43.1) 7.0 (41.6) 4.2 (13.4) 14.8 (205.1) 0.27 0.77 Sedge weed Density (no/m2) 2.6 (4.3) 2.5 (4.1) 2.5 (3.8) 3.1 (6.7) 2.9 (6.0) 2.8 (5.2) 2.5 (3.9) 2.4 (3.6) 2.4 (3.4) 2.5 (4.1) 2.5 (3.8) 2.4 (3.6) 2.9 (5.9) 2.4 (3.5) 3.5 (8.9) 0.08 0.23 Total weed density (no/m2) 12.3 (139.1) 11.9 (130.9) 11.4 (118.9) 13.4 (165.5) 13.1 (157.9) 12.6 (147.0) 9.8 (87.3) 8.3 (60.3) 6.9 (41.5) 10.5 (100.5) 9.2 (75.9) 8.4 (62.0) 8.9 (71.2) 5.3 (23.1) 16.4 (252.4) 0.34 0.97 Total weed biomass (g/m2) 105.7 93.0 87.4 104.4 99.9 96.2 55.8 48.4 36.8 56.1 47.4 37.2 96.6 17.3 161.8 4.23 11.86 *Data subjected to square root ( x 0.5 ) transformation and figures in parentheses are original values WCE (%) 44.9 48.1 52.9 34.4 37.4 41.8 65.4 76.1 83.6 60.2 69.9 75.4 71.8 90.9 1.36 3.82 Weed dry matter(kg/ha) at 80 DAS 1057 930 874 1044 999 963 558 484 376 561 474 372 215 173 1617 42.81 118.6 Weed index (%) 39.7 37.3 35.2 41.0 38.2 36.3 23.5 12.3 04.2 24.4 20.1 13.4 15.4 54.4 1.02 2.84 308 Journal of Food Legumes 25(4), 2012 Imazethapyr effectively controlled germinated broad leaved as well as grassy weeds either through directly killed or suppressed these (smothering effect) and thus, resulting in least weed biomass and higher crop growth. Similar, findings were reported by Godara and Deshmukh (2002) in soybean and Ram et al. (2011) in field pea. On the contrary, quizalofop ethyl was effective against grassy weeds only as it failed to curb the population of broad leaved weeds in comparison to imazethapyr. Nevertheless, hand weeding twice at 20 and 40 DAS recorded the lowest weed biomass (17.3 g/m2) at 80 DAS of all the herbicide treatments including weedy check by controlling weed population to the extent of 90.9% (Table 1). On efficiency factor, imazethapyr 50 g/ha at 20 DAS had maximum weed control efficiency (83.6%) recorded at 80 DAS and was followed by imazethapyr 50 g/ha at 30 DAS (75.4%) whereas, it was the least under quizalofop ethyl 40 g/ha at 30 DAS. This might be due to lower weed biomass and higher efficiency of weed control under imazethapyr against both broad leaved and grassy weeds (Table 1). Application of pendimethalin 1.0 kg/ha as pre-emergence herbicide was also superior over the graded dose of quizalofop ethyl at 20 and 30 DAS and weedy check. Ram et al. (2011) reported highest weed control efficiencywith imazethapyr 50 g/ha at 20 DAS in field pea. Use of post emergence herbicides was also found to be superior over pre-emergence and pre-plant incorporation applied herbicides as weeds were killed in their active growth stage bearing 2-3 leaves (Chen et al. 1998). Similarly, minimum weed index (4.2%) was recorded with imazethapyr 50 g/ha at 20 DAS over rest of the herbicide treatments and weedy check (Table 1) as the treatment effectively controlled both broad leaved and grassy weeds. Crop growth and yield: Post emergence herbicides had signi ficantly higher values of crop gro wth and yield contributing characters over the weedy check. Among the herbicide treatments, tallest plants (43.0 cm) and highest branches/plant (3.7), pods/plant (18.2) and seeds/pod (4.2) were recorded with imazethapyr 50 g/ha at 20 DAS; and was statistically on par with imazethapyr 50 g/ha at 30 DAS and pendimethalin 1.0 kg/ha in respect of plant height and seeds/ pod, respectively. Because of poor weed control efficiency and higher weed competition index among the crop and weeds, quizalofop ethyl was least effective for raising crop growth and yield contributing characters of rajmash (Table 2). On the contrary, hand weeding twice at 20 and 40 DAS (weed free) recorded significantly higher plant height (47.2 cm), branches/ plant (5.0), pods/plant (20.9), seeds/pod (4.8) and seed index (28.5 g) over weedy check and most of the herbicide treatments. Seed yield of rajmash varied significantly with weed control treatments. Maximum seed yield (741 kg/ha) was obtained with hand weeding twice at 20 and 40 DAS and was statistically on par with imazethapyr 37.5 and 50 g/ha at 20 DAS. The seed yield registered under the above was also significantly higher over rest of the herbicide treatments and weedy check. Among the herbicides, imazethapyr 50 g/ha at 20 DAS recorded maximum seed yield (710 kg/ha) which was obviously due to its higher values of yield attributes, weed control efficiency (83.6%) and lowest weed index (4.2%) compared to the rest of the herbicide treatments. In addition, imazethapyr 50 g/ha at 20 DAS also increased the seed yield to the tune of 110% while pendimethalin at 1.0 kg/ha as preemergence raised seed yield by 85.5% over weedy check. Effectiveness of these treatments could be attributed to better controls of weeds during critical period of crop-weed competition under moist soil condition which in turn reduced Table 2. Effect of post emergence herbicides on growth, yield attributes, seed yield and economics of rajmash (pooled) Treatments Quizalofop Ethyl 40 g/ha at 20 DAS Quizalofop Ethyl 50 g /ha at 20 DAS Quizalofop Ethyl 60 g/ha at 20 DAS Quizalofop Ethyl 40 g/ha at 30 DAS Quizalofop Ethyl 50 g/ha at 30 DAS Quizalofop Ethyl 60 g/ha at 30 DAS Imazethapyr 25 g a/ha at 20 DAS Imazethapyr 37.5 g/ha at 20 DAS Imazethapyr 50 g/ha at 20 DAS Imazethapyr 25 g/ha at 30 DAS Imazethapyr 37.5 g a/ha at 30 DAS Imazethapyr 50 g/ha at 30 DAS Pendimethalin 1.0 kg/ha PE Hand Weeding at 20 & 40 DAS Weedy check SEm (±) CD (P= 0.05) Plant height (cm) Branches/ plant Pods/ plant 32.6 33.5 34.7 34.7 36.1 36.9 36.9 40.1 43.0 37.5 39.9 41.2 41.2 47.2 26.6 1.0 2.8 3.2 3.3 3.4 3.1 3.2 3.3 3.4 3.6 3.7 3.3 3.6 3.7 3.7 5.0 2.6 0.14 0.38 9.7 10.2 10.7 9.4 9.6 10.1 12.6 14.8 18.2 11.7 12.8 15.2 13.9 20.9 6.1 0.78 2.17 Seeds/ Seed index Seed yield *Cost of (g) pod (kg/ha) cultivation (`/ha) 3.3 27.6 447 14810 3.5 27.8 465 15200 3.5 27.7 480 15575 3.2 27.9 437 14810 3.3 27.9 458 15200 3.4 28.0 472 15575 3.7 28.1 567 14145 3.9 28.2 650 14398 4.2 28.5 710 14590 3.3 28.0 560 14145 3.7 28.1 587 14398 3.9 28.2 642 14590 3.8 28.2 627 14925 4.8 28.5 741 18900 2.6 26.8 338 13400 0.14 0.21 23.6 0.39 0.59 65.6 - *Net return (`/ha) 5305 5725 6025 4855 5410 5665 11370 14852 17360 11055 12017 14300 13290 14445 1810 1067 2966 B: C ratio 0.36 0.37 0.39 0.33 0.36 0.36 0.80 1.03 1.19 0.78 0.83 0.98 0.89 0.76 0.13 0.08 0.24 *The price of imazethapyr and quizalofop ethyl were `1680 and 1380/litre, respectively, whereas, the cost of two hand weedings (20 and 40 DAS) were `7500 for 60 mandays and the sale price of rajmash was `45/kg Ram et al.: Efficacy of post emergence herbicides in rajmash biotic stress (due to weed competition) and thus, provided a weed free environment for a better growth and development of rajmash. These findings are in close proximity with that of Billore et al. (1999) with imazethapyr on soybean and Ram et al. (2011) with imazethapyr on field pea. Irrespective of its doses and time of application, application of imazethapyr was superior to weedy check. Lower seed yield under quizalofop ethyl could be attributed to its poor weed control efficiency and higher weed index against broad leaved weeds. Economics: The highest net return (` 17,360/ha) and benefit: cost ratio (1.19) was fetched with imazethapyr 50 g/ha at 20 DAS owing to lower cost of cultivation and effective control of broad leaved as well as grassy weeds (Table 2); and was followed by imazethapyr 37.5 g/ha at 20 DAS and hand weeding twice at 20 and 40 DAS. Excellent control of dominant broad leaved as well as grassy weeds without any adverse effect on crop growth resulting in higher seed yield might have caused superior economic indices in these treatments. Least net return (` 1810/ha) and B: C ratio (0.13) was recorded with weedy check due to both poor weed control and low crop yield. Thus, it was inferred from the above that post emergence application of imazethapyr 50 g/ha at 20 days after sowing could be recommended for effective control of broad leaved as well as grassy weeds in rajamsh for getting higher productivity and profitability under the existing condition. 309 REFERENCES Billore SD, Joshi OP and Ramesh A. 1999. Herbicidal effect on nodulation, yield and weed control in soybean [Glycine max (L.) Merill]. Indian Journal of Agricultural Sciences 69: 329-331. Chen TiebBao, Yong ShaoYi, Huang, ChunYan, Wang Yu, Zhang ShanYing, Chen TB, Yang CY, Wang Y and Zhang ZY. 1998. Study on weed control for soybean field with Jindou (AC 299263). Soybean Science 17: 271-275. Godara SP and Deshmukh SC. 2002. Weed biomass, weed control efficiency and yield of soybean (Glycine max L.) as influenced by various weed control measures. In: Proceedings of 2 nd International Agronomy congress on Balancing Food and Environment SecurityA continuing challenge, 26-30 November 2002, IARI, New Delhi. Pp. 1198-1200. Kandasamy OS. 2000. Cost effective weed management strategies in pulse production. In: Proceedings of CAS on Recent Advances in Pulse Production Technology, 13 th September-30 t h October, Coimbatore, Tamilnadu. Pp. 116-119. Mishra JS. 2006. Efficacy of post emergence herbicides against wild oat in field pea. Indian Journal of Weed Science 38: 140-142. Mishra JS and Chandra Banu. 2006. Effect of herbicides on weeds, nodulation and growth of Rhizobium in summer blackgram (Vigna mungo). Indian Journal of Weed Science 38: 150-153. Ram Baldev, Punia SS, Meena DS and Tetarwal JP. 2011. Bio-efficacy of post emergence herbicides to manage weeds in field pea. Journal of Food Legumes 24: 254-257. Tiwari DK, Kewat ML, Khan JA and Khanparia NK. 2007. Evaluation of efficacy of post emergence herbicides in soybean (Glycine max L.). Indian Journal of Agronomy 52: 74-76. Journal of Food Legumes 25(4): 310-313, 2012 Enhancing water use efficiency and production potential of chickpea and fieldpea through seed bed configurations and irrigation regimes in North Indian Plains J.P. MISHRA, C.S. PRAHARAJ1 and K.K. SINGH1 NRM Division, Krishi Anusandhan Bhawan-II, ICAR, New Delhi- 110012, India; 1Crop Production Division, Indian Institute of Pulses Research, Kanpur-208 024, U.P., India; E-mail: [email protected] (Received: October 27, 2011; Accepted: December 12, 2012) ABSTRACT Chickpea and fieldpea are grown during fall in India under diverse production systems including both rainfed and irrigated. Moisture scarcity especially at terminal stages of these crops results in low productivity and less farm income despite the fact that these crops have unique adaptive mechanisms to moisture stress. Therefore, one or two life saving irrigations at the most critical stages is of immense relief for maintaining plant water status in addition to other in situ water saving measures such as land configuration and mulch. Thus, a field trial was carried out in these pulses to study the effect of seed bed configurations, mulch and irrigation regimes on seed yield and water use efficiency (WUE) under North Indian Plains during 2007-08 and 2009-10. The study revealed that pre-sowing irrigation followed by a post-sowing irrigation depending on critical stage of crop was optimum for realization of optimum yield and WUE. Irrigation combined with straw mulch @ 6 t/ha was also useful in getting maximum seed yield. Planting on a raised bed wa s superior to flat planting. Subsequent confirmatory field trial also revealed the yield advantage of furrow irrigated raised beds (FIRB) planting at 60 cm with two rows of field pea on beds over both flat and FIRB at 90 cm with three rows of field pea on beds. Water use efficiency was however, improved with 90 cm wide FIRBs in chickpea in comparison to 60 cm wide FIRBs in field pea. Both chickpea and field pea responded to single irrigation at branching only. Key words: Chickpea, Fieldpea, Mulch, Seed bed configurations, Seed yield, Water use efficiency The intrinsic capacity of pulses to sustain soil health and fertility through addition of organic matter (roots and fallen leaves) and assimilation of atmospheric nitrogen into ammonia and consequent economy in N fertilizer application following BNF play a vital role in sustenance in cultivation of pulses and its popularization among growers (Mishra et al. 2012). Furthermore, pulses do offer an attractive opportunity through their vertical diversification for varied economic uses such as feed, fuel and fodder (Panwar and Basu 2003, Praharaj et al. 2011). Besides improving soil conditions, pulses have become more remunerative in view of the recent price regime triggered due to higher minimum support prices (MSP) and changing domestic economy in the rural areas resulting in increased demand for the commodity. Chickpea and field pea are important pulses of North Indian Plains during fall or Rabi season wherein these are cultivated under diverse production systems including rainfed condition. Chickpea (Cicer arietinum L.) is the premier pulse crop in India which accounted about 44.5% of country’s total pulse production of 18.09 million tonnes during 2010-11 (Anonymous 2011). On acreage and production front, chickpea is grown in India in 6.67 Million hectares (Mha) with production of 5.3 million tonnes (Mt) while field pea is grown in 0.72 Mha with production of 0.6 Mt during 2011-12 (IIPR 2012). Being mostly grown under rainfed condition with cessation of monsoon rain many a time early in the season, moisture scarcity especially at terminal stages of these crops is conspicuous and more-frequent that results in low productivity/production despite the crops have a unique adaptive mechanisms to moisture stress. Thus, one or two life saving irrigations at the most critical stages of the crop is of immense relief for maintaining plant water status (Ali 2009, Ali et al. 2008). In addition, resource conservation for higher input use efficiency through appropriate seed bed configurations in conjunction of appropriate irrigation scheduling have significant role in enhancing both production and productivity of pulses especially under Rabi season that usually thrived better under limited soil moisture availability (Chaudhury et al. 2005). Moreover, potential morpho-physiological traits in plants viz., water use efficiency (WUE), deep root system, higher relative biomass and harvest index, osmotic adjustment of chickpea are advantageous under water scarce situation (Chaudhury et al. 2005). Despite all this, crop experiences terminal drought during seed development stage as it is invariably grown on residual soil moisture after a preceding rainy crop(s), thereby making the terminal moisture stress as the major constraint in achieving potential productivity of chickpea (Singh et al. 2010). Under such sit uations, photosynthetic activity of leaves is hampered for the want of nitrogen and thus, seed filling is affected (Davies et al. 2000). Therefore, a judicious management of available soil moisture through in-situ conservation either by a mulch practice or a suitable land configuration viz., furrow irrigated raised bed (FIRB) improves crop productivity (Panwar and Basu 2003). These water saving measures may possibly render the crop in part to get rid of water stress at least for sometime without appreciable loss in biomass production leading to probably higher productivity. Since the information on growing Rabi Mishra et al.: Performance of Rabi pulses under seed bed configurations and irrigation pulses on raised bed vis-a-vis flat system coupled with irrigation regimes and mulches is limited, hence the present study was undertaken on two important Rabi pulses (chickpea and fieldpea) under modified seed bed configuration, mulch and limited irrigation for determining the efficacy of these on higher productivity and efficient water use under North Indian Plains. MATERIALS AND METHODS A field experiment was conducted at Indian Institute of Pulses Research, Kanpur, India (26o 27/ N, 80o 14/ E and 152.4 m above msl) under Indo-Gangetic Plains during Rabi 2007-08 and 2009-10 to evaluate the productive performance of ‘DCP 92-3’ chickpea (a small seeded semi erect and medium duration desi variety) and ‘Akash’ field pea (a bold seeded and dwarf variety) under different seed bed configurations, irrigation regimes and rice straw mulch. The trial involving two seed bed configurations (flat bed versus raised bed) in main plot and four irrigation schedules (one irrigation, one irrigation + rice straw mulch @6t/ha, two irrigations, and two irrigations + rice straw mulch @6t/ha) in sub plot was carried out during 1st year in a split plot design with three replications. During 2nd year, precisely three seed bed configurations (flat bed, raised bed of 60 cm width and raised bed of 90 cm width) in main plot along with 3 irrigation schedules (rainfed, one irrigation at branching and two irrigations at both branching and pod formation) in sub plot were taken up at the same site in a split plot with three replications. The climate of the region is tropical sub-humid receiving an annual rainfall of 722 mm with mean annual maximum and minimum temperature of 33°C and 20°C, respectively. The climatic situation for the representative year (2009-10) was also given in Table 1. The soil was sandy loam (Typic Ustochrept) with 8.1 pH, 1.43 g/cc bulk density and low in organic carbon (SOC, 0.28 %) at the time of initiation of field experiment. On soil fertility account, it was low in available N (215 kg/ha), medium in P (10.5 kg/ha) and K (230 kg/ha) and S (15.0 kg/ha). The field was prepared well and furrow irrigated raised beds (FIRB) of 60 and 90 cm width were prepared with tractor drawn modified conventional bed maker. Two rows of chickpea and field pea were planted on 60 cm FIRBs (at an inter-row spacing of 20 cm) while three rows of both the pulses 311 were planted on 90 cm FIRBs (at an inter-row spacing of 22.5 cm). Recommended seed rate was used under both system of planting along with recommended package of practices including use of fertilizers and appropriate Rhizobium inoculation. The rainfall received during the crop period was optimum and well distributed during both the years. The moisture studies were carried out in four depths (0-15, 15-30, 30-45 and 45-60 cm) using gravimetric method; and soil moisture depletion, water use and its efficiency (from seed yield data) were calculated using standard procedures. Normal practice of crop husbandry for successful crop raising was also followed. RESULTS AND DISCUSSION Seasonal variation: The distribution and intensity of rainfall during fall in 2007-08 and 2009-10 are near optimal although it was the most congenial during 2009-10 (Table 1) as it coincided with active branching and late flowering stages, thereby improving crop productivity yet nullifying the impact of tillage and mulch treatments. The data could not be pooled due to differences in treatments and differential trends observed for seed yield during the seasons with different rainfall pattern. Crop year 2007-08: During the year 2007-08, the objective was to efficiently manage the irrigation water in chickpea through seedbed configurations, irrigation regimes and mulch for improving WUE in chickpea through conjunctive use of mulch and irrigation under improved agronomic management (FIRB i.e., furrow irrigated raised bed). Crop performance under different land configurations indicated that FIRB planting (2439 kg/ha) out yielded flat planting (2052 kg/ha) significantly to the extent of 18.8% and was mainly attributed to higher crop growth (in terms of plant height, nodule/plant, nodule dry weight and branches/plant) and yield attributes (pod/plant) (data not given). Though maximum seed yield was recorded with two irrigations + mulch (2490 kg/ha), it was at par with two irrigations alone (2450 kg/ha). However, seed yield response to mulch in chickpea was significant up to one irrigation only. Yet, the WUE was maximum both under raised bed planting and one irrigation + mulch as mulch enhanced WUE significantly (Fig. 1). Low soil moisture availability was mainly attributed to Table 1. Climatic situation at the location during the year 2009-10 Months Temp max Tempmin RH max RHmin Oct, 09 Nov, 09* Dec, 09 Jan, 10 Feb, 10 Mar,10* Apr,10 32.3 28.2 24.9 18.4 26.6 35.7 42.1 19.5 14.5 09.8 07.8 12.2 17.9 24.3 68.6 70.5 75.7 92.0 75.0 57.7 35.2 55.2 61.1 65.9 76.8 55.5 33.2 21.2 *Planting date was early Nov, 2009 and the crop was harvested during March, 2010 R.F. (mm) 95.8 3.8 10.0 4.4 15.6 0.0 0.0 Cum. R.F. (mm) 3.8* 13.8 18.2 33.8 33.8* - Avg. Evap. (mm/day) 4.3 2.5 1.9 1.3 3.4 7.7 12.2 312 Journal of Food Legumes 25(4), 2012 higher soil moisture depletion due to evaporation from undisturbed soil capillary under no mulch treatment (data not included). Similarly, no mulch recorded higher consumptive use over paddy straw mulch. Thus, water use efficiency was higher in mulch treatment (with one or two irrigations at crop critical stages). WUE, being the product of economic yield to consumptive use of water, reflects the efficacy of a given treatment of transforming the water used into economic produce i.e., seed yield per unit area. The efficiency of water use was therefore, enhanced following mulch and rice straw; and was more conspicuous up to one irrigation at branching only (Fig. 1). apart as recommended) for chickpea and field pea was further compressed to 22.5 cm on FIRBs of 90 cm containing three rows of pulses. Under 60 cm furrow irrigated raised beds, two rows of crops were planted so that these could get an adequate space for horizontal spread in the left over space between two ridges, while under 90 cm raised beds, three rows of these crops were planted on raised beds and the middle row suffered adversely due to increased inter- and intra-plant competitions for resources. This could result in lower yield in 90 cm raised beds (1.91 t/ha in chickpea and 2.01 t/ha in field pea) in comparison to 60 cm FIRB (1.99 t/ha in chickpea and 2.68 t/ha in fieldpea). Crop year 2009-10: Both the rabi pulses responded differentially to seed bed configurations as well as irrigation scheduling (Table 2). The seed yield of chickpea did not differ significantly due to manipulation in seed bed configurations whether it was a flat bed or FIRB. Contrary to this, fieldpea yielded significantly higher (2.68 t/ha) when planted on 60 cm FIRB as it registered an increase in seed yield to the tune of 20% over flat bed and 33% over 90 cm wide FIRBs. The lowest seed yield of both crops was recorded in 90 cm FIRBs. It was due to the fact that normal planting distance (30 cm rows The consumptive use of water was also the highest in flat bed sowing under both the crops that reduced further in furrow irrigated raised bed planting. Thus, water use efficiency was increased under raised bed systems with the higher value of 11.1 kg/ha-mm under 60 cm FIRBs in chickpea (and 11.8 kg/ ha-mm under 90 cm FIRBs) and 17.7 kg/ha-mm in field pea. This indicated the fact that furrow irrigated raised bed systems could produce higher seed yield with low water use or application of less water (Anwar et al. 2003, Praharaj et al. 2011). Of the two crops, field pea proved more efficient in water use under all the planting configurations over chickpea (Table 2). Yield WUE 3000 20 Yield (kg/ha) 2500 18 2000 16 1500 14 1000 12 500 Fl at 0 d se ai R d be ne O i ir r O Fig 1. g. ne ig irr ch ul +m o Tw ig irr 10 . o Tw i rr ig h ulc +m Seed yield and WUE as influenc ed b y planting techniques and irrigation schedules The wetter moisture regimes under two irrigations at branching and pod formation also proved counterproductive for chickpea as only a marginal increase in the seed yield of field pea was evident (Table 2). During rabi 2009-10, the winter rains were well distributed coinciding with the late flowering and initiation of pod formation stages of chickpea and field pea (Table 1) that led to no response for two irrigations in both the crops. The consumptive use was also the highest under two irrigations (21.3 cm in chickpea and 21.1 cm in field pea). However, no irrigation (rainfed condition) was more conducive for higher water use efficiency in chickpea (12.8 kg seed/ha/mm water) and field pea (16.4 kg seed/ha/mm water). Relatively higher water use efficiency in field pea over chickpea was due to higher seed yield in case of former. Thus, lower water use efficiency in wetter moisture regimes was due to Table 2. Effect of seed bed configurations and irrigation schedules on water use and seed yield of chickpea and fieldpea Treatments Seed bed configurations Flat 60 cm bed FIRB 90 cm bed FIRB C.D. (0.05) Irrigations schedules No irrigation one irrigation Two irrigation C.D.(0.05) *CU: Consumptive use of water Seed yield (t/ha) Chickpea CU* (cm) WUE (kg/ha-mm) Seed yield (t/ha) Field pea CU* (cm) WUE (kg/ha-mm) 2.03 1.99 1.91 NS 19.6 18.2 16.3 - 10.7 11.1 11.8 - 2.24 2.68 2.01 0.28 18.7 15.6 16.6 - 12.4 17.7 12.3 - 1.90 2.04 1.99 0.11 14.8 18.0 21.3 - 12.8 11.4 9.4 - 2.14 2.38 2.40 0.21 13.0 16.8 21.1 - 16.4 14.5 11.5 - Mishra et al.: Performance of Rabi pulses under seed bed configurations and irrigation disproportionate increase in seed yield as compared to water use. Thus, the role of single irrigation at the critical stage of crop was evident from the increase in both seed yield and water use efficiency up to the maximum level of compensation in use of precious water as an input for crop use (Table 2). Thus, it was inferred from the above that furrow irrigated raised bed (60 cm width FIRBs accommodating 2 rows) could be an effective land configuration measure in conserving both soil moisture and enhancing productivity of chickpea and field pea. In case of terminal moisture stress, single irrigation at branching could be advocated for realizing higher yield and input use efficiency. REFERENCES Anonymous. 2011. IV Advance Estimates, 2010-11, Directorate of Economics & Statistics, Department of Agriculture & Cooperation, Ministry of Agriculture, Government of India. Anwar MR, Mckenzie BA and Hill GD. 2003. The effect of irrigation and sowing date on crop yield and yield components of Kabuli chickpea (Cicer arietinum L.) in a cool-temperate subhumid climate. The Journal of Agricultural Science 141:259-271. Chaudhury J, Mandal UK, Sharma KL, Ghosh H and Mandal B. 2005. Assessing soil quality under long-term rice-based cropping system. Communications in Soil Science and Plant Analysis 36: 1141-1161. Davies SL, Turner NC, Palta JA, Siddique, KHM and Plummer JA. 313 2000. Remobilization of carbon and nitrogen supports seed filling in desi and kabuli chickpea subject to water deficit. Australian Journal of Agricultural Research 51: 855-866. IIPR 2012. All Indian Coordinated Projects on Chickpea and MULLaRP, Indian Institute of Pulses Research, Kanpur, India. Ali Masood. 2009. 25 years of pulses research at IIPR. Indian Institute of Pulses Research, Kanpur. Pp. 211. Ali Masood, Ganeshamurthy AN, Singh KK and Sekhon HS. 2008. Integrated nutrient and water management in food legumes in semiarid tropics, Vol 1: 485-502. In: Food legumes for nutritional security and sustainable agriculture (Ed M.C.Kharwal), M/s Kamala Print-npublish, New Delhi, India. Mishra JP, Praharaj CS, Singh, KK and Narendra Kumar 2012. Impact of conservation practices on crop water use and productivity in chickpea under middle Indo-Gangetic plains. Journal of Food Legumes 25: 41-44. Panwar JDS and Basu PS. 2003. Improving drought tolerance and water use efficiency in chickpea. In: Masood Ali, B.B. Singh, Shiv kumar, Vishwa Dhar (Eds), Pulses in new perspective. Indian Institute of Pulses Research, Kanpur, India. Pp 480-488. Praharaj CS, Sankaranarayanan K, Narendra Kumar, Singh KK and Tripathi AK. 2011. Low-input technologies for increasing crop productivity and sustainability. Current Advances in Agricultural Sciences 3: 1-12. Singh AK, Singh SB, Singh AP, Singh AK, Mishra SK and Sharma AK. 2010. Effect of different soil moisture regimes on biomass partitioning and yield of chickpea genotypes under intermediate zone of J & K. Journal of Food Legumes 23: 156-158. Journal of Food Legumes 25(4): 314-320, 2012 Variability in the nutrients, antinutrients and other bioactive compounds in soybean (Glycine max (L.) Merrill) genotypes REETI GOYAL, SUCHETA SHARMA and B.S. GILL1 Department of Biochemistry, 1Department of Plant Breeding and Genetics, Punjab Agricultural University, Ludhiana141004, India; E-mail: [email protected] (Received: February 16, 2012 ; Accepted: October 09, 2012) ABSTRACT One hundred fourty soybean genotypes were evaluated for their physicochemical properties and biochemical diversity in contents of nutrients (total proteins, oil, total sugars and sucrose), antinutrients (trypsin inhibitor activity and phytic acid) and bioactive compounds (tannins, saponins, phenolics and tocopherols) with the aim to identify the improved genotypes having low antinutrient and high nutrient traits for human/livestock consumption. Physicochemical characterization indicated that exotic genotypes had highest mean values for water absorption, volume expansion, hydration capacity, swelling capacity and their indices. Soybean genotypes exhibited 38.4-46.5% protein, 20.8-23.6% oil, 1.3-13.9% total soluble sugars, 0.24-13.8% sucrose, 2.50-33.5 mg/g tocopherol, 8.9-20.5 mg/g tannins and 11-38.3 mg/g phenols. Genotypes SL 1000, SL 716 and SL 990 recorded minimum trypsin inhibitor activity, phytate and tannin content respectively. Soybean genotypes investigated differed in their nutritional and antinutritional characteristics. Such information will be useful for breeding purposes as well as in selecting soybean varieties to manufacture various soy food preparations. Key words: Oil, Phytic acid, Protein, Soybean genotypes, Total soluble sugars, Trypsin inhibitor Soybean is the most important feed grain legume with a total world production of 234.65 million tonnes and harvested area of 94.89 million hectare (FAOSTAT 2009). In India, it is grown on an average area of 9.79 million hectare with production and productivity of 99.65 mt and 1,026 kg/ha, respectively (Ram et al. 2011). The seeds of soybeans contain a moderately high amount of calories (calorific value of 400g/ 100g), protein (40%), lipid (20%), relatively high insoluble carbohydrate content (11%), crude fibre (9%) and ash (5%) (Varsha and Grewal 2009). In addition, soybean oil contains approximately 14% saturated fats on an average. Reduction of palmitic acid and stearic acid would be desirable for lowering saturated fat content in human diet to improve cardiovascular health (Spencer et al. 2003). Soybean oil is also a good source of vitamin E and its content varies with variety (Lokuruka 2010). Tocopherols are known to reduce incidence of prostate cancer and coronary heart disease (Evans et al. 2002) and improve oil stability (Stone and Papas 2003). Soy seeds contain high valued proteins that are used as food/feed for human and animals. Although soybeans are deficient in methionine but contain sufficient lysine to overcome the lysine deficiency of cereals (Neus et al. 2005). Carbohydrate content is one of the important quality traits in soybean. The major sugars in soybean seeds are glucose, fructose, sucrose, and galactooligosaccharides (Blochl et al. 2007). Although soybean is rich in nutrients but its acceptability as raw food is limited due to the presence of anti-nutritional factors which decrease nutritive value of grain legumes and cause health problems to both human and the animals when taken in large amounts (Mikic et al. 2009). Protease inhibitors represent 6% of the protein present in soybean seed. Trypsin inhibitors cause enlargement of the pancreas in rodents, hyper secretion of digestive enzymes that leads to a loss of trypsin and chymotrypsin, and reduce the hydrolysis of dietary protein, thereby decreasing amino acid absorption and protein synthesis (Roy et al. 2010). However, these inhibitors are effective in preventing or suppressing carcinogen induced transformation in vitro and demonstrates potent antiinflammatory properties (Roy et al. 2010). Soybean seed contains phytic acid (myo-inositol hexakis phosphate) to the extent of 1-1.5% DM and 65-80% of total phosphorus from soybean seeds is bound to phytic acid (Raboy et al. 2000). It chelates mineral nutrients such as copper, zinc, manganese, iron and calcium thus reducing their availability (Ramakrishna et al. 2006). Beside above mentioned anti-nutri tional facto rs, soybean cont ains bio acti ve compounds with small or unknown effects, such as tannins, saponins, lectins, antivitamins and isoflavones (Jain et al. 2009). Raw soybeans contain between 2 to 5 g saponin/100g. Because of presence of both hydrophobic and hydrophilic regions, saponins are excellent emulsifiers and foaming agents and provide functional role in foods (MacDonald et al. 2005). The tannins and phenolic constituents are known to adversely affect the utilization of proteins in animal and human diet due to t heir abi lity to bind wi th and precipit ate prot eins (Khandelwal et al. 2010). The current focus of the breeders should be on screening the soybean cultivars for decreasing the content of ant inutrienal factors to a safe extent. In the present communication, an attempt has been made to evaluate soybean genotypes for their physical, nutritional and antinutrient factors so as to identify the diversity in the available germplasm in relation to their nutritional quality. Such Goyal et al.: Variability in the nutrients, antinutrients and other bioactive compounds in soybean information will be useful in selecting soybean varieties for manufacture of various soy food as well as medicinal preparations. MATERIALS AND METHODS One hundred and forty soybean genotypes including 74 from PAU, Ludhiana collection, 51 from other parts of India and 15 exotic genotypes were used in the present study (Table 1). The experiment was conducted in 2009 and all the genotypes were grown in the field of Department of Plant Breeding and Genetics, PAU, Ludhiana by following the recommended package of practices in a randomized complete block design with three replications. The seeds were sown in rows keeping 45 cm distance between the rows and plant to plant distance was kept at 5 cm. The row length was 2 meter with 2 rows per entry. The experimental soil was sandy loam in texture, pH 7.24, electrical conductivity 0.30 ds/m, organic carbon 0.34%, available P and K (58.5 and 57.5 kg/ha, respectively). The seeds were collected at maturity, cleaned by hand to remove dirt and broken grains and then stored in air tight plastic containers till further analysis. Later, seeds were crushed into fine powder by Cemotec 1090 sample mill and then stored for further use. The seed yield (kg/plot) for each genotype was recorded. Physical seed characteristics like seed weight (g), seed volume (ml), water absorption (%), volume expansion (%), swelling capacity (g/seed) and hydration capacity (ml/seed) and their indices were determined by the methods of Santhan and Shivshankar (1978). The contents of moisture, protein and lipid in seed powder were determined by NIR Method using Infratec TM 1241 Grain analyzer from Foss (North America). Soluble sugars were extracted from the soybean seed powder with 80% ethanol followed by 70% ethanol. Total 315 soluble sugar levels in the pooled extract were determined with the phenol-sulfuric acid reagent (Dubois et al.1956) using glucose as standard. Sucrose content was determined after destroying the free fructose with 30% KOH by resorcinol-HCl procedure (Roe 1934). Trypsin inhibitor (TI) activity was determined as described by Kakade et al. (1974). The powdered seed (1g) was homogenized with 1% NaCl. The TI activity of the extract was determined using casein as a substrate. One unit of trypsin inhibitor activity is defined as the quantity of inhibitor which reduces the activity of trypsin by one unit at 37°C. Phytic acid in the powdered seeds was determined by the procedure described by Vaintraub and Lapteva (1988). Five hundred mg seed powder was stirred using a magnetic stirrer in 3.5% HCl. The contents were centrifuged at 3000g for 10 min to obtain supernatants. A suitable aliquot of the supernatant was taken and 1 ml of Wade reagent (0.03% FeCl3 containing 0.3% sulfosalicylic acid) was added to it and again centrifuged. The absorbance was measured at 500 nm using spectrophotometer. Phenolic compounds were extracted by refluxing the seed powder with 80% aqueous methanol at 60oC in a water bath with continuous shaking for 2h. The refluxed material after filtration was used for the estimation of total phenols (Swain and Hillis 1959). A standard curve of gallic acid (10-100 µg) was simultaneously prepared and the amount of the phenols was calculated and expressed as mg/g seed. Tannins were extracted from the powdered seeds and estimated. Using a Folin–Denis Reagent, the intensity of the colour developed was measured at 700 nm (Sadasivam and Manickam 1992). A standard curve of tannic acid (10–100 ìg) was simultaneously prepared. Table 1. Soybean genotypes used in the present study PAU, Ludhiyana collections SL 137, SL 202, SL 255, SL 290, SL 313, SL 525, SL 568, SL 587, SL 592, SL 688, SL 707, SL 716, SL 744, SL 773, SL 778, SL 790, SL 791, SL 793, SL 799, SL 806, SL 834, SL 871, SL 878, SL 894, SL 900, SL 903, SL 907, SL 914, SL 917, SL 925, SL 926, SL 955, SL 958, SL 967, SL 973, SL 976, SL 977, SL 978, SL 979, SL 980, SL 981, SL 982, SL 983, SL 984, SL 985, SL 986, SL 987, SL 988, SL 989, SL 990, SL 991, SL 992, SL 995, SL 996, SL 997, SL 998, SL 999, SL 1000, SL 1001, SL 1002, SL 1003, SL 1004, SL 1005, SL 1006, SL 1007, SL 1008, SL 1009, SL 1010, SL 1012, G-237XSL 295, 11-4-1 Genotypes from other parts of India DS-76-1-139, DS-98-1, DS-98-2, DS 2613, DS 2614, DS 143, DS 200 Origin IARI, New Delhi JS 72-45, JS 81-340 , JS-84-16, JS-89-67 JNKVV, Jabalpur Exotic Genotypes BRAGG, EC-457156, EC-457159, EC-457161, EC-457286,EC-457466, EC-457471, EC-230143 Origin USA PK 1026, PK-1042, PS 1414, PS 1042, PS 1420, PS 1437, PS 1444 GB Univ. of Agri EC-103332,TG-849-309, Sci & Tech, TGX825-3FF Pantnagar Phillipines IC-49865, IC- 437079, IC 15977, IC 100497 Himalayan Region EC-280148 Taiwan G 114, GB 1587, G-18, YMV- 25, YMV-35, YMV-36 DSR, Indore EC-251401, EC-251498 Argentina EC-309541 Brazil Local collections SEL P, SEL-37, SEL 40, SEL 41, SEL 46, SEL 174, K-88-2629, K-B-65, UCM 47, UPSM 124, DCB 194, DM -51336, HM 1, AK 99-67, B 86-24, R-5, R-11, GP 650, GP 1036,GP 1037, F-67-3975, NRC-05-976 316 Journal of Food Legumes 25(4), 2012 For the extraction and estimation of saponins, 500 mg of seed powder was homogenized with acetone and later with methanol. The saponin content of soybean seeds was estimated from the methanolic extract by the method of Fenwick and Oakenfull (1983). A standard curve (10-40 µg) of saponin (Himedia, Mumbai) was simultaneously prepared. Tocopherols were extracted from the powdered seeds by ethanol. Purified xylene was added to the supernatant and centrifuged. Xylene layer was pipetted out and the content was estimated using bathophenanthroline reagent, ferric chloride and ortho-phosphoric acid (Kayden et al. 1973). The amount of tocopherols was calculated from the standard curve prepared by using dl-á-tocopherol(Sigma-Aldrich Corporation, Bangalore) as standard (0.02mg/ml ethanol). Statistical analysis: All results in this study are reported as means of three replicates. Means, standard deviation and correlation coefficients for different nutrients and antinutrients were calculated using software Statgraphics Centurion Version XV: II (Statpoint, Inc.). One way analysis of variance (ANOVA) was carried out to determine the significant differences between means among the different groups of genotypes at P<0.05. The genotypes were clustered on the basis of their biochemical similarity. Standardized matrix was used for the Unweighted pair group method using arithmetic averages (UPGMA method) to generate the cluster tree using NTSYS pc 2.0 software (Rohlf 1998). RESULTS AND DISCUSSION Nutritional composition of soybean seeds: The nutritional composition of soybean genotypes was estimated and the mean values of the contents are represented in Table 2. The moisture content of the seeds of 140 different soybean genotypes ranged from 7.60-12.9%. The values are comparable to the moisture content of soybean genotypes from other sources (9.6-13.2%) (Awadesh et al. 2003). Soybean contained protein with a range of 38.4-46.5%, with an average value of 42.9%. Genotype PS 1444 showed the highest protein content of 0.46 mg/g seeds. Oil content was in the range of 20.8 (EC309541) to 23.6% (SL 793). Among various genotypes studied, 5 genotypes exhibited protein content >45% and 2 genotypes recorded lipid content >23%. Genotypes PS 1444 and EC 309541 exhibited maximum protein and minimum lipid content, respectively as compared to other genotypes. The mean values of protein and lipid content of seeds of various soybean genotypes from different categories did not differ significantly. Total soluble sugars and sucrose content ranged from 1.3013.9% and 0.24-13.8% respectively. 15 genotypes contained sucrose content more than 10% and genotype K-B-65 showed Table 2. Mean, standard deviation (SD), ranges of various nutrients (%), antinutrients (mg/g) and bioactive compounds (mg/g) in soybean genotypes Total Genotypes Local Genotypes Genotypes from other Parts of India Exotic Genotypes Oil 20.8-23.6 21.7 0.45 21.0-23.6 21.7 0.48 21.0-22.7 21.7 0.38 20.8-22.9 21.7 0.53 0.43 Range Mean SD Range Mean SD Range Mean SD Range Mean SD CD (P=0.05) Nutrients Protein 38.4-46.5 42.9 1.23 38.4-45.3 42.7 1.16 40.0-46.5 43.2 1.22 39.8-44.5 43.1 1.49 0.20 Sucrose 0.24-13.8 5.9 3.01 0.24-13.6 6.8 2.71 0.60-13.8 4.3 2.92 4.00-13.8 7.0 2.35 0.48 Total Sugars 1.30-13.9 5.9 2.78 1.8-13.9 6.9 2.73 1.30-13.7 4.5 2.43 1.30-8.9 5.7 2.01 0.38 Moisture 7.60-12.9 11.0 1.42 7.8-12.9 11.3 1.37 7.60-12.7 10.5 1.42 8.30-12.6 11.0 1.30 0.46 Antinutrients and bioactive compounds Total Genotypes Range Mean SD Local Range Genotypes Mean SD Genotypes Range from other parts of Mean India SD Exotic Range Genotypes Mean SD CD (P=0.05) Phytate 1.2-28.5 10.5 6.10 1.2-28.5 10.5 7.22 2.1-21.2 10.8 4.62 2.3-19.0 10.3 4.57 2.27 Trypsin inhibitor 11.3- 142.5 68.8 34.8 11.2-138.7 68.9 33.5 11.3-142.5 62.3 35.9 33.8-123.7 90.8 30.1 2.42 Phenols 11.0-38.3 22.7 6.24 11.0-38.3 20.2 6.45 15.6-35.6 25.6 4.82 22.5-35.5 25.5 3.82 2.74 Tannins 8.9-20.5 14.3 3.07 8.9-20.5 15.1 2.81 10.2-20.0 13.4 3.21 9.6-19.5 13.7 2.99 0.85 Saponins 11.0-35.6 19.2 5.60 11.0-29.5 16.8 4.97 12.3-35.6 22.7 5.14 12.5-25.6 19.3 3.70 0.82 Tocopherols 2.5-33.5 14.8 5.94 2.5-33.5 14.1 6.59 9.5-28.4 15.4 4.89 8.5-26.5 16.1 5.80 1.23 Goyal et al.: Variability in the nutrients, antinutrients and other bioactive compounds in soybean the highest sucrose content of 138 mg/g seeds. Local genotypes exhibited significantly higher mean total sugars and sucrose content as compared to mean values for these parameters for total genotypes and genotypes collected from other parts of India and exotic genotypes (only total sugars content). These genotypes can be preferred in terms of their nutritional attributes. Antinutrients and bioactive compounds: Large variation was seen in soybean genotypes for antinutritional factors studied (Table 2). Soybean genotypes were found to have a significant variation in the phytic acid content as it varied from as low as 1.2 (SL 716) to as high as 28.5 mg/g seeds (SL 137). The mean content of phytic acid was found to be less for exotic lines as compared to genotypes developed locally or within India. Mean Trypsin Inhibitor Activity (TIA) was significantly higher in exotic genotypes as compared to those collected locally or from other parts of India. SL 1000 had the lowest trypsin inhibitor activity of 11.3 mg/g seeds and PS 1414 had the highest activity of 142.5 mg/g seeds. Different authors have reported varied ranges of TIA in soybean. Guillamon et al. (2008) reported TI values of 43-84 TIU mg -1 sample whereas other repo rted values for TI are 7 6.52 TIU/mg seed (Rameshbabu and Subrahmanyam 2011) and 15.35 mg/g (Peric et al. 2009). Variation reported in the trypsin enzyme inhibitory activities by different authors might be because of differences in the methods and units used. Phenolic content of the genotypes varied from 11.0 to 38.3 mg/g with a mean value of 22.7 mg/g seeds. Among the local genotypes, SL 987 was found to have the lowest content of total phenols. The phenolic content ranged from 11.0 to 38.3, 15.6 to 35.6 and 22.5 to 35.5 mg/g seeds for genotypes collected from Ludhiana, other parts of India and exotic lines, respectively. The local genotypes showed significantly lower mean phenolic content values as compared to mean values for genotypes collected from other places within or outside India. Genotypes with low phenolic content are preferred for nutritional purpose, but genotypes with high phenols are beneficial to plant against insect/pest resistance and also as a source of bioactive compounds (Xu and Chang 2008). Tannin content of 140 soybean genotypes was found to be in the range of 8.9 (SL 990) to 20.5 mg/g seeds (SL 894) with the mean value of 14.3 mg/g. Tannins and phenolic constituents bind with the proteins of saliva and the mucosal membrane of the mouth during mastication of food and adversely affect the utilization of proteins in animal and human diets (Akinyede et al. 2005). Large variation was observed in the saponins and tocopherols contents of 140 soybean genotypes (Table 3). Saponin content varied from 11.0 to 35.6 mg/g seeds with mean value of 19.2 mg/g seeds. Dehulled soybean and seeds are reported to contain saponin content between 0.08% and 0.25% (Dandanell et al. 1995). Lower saponin content of a genotype is desirable from nutritional point of view as these 317 compounds are toxic (Campos-Vega et al. 2010) and result in retarded growth (Golawaska 2007). Tocopherol content of 140 soybean genotypes varied from 2.5-33.5 mg/g seeds. The mean tocopherol content of exotic lines was significantly higher than the local genotypes/total genotypes studied. Maximum tocopherol content was observed in genotype SL 790. Tocopherols are reported to have antioxidant properties (Evans et al. 2002) and also result in oil stability (Stone and Papas 2003). Manipulating seed tocopherol biosynthetic pathway in soybean to convert the less active tocopherols to the most active á- tocopherol could have significant human health benefits and make this crop an attractive target for the improvement of tocopherol composition (Van Eenennaam et al. 2003). Cluster analysis: The genotypes were grouped into clusters on the basis of biochemical similarity (Fig. 1-2). Local genotypes were divided into three clusters A to C (Fig. 1). Cluster A was composed of 41 genotypes with protein content and TIA in the range of 39.3-45.3 % and 11.3-138.8 mg/g, respectively. Further, the mean protein and TIA content for cluster A was 42.6 % and 94.4 mg/g respectively. The sucrose content of cluster A ranged from 0.24-13.6 g/100g with mean value of 7.2 g/100g. Cluster A was further subdivided into six sub clusters: A-I to A-VI with A-I forming the largest sub cluster having 9 genotypes. The genotypes in sub clusters A-I to A-VI had mean protein content of 43.4, 42.6, 42.3, 42.8, 42.9 and 41.6 %, respectively. Mean TIA of sub cluster A-I was highest (113.3 mg/g), whereas sub cluster A-V showed minimum TIA (68.8 mg/g) in this group. Cluster B contained 27 genotypes most of which had comparatively higher protein content (mean 43.2%) and lower TIA (39.6 mg/g) than members belonging to cluster A. Cluster B was subdivided into five sub-clusters B-I to B-V. Sub-cluster B-III consisted of 3 genotypes with a higher mean TIA (51.3 mg/g) than other four sub-clusters. The genotypes from sub-cluster B-IV had the higher protein, phytate and tocopherol content than subclusters B-I, B-II, B-III and B-IV. Cluster C contained 6 genotypes having average protein content of 42.4 %. The genotypes belonging to this cluster had the lowest mean TIA (20.6 mg/g) & sucrose content (4.9g/100g) and comparatively higher phytate content (12.1 mg/g) than those in clusters A and B. The genotypes of all the three clusters had almost similar mean oil content of about 21.8 %. Genotypes from other parts of India were grouped into two clusters A & B having 29 and 22 genotypes, respectively (Fig. 2). Both clusters had almost similar mean protein content of 43.2 % except genotypes PS 1444 in cluster A and PK 1026 in cluster B which had high protein content of 46.5% and 45.1%, respectively. The TIA of cluster A ranged from 11.3143.6 mg/g. Cluster A was divided into 4 sub-clusters A-I to A-IV. The sub-clusters did not differ much for their mean protein and oil contents. However, members of sub cluster A-I had a much higher TIA (116.9 mg/g) and sucrose content (6.6 g/ 318 Journal of Food Legumes 25(4), 2012 100g) in comparison to other three sub-clusters. YMV-25 forms an outlier as it has higher protein, lower TIA and sucrose content (44.1, 30 and 1.3, respectively). Cluster B was also divided into two sub clusters B-I & B-II composed of 6 and 10 genotypes, respectively. Both the sub clusters had almost similar protein, oil and tocopherol contents. Mean TIA of two sub-clusters was 41.3 & 51.4 mg/g, respectively. The genotypes in sub cluster B-II had higher mean phytate content (10.7 mg/g) and lower sucrose content (3.2g/100g) as compared to sub cluster B-II. Sel-46 was an outlier with higher TIA (101.3 mg/g) and lower phytate content (2.3 mg/g). The oil content of the outlier was also high (22.1%) as compared to mean oil content of clusters A and B. Exotic genotypes were also grouped into two clusters A & B (Fig. not given). Cluster A was composed of 11 genotypes with a higher mean protein content (43.7%) and a lower oil content (21.5%) than cluster B (41.0 & 22.4%, respectively). The mean TIA and tocopherol content of members of cluster A (96.5 & 26.1 %, respectively) was higher Fig. 1 than that of cluster B (75.6 & 23.1% respectively) whereas phytate content exhibited reverse trend (14.9 mg/g in cluster B as compared to 8.9 mg/g in cluster A). The members of cluster B also had a higher mean sucrose content of about 7.6 g/100g as compared to cluster A (6.8 g/100g). Significant variation was observed with respect to various nutritional and antinutritional characteristics among different soybean genotypes investigated. The mean grain yield for 140 soybean genotypes was 3.29 kg/plot. R-5 and DS-98-2 exhibited the maximum and minimum yield (5.0 and 1.47 kg/plot, respectively). Genotypes SL 992 and PK 1026 contained protein content of 44.6 and 45.1 %, respectively and a lower trypsin inhibitor activity and phytic acid content. Some of the genotypes studied had very low levels of different antinutritional factors. This diversity in soybean germplasm will be useful to the plant breeders for using genetic resources for the development of new cultivars with improved quality traits, enhancement of germplasm and commercialization of the end-products. Dendrogram of 74 local soybean genotypes by UPGMA clustering method based on standardized matrix derived from eleven seed quality traits. A, B, C indicate clusters; I, II, III, IV, V and VI are subclusters Goyal et al.: Variability in the nutrients, antinutrients and other bioactive compounds in soybean Fig. 2 319 Dendrogram of 51 soybean genotypes collected from other parts of India using UPGMA clustering method based on standardized matrix derived from eleven seed quality traits. A and B indicate clusters; I, II, III, IV, V and VI are subclusters ACKNOWLEDGEMENT We gratefully acknowledge Dr M. Javed, Associate Professor of Department of Mathematics, Statistics and Physics and Mr. D. Bhatia, Research fellow from School of Agricultural Biotechnology for their assistance in statistical analysis. We are also grateful to University Grants Commission for funding this project. Food Research International 43: 461-482. Dandanell DY, Aman P, Betz JM and Obermeyer R. 1995. Saponin in dehulled seeds from peas (Pisum sativum L.). In: Proceedings of the 2nd European conference of grain legumes , Copenhagen, Denmark. Pp. 310-311. Dubois M, Gules KA, Hamilton JK, Roberts PA and Smith F. 1956. Colorimetric method for the determination of sugars and related substances. Analytical Chemistry 28: 350-356. REFERENCES Evans JC, Kodali DR and Addis PB. 2002. Optimal tocopherol concentrations to inhibit soybean oil oxidation. Journal of American Oil Chemists’ Society 79: 47-51. Akinyede AI, Amoo IA and Eleyinmi AF. 2005. Chemical and functional properties of full fat and defatted Dioclea reflexa seed flour. International Journal of Food Agriculture and Environment 3: 112115. Faostat 2009. ProdSTAT: Crops. FAO Statistical Databases (FAOSTAT), Food and Agriculture Organization of the United Nations (FAO): http:/faostat.fao.org. Awadesh K, Singh G, Kumar D and Agrawal K. 2003. Physico-chemical characteristics of some new varieties of soybean. Journal of Food Science and Technology 40: 490-492. Blochl A, Peterbauer T and Richter A. 2007. Inhibition of raffinose oligosaccharide breakdown delays germination of pea seeds. 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Engineering vitamin E content: From Arabidopsis mutant to soy oil. Plant Cell 15: 3007-3019. Varsha R and Grewal RB. 2009. Carbohydrate profile, dietary fibre, antinutrients and in vitro digestibility of nine cultivars of soybean (Glycine max L.) Merr. Legume Research 32: 31-35. Xu B and Chang SK. 2008. Antioxidant capacity of seed coat, dehulled bean, and whole black soybeans in relation to their distributions of total phenolics, phenolic acids, anthocyanins and isoflavones. Journal of Agricultural and Food Chemistry 56: 8365-8373. Journal of Food Legumes 25(4): 321-325, 2012 Effect of presoak treatment on cooking characteristics and nutritional functionality of rice bean V.D. PAWAR , M.K. AKKENA, P.M. KOTECHA, S.S. THORAT and V.V. BANSODE1 Department of Food Science and Technology, Post Graduate Institute, Mahatma Phule Krishi Vidyapeeth, Rahuri 413722, India; 1Rajiv Gandhi College of Food Technology, Marathwada Agricultural University, Parbhani 431401 (M.H.), India; E-mail: [email protected] (Received : May 25, 2012 ; Accepted : December 04, 2012) ABSTRACT The effect of soaking rice bean on proximate composition, cooking characteristics, phytic acid, polyphenols content and functional properties were investigated. This study aimsed at comparing the changes that occured on soaking of blanched rice bean seeds in distilled water for 12 h and in salt solution (1.5% NaHCO3, 0.5% Na2CO3 and 0.75% citric acid, pH 7.0 ± 0.1) for 3, 6, 9 and 12 h each. Soaking, significantly (P < 0.05) increased the water absorption and dispersal of solids while it decreased the cooking time following increase in time of soaking. The anti-nutrients viz., polyphenols and phytic acid significantly decreased during soaking. The decrease in phytic acid was higher in salt solution soaking as compared to distilled water soaking. Soaking of beans for 12 h in different solutions did not show remarkable changes in proximate composition of rice bean flours. But foaming and emulsifying properties, nitrogen solubility and PCMP number showed a negative correlation, whereas, bulk density, water and oil absorption capacity showed a positive correlation with soaking time. Key words: Anti-nutrients, Cooking characteristics, Functional properties, Presoak treatment, Rice bean Rice bean is a rich source of nutrients. There are many reports although indicate that the protein content in rice bean is in lower range as compared to other pulses yet its bioavailability is high. As in other pulses, an important problem with rice bean is that it contains various anti-nutrients, notably phytic acid and polyphenols that reduce the uptake of several micronutrients. Kaur and Kapoor (1992) reported in rice bean that the poylphenolic content varied between 1279 and 1587 mg and phytic acid content between 1875 and 2270 mg/100 g. Saikia et al (1999) observed phytic acid ranged from 1998 to 2170 mg/100 g in five varieties of rice bean. Soaking of beans facilitates quicker cooking. Soaking and cooking of legumes result in significant reduction in phytic acid and tannin contents. Maximum reduction of phytic acid (78.05%) and tannin (65.81%) was found for sodium bicarbonate soaking followed by cooking. These treatments also result in a slight reduction in nutrients such as protein, minerals and total sugars (Iyer et al. 1980). The effects of presoaking of pulses in the salt solution of several chemicals in reducing the cooking time of pigeonpea splits (Narsimha and Desikachar 1978a), peas (Bongirwar and Sreenivasan 1977), beans (Rockland and Mertzler 1967, Iyer et al. 1980), winged bean (Narayana 1981), pigeon pea, chickpea, black bean, mung bean and lentil splits (Chavan et al. 1983) and moth bean (Pawar and Ingle 1986) were also reported. Thus, the current investigation was carried out to study the effect of presoak treatment with distilled water vis-à-vis salt solution on proximat e co mpositio n, cooki ng characteristi cs, antinutrients and nutritional functionality of rice bean. MATERIALS AND METHODS In the present investigation, seeds of rice bean (Vigna umbellata) were obtained from the Department of Agricultural Botany, MPKV, Rahuri. These were cleaned for extraneous matter and stored in clean glass bottles at 40C, until use. Quick cooking beans were prepared as per the procedures described by Rockland and Mertzler (1967). One lot was soaked in distilled water for 12 h and the unsoaked sample was taken as the control. The soaked beans were cooked traditionally in distilled water at 100 0C (beans to water ratio was 1:4 in terms of weight/ volume) until these were softened to a uniform mass when pressed between thumb and forefinger (Sharma et al. 1977) for determination of cooking time. The proximate composition of unsoaked rice bean such as moisture, protein, fat, ash and carbohydrates of the bean was determined as per AOAC (2000) procedures. The rate of hydration, dispersed solids and cooking time were determined as per the procedures of Narasimha and Desikachar (1978). The PCMP number was calculated by the following formulae. PCMP = pectin (Ca + Mg/2) /phytin or pectin + (Ca +Mg/2) – phytin. The pectin, calcium and magnesium content were estimated according to standard procedure of AOAC (2000). Anti-nutrient factors like, polyphenolic content was estimated according to the method of AOAC (2000). Extraction and precipitation of phytate phosphorus were performed according to the method of Wheeler and Ferrel (1981). Phytate phosphorus estimation was carried out according to the method of Makower (1970). The iron content was measured by the AOAC(2000) method using O-phenanthroline reagent. On the basis of phytate phosphorus contents, the phytic acid was calculated assuming 28.20% phosphorus in the molecule. Functional properties such as water and oil absorption 322 Journal of Food Legumes 25(4), 2012 capacities were determined by the procedure of Beuchat et al. (1977).The least gelation concentration was determined by method of Coffman and Garcia (1977) with slight modifications as described by Deshpande et al. (1982). Nitrogen solubility was determined as per standard procedure. The foaming properties such as foaming capacity was determined according to method of Coffman and Garcia (1977) while specific volume of foams was determined as an index of air uptake during whipping and weights were taken before and after whipping and specific volumes calculated according to method by Baldwin and Sinthavalai (1974).The emulsion properties such as emulsifying activity and emulsifying stability of the samples were determined by the method of Yasumatsu et al. (1972) with slight modifications as described by Deshpande et al. (1982). The bulk density was determined according to Okezie and Bello (1988). Statistical analysis: The analysis of triplicate data as a function of treatment levels was done to determine significant difference (P < 0.05) by computing standard error and critical difference by using methods of Snedecor and Cochran (1980). RESULTS AND DISCUSSION Change in proximate and chemical composition of flours: A net loss of dry weight occurred as a result of soaking and oxidation of stored compounds with a gradual increase of moisture (Table 1). At the end of 12 h soaking period, per cent moisture content of the bean increased from the initial 10.3 to 11.4 and 11.7 for distilled water and salt solutions, respectively. There was a significant decrease in total carbohydrate content during soaking of rice bean. Similar observations were reported by Zacharie and Ronald (1995) on common beans. The total ash content also gradually decreased throughout the soaking period. Rao and Deosthale (1983) also reported loss in ash content of mungbean and urdbean during overnight soaking in distilled water due to leaching of total minerals in the soaking medium. Per cent protein content of bean also decreased significantly from 17.5 to 15.4 and 15.2 during soaking for 12 h in distilled water and salt solution, respectively. Little changes in the concentration of pectin, Ca and Mg were observed due to soaking while greater change in the concentration of phytate was found. Sample soaked in salt solution for 12 h showed a significant decrease in pectin, Ca, Mg, phytate and PCMP number from 3.82 to 2.92 mg/g, 2.58 to 1.79 mg/g, 0.72 to 0.51 mg/g, 21.72 to 12.39 mg/g and 0.514 to 0.479, respectively. The hard-to-cook defect in legume seeds is based on PCMP number which represents pectin insolubilization via binding with divalent cations (e.g. Ca2+, Mg2+) resulting from phytate breakdown by phytase. The effect of soaking on rice bean in a salt solution containing Na+ cations or distilled water decreased the amount of pectin and was associated with divalent cations that led to insoluble form reducing the PCMP Table 1. Effect of Soaking on Proximate composition, pectin, calcium, magnesium, phytate and PCMP number of rice bean flours Treatment Moisture (%) 10.3 11.4 Unsoaked Soaking a Soaking b 3h 6h 9h 12 h SEm(±) CD(P=0.05) Soakinga: Soaking in Protein (%) 17.5 15.4 Fat (%) 0.51 0.49 Ash (%) 4.5 4.4 Carbohydrate (%) 61.6 60.1 Pectin Calcium (mg/g) (mg/g) 3.82 2.58 3.05 1.83 Magnesium (mg/g) 0.72 0.54 10.5 16.9 0.48 4.4 61.0 3.63 2.35 10.9 16.4 0.46 4.4 60.5 3.42 2.13 11.5 15.8 0.48 4.4 60.2 3.22 1.95 11.7 15.2 0.47 4.3 60.0 2.92 1.79 0.07 0.05 0.005 0.007 0.08 0.03 0.02 0.21 0.17 0.01 0.02 0.2 0.10 0.07 distilled water for 12 h at 27 0C; Soakingb: Soaking in mixed salt solution at 27 0 C 0.69 0.64 0.58 0.51 0.01 0.04 Phytate (mg/g) 21.72 19.56 PCMP number 18.97 16.76 14.52 12.39 0.09 0.28 0.511 0.497 0.493 0.479 0.007 0.022 0.514 0.322 Table 2.Effect of soaking on cooking characteristics and functional properties of rice bean flours Treatment Rate of hydration (g/100g) Unsoaked Soaking a Soaking b 3h 6h 9h 12 h SEm(±) CD(P=0.05) 166.6 49.5 62.9 101.4 142.7 1.22 3.84 Dispersed solids Cooking time Reduction in cooking time (g/100g) (min) (%) 30 21.8 10 66.7 9.7 13.7 17.6 21.6 0.787 2.48 25 20 15 8 0.47 1.45 16.6 33.3 50.0 73.3 - Water Oil absorption absorption capacity capacity (g/g) (g/g) 2.40 1.45 2.41 1.45 2.39 2.41 2.44 2.50 0.03 0.10 Soakinga: Soaking in distilled water for 12 h at 27 0C; Soakingb: Soaking in mixed salt solution at 27 0C 1.45 1.50 1.55 1.58 0.007 0.02 Foaming capacity (%) 25 21 Specific volume (ml/g) 1.27 1.21 24 22 21 19 0.47 1.45 1.25 1.22 1.21 1.18 0.05 0.01 Pawar et al.: Presoak treatment on cooking and nutritional characteristics of rice bean number and hardness of the bean. Bicarbonate and carbonate of sodium salt solution have solubilizing effect on the pectic substances that facilitates easy penetration and faster hydration on interior starch and protein molecules resulting into quick cooking beans. Cha nge in cooki ng cha racteristics and functional properties: It was observed that rate of water uptake and leaching losses of total solids were higher when beans were soaked in distilled water, but lower when soaked in salt solution (Table 2). However, the rates of water uptake and leaching losses of total solids increased progressively when beans were soaked in salt solution for 3, 6, 9 and 12 h. Rate of hydration and dispersed solids were also found significant in case of 12 h distilled water soaked sample than that of salt solution. Iyer et al. (1980) also observed increased rate of both hydration and leaching losses with increased soaking time with mixed salt solution from 6 to 24 h in Great Northern, Kidney and Pinto beans. Moreover, time required for cooking blanched rice bean was 30 min (Table 2) when the beans to cooking water ratio were 1.4 (wt/vol). When the beans were soaked in distilled water for 12 h or in salt solution for 3, 6, 9 and 12 h, the time required for cooking significantly decreased from 30 min to 10, 25, 20, 15 and 8 min, respectively. A significant decrease in cooking time was found in 12 h salt soaked sample compared to 12 h distilled water soaked sample. The beans soaked in distilled water and salt solution for 12 h caused reduction in cooking time by 66.7% and 73.3%, respectively. In the present study, the pretreatment of seeds, duration of soaking and type of soaking solution influenced the cooking time. Increased reduction in cooking time observed by soak treatment of blanched seeds in salt solutions than in distilled water might be due to the presence of cations in the soaking medium that increased the softening rate (as a result of ion exchange and probably by chelation of ions resulting in solubilization of pectin substances). Kadam et al. (1981) reported 67% reduction in cooking time of moth bean when soaked in a mixed salt solution for 12 h. Narayana (1981) reported that soaking the winged bean splits in salt solution for 2 h reduced the cooking time by nearly 50%. 323 Similarly, soaking in salt solution for 12 h had a significant water absorption capacity (2.5 g/g flour) than soaking in distilled water (Table 2) for 12 h (2.4 g/g flour). Although there was apparent decrease in protein content in salt soaked flours, yet the water absorption capacity increased significantly due to dissociation of proteins during blanching of rice bean before soaking in salt solution. Change in nitrogen solubility: Unsoaked rice bean flour had minimum N solubility of 12% at pH 5 (Fig 1). However, at pH 3.0, about 60% of nitrogen was soluble and at pH 12 it was about 90%. In the present investigation, N solubility of raw and 12 h distilled water soaked rice bean flours was increased even beyond pH 10 although there was no significant improvement in N solubility in rest of the cases (salt soaked samples). However, it remained more or less constant up to pH 12.0. Moreover, as the time of soaking in salt solution increased the solubility of N decreased at all pH levels in all soaked flours. Decrease in N solubility of 12 h water soaked rice bean flour might be due to decrease in protein content. Changes in foam stability: The decrease of total volumes after soaking rice bean in distilled water and salt solution for 12 h was 9.09 and 9.2%, respectively compared to 7.2% of raw rice bean flour (Table 3) while the corresponding decrease in Raw Nitrogen solubility Fig 1. Effect of soaking on the nitrogen solubility (%) of rice bean flours Table 3. Foam stability of soaked rice bean flours Treatment Raw Soaking a Soaking b 3h 6h 9h 12 h SEm(±) CD(P=0.05) 0 min Total Foam 125 33 121 29 124 122 121 119 0.47 1.45 32 30 29 27 0.47 1.45 30 min Total Foam 124 31 118 27 122 120 119 116 30 28 27 25 60 min Total Foam 121 29 116 25 120 116 115 113 26 26 25 23 90 min Total Foam 118 27 113 21 118 113 113 111 24 22 21 21 120 min Total Foam 116 23 110 18 % decrease in volume Total Foam 7.20 30.3 9.09 37.9 114 111 110 108 0.52 1.62 8.06 9.01 9.09 9.20 Soakinga: Soaking in distilled water for 12 h at 27 0C ; Soakingb: Soaking in mixed salt solution at 27 0C 21 19 18 16 0.52 1.62 34.3 36.6 37.9 40.7 324 Journal of Food Legumes 25(4), 2012 Table 4. Effect of soaking on emulsion activity and stability of rice bean flours Treatment Unsoaked Soaking a Soaking b 3h 6h 9h 12 h SEm(±) CD(P=0.05) Emulsifying activity (%) 52.40 42.44 Emulsifying stability (%) 49.22 40.44 Bulk density (g/ml) 0.582 0.642 % increase of bulk density 46.58 44.76 43.24 41.82 0.47 1.46 44.23 42.46 41.28 38.77 0.08 0.27 0.592 0.614 0.638 0.651 0.005 0.012 1.72 5.17 8.62 12.06 10.34 Soakinga: Soaking in distilled water for 12 h at 27 0C; Soakingb: Soaking in mixed salt solution at 27 0C foam volume was 37.9, 40.7 and 30.3%, respectively. Thus, the foam stability was comparatively more in raw rice bean flours than water or salt soaked rice bean flours. Change in emulsion properties and bulk density: Emulsifying activity of unsoaked dry rice bean flour sample was 52.4% which decreased significantly (P < 0.05) on soaking rice bean either in distilled water or salt solution (Table 4). As the time of soaking in salt solution increased from 3 to 12 h, the emulsion properties decreased significantly. Bulk density of raw rice bean flour was also increased from 0.582 to 0.651 and 0.642 g/ ml following 12 h soaking with salt solution and distilled water, respectively. Effect of soaking on anti-nutrients of rice bean flours: The polyphenols expressed as tannic acid decreased significantly from 9.62 to 6.37 and 5.77 mg/g in rice beans soaked in distilled water and salt solution for 12 h respectively. The reduction of pol yphenols on soaking in salt solu tion increased progressively with time of soaking. Kaur and Kapoor (1992) observed a remarkable reduction (27.2-36.8%) in the polyphenolic contents of rice bean when soaked for 6 h and 8 h in distilled water. The phytic acid content also decreased significantly Table 5. Effect of soaking on polyphenolic content, phytate phosphorus and phytic acid of rice bean flours Treatment Polyphenols Reduction Phytate Phytic Reduction in phosphorus acid of phytic (mg/g) polyphenols acid (mg/g) (mg/g) (%) (%) Unsoaked 9.62 6.12 21.72 Soaking a 6.37 33.7 5.51 19.56 9.94 Soaking b 3h 8.12 15.5 5.34 18.97 12.66 6h 6.62 31.1 4.72 16.76 22.83 33.14 9h 6.12 36.3 4.08 14.52 12 h 5.77 40.0 3.48 12.39 42.95 SEm(±) 0.04 0.09 CD(P=0.05) 0.13 0.28 Soakinga: Soaking in distilled water for 12 h at 27 0C; Soakingb: Soaking in mixed salt solution at 27 0C from 21.72 mg/g to 12.39 mg/g and 19.56 mg/g when rice beans were soaked for 12 h in salt solution and distilled water, respectively (Table 5). The rice beans soaked in salt solution for 3 h also showed a remarkable decrease in phytic acid (12.66%) when compared with beans soaked in distilled water for 12 h. Thus, the beans soaked in salt solution for 12 h showed a significant decrease in phytate content compared to other salt soaked and distilled water soaked samples (Kaur and Kapoor 1992, Deshpande and Cheryan (1983). This decrease in phytic acid in legume seeds during soaking can be attributed to leaching of phytate ions into soaking water under the influence of concentration gradient. The study suggested that the anti-nutrients such as polyphenols and phytic acid (along with N solubility, foaming and emulsifying properties) were decreased with soaking time whereas, the water and oil absorption capacity and bulk density of rice bean flours increased. The PCMP number also decreased with increase of soaking of rice bean in salt solution and had an advantage of improving the cooking quality characteristics, removal of antinutrients and functional properties over rice beans either unsoaked or soaked in distilled water. REFERENCES AOAC. 2000. Official Methods of Analysis, 17 th Edition. Association of Official Analytical Chemists. Washington, DC. Baldwin RE and Sinthavalai S. 1974. Fish protein concentrate. Journal of Food Science 39: 880-882. Beuchat LR, Cherry JB and Quinn MR. 1977. Physicochemical properties of peanut flour as affected by proteolysis. Journal of Agricultural and Food Chemistry 23: 616-620. Bongirwar DR and Sreenivasan A. 1977. Development of quick cooking peas. Journal of Food Science and Technology 4: 17-23. Chavan JK, Jawale HK, Shere DM, Jadhav SJ and Kadam SS. 1983. Effect of presoak treatments on the cooking time of legume dhal. Indian Food Packer 37: 78-81. Coffman AW and Garcia VV. 1977. Functional properties and amino acid content of protein isolate from mung bean flour. Journal of Food Science 48: 1654-1662. Desphande SS, Sathe SK, Cornforth D and Salunkhe DK. 1982. Effects of dehulling on functional properties of dry bean flours. Cereal Chemistry 59: 396-401. Iyer V, Salunkhe DK, Sathe SK and Rockland LR. 1980. Quick-cooking beans: Investigations on quality. Qualitas Plantarum Plant Foods for Human Nutrition 30: 27-31. Kadam SS, Satwadhar PN and Salunkhe DK. 1981. Effects of germination and cooking on polyphenols and in vitro protein digestibility of horse gram and moth bean. Qualitas Plantarum Plant Foods for Human Nutrition 31: 71-76. Kaur D and Kapoor AC. 1992. Nutrient composition and antinutritional factors of rice bean. Food Chemistry 43: 119-124. Makower RU. 1970. Extraction and determination of phytic acid in beans. Cereal Chemistry 47: 288-295. Narasimha HV and Desikachar HSR. 1978. Simple procedures for Pawar et al.: Presoak treatment on cooking and nutritional characteristics of rice bean 325 reducing the cooking time of split red gram. Journal of Food Science and Technology 15: 149-152. antinutritional factors and effect of cooking on nutritional quality of rice bean. Food Chemistry 67: 347-352. Narayana K. 1981. Cooking characteristics of winged bean. Journal of Food Science and Technology 18: 32-33. Sharma YK, Tiwari AS, Rao KC and Mishra A. 1977. Studies of chemical constituents and their influence on cookability of pigeon pea. Journal of Food Science and Technology 14: 38-42. Okezie BO and Bello AE. 1988. Physicochemical and functional properties of winged bean flour and isolate compared with soy isolate. Journal of Food Science 53: 450-455. Pawar VD and Ingle UM. 1988. Functional properties of raw and cooked moth bean flours. Journal of Food Science and Technology 25: 186-189. Rao PV and Deosthale YG. 1983. Tannin content of pulses: varietal differences and effect of germination and cooking. Journal of Science of Food and Agriculture 33: 1013-1016. Rockland LB and Mertzler EA. 1967. Quick cooking lima and other dry beans. Food Technology 21: 345-349. Saikia P, Sarkar CR and Borua I. 1999. Chemical composition, Snedecor GW and Cochran WG. 1980. Statistical Methods, 7 th Ed. Iowa State University Press, Ames, IA. Pp. 358-360. Wheeler EL and Ferrel RE. 1981. A method for phytic acid determination in wheat and wheat fractions. Cereal Chemistry 48: 312-316. Yasumatsu K, Sawada K, Moritaka S, Misaki M, Toda J, Wada T. and Ishil K. 1972. Whipping and emulsifying properties of soyabean products. Agricultural and Biological Chemistry 36: 719-727. Zacharie B and Ronald ES. 1995. Effects of soaking, cooking and fermentation on composition, in-vitro starch digestibility and nutritive value of common beans. Qualitas Plantarum Plant Foods for Human Nutrition 48: 349-365. Journal of Food Legumes 25(4): 326-329, 2012 Factors associated with economic motivation of legume growers in desert area of Rajasthan SUBHASH CHANDRA, P.SINGH1 and J.P. LAKHERA2 Krishi Vigyan Kendra; 1Agricultural Research Station, Beechwal, 2Directorate of Extension Education, SKRAU, Bikaner-334006, Rajasthan, India; E-mail: [email protected] (Received: October 20, 2012; Accepted: November 23, 2012) ABSTRACT The study was conducted during Kharif 2010 in arid region of Rajasthan involving 108 legume growers to find out the association of socioeconomic attributes with economic motivation of legume growers. The independent variables, such as age, education, land holding, farm power, social participation socioeconomic status and economic motivation were measured by the standard scale developed for this purpose. The study revealed that about half of respondents were middle aged with large land holding and educated upto middle standard. Majority of them belonged to medium (66.66%) and high (17.59%) socioeconomic status. It was observed that socioeconomic attributes such as age, land holding, farm power and socioeconomic status significantly associated with economic motivation whereas education and social participation were not associated with it. The information sources mostly utilized by farmers were the village level worker/ agriculture supervisor/ krishakmitra followed by radio, neighbours, input dealers and field demonstrations. Key words: Arid legume, Co mmunication behaviour, Demonstration, Economic mo tivation, So cio economic status Pulses are grown in scattered and specific agro-climatic input situations all over India to meet food needs of inhabitants and fodder demand for livestock. The legumes like cluster bean, cowpea, moth bean, mungbean and horse gram have pivotal and unparallel role in harsh farming conditions. These annual legumes are categorized as arid legumes and are specially known for their sustained production under extreme arid ecosystems, frequently encountered with harsh and hostile growing environments with unpredicted intensity and interval. Need-based adaptations of these legumes towards inclement weathers have recognized them as the source of livelihood for farmers surviving on resource constraint arid farming. Contribution of arid legumes in combating severe droughts, improving soil health, diversification of agriculture and as a source of organic foods have pushed them from their secondary status to major ones. The status of these arid legumes is ascertained from their vast acreage covering 4.9 million hectares (mha) in India including Jammu and Kashmir in interior north to deep in Kerala and from western states of Rajasthan and Gujarat to rear eastern states of Orissa and West Bengal. The Hyperarid partially irrigated western plain (Zone-1C) has maximum area (1.51 mha) in Rajasthan with only 0.56 million tonnes (m t) annual production because of very low productivity of these crops (40.0 Kg/ha). The hot arid regions of the country are characterized by hostile agro-climate and fragile eco-system with an annual rainfall of 100-150 mm inclusive of both rainfall and temperature related extreme events, low relative humidity and high potential evapo-transpiration of 1600 to 1800 mm especially in western part of region. Despite various biophysical constraints, the hot arid areas of Rajasthan like, Bikaner, Jaisalmer and Churu districts of western Rajasthan offers very good opportunities for cultivation of these legumes. Thus, location specific technological backup are key to sustainability of this region. These interventions are expected to stimulate a definite shift in cultural practices on a farm but may encourage a shift in investment layout, farm inventory and farm plan etc. The economic gain of farmers also depends upon their age, education, size of holding, socioeconomic status and their progressiveness as dynamics in their outlook motivates them to adopt new ideas or agricultural technology for economic gains. Keeping these in view, the present study was conducted to study both personal and socioeconomic characteristics of legume growers and also to find out the association of socioeconomic attributes with the level of economic motivation to them. The economic motivation refers to the occupational success in terms of profit maximization and relative values placed by the farmers on the economic ends (Supe 1969). MATERIALS AND METHODS The study was conducted in Hyper arid partially irrigated Western Plains (Agro-climatic Zone-Ic) of Rajasthan during kharif 2010. Bikaner and Churu districts were selected on the basis of higher area and production of arid legumes viz., cowpea, moth bean, cluster bean and mung bean. From each of the selected district, two panchayat samities were selected randomly from which three gram panchayats were again selected randomly from each of them. One revenue village was selected from each gram panchayat, randomly making a total of 12 villages for conducting the study. A comprehensive list of farmers who were growing arid legumes like, mungbean, mothbean, clusterbean and cowpea at least Chandra et al.: Factors associated with economic motivation of arid legume growers for the last three years was prepared with the help of agricultural supervisors. Thus, from each village, nine farmers were selected randomly and accordingly 108 contact farmers constituted the sample for the study. Independent variables, such as age, education, land holding, farm power, social participation and socioeconomic status were measured with the help scale developed by Trivedi (1963) while for computing economic motivation the scale developed bySupe (1969) was used. Farmers were categorized into three groups viz., high, medium and low on the basis of mean score and standard deviation. Communication sources utilized was measured using the scale developed under standard procedure. The data were tabulated and analyzed with the help of statistical tools viz., frequency, percentage and chi-square. RESULTS AND DISCUSSION Socioeconomic attributes: The study revealed that about half of the farmers (47.3%) were middle aged while 25% were young and the rest (25.9%) were old aged (Table 1). On educational qualification, one-third (32.4%) farmers were educated up to high school while one-fourth (26.8%) had passed middle school and one-fifth up to primary (19.4%). Regardi ng l and holding, more than half (51.8%) of respondents were large farmers, while 41.6% were small farmers Table 1. Distribution o f fa rme rs a s pe r pe rsonal socioeconomic attributes Characteristics Age (Years) Young (20-35) Middle(36-50) Old(above50) Education Illiterate Can read only Can read & write Primary schooling Middle school High school Graduate and above Land holding Marginal (< 1.0 ha.) Small (1.0-2.0 ha.) Medium and Big (> 2.0) Farm power No any farm power Camel/Bullocks Diesel engine/Electric motor Tractor (with implements) Sprinkler sets Social participation No Social participation Member of one organization Member of >one organization Office bearers Socio-economic status High (score above 36) Medium(score 20-35) Low(score up to 20) Frequency Percent 29 51 28 26.8 47.2 25.9 07 09 04 21 29 35 03 6.4 8.3 3.7 19.4 26.5 32.1 2.8 07 45 56 6.4 41.6 51.8 24 42 14 13 15 22.2 38.8 12.9 12.0 13.8 44 43 14 07 40.7 39.8 12.9 6.4 19 72 17 17.5 66.6 15.7 327 and only 6.4% were marginal farmers. However, on farm power front, less than half of the respondents (38.8%) had camel/ bullock power while 12.9% had diesel engine/electric motor and only 12.0% had tractor for effective farm mechanization. Similarly, only 13.8% farmers had sprinkler set for irrigation while 22.2% respondents did not have any source of farm power. As far as social participation is concerned, 39.8% farmers were member of one organization while 12.9% were member of more than one organization, thereby enabling half of them to have a direct contact/role in village level organization. These type of normal socioeconomic attributes were also reported by Singh et al. (2009) and Singh et al. (2011). In case of socioeconomic status (Table 1), majority of farmers possessed medium socioeconomic status (66.6%) while one-sixth belonged to high socioeconomic status (17.5%). Similar findings were made by Trivedi (1963), Singh et al. (2009) and Singh et al. (2011). Socio economic status vis-à-vis economic motivation: The age of the farmers, their land holding and socioeconomic status and availability of farm power were significantly associated with economic motivation. However, education and social participation were not associated with it (Table 2). Moreover, the age and economic motivation were dependent on each other. Age was an influencing and important factor in the pursuit of economic motivation of a person because of the fact that need and requirements were increased with the age of individual which motivated oneself to earn more and more (Singh and Sohal 1969 and Singh et al. 2009). However, the education had no positive bearing on economic motivation as both were independent attributes. The study also revealed uneven distribution of farmers regarding their education level. However, land holding seemed to have positive and significant association with economic motivation as the size of land holding affected the state of economic motivation. It may be due to the fact that almost all respondents were having medium and large land holdings and were engaged themselves in intensive cultivation so as to earn more income from farming by adopting new farm technologies. Farm power was found to be significantly associated with economic motivation as sufficient number of farm equipments and their accessories states the socioeconomic condition of the farmers. Moreover, power showed the progressiveness and innovativeness of the farmers and the innovative farmers were engaged in intensive farming so as to earn more profit from farm by using improved agriculture through equipments (farm mechanization). The social participation and economic motivation had no positive and significant association with each other and thus had no impact on economic motivation. However, the socioeconomic status was significantly associated with 328 Journal of Food Legumes 25(4), 2012 economic motivation. Since, majority of farmers were socioeconomically sound, thus acted as supplementary factor influencing level of economic motivation. Furthermore, farmers were unevenly distributed among various socio-economic status, yet they had difference in their perception from time to time regarding other factors of socioeconomic status. The overall situation reflected that the variables mentioned as socioeconomic attributes (Table 2) complimented and supplemented to socioeconomic motivation. These findings are in conformity with the findings obtained by Supe (1969), Singh et al. (2009) and Singh et al. (2011). Communication behaviour: The study also showed that the majority (75%) of farmers had access to VLW/Agricultural supervisors/Krishakmitra as they met them frequently. Moreover, radio and neighbours were next to VLW as the known source for information by the farmers (Upadhyay and Hansra 1986). The input dealers and demonstration jointly (37.04%), friends (30.55%) and cooperative societies (29.63%) were also important sources of information utilized by farmers. However, extension officers, B.D.O, scientists, AAOs, television, film show, leaflet, folder, farm magazine and bulletins were not preferred as a source of information frequented by the farmers. The occasional utilized sources included AAOs (80.55%) followed by block officials (63.89%), cooperative societies (53.71%), relatives (52.78%) and local leaders (50%), input dealers (46.30%), television and field demonstrations (35.19%). The different sources utilized by the farmers included film shows (92.60%), scientists (91.66%), farmers fair/ Ki san Gost hi (88.3 4%), members of Panchayat and Cooperatives, telephone talks/help lines, (79.63%), news papers (77.78%), group meetings (75.0%), television (64.81%) and krishi upaj mandi (50%), respectively. Thus, the primary source of information for the respondents were village level workers/agricult ural supervisors/krishakmitra, radio, neighbours, input dealers and field demonstrations. These findings are in accordance with the findings of Upadhyay and Hansra (1988), Saravanan et al. (2009) and Meena et al. (2010). Conclusively, the study suggested that socioeconomic att ributes viz., age, land holding, farm power and socioeconomic status were associated with economic motivation. The information source mostly utilized by farmers included village level workers/supervisors and was followed by inter-personal cosmopol ite sources such as radio, neighbours, input dealers and field demonstrations. Table 2. Association between socioeconomic attributes and economic motivation of farmers Attributes Age 20-35yrs 36-50yrs Above-50yrs Education Illiterate Can read only Can read & write Primary schooling Middle schooling High schooling Graduate and above Land holding Marginal Small Medium and Big Farm power No any farm power Camel/ Bullocks Diesel engine and Electric motors Tractor Sprinkler set and drip system Social participation No Social participation Member of one organization Member of > one organization Office bearers Socio-economic status(SES) Low Medium High *Significance at 0.05 level of probability, NS: Non-significant Economic motivation total Low (6-13) High (13-20) 9 20 7 44 13 15 0 07 02 07 02 02 04 17 11 18 12 23 0 03 02 05 8 37 14 42 11 13 13 29 06 08 04 09 05 10 27 17 18 25 03 11 05 02 08 09 15 52 05 19 Total 29 51 28 07 09 04 21 29 35 03 07 45 56 24 42 14 13 15 44 43 14 07 17 67 24 Chi-square value 11.21* 4.03 (NS) 6.39* 6.56* 4.05 (NS) 6.67* Chandra et al.: Factors associated with economic motivation of arid legume growers 329 Table 3. Distribution of farmers as per communication behavior* Source of Information/Channels Frequently Personal/ local source Neighbours Friends Relatives Progressive farmers Local leader Member of panchayat & cooperative Telephone talk(help line) Personal cosmopolite source Village level workers, Agrl. Sup.& krishakmitra Assistant Agriculture officer Group meeting Demonstration Farmers fairs & Kishan Gosthi Block dev. Officials (BDO) Scientist of ICAR & University Impersonal cosmopolite source Radio Television News paper Film shows Folder, farm magazine & bulletins Commercial agencies and NGOs Krishi Upaj Mandi Input dealers Cooperative societies Extension contact Occasionally Never 60(55.56) 33(30.55) 23(21.30) 17(15.74) 12(11.12) 9(8.34) 9(8.34) 20(18.52) 49(45.37) 57(52.78) 40(37.04) 54(50.00) 13(12.03) 13(12.30) 28(25.92) 26(24.08) 28(25.92) 51(47.22) 42(38.88) 86(79.63) 86(79.63) 81.0(75.00) 0.00 6(5.55) 40(37.04) 8(7.40) 0.00 0.00 18.0(16.66) 87.0(80.55) 21(19.45) 38(35.18) 10(9.25) 69(63.89) 9(8.34) 9.0(8.34) 21.0(19.45) 81(75.00) 30(27.78) 90(83.35) 39(36.11) 99(91.66) 75(69.45) 0.00 7(6.48) 0.00 0.00 20(18.52) 38(35.19) 17(15.74) 08(7.40) 02(1.85) 13(12.03) 70(64.81) 84(77.78) 100(92.60) 106(98.15) 23(21.30) 40(37.04) 32(29.63) 31(28.70) 50(46.30) 58(53.71) 54(50.00) 18(16.66) 18(16.66) *Figures in the parenthesis indicates percentage REFERENCES Meena SR, Sisidia SS, Punjabi NK and Sharma Chitranjan. 2010. Information seeking behaviour of farmers about guava Production Technology. Rajasthan Journal of Extension Education 17&18:5255. Saravanan R, Raja P and Tayeng Sheela 2009. Information input pattern and information needs of tribal farmers of Arunachal Pradesh. Indian Journal of Extension Education 45: 51-54. Singh DK, Singh AK, Yadav VP, Singh RB, Baghel RS and Singh Mayank. 2009. Association of socioeconomic status with economic Motivation of the farmers. Indian Research Journal of Extension Education 9: 53-56. Singh P, Sharma SK and Singh Sangram 2011. Information seeking behaviour of mothbean growers in western Rajasthan. Indian Journal of Agricultural Research and extension IV: 97-100. Singh R and Sohal T. 1969. Size of holding as related to acceptance of crop plans, extension contacts and education of farmers. Indian Journal of Extension Education 5: 42-48. Supe SV. 1969. Factors related to different degree of rationality in decision making among farmers. Ph.D. thesis, Division of Agricultural Extension, I.A.R.I., New Delhi. Trivedi G.1963. Measurement and analysis of socioeconomic status of rural families. Ph.D. Thesis, Division of Agricultural Extension, I.A.R.I., New Delhi. Upadhyay KP and Hansra BC. 1988. Evaluation of agricultural broadcast of radio Nepal in adoption of improved agricultural technology by Nepalese farmers. Journal of Research 23: 143-145. Journal of Food Legumes 25(4): 330-333, 2012 Farmers participatory approach in seed multiplication of pulses in Bundelkhand region - A case study PURUSHOTTAM, S.K. SINGH, C.S. PRAHARAJ and LAKHAN SINGH1 Indian Institute of Pulses Research, Kanpur-208 024; 1Zonal Project Directorate (KVK), Kanpur, U.P., India; E-mail: [email protected] (Received: October 20, 2012; Accepted: December 06, 2012) ABSTRACT A farmers participatory action research programme (FPARP) was carried out in seed production and multiplication of major pulses through modern seed plot techniques (SPT) to generate awareness about SPT and ensure availability of quality seed to farmers during 2009-2011 in Baank and Bannki villages in Hamirpur district of Bundelkhand region of Uttar Pradesh, India. A total of 161 farmers with 61 ha of net cultivated area participated in this FPARP programme by sharing half of the seed cost in three major pulses viz., chickpea, lentil and pigeonpea. Farmers were trained through one institutional, six field level and one special training on “know your crops (KYC)” for acquiring initial know-how and subsequent skill/ expertise development. The study amply demonstrated that yield advantages to the tune of 37.3, 24 and 51% in improved varieties of chickpea, lentil and pigeonpea, respectively over their counterparts (local variety) were obtained following above practice. The highest yields of 1320, 1000 and 1370 kg/ha were realized in chickpea, lentil and pigeonpea, respectively under farmers’ condition. The average cost of cultivation improved varieties of chickpea, lentil and pigeonpea were also reduced to the tune of ` 7500, 9386, 7287/ha, respectively resulting in higher benefit cost ratios (BCR) in chickpea (2.20), lentil (2.32) and pigeonpea (3.11) over the local (1.23, 1.09 and 1.17, respectively). Due to FPARP, farmers could able to produce 19.3, 9.0 and 14.2 tonnes of truthful level seed for the chickpea, lentil and pigeonpea crops, respectively. In addition, significant quantity (73.4%) of this produce could also enter into the seed chain (31.2 t of seed materials out of 42.5 t total produce) by the adopted farmers themselves. Chickpea seed was diffused fastest (from 60 adopted farmers to 119 other farmers in the very first season) and farthest (from the adopted villages to other 18 villages in a radius of 24 km). The factors affecting the overall yield levels and variations in the yields included soil based (nature of soil, moisture level and water stagnation), plant based (plant population and timely weeding), climate based (continuous fog, frost at flowering and winter rains) and pest and diseases based (aphid, root rot and pod borer) i.e., both abiotic and biotic in nature. Thus, the study suggested that with awareness and knowledge upgradation through FPARP, the farmers could ensure quality seed production and its multiplication and thereby, making seed production system viable and remunerative. Key words: Bundelkhand region, Farmers participation, FPARP, Pulses, Quality seed production Pulses are rich source of vegetative protein and play an important role in nutritional security of majority of vegetarian population in India. The country is the largest producer and consumer of pulses occupying 33% of the world’s area and 22% of the production (FAO 2008). Pulse production in the country has fluctuated widely between 13 and 15 million tonnes (m t) with no significant growth trend between 1991 and 2010. The latest estimate indicates that the present production of pulses has reached 14.7 million tons (mt) with productivity of 637 kg/ha although the projected pulse requirement by the year 2030 (32 mt) is estimated to be more than double the current production level (Anonymous 2011). Thus, increasing pulses production through either productivity increase or allocating non-traditional areas by cultivating pulses in specific pulses growing regions is becoming indispensable and crucial to make the Indian subcontinent self-sufficient in pulses. Bundelkhand region is considered to be the pulse bowl of Uttar Pradesh as it shares about 50% area and 45% of total pulse production of the state. The average pulses productivity in the Bundelkhand region was low (657 kg/ha) against 725 kg/ha as the state average. The reasons are biotic, abiotic, and socio-economic constraints causing low productivity in pulses in this region. In addition, lack of improved varieties is reported as the most serious constraint among all biophysical constraints in pulses production (Roy Burman et al. 2006). Moreover, unavai lability o f quality seed and l ack of technological awareness as revealed by 94.2 and 74.2% respondents from the farming community, also contributes much towards both low production and productivity of pulses (Purushottam et al. 2011). In the existing scenario, seed village could be the best approach to ensure better SRR and availability of seeds at the farmers’ door (Chaturvedi et al. 2010). A study revealed that ado ption of i mproved varieti es it self could enhance productivity by 20 to 25% in rabi pulses (Samra et al. 2011). Moreover, biotic factors including some soil borne fungal pathogens could cause extensive damages through Fusarium wilt and root rot complex through reduction in seed yield to the tune of 10-50% at farmers’ field in Bundelkhand region. It was also reported that low yield of chickpea due to wilt was the major problem in district Mahoba of Bundelkhand (Singh 2009). Thus, improved varieties of pulses could come to the farmers’ rescue in the event of crop failure as these were Purushottam et al.: A case study for seed multiplication in pulses on participatory mode basically resistant to major diseases (wilt and rust) and were mostly high yielder. Therefore, adoption of disease resistant improved varieties of rabi pulses could increase seed yield between 25-70% in rainfed mono-cropping and partially irrigated double cropping situations in Hamirpur district (Singh et al. 2005a). Since there was meager information available on these lines especially on remunerative seed multiplication programme in Bundelkhand region, hence a case study was undertaken with the objective of generating awareness and knowledge on quality seed production and ensuring its availability in major pulses in these region. MATERIALS AND METHODS A farmers’ participatory action oriented research programme was carried out in selected villages in Hamirpur district in Bundelkhand region of Uttar Pradesh, India during 2009-2011. The district has 2 tehsils, 7 blocks and 926 villages and the average (land) size of holding is 1.95 ha. Socioeconomic analysis of the farmers in the district revealed that there are 46% marginal, 23% small and 31% large farmers living in the district. Two representative pulse growing villages namely, Baank and Baanki were selected under Bharuwa Sumerpur block as majority of the farmers (based on land holdings) were involved in cultivation of pulses. The total population of the villages was 5022 with 1060 families and the cultivable area constituted about 80.6% (1052 ha) of the total geographical area (1306 ha). However, the cultivated area during rabi (894 ha) was much higher over that in kharif (357 ha). Chickpea, lentil and pigeonpea were the major pulses grown in an area of 312, 77 and 238 ha, respectively in diverse soil types viz., Mar, Kabar, Paruwa and Rakar during 200708. Mixed cropping was usually the most common practice adopted in pulses; and use of costly inputs like fertilizers, insecticide and herbicide was limited. Although line sowing was practiced in rabi pulses yet broadcasting was extensively adopted during kharif resulting in poor plant stand and low yield. Farmers’ participatory action research programme (FPARP) for seed production and multiplication was carried out by supplying quality foundation seed (2000 kg) of chickpea, certified seed (720 kg) of lentil and breeder seed (390 kg) of pigeonpea from registered/certified sources viz., IIPR Kanpur, CSAUA&T Kanpur, NDUA&T Faizabad and NSC, Kanpur. Recommended cultivars viz., ‘DCP 92-3’, ‘JG 16’ and ‘KGD 1168’ (chickpea), ‘Narendra Arhar 1’ (pigeonpea) and ‘DPL 62’ (lentil) were used. The areas planted with chickpea, lentil and pigeonpea were 20, 15 and 26 ha, respectively with respective 60, 36 and 65 farmers (a total of 161) participation. Farmers shared only half (50%) of the seed cost out of total cost of `88,875 (`49725, 21600, 17550 for chickpea, lentil and pigeonpea, respectively) as per project guidelines. For farmers’ own involvement and commitment for successful adoption of the concept, all other inputs except 331 the quality seed were managed by the farmers themselves. The crop was sown under rainfed condition. A base line survey indicated that farmers were not aware of seed pro duct ion/ mult ipli cati on t echniques and market ing intelligence for disposal of quality produce with a premium. Therefore, albeit their own understanding, skill (along with awareness/knowledge) enhancements were further improved through diverse hands on trainings both at field (six numbers) and institutional level (one number). Individual communication through face to face and over telephone was also maintained to have a special training on perfect transfer of “know your crops (KYC)” for effective FPARP. It was further strengthened through linkage and coordination with associated line departments viz., District Agriculture Departments, Input agencies and NGOs. The crop was inspected periodically and the seed produced was registered through Uttar Pradesh Seed Certification Agency, Jalaun. Project monitoring was made through the funding agency (NABARD) through a separate Project Implementation and Monitoring Committee (PIMC). To have a further boost to seed production programme, marketing of quality produce on community/village level and for further follow ups, a farmers’ club namely Harit Kisan Club was constituted in association with a Government Banking Agency (Allahabad UP- Gramin Bank). RESULTS AND DISCUSSION Gain in seed yield: Despite adverse climatic condition (continuous fog and winter rains) causing flower drop, poor seed setting and subsequent yield reduction, 37.3% gain in seed yield was realized as a result of improved varieties over local cultivars of pulses (with an average yield of 590 kg/ha). On an average, higher seed yield (1000 kg/ha) in chickpea was realized in ‘KGD 1168’ followed by ‘DCP 92-3’ and ‘JG 16’ (Table 1) whereas, the highest yield of 1500 kg/ha under farmers’ condition was recorded under ‘DCP 92-3’ followed by ‘KGD 1168’ and ‘JG 16’. The incidences of root rot and wilt was almost absent in case of ‘DCP 92-3’ and ‘KGD 1168’ chickpea. The study also revealed higher yield losses (2530%) in case of local chickpea varieties due to the above diseases. Even the incidences of root rot and wilt in JG 16 was Table 1. Yield levels of chickpea, lentil andpigeonpea through FPARP Crop Variety Chickpea DCP 92-3 KGD 1168 JG 16 Av. Lentil DPL 62 Pigeonpea Narendra Arhar 1 * Stalk yield Crop Average seed yield Increase Highest area (kg/ha) in yield yield (ha) Improved Local (%) (kg/ha) cultivars cultivars 10.0 980 590 36.7 1500 5.0 1000 590 40.9 1320 5.0 900 590 34.3 1150 960 37.5 1320 15.0 590 450 24.0 1000 26.0 550 270 51.0 1370 (3540)* (2320)* (34.0)* (45.0)* 332 Journal of Food Legumes 25(4), 2012 low and varied (5-20%) from one to another field. Thus, the role of improved variety over local or desi was established for realization of higher yield under farmers’ condition. Similar to chickpea, pigeonpea farmers also had a perceptible additional yield advantages through growing of improved varieties over local or desi varieties. Thus, visible differences between local and improved variety were apparent in terms of both average seed yield (270 kg/ha versus 550 kg/ ha) and stalk yield (2320 kg/ha versus 3540 kg/ha) resulting in yield gain to the tune of 51 and 34%, respectively by growing improved variety over the local. The stu dy also revealed variabl e seed yield in pigeonpea (300-1370 kg/ha) due to combination of several factors including sowing in light soil with moisture stress, water stagnation after sowing due to rainy months, poor germination, lack of timely weeding as a result of traditional practice of broadcasting and higher plant population, lack of appropriate insect control practice (mostly lack of insecticide for pod borer control), frost at low elevation and rainfed condition with no life saving irrigation. It was observed that a supplemental irrigation at flowering and use of insecticide against pod borer was beneficial in terms of realization of higher yield by the participating farmers over others. Under Indian condition as pulses are grown in rainfed areas, mostly under poor management conditions, the crop faces many biotic and abiotic stresses (Ali and Kumar 2009). Thus, substantial yield gain is possible when package of practices with modern technology is adopted by the farmers for raising crops. Quite similar observation was recorded in case of lentil as on an average higher seed yield (590 kg/ha) was obtained with improved variety in comparison to local variety (450 kg/ ha). Apparently, a yield gain to the tune of 24% (140 kg/ha) was observed in improved variety over local. In case of the yield level of lentil under farmers’ condition ranged from 350 to 1000 kg/ha. Moreover, large and lustrous seeds were obtained with the improved variety of lentil over local. Lower seed yield and higher variation in seed yield were again due to both biotic (incidence of aphid and root rot) and abiotic factors (soil type, poor land preparations, delayed sowing, lack of winter rains, low soil moisture at critical stages and frost at flowering). Thus, modification in abiotic stresses through appropriate remedial measures (viz., low soil moisture by life saving supplemental irrigation) and biotic stresses by control measures could increase seed yield significantly as observed in case of adopted farmers over others. Thus, possibility of higher yield realization could be explored by the farmers provided t hey adopted bett er recommended management practices. Although they had a mind set for low cost of cul tivation because of harvesting poor yield consistently due to associated risk factors, yet change in attitude and ability to learn and adopt newer technologies could land them in progressive mode, avert risks associated with crop husbandry and exploit yield potential of the crop(s). This was possible in case of project farmers in the adopted villages at least to some extent. Study also suggested that ‘DCP 92-3’chickpea was the best variety under the existing situation of clay loam and clay soils as single pre-sowing irrigation could yield 400 kg/ha more over local (951 kg/ha). Similarly, ‘DPL 62’ lentil and ‘Narendra Arhar 1’ pigeonpea grown under rainfed condition yielded 32% (1100 kg/ha) and 52% (918 kg/ha) higher over local check(s), respectively as reported by Dubey et al. 2011. Singh et al. (2005) also reported that improved varieties of chickpea, lentil and field pea had increased their seed yields to the tune of 45, 70.4 and 61.5% over local checks, respectively. Table 2. Eco nomics o f improv ed practice of seed multiplication in pulses* Crop Variety Chickpea Improved Local Lentil Improved Local Pigeonepa Improved Local Average Improved Local CC (`/ha) 7500 5300 9386 8654 7287 5482 8057 6478 Gross Income (`/ha) 24000 11820 20450 18093 29958 11940 24802 13951 Net return (`/ha) 16500 6520 21840 9440 22671 6458 20337 7472 BCR Additional Additional CC net return (`/ha) (`/ha) 2.20 2200 9980 1.23 2.32 732 12400 1.09 3.11 1805 16213 1.17 2.54 1579 12864 1.16 *Market price (improved versus local): Chickpea (`25 and `20/kg), Lentil (`33 and `29/kg) and Pigeonepa (`35 and `30/kg; by product/ stalk (`1300/ha for lentil and `400/t for pigeonpea) Gain in net return: Based on prevailing prices of inputs and outputs, improved varieties were economically viable over local varieties grown by the farmers although the cost of cultivation (CC) in case of former was on higher side (ranging from `732 to 2200/ha) primarily due to qualityseed cost. The average cost of cultivation for improved varieties of chickpea, lentil and pigeonpea were `7500, 9386, 7287/ha, respectively while the corresponding values for local varieties were `5300, 8654, 5482/ha in chickpea, lentil and pigeonpea, respectively. However, additional net return was increased to higher order (ranged from ` 9980 to 16213/ha with mean return of `12864 / ha) over local. Thus, seed multiplication resulted in higher benefit cost ratio (BCR) of 2.20, 2.32 and 3.11 in improved varieties as compared to 1.23, 1.09 and 1.17 in local checks in chickpea, lentil and pigeonpea, respectively. Thus, there was possibility of higher income per unit area and rupee invested through seed production programme over normal crop raising programme in pulses (meant for consumption). Similar work of Tomar et al. (2009) revealed higher mean net income of `9856/ha with a BCR of 2.13 in urdbean under improved technology package as compared to local practice (with net income of `3357/ha and BCR of 1.64) in Tikamgarh district of Bundelkhand. Disposal of quality Seed: The quantum of quality seed produced by the farmers as a result of improved technology were to the tune of 19.3, 9.0 and 14.2 t (Table 3) under chickpea, lentil and pigeonpea, respectively (a total of 42.5 t seed). This Purushottam et al.: A case study for seed multiplication in pulses on participatory mode not only met their own requirements (73.4%) for seed (31.2 t) but also fulfilled partial requirements (11.8 t) of their neighbours, relatives, land share holders and NSC besides meeting a few personal need for miscellaneous purposes (for exchange and loan). More importantly, quality chickpea seed was diffused from the adopted two villages to other 18 villages in a radius of 24 km (from a mere 60 adopted farmers to 119 in the very first season only). Since farmers usually do not open the stored seed during rainy season therefore, majority of seed was disposed off either just after harvesting or at sowing in the next season. Additional benefit included covering the majority of area of chickpea, lentil and pigeonpea (grown in 312, 77 and 238 ha in adopted villages) in due course of time by improved varieties that requires quality seed (24.9, 3.8, 3.5 t, respectively) through SRR. Thus, farmers became more progressive and their entrepreneurship behaviour led to constitution of a Farmers’ Club under project linking to International Traceability System Limited (ITS) for welfare of their locality. ITS helped the farmers club in lifting 2.3 t of lentil and 0.3 t of pigeonpea seed pooled from 20 farmers. ITS agency is an agency working with coordination of NSC for quality seed production and its safe disposal. Table 3. Disposal of quality seed of pulses from the villages Crop Variety Chickpea DCP 92-3 KGD 1168 JG 16 Sub-total Lentil DPL 62 Pigeonpea Naraendra Arhar 1 Total * Stalk yield Total Farmers’ own use (t) produce (t) 9.8 7.2 5.0 2.5 4.5 3.5 19.3 13.2 9.0 6.2 14.2 11.8 (95.0)* 42.5 31.2 Quantity for sale (t) 2.6 2.5 1.5 6.6 2.8 2.4 11.8 Marketing constraints in quality seed disposal: The study also showed that lack of seed processing plant in nearby locality, dissatisfaction of farmers on sample checking for quality, (delayed) payment in installments and miscellaneous handling charges influenced speed of quality seed disposal to Government Agencies. Due to these constraints, farmers were often forced to sell off their produce in local market meant for consumption resulting in jeopardizing actual seed replacement rate (SRR). Even small and marginal farmers initially followed their counterparts (other farmers) where they would receive the payments after lifting the seed from the villages that also resulted in delayed disposal of their produce. Therefore, appropriate marketing strategies such as earliest lifting of first lot just after threshing, disposal in small lots instead of pooling from large number of farmers and such other measures would boost their morale for sustenance in farming enterprises. Thus, it was inferred from the study that improved 333 varieties had adequate potential in enhancing quality seed multiplication through FPARP and thus, enabling adequate SRR for economic development in Bundelkhand region of Uttar Pradesh. Integration of pulses technologies played a greater role for higher social acceptability and enhancing profitability. This was possible through various confidence building measures for improving their awareness/knowledge through qu alit y traini ngs, KYCs, development of entrepreneurship and effective marketing strategy. ACKNOWLEDGEMENT The authors acknowledge financial support received from NABARD for implementing the project “Farmers’ participatory seed production of major pulse crops in selected villages of Hamirpur district in Bundelkhand region of Uttar Pradesh” at IIPR, Kanpur. REFERENCES Ali M and Kumar S. 2009. Major technological advances in pulses: Indian scenario (Eds), Indian Institute of Pulses Research, Kanpur, India. Pp 1. Anonymous 2011. Vision 2030, Indian Institute of Pulses Research, Kanpur. Pp iii-vii. Chaturvedi SK, Nadarajan N, Singh SK and Mishra JP. 2010. Strategies for enhancing pulses production in Bundelkhand tracts of U.P. and M.P. In: Extension strategy for Bundelkhand region, ZPD, ZoneIV, ICAR, Kanpur. Dubey SK, Sah Uma and Singh SK. 2011. Participatory impact assessment of technological interventions disseminated in Bundelkhand region of Uttar Pradesh. Journal of Food Legumes 24: 36-40. FAO 2008. Diagnosis of pulses performance of India. In: Srivastava SK, Sivaramane N. and Mathur VC (Eds.), Agricultural Economics Research Review 23: 137-148. Purushottam, Kumar Rajesh and Kumar Hemant 2011. Pulse production issues in Bundelkhand region of Uttar Pradesh. Agriculture Situation in India. LXVII: 661-666. Roy Burman R, Singh SK, Singh Lakhan and Singh AK. 2006. Adoption of improved pulses production technologies and related constraints in Uttar Pradesh. Indian Journal of Pulses Research 19: 104-106. Samra JS. 2009. A report on drought mitigation strategies for Bundelkhand region of U.P. and M.P. Rainfed Authority of India, New Delhi. 12 pp. Singh Atar and Singh AK. 2009. Yield advantages in pulses at farmers’ field. Journal of Food Legumes 22: 198-201. Singh Atar, Lakhan Singh and Singh NP. 2005. Performance analysis of pulses in frontline demonstrations. Indian Journal of Pulses Research 18: 202-205. Singh SK, Roy BR, Chaudhary RG, Singh KK and Ansari S. 2005a. Impact of usable technologies identified under pulse based rainfed agro ecosystem. Indian Journal of Pulses Research 18: 60-63. Tomar RKS, Sahu BL, Singh RK and Prajapati RK. 2009. Productivity enhancement of blackgram through improved production technologies in farmers’ field. Journal of Food Legumes 22: 202204. Journal of Food Legumes 25(4): 334-339, 2012 Tropical Legumes 2 pigeonpea seed system in India: An analysis M.E. HOLMESHEORAN, M.G. MULA, C.V.S. KUMAR¹, R.P. MULA and K.B. SAXENA International Crops Research Institute for the Semi-Arid Tropics, Patancheru 502 324, Andhra Pradesh, India; ¹Acharya N.G. Ranga Agricultural University (ANGRAU), Hyderabad, Andhra Pradesh, India; E-mail: [email protected] (Received : January 30, 2012 ; Accepted : October 01, 2012) ABSTRACT Pigeonpea farmers in India have historically relied on selfsaved seed of local varieties as their seed source for upcoming growing seasons. As improved varieties for disease resistance and yield have been consistently developed, the challenges have been to help farmers gain and retain access to these improved varieties. The above objectives were tried to be accomplished through improved agronomic practices, promoting the seed village concept to minimize the effects of out-crossing, and developing local seed production capacity under the aegis of the Bill Melinda Gates Foundation funded Tropical Legumes 2 (TL 2) project operational with the pigeonpea farmers for the last 4 years. The project was implemented in Tandur, Ranga Reddy District of Andhra Pradesh, India, a region where the pigeonpea is cultivated as monocropped or intercropped with other crops. Ahandful of farmers have become truthfully labeled seed producers, but educational programs and improved seed have not yet reached the majority of individuals in the communities targeted, creating a gap both in understanding and in meeting project goals. Small hold farmers because of their subsistence level are usually not involved in seed production. However, improved varieties should be made available to them for meeting the above objectives. The focus on continuing increasing opportunity for small holders through seed system improvement would yield more innovative methods for community involvement and accessibility so that the gaps in understanding can be bridged up for the welfare of the society as a whole. Key words: Pigeonpea, Seed village system, Tropical legumes 2, Truthfully labeled seed The process through which viable seed is produced, stored, marketed and used is known as a seed system or seed chain. Thus, seed system includes all the channels through which farmers acquire genetic materials, both outside of, and in interaction with, the commercial seed industry (Tesfaye et al. 2005). Seed systems vary widely depending on locality, market availability, and farmer knowledge, and can be informal, formal, or a combination of the two. An informal seed system functions primarily through farmer’s saving and storing their own seed for the next season. New infusions of seed stock may be purchased every few years, but usually from a local supplier. The formal seed sector is defined as any seed supplied through companies or government agencies that is registered and certified for quality. These entities usually exist at the regional and national level. Any farmer receiving seed supply through the formal seed channel is using the formal seed system for cropping. In India, self-saved seed accounts for roughly 80% of seed cultivated for food crops in any given growing season (Ravinder Reddy et al. 2007) indicating the fact that lower quality seed is being grown by the majority of the farmers, and accordingly yields are not as high as with improved varieties. In pulses also, low seed replacement rate (SRR) similar to cereals is a major problem. Though pigeonpea production has seen an increase in the seed replacement rate in recent years, farmers still self-produce and meet more than 85% of their own seed need. An attempt is made for small holders through improved seed system involving their community so as to bridge the gaps in understanding and operating the system for the welfare of the society. The seed replacement rate expresses the percentage of seeds for a specific crop purchased for a given season and indicates the status of improved seed replacement over the season. MATERIALS AND METHODS The Tropical Legumes 2 : The Tropical Legumes 2 project was a four year project (2008-2011) funded by the Bill Melinda Gates Foundation and has the goal of ‘improving the livel ihoods o f small hold farmers through improved productivity and production of tropical grain legumes in SubSaharan Africa and South Asia’. Small hold farmers in India are ‘marginal and sub-marginal farm households that own or/ and cultivate less than 2.0 hectare of land’ (Singh et al., 2002), which translates into 2 hectares per farm family. ICRISAT works in collaboration with Acharya N.G. Ranga Agricultural University (ANGRAU) located in Hyderabad in Andhra Pradesh (India) to implement the TL 2 project in two districts (Ranga Reddy and Mahaboobnagar) of Andhra Pradesh. Targeted areas in both districts are historically famous for pigeonpea production with a reputation of excellence in high dal quality produce. The TL 2 project has also provided the opportunity for the Agriculture Research Station, Tandur (ARS) to develop a few local seed producers, including one in Kolkat, a village included in the assessment. Free seed pigeonpea packets of 4-5 kg have also been given out to 783 farmers for 4 growing seasons of the TL 2 project. In order to assess the impacts of the TL 2 project so far, three villages were selected in Ranga Reddy of Andhra Pradesh, two villages that the project has been implemented Holmesheoran et al. : Tropical Legumes 2 pigeonpea seed system in India: An analysis in, and one outside of the project area for data collection in April 2011 for two weeks for comparison. Large group farmer interviews were conducted in each village followed up by individual interviewing of key informants including village sarpanchs (village president)] by using guide questionnaires that included the followings key points. 1. What were the past and current seed systems for the village? 2. Do farmers receive any training, and if so, by whom? 3. What is the total economic benefit of pigeonpea cultivation? 4. Are communities functioning on the seed village concept? 5. What are FPV’s for pigeonpea? 6. How are farmers selected as project beneficiaries? 335 food and non-food crops wherein 35,000 hectares is devoted to pigeonpea. The Tandur area that lies approximately 160 km west of Hyderabad is famous for pigeonpea cultivation (with a total area of 10,000 hectares) and processing of blue slate tiling used for home construction and concrete production. Two of the three villages studied were in the Tandur area (Kolkat and Gopalpur) for TL 2 project site and one in nonproject site at Godamguda for establishing comparative values between project and non-project areas. All three villages cultivated pigeonpea as roughly half of their total land area, either as sole or intercropping with sorghum, black gram, or green gram, depending on seasonal rains for the year. Yield of pigeonpea varied substantially with new varieties as compared to the traditional varieties (Table 1). The average land area owned was 0.8-1.6 hectares classifying the majority of farmers in these villages as smallholders (Table 2). Pigeonpea was primarily used as a staple food, and all farmers surveyed stated that they would first save food (preferred) stock and seed for the year and then sell the surplus stock in market. This assessment tried to ascertain what seed types were currently being used in the Tandur area of Ranga Reddy district, and also to understand seed saving systems in the past, both before seeds available in market as well as before implemented TL 2 intervention. The researcher mapped out ANGRAU’s recommendations for seed system change. Training materials and planned programs were also evaluated. Cu rrent levels of true adopti on o f interventi on recommendations was assessed and analyzed for gaps, indicating areas of knowledge not covered and specific populations not reached. Pigeonpea availability was also assessed for the area in order to understand the usage of crop at harvests both as a food and as a cash crop. Based on this baseline data, information was gathered from the community regarding potential improvements that could be made in implementation frameworks for the recommended food system. RESULTS AND DISCUSSION Existing Seed System Model: Farmers all over India have traditionally relied on saved seed as their primary mode to seed access. For pigeonpea specifically, farmers in the past depended on cultivation of four local varieties and would trade seed amongst themselves or between the villages when their seed became unviable after 3-4 years of successive cultivation (Fig. 1). The trade between the farmers and then with neighboring villages helped to give new exposure to existing variety in the village. When a new seed was cultivated then new genetic material obviously entered the cycle due to natural outcrossing which strengthened the all rounded varietal performance. All three villages surveyed noted that seed saving was still their primary method of retaining access to variety and that they only went for new seed every 2-3 years when the The Project Site (Tandur Village, Rangareddy District) : In Ranga Reddy District, 30.7% of land is cultivated for both Table 1. Agro-demographic survey across study villages Village Name Kolkat (4 years in project) Gopalpur (2 years in project) Godamguda (non-project) Soil Type Cropping Pattern Red-Brown soil (Alfisol) Red-Brown soil (Alfisol) Heavy black (Vertisol) Row cropping with sole and mixed pigeonpea cultivation either with sorghum, black gram, green gram Yield (New Varieties) (kg/ha) 800-1000 Yield (Local Varieties) (kg/ha) 500-700 Potential Difference in Yield (kg/ha) 100-500 800-1000 500-700 100-500 N/A 600-800 N/A Table 2. Percentage of farmers’ agricultural land use across sample villages Village Kolkat Gopalpur Godamguda (Non-project) Total area (ha) Farmers Interviewed (no.) 1,862 486 19 24 < 2 ha (Smallholder Farmers) 6 (32%) 17 (70%) 445 15 12 (80%) % distribution of landholdings 2.5-3.5 ha 4-8 ha > 8 ha 4 (21%) 2 (8%) 5 (26%) 1 (4%) 4 (21%) 4 (16%) 3 (20%) 0 0 336 Journal of Food Legumes 25(4), 2012 Fig 1. A flow chart of Tandur seed system model variety of their saved seed had digressed to the point of being uncultivable. Additionally, interviewed farmers reported that they preferred complete self-sufficiency and almost never got new variety from outside the village. It was primarily the larger landholders who purchased seed from outside the village. Normally the seed system cycle operates with a certified seed production agency (corporate seed company, ICRISAT or ANGRAU-ARS) producing breeder and foundation seed. The Department of Agriculture or local seed traders buy it and then sell it to large farmers (with > 10 ha) who can afford to purchase new seed although they constitute 10% of the overall farming population. Neighboring smallholder (90% of farming population) who see a decline in seed yield of their saved seed from previous years trade go for the second generation of new seed cultivated by above large farmers by replacing their seed (food) at the rate of 2 kg for 1 kg improved seed. They trade with the farmers who have a high yield in the previous season (notably the large farmers) who could afford to purchase first generation breeder or foundation seed. In the surveyed project area, there were some additional players in the seed system. ICRISAT provides breeder seed to ANGRAU-ARS for seed producti on, and farmer multiplication to provide additional local source for seed (Fig. 1) to farmers, a very helpful option as seed traders’ prices are high and the Department of Agriculture frequently has a shortage of seed. The ARS has also developed a few seed producers who grow truthfully labeled seed. Production of Improved varieties: During 2009-2010 per hectare average pigeonpea yield was 510 kg for Andhra Pradesh which was lower than that of states viz., Maharashtra, Karnataka, and Uttar Pradesh (Gopal and Babu, 2010). The yield level was more closely related to average yields for local varieties although new (improved) varieties could potentially create a seed yield differentiation of 100-500 kg/season (Table 1). Additionally, the average area cultivated for pigeonpea was 1.8 ha i.e.,the average pigeonpea plot is categorized as smallholder farming (Gopal and Babu, 2010). Proposed Seed Village System An alternative to creating isolation through distance is to encourage the majority of neighbour farmers to cultivate the same variety of seed, thus Holmesheoran et al. : Tropical Legumes 2 pigeonpea seed system in India: An analysis 337 eliminating the danger of outcrossing with other varieties. If an entire village or a large section of a village can be motivated through extension education and community organization to plant the same variety, yields will be maintained from season to season and the number of year’s seed can be repetitively saved for re-cu ltivatio n wi thou t lo ss o f desirable characteristics. Intensive community organization is needed to reap these highly desirable benefits. Additionally, the development of cooperation between informal and formal seed systems will help to maintain a system that allows the longterm benefits of improved varieties to be gleaned by smallholder farmers for longer periods of time (Nagarajan et al, 2007). Seeds can then be sourced according to the community’s preference from a variety of suppliers, and any seed traded within the community will be of the same variety, which will remain unsullied by outcrossing (Diagram 2). absence of these markers makes any assessment very challenging. Assessing the impact: At the inception stage, a clear picture must be formed about the project targets and changes hoped for in the targeted population. Based on these changes, the conceptualizing team should build a set of markers which can be easily evaluated throughout the duration of the project at regular intervals in order to keep the project on track. The Successes in approach and methodology: The Tandur ANGRAU-ARS engaged in some extension education and meeting of farmers in villages for the first time during the rollout of the TL 2 project. This exposure was invaluable for the fu ture as they develo p bo th a more co mplete understanding of farmers’ needs as well as the techniques Fig 2. A flow chart of the proposed seed village system Thus, regular evaluation and monitoring should take place among all partnering organizations, with optional outside assessment as well to ensure that project goals are being met and that adjustments in targets and approach can be made at the appropriate times. Tesfaye et al. (2005) suggested that in assessing the information pathways through which transfer of innovation occurs, source of introduction, frequency of visits by extension agents, availability of and membership in local agricultural institution’s, and presence of local leaders who will advocate for the innovation can all be used as indicators of community acceptance. The crucial ingredient in all these pathways is the presence of an individual or set of individuals who can spend the time to identify and build local capacities. 338 Journal of Food Legumes 25(4), 2012 needed to transfer innovative technology to farmers. One Farmers’ Day on the topic of pigeonpea was held at the ARS in 2009 with more than 2000 farmers participating giving the community an opportunity for more exposure to new techniques and seed varieties. The ARS had also distributed free seeds of 4-5 kgs/pack to 783 farmers, infusing the villages with new germplasm that would give high yields for the first season, and might add new genetic material into the local germplasm base. Gaps in approach and methodology 1. Improved understanding of seed village concept and farmer perspectives and needs by project staff is necessary. Staff should receive training to sensitize them to the situation and needs of the population they are meant to work with as well as human research and development skills. Many times, unfortunately, people are not usually conscious of their perceptions, beliefs, attitudes, and behaviour and are generally unaware of ho w these determine and influence their participation in social and economic activities and the benefit they derive. This lack of consciousness has important implications and serious consequences for the outcomes and impacts of development projects (Ellis 1997). 2. More clearly developed targeting criteria will help resources flow towards intended beneficiaries and achieve intended goals. There is currently a lack of a clear rubric for selecting sites, the amount of time to be spent in farmer visits, and follow-up methods to ensure that full education has been given. A rigorous procession system should be developed to make sure that each beneficiary moves through a preprescribed set of steps to ensure maximization of benefit. Free seed is currently given to farmers who are referred to as ‘progressive’, defined by ARS staff as those who own large areas of land and are deemed to be cooperative by the Department of Agriculture. It is important to remember that progressiveness encompasses much more than the size of land area, and also those smallholders, who may just as well be willing to experiment with a new practice, is less likely to have contact with the Department of Agriculture as they more rarely seek out seed from outside their village (Ellis 1997). 3. Lack in needed exclusive focus through spreading of staff to both jobs and projects. Farmer education in such a large spread of villages is a specialization that must be given full time and concentration. Age appropriate non-traditional educational styles required for positive communication with farmers require a certain level of expertise in human interaction, and demand the full attention of the staff person assigned to them. 4. More precise documentation should be provided at every step. A complete database with detailed information about each beneficiary farmer, their cropping patterns, and input and return costs for the year should be maintained in order to assess whether the target of poverty reduction in smallholders is being reached. According to Weinberger and Lumpkin (2007), poverty alleviation in an agricultural setting is based on the combination of both market prices and input costs of the crop cultivated. A fully populated data set will enable TL 2 staff to accurately express successes in terms of this relationship. 5. Seeds are sometimes given for free with no training for usage and no follow-up. For a farmer to receive such high quality seed for free, demands institutional stewardship of the opportunity they are giving along with the seed. Farmers must understand completely the value of the seed they are given and the intended functionality of that seed. The TL 2 project has made some progress towards its project goal of improving smallholder farmer access to improved seed. The project enabled ARS Tandur to organize Farmers Participatory Varietal selection experiments in farmers’ fields which are unique and first of its kind from this centre. The training programs conducted, field days organized and literature in local vernacular language distributed to the farmers in target areas has benefited the farming community and lead to progress towards the targeted goals of the TL 2 project in a great way. While not all smallholders are capable of taking the types of risk needed to try new seed varieties, and more effort and time spent in extension education can help at least some smallholders get access to varieties that have already been proven to perform well. Smallholders cannot be expected to be involved in experiments on germplasm or in seed production because they live much closer to a subsistence level. In spite of this, the already developed high quality seed for pigeonpea should be made available to them for immediate economic relief. Additionally, projects have shown the economic benefit of improving distribution and marketing capacities for small farmers in tandem with the provision of seed that will ensure higher yields (Jones et al. 2002). Continu ing to focus on increasi ng o pportuni ty for smallholders through seed system improvement at all levels will yield more innovative methods for cultivating community involvement and improving accessibility. REFERENCES Dewey KG. 1981. Nutritional consequences of the transformation from subsistence to commercial agriculture in Tabasco, Mexico. Human Ecology 9:151-187. Ellis P. 1997. Gender sensitive participatory impact assessment: Useful lessons from the Caribbean. Knowledge and Policy: The International Journal of Knowledge Transfer and Utilization 10: 71-82. Gilespie S and Kadiyala S. 2011. Exploring the agriculture-nutrition disconnect in India. IFPRI. Gopal B and Babu KK. 2010. Agricultural Statistics at a Glance: Andhra Pradesh. Directorate of Economics and Statistics, Government of Andhra Pradesh, Hyderabad. Jones R, Freeman HA and Lo Monaco G. 2002. Improving the access of small farmers in eastern and southern Africa to global pigeonpea markets. Agricultural Research & Extension Network, Network Holmesheoran et al. : Tropical Legumes 2 pigeonpea seed system in India: An analysis Paper No. 20, January 2002. Nadarajan N and Chaturvedi SK. 2010. Genetic options for enhancing productivity of major pulses- retrospect, issues, and strategies. Journal of Food Legumes 23: 1-8. Nagarajan L, Audi P, Jones R, and Smale M. 2007. Seed provision and dryland crops in the semi-arid regions of eastern Kenya. IFPRI Discussion Paper 00738. Nagarajan L, Smale M and Glewwe P. 2007. Determinants of Millet Diversity in the Household-Farm and Village-Community Levels in the Drylands of India: The Role of Local Seed Systems. Agricultural Economics 36: 157-167. Ravinder Reddy Ch, Tonapi VA, Bezkorowajnyj PG, Navi SS and Seetharama N. 2007. Seed System Innovations in the semi-arid Tropics of Andhra Pradesh, International Livestock Research Institute (ILRI), ICRISAT, Patancheru, Andhra Pradesh, India. Pp 339 224. Singh A and Singh AK. 2009. Yield advantage in pulses at farmers’ fields. Journal of Food Legumes 22: 198-201. Singh RB, Kumar P and Woodhead T. 2002. Smallholder Farmers in India: Food Security and Agricultural Policy. FAO Regional Office for Asia and the Pacific, Bangkok, Thailand. Swaminathan MS. 2006. Restoring farmers’ faith in farming. Indian Journal of Pulses 19: 1-6. Tesfaye A, Jemal I, Ferede S and Curran MM. 2005. Technology Transfer Pathways and Livelihood Impact Indicators in Central Ethiopia. Tropical Animal Health and Production 37: 101-122. Weinberger K and Lumpkin T. 2007. Diversification into Horticulture and Poverty Reduction: A Research Agenda. World Development 35: 1464-1480. Journal of Food Legumes 25(4): 340-343, 2012 Dissemination of pulse production technologies in Uttar Pradesh : A micro-level analysis RAJESH KUMAR, S.K. SINGH, PURUSHOTTAM and UMA SAH Division of Social Science,Indian Institute Pulses Research, Kanpur-208024 (U.P.), India; E-mail: rkm_13t2yahoo.com (Received: December 13, 2011; Accepted: December 11, 2012) ABSTRACT A survey was conducted in Lakhimpur Kheri and Bahraich districts of Uttar Pradesh, India during 2010 on 100 farmers to ascertain the methods used for dissemination of pulse production technologies (PPT) by extension worker, mode of dissemination, adoption of these improved technologies and constraints perceived in pulse production. About 24% farmers informed that excursion trip was conducted by extension workers for dissemination of PPT followed by group discussion and meeting. Most of these technologies were disseminated through progressive farmers in both the districts. Study also indicated that growing of pulses was profitable, environment friendly and also useful in way of improving the soil health although adoption of PPT was found low in both districts. The first and foremost constraint faced by these farmers was nonavailability of quality seed. Key words: Constraint analysis, Pulse production technologies, Technology adoption, Technology dissemination process Pulses are the group of food legumes which have the unique built-in ability of fixing atmospheric nitrogen through symbiosis and are beneficial for enriching soil fertility. Besides, they also make a significant contribution to human and animal nutrition through protein supplements. On account of these qualities, there will be no exaggeration if these crops may be referred as “unique jewels” of Indian crop husbandry. In spite of this importance, their acreage (20.2 to 25.5 m ha), production (14 o 18 m t) and productivity (632 kg/ha) remained nearly unchanged during the last several years. On the other hand, the population has increased tremendously which has resulted in declining intake of pulses from 75 g/capita/day during 1960 to a mere 26 g/capita/day during 2010. The distribution of acreage under pulses in different states/regions is also quite variable. Among 19 divisions of the state of Uttar Pradesh, Jhansi division contributes maximum to its production and acreage (>50%) and is followed by Allahabad and Faizabad divisions. The area, production and average productivity of pulses in U.P. state are 2.45 m ha, 2.43 m tonnes and 991 kg/ha, respectively. The farmers are mainly cultivating pigeonpea, urdbean, chickpea, lentil and pea in UP. However, pulses are cultivated on limited scale in irrigated areas with less external input application. Various constraints of pulse production are vagaries of weather, insufficient irrigation, insufficient use of N and P fertilizers, availability of rhizobium culture, proper management of pests and diseases and use of poor quality (both yield and pests tol erance) seeds. Technolo gy generati on, t echnol ogy assessment and refinement, technology transfer/dissemination and technology utilization process also contribute for reducing the time lags between generation of pulses production technologies (PPT) and their adoption on large scale by farming communities. The PPT has been disseminated through transfer of technology by KVKs, main extension agency (Department of Agriculture), NGOs and private organization etc. Thus, to analyze PPT disseminated in Uttar Pradesh, the current studies was carried out. MATERIALS AND METHODS The study was conducted in selected two districts (Lakhimpur Kheri and Bahraich) of Uttar Pradesh, India. From each district one block was identified; and from each identified block, 50 farmers were selected using simple random sampling technique. The list of pulses growers was collected from Village Pradhan. The data was collected from farmers through personal interview by pre-tested structured schedule and the total sample size was 100 farmers. The collected data was analyzed by using suitable statistical tools for arriving at valid conclusion. Studies were made with reference to socioeconomic profile of pulse growers, methodology used for dissemination and mode of dissemination of pulse production technologies, opinion of the farmers about the pulse production, adoptions behavior and constraints perceived in pulses. The variables used in this study included age, education, family size, type of family, land holding, land ownership, crops grown, social participation, extension contact and mass media exposure. RESULTS AND DISCUSSION Socio Economic Profile: The surveycarried out in Lakhimpur Kheri district (Table 1) revealed that 60 per cent farmers belonged to middle age category while 42 % of them were educated up to standard 5 (primary school). Two per cent of them were graduate who were engaged in farming while majority of them were having medium family size (50%) and joint family system (70%). Further, it was observed that 64% farmers had medium size land holding while 18% of them had leased in their land for expansion of their farming activity and Kumar et al.: Dissemination of pulse production technologies in Uttar Pradesh : A micro-level analysis only 4% of them had leased out their land to others to optimize the agricultural activities. In case of Bahraich, 44% farmers belonged to middle age category while 52% of them were educated up to standard 5. A majority of them (60%) had medium size family and joint family (82%). It was also observed that 48% farmers belonged to medium land holding category with cent per cent ownership of land. All the farmers were cultivating wheat, rice, lentil, pigeonpea, chickpea, urdbean and mungbean. However, the farmers had low social participation, extension contacts and mass media exposure (Table 1). Methods of Dissemination: It was opined by 10% farmers that extension workers used the individual approach for dissemination of improved PPT in Lakhimpur Kheri district, whereas 16% farmers agreed that extension workers did visit 341 their field and home to provide the knowledge about PPT (Table 2). Extension workers were not making contact through individual mode either through letter, telephone or mobile except in Bahraich where only 4% used these individual modes. This could be attributed to inadequate representation of extension workers per village and unavailability of modern communication links (mobile or telephone facility) with the farmers themselves. It was expressed by 24% farmers that excursion trip by the extension workers was the major mode for dissemination of PPT followed by group discussion and meeting in Lakhimpur Kheri (Table 2). Similarly, 20 % farmers in Bahraich district informed that extension workers were either using group approach or meeting/group training. Participatory approach was almost negligible in both these districts. Mode of dissemination: It was indicated that most of Table 1. Socioeconomic profile of farmers of Lakhimpur Kheri and Bahraich districts Variables Category Age (year) Young up to 30 Middle (31-49) Old (50 & above) Illiterate Up to class 5 Up to class 10 Up to class 12 Graduation & above Small (upto 5) Medium (5-10) Large (10 & above) Nuclear Joint Small (2) Medium (3-5) Large (5 & above) Owned Leased in Leased out Wheat Rice Arhar Urdbean Mungbean Chickpea Lentil Field pea Sugarcane Maize Ground nut Mustard Low Medium High Low Medium High Low Medium High Education Family size Type of family Land holding (in acres) Land ownership Crops grown Social participation Extension contact Mass media exposure Lakhimpur (n = 50) Frequency % 12 24 20 40 18 36 11 22 21 42 09 18 05 10 02 04 05 10 25 50 20 40 15 30 35 70 10 20 32 64 08 16 50 100 09 18 02 04 50 100 50 100 05 10 15 30 01 02 05 10 35 70 01 02 13 26 00 0 10 20 18 36 26 52 12 24 12 24 35 70 13 26 02 04 25 50 20 40 05 10 Bahraich (n = 50) Frequency % 14 28 22 44 14 28 15 30 26 52 08 16 01 02 04 08 08 16 30 60 12 24 09 18 41 82 08 16 24 48 18 36 50 100 04 08 00 00 50 100 50 100 20 40 07 14 05 10 13 28 50 100 02 04 17 34 30 60 00 0 20 40 20 40 18 36 12 24 30 60 15 30 05 10 30 60 14 28 06 12 Pooled(N=100) Frequency % 26 26 42 42 32 32 26 26 47 47 17 17 6 6 6 6 13 13 55 55 32 32 24 24 76 76 18 18 56 56 26 26 100 100 13 13 2 2 100 100 100 100 25 25 22 22 6 6 18 18 85 85 3 3 30 30 30 30 10 10 38 38 46 46 30 30 24 24 65 65 28 28 7 7 55 55 34 34 11 11 342 Journal of Food Legumes 25(4), 2012 Table 2. Methods of Dissemination of pulse production technology Variables Lakhimpur (n = 50) Frequency % Individual approach Farm & home visit Mobile Group approach Meeting Group discussion Cooperative society Group training Excursion trip Participatory approach Passive participation Active participation Interactive participation Bahraich (n = 50) Frequency % 05 0 10 0 08 02 16 04 06 10 32 04 12 12 20 64 8 24 10 12 28 04 10 20 24 56 8 20 04 01 01 8 2 1 05 02 01 10 4 2 technologies were disseminated through progressive farmers in both the districts as revealed by the extension functionaries and was followed by direct contact and Village Pradhan (Table 3). Table 3. Mode of dissemination of technologies Variables Direct Village Pradhan Progressive farmers Lakhimpur (n = 50) Frequency % 16 32.0 8 16.0 32 64.0 Bahraich (n = 50) Frequency % 30 60.0 10 20.0 35 70.0 Table 4. Opinion of farmers about pulse production technology Variables Lakhimpur (n = 50) Bahraich (n = 50) Frequency % Frequency % Suitability for growing of pulses 50 100 50 100 Profitability in pulse production 30 60 35 70 Environment friendliness 12 24 10 20 Soil health improvement 05 10 05 10 Enterprise possibility 0 0 02 20 Employment generation 0 0 01 2 Table 5. Adoption of improved pulse production technologies Practices Seed Fertilizer Bold seeded Small seeded Improved seed Local seed DAP Urea Rhizobium Trichoderma Implement Local Improved Processing Local Improved Postharvest Local technology Improved Soil testing Lakhimpur Bahraich (n = 50) (n = 50) Frequency Percent Frequency Percent 0 0 0 50 100 50 100 05 10 03 6 45 12 0 0 0 50 0 50 0 50 90 24 0 0 0 100 0 100 0 100 47 08 0 0 1 50 0 50 0 50 94 16 0 0 0 100 0 100 0 100 0 0 0 0 0 0 0 0 Opinion of farmers about PPT: There was consensus on the fact that pulses were suitable and profitable for farming. It was also revealed from the study that these were both environment friendly and were useful in improving soil health (Table 4). Adoption of improved PPT: The study indicated that only 10% farmer used improved seed and the rest used local seed for farming (Table 5). Farmers preferred small seeded lentil in both districts. In Bahraich, only 6% farmers used improved seed due to non availability of quality seed in time. Most of the farmers used DAP in pulses at the time of sowing, while use of rhizobium and trichoderma was absent. All the farmers were only using local implements and pest management schedules. Farmers were not able to test their soil due to lack of knowledge about laboratory/facility. These findings were supported by result of Khatiwada (1986). Table 6. Constraints perceived by the pulse growers Constraints Seed management Lack of quality seed Lack of knowledge about quality seed Non availability of seed timely Lack of quality seed of lentil Fertilizer and manures Lack of knowledge about use of Rhizobium Lack of knowledge about use of Trichoderma Nonavailability of Rhizobium of different pulse crops Nonavailability of fertilizer High cost of fertilizer No visible effect of fertilizers No timely & sufficient fertilizer supply by Govt.bodies Nutrient Management Deficiency of iron in soil Plant protection management Problem of wilt Problem of yellow mosaic disease Problem of pod borer Non availability of fungicide/ insecticide Poor effect of fungicides on crop Weed management Problem of weed management Marketing Bold seeded lentil has no demand in market Bold seeded lentil has no preference for consumption No knowledge about support price Forced sale of lentil at lower rates Other management Problem of blue – bull Problem of wild animals Natural calamity- draught, flood , hailstone Bahraich Lakhimpur (n = 50) (n = 50) Frequency % Frequency % 42 32 45 30 84 64 90 60 44 36 48 45 88 72 96 90 41 82 40 80 50 100 49 98 50 100 50 100 30 16 12 16 60 32 24 32 32 20 10 20 64 40 10 40 8 8 42 82 50 30 20 22 100 60 40 44 50 30 10 35 100 60 20 44 10 20 9 18 5 10 5 10 8 21 42 16 42 84 45 90 11 22 10 2 20 4 3 4 6 8 5 0 0 10 0 0 50 8 20 100 16 40 Kumar et al.: Dissemination of pulse production technologies in Uttar Pradesh : A micro-level analysis Constraints analysis: The study revealed that non-availability of seed followed by lack of quality seed of lentil in Lakhimpur Kheri and Bahraich were the major constraints for adoption of PPT (Table 6). In nutrient management, nonavailability of Rhizobium culture of different pulse crops followed by lack of knowledge about use of trichoderma and Rhizobium were the major constraints. Problem of wilt was again the common most problem for all the farmers of both the districts followed by yellow mosaic diseases, non availability of pesticides. Some farmers reported about the problem of weed management in pulses. These finding were supported by Masood et al. (2008) and McWilliam and Dillon (1986). Marketing is also one of the major problems and was associated with less preference for bold seeded lentil. There were also some common problems including the blue bull menace in both the districts and problem of wild animals in Bahraich. It could be inferred from the above that progressive farmers played the key role in dissemination of pulse 343 production technologies. Adoption of improved pulse technologies was poor due to non-availability of quality seed in time. There is also need to provide the inputs/trainings in time about successful management of constraints in adoption of these improved technologies. REFERENCES Masood Ali, Singh SK and Singh BB. 2008. Half Yearly Report of ISOPOM Project Development and Popularization of Model Seed System for Quality Seed Production of Major Legumes to ensure Seed Sufficiency at Village Level, Indian Institute of Pulses Research, Kanpur, U. P. Khatiwada MK 1986. The production of food legumes in the Himalayan range.In: Proceedings of the workshop on Food Legume Improvement for Asian farming system, Khon Khaen, Thaialand. McWilliam JR and Dillon JL. 1986. Improvement of food legumes: progress and constraints. In: Proceedings of the workshop on Food Legume Improvement for Asian farming systems, Khon Khaen, Thaialand. Journal of Food Legumes 25(4): 344-347, 2012 Pigeonpea (Cajanus cajan L.) price movement across major markets of India D.J. CHAUDHARI and A.S. TINGRE Department of Agricultural Economics and Statistics, Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola- 444 104, India; E-mail: [email protected] (Received: March 03, 2012;Accepted: November 03, 2012) ABSTRACT Pigeonpea is one of the major pulse crops grown across India. The last decade witnessed large fluctuations in prices of pigeonpea. The present study aimed to study price movement of pigeonpea, i.e., seasonal variation, price volatility and cointegration among the major pigeonpea markets in India. Data related to monthly average prices of pigeonpea were collected from major markets for the period 2003-2011across different States, viz., Akola and Latur (Maharashtra), Alwar (Rajasthan), Sedam (Karnataka) and Thandur (Andhra Pradesh). Moving average method was used to study seasonal variation. The econometric tools like ADF test, Johansen’s multiple cointegration test, Granger causality test and ARCH-GARCH model were used to analyse integration of markets across locations. The results of the study showed that the prices of pigeonpea were higher in the months from June to August in all selected markets. All the price series were stationary at first difference. The selected markets showed long run equilibrium relationship and co-integration between them. Most of the markets showed bidirectional influence on pigeonpea prices of each other. Alwar, Sedam and Latur markets were influenced by their own lag, while Akola, Latur and Thandur markets showed high price volatility. Key words: ADF test, ARCH- GARCH, Granger causality test, Co-integration, Pigeonpea, Price movement, Seasonal variation Pigeonpea (Cajanus cajan L.) is an important pulse crop of India and is grown in the tropical and subtropical regions of the world. Being rich in protein and relatively cheaper on price, a large section of vegetarian population of the country consumes it as ‘Dal’ in cooked form. The price of pigeonpea showed instability and fluctuations in the last decade. Pigeonpea displayed price volatility depending on monsoon, production in various regions of the country, import and the stock available with the traders. The prices of pigeonpea showed seasonal variations. Most of the pigeonpea markets in India are co-integrated that affected on the prices of each other. The instability in prices of agricultural commodities like, pigeonpea was influenced by a number of factors such as variation in production, low price elasticity of demand and seasonality in terms of price level, volatility and co-integration among the prices of different markets etc. These were the most important factors in determining competitiveness of the commodity and to formulate long term trade strategy. Thus, analyzing the past trend in the price of commodities was also useful in understanding the present scenario and to formulate appropriate strategies to improve the existing marketing system. Therefore, the present study was undertaken with the objectives of studying both seasonal variations in prices of pigeonpea and to assess the price volatility and cointegration among the major pigeonpea markets in India. MATERIALS AND METHODS The study was conducted in major pigeonpea markets fro m di fferent states sel ected vi z., Akola and Lat ur (Maharashtra), Alwar (Rajastan), Sedam (Karnataka) and Thandur (Andhra Pradesh). As per the records available the time series data on monthly average prices of pigeonpea for the period from 2003 to 2011 were collected from Agricultural Produce Market Committee of respective market. The method of moving average is the most widely used method of measuring seasonal fluctuations and the seasonal indices were obtained from the following steps: 1. Twelve month centered moving average value for arrivals and prices for the markets were obtained. 2. The original value as a percentage of centered moving average values for all months were expressed except for first six month and six month at the end. 3. These percentages were arranged according to the years and month. Primarily seasonal indices were obtained on eliminating the irregular component by averaging these percentages for each month. The average was taken over different year. The econometric tools like ADF test, Johansen’s multiple co-integration test, Granger Causality Test and ARCH-GARCH model were used to analyze integration of markets across locations. In addition, various parameters were estimated by using E-views-7 software. Augmented Dickey-Fuller test (ADF): Before analyzing any time series data testing for stationarity is pre-requisite. At first, the test for stationarity of time series data on pigeonpea prices was conducted. An Augmented Dickey-Fuller test (ADF) is the test for the unit root in a time series sample. A stationary series is one whose parameters are independent of time, exhibi ting constant mean and variance and havi ng autocorrelations that are invariant through time. If the series is found to be non-stationary, the first differences of the series are tested for stationarity. The number of times (d) a series is Chaudhari & Tingre: Pigeonpea (Cajanus cajan L.) price movement across major markets of India differenced to make it stationary is referred to as the order of integration i.e., I(d). ADF unit root test are based on the following three regression forms: Without constant and trend, Yt = Yt-1 + ut With constant, Yt = + Yt-1 + ut With constant and trend, Yt = + T + Yt-1 + ut The hypothesis is : H0: = 0 (unit root) H0: 0 t* > ADF critical value, then accept the null hypothesis, i.e. unit root exists. t* < ADF critical value, then reject the null hypothesis, i.e. unit root does not exists. Johansen’s multiple co-integration test: Johansen’s multiple co-integration test is employed to determine the long run relationship between the price series. The test shows whether the selected pigeonpea markets are integrated or not. Johansen (1988) has developed a multivariate system of equations approach, which allows for simultaneous adjustment of both or even more than two variables. The multivariate system of equations approach is more efficient than single equation approach as it allows estimating the co-integration vector with smaller variance. The second advantage of the multivariate approach is that in the simultaneous estimation it is not necessary to presuppose erogeneity of either of the variables. Granger causality test: In order to know the direction of causation between the markets Granger Causality test was employed. When a co-integration relationship is present for two variables, a Granger Causality Test (Granger 1969) can be used to analysis the direction of this co-movement relationship. Granger causality tests come in pairs, testing weather variable xt Granger-causes variable yt and vice versa. All permutations are possible viz., univariate Granger causality from xt to yt or from yt to xt , bivariate causality or absence of causality. The Granger causality test analyses weather the unrestricted equation, yt = 0 + Ti = 1 1i yt-i + Tj = 1 2i xt-j + t with 0 i , j T Yield better results than the restricted equation. yt = 0 + Ti =1 1i yt-i + t with Tj = 1 2j xt-j = 0 (The null hypothesis) i.e. if H0, in which 21= 22= ……= 2T = 0, is rejected then one can state “variable xt Granger causes variable yt ” ARCH-GARCH model: To access the presence of price vo lati lity, ARCH-GARCH analysi s was carried ou t. Autoregressive Conditional Heteroscedastcity (ARCH) models are specifically designed to model and forecast conditional variances. ARCH model was introduced by Engel (1982) and generalized as GARCH by Bollersllev (1986). The ARCH model have two distinct specifications, one for the 345 conditional vari ance and the standerd GARCH (1 ,1) specification is presented as under: Yt = 0 + 1 X1t +……..+ k Xkt + e 2 2 t-1 2 t-1 t = + e + …………………1 ………………..2 Equation (1) is the mean equation and equation (2) is the conditional variance equation. The ARCH component () indicate the lag of the squared residual from the mean equation and the GARCH term () the last period’s forecast variance and the resultant sum of these co-efficient ( + ) are presented. The sum of co-efficient very close to 1 indicates that the volatility shocks are quite persistent in the series. RESULTS AND DISCUSSION Seasonal variation: In major producing areas, the market arrivals of pigeonpea started in the month of December and lasted for four months. During the peak arrivals period, the prices are generally low. The prices were also recorded higher from June to August. Most of the traders released the stored stock of pigeonpea during this period in anticipation of making the profit. The seasonal indices of monthly average prices of pigeonpea in different markets of India were worked out to study seasonal variation (Table 1). It depicted that the prices of pigeonpea were higher in the months from June to August in Akola and Latur markets of Maharashtra and Alwar market of Rajasthan while from July to September in the markets of Southern India viz., Sedam (Karnataka) and Thandur (Andhra Pradesh). The prices were highest by 7.52, 6.18, 6.26 and 7.19% in the month of July in Akola, Latur, Alwar and Thandur market, respectively and by 10.11 per cent in Sedam market. It was observed that the prices were lowered by 5.2 and 3.6% in December in Akola and Alwar market, by 5.25 and 6.54% in January in Latur and Thandur market and by 7.13% in March in Sedam market. Thus, the largest arrivals in these months lowered down the prices of pigeonpea. Chaudhari and Pawar (2010) found that the price indices were higher in the month of July in the Osmanabad and Paranda markets. Table 1. Seasonal indices of monthly average prices of pigeonpea in different Indian markets Month January February March April May June July August September October November December Akola (MS) 96.21 99.24 97.87 102.39 98.62 101.66 107.52 103.95 99.83 99.68 98.25 94.80 Latur (MS) 94.75 96.44 98.87 100.05 96.62 103.18 106.18 103.49 101.84 101.68 97.60 99.29 Sedam (KA) 97.31 95.31 92.87 94.52 94.49 98.59 105.54 106.38 106.07 110.11 100.98 97.83 Alwar (RJ) 97.77 96.86 102.68 99.46 99.79 100.94 106.26 101.18 99.00 99.03 100.63 96.40 Thandur (AP) 93.46 96.73 98.32 98.86 96.88 97.35 107.19 102.67 102.49 103.70 99.42 102.92 346 Journal of Food Legumes 25(4), 2012 Aug ment ed Di ckey-Full er t est (ADF): The test for stationarity of time series data on pigeonpea prices was conducted. The results of ADF test (Table 2) indicated that prices in level with lag 1, the ADF values were less than the critical value at 1% level indicating the existence of unit root implied non stationary nature of price series in all markets. In first difference with lag 1, the ADF values were higher than critical values at 1% level indicated that the price series are free from the consequences of unit root. This implied that the price series were stationary at first difference level. Ghosh (2011) found the prices of rice and wheat were non-stationary in levels but stationary in first-differences which implied that all the series of rice and wheat prices contain a single unit root and were integrated of order one, I (1) for both the periods. Similarly Reddy and Reddy (2007) for groundnut and Reddy (2007) for rice also concluded that prices were stationary at first difference level. Table 2. ADF test results of pigeonpea prices Name of Market Akola Alwar Latur Sedam Thandur Level First difference -2.173171 -9.324023 -1.570010 -13.39667 -2.122809 -9.005828 -3.098783 -9.572111 -2.641566 -10.46072 Table 4. Results of pair wise Granger casuality test -4.057528 Table 3. Results of multiple co-integration analysis 0.47 0.35 0.21 0.06 0.02 Trace Critical statistics value (5%) 132.9 71.8 30.6 8.4 2.4 88.8 63.8 42.9 25.8 12.5 Price volatility: To assess the presence of price fluctuations in the prices of pigeonpea in Akola, Alwar, Latur, Sedam and Thandur market, ARCH-GARCH analysis was carried out (Table 5). Among the markets, the sum of alpha and beta were near to 1, viz., 0.97, 1.46 and 0.92 as for Akola, Latur and Thandur market respectively, which indicated the presence of price fluctuations in the pigeonpea prices in selected markets during the study period. Similar results were found by Sekhar (2003) for rice and wheat, and by Lavanya (2011) for turmeric. Critical value (1%) Johansen’s multiple co-integration test: To determine the long run relationship between the price series Johansen’s multiple co-integration test was employed to all the markets. The test showed whether the selected pigeonpea markets were integrated or not. Eigen value showed that there was a bidirectional influence on prices of Akola and Alwar, Akola and Thandur, Alwar and Latur, Thandur and Alwar and Sedam and Alwar market. The prices at Akola market also influenced Latur and sedam market. Thandur and Sedam market price were also affected by Latur market prices. It was observed that Sedam market price influenced Thandur market. Similar results were obtained by Ajjan et al. (2009) for Red chilli in Tamil Nadu Moe et al. (2008) for the pigeonpea and green gram in Myanmar. Hypothesized Number of co-integration equation None* At most 1* At most 2 At most 3 At most 4 Number of co-integration equation Two The results showed that at least two co-integration equations were significant at 5% level of significance (Table 3). Thus, the selected pigeonpea markets were having long run equilibrium relationship and there existed a co-integration between them. Mukim et al. (2009) found that the whole sale prices of wheat were co-integrated in the long run. Similar results were recorded by Gandhi and Koshy (2006) for wheat and by Ghosh (2011) for rice and wheat markets in India. Granger causality tests: Granger Causality test was employed to know the direction of causation between the markets. Theoretically, a variable is said to Granger-cause another variable, if the current value is conditional on the past value. The results of Pairewise Granger Causality test (Table 4) Null hypothesis Observed F-statistic Probability LTR does not Granger Cause AKL 96 0.10222 0.9029 AKL does not Granger Cause LTR 19.7226 8.E-08 TND does not Granger Cause AKL 96 2.84679 0.0632 AKL does not Granger Cause TND 25.6474 1.E-09 SDM does not Granger Cause AKL 96 0.00102 0.9990 AKL does not Granger Cause SDM 3.21939 0.0446 ALR does not Granger Cause AKL 96 5.89082 0.0039 AKL does not Granger Cause ALR 12.7987 1.E-05 TND does not Granger Cause LTR 96 0.73444 0.4826 LTR does not Granger Cause TND 12.8264 1.E-05 SDM does not Granger Cause LTR 96 0.80564 0.4500 LTR does not Granger Cause SDM 2.84841 0.0631 ALR does not Granger Cause LTR 96 11.7768 3.E-05 LTR does not Granger Cause ALR 18.7670 1.E-07 SDM does not Granger Cause TND 96 4.43699 0.0145 TND does not Granger Cause SDM 2.06703 0.1325 ALR does not Granger Cause TND 96 11.2190 4.E-05 TND does not Granger Cause ALR 20.6234 4.E-08 ALR does not Granger Cause SDM 96 5.76220 0.0044 SDM does not Granger Cause ALR 3.51057 0.0340 Note : Sample: 2003M04 2011M05 Series ; AKL, LTR, ALR, SDM and TND where, AKL : Akola market, LTR : Latur market, ALR: Alwar market, SDM: Sedam, TND: Thandur Table 5. Results of ARCH-GARCH analysis Parameter Alpha (α) Beta (β) Sum of α & β Akola Alwar Latur Sedam 0.982153 0.970484 0.979363 0.895793 -0.010690 -1.002528 0.487176 -0.129860 0.971463 -0.032044 1.466539 0.765933 Thandur 0.970944 -0.049272 0.921672 Policy implication: The study examined the price movement of pigeonpea across the main markets in major pigeonpea producing states of India. The prices of pigeonpea showed seasonal fluctuations and were recorded higher in the months from June to August in all selected markets. The results of ADF test showed that all the markets having the ADF values Chaudhari & Tingre: Pigeonpea (Cajanus cajan L.) price movement across major markets of India higher than the critical values at 1% level, and thus, the price series were stationary at first difference level. The analysis of multiple co-integration depicted that the selected markets having long run equilibrium relationship and their existed cointegration between them. Most of the markets showed the bidirectional influence on prices of each other. As the sum of Alpha and Beta worked out nearer to 1 for Akola, Latur and Thandur market, this indicated high price volatility in pigeonpea prices in these markets. The results for other legume crops like groundnut (Reddy and Reddy, 2011) also showed that the prices are co-integrated in the long run, however in short run adjustment to market disequilibrium took longer time. It is inferred from the study that it is important to invest in modernization of commodity markets, development of road connectivity and marketing related infrastructure like, warehouses etc. so as to adjust the demand and supply gap across regions within a short period of time. 347 Agriculture Update 5: 158- 162. Engle RF and Granger CWJ. 1987. Co-integration and Error Correction: Representation, Estimation and Testing. Econometrica 55: 251276. Gandhi Vasant P and Abraham Koshy 2006. Wheat Marketing and its Efficiency in India. Working Paper No. 2006-09-03 (Indian Institute of Management). Gosh Madhusudan 2011. Agricultural policy reforms and spatial integration of food grain markets in India. Journal of Economic Development 36: 15-36. Lavanya M. 2011. Price behaviour and marketing practices of turmeric in erode district of Tamilnadu. M.Sc. Thesis, TNAU, Coimbatore, India. Megha Mukim, Singh Karan and Kanakaraj A. 2009. Market integration, Transaction costs and the Indian wheat market: A systematic study. Economic & Political Weekly XLIV: 149-155. Moe AK, Yutaka T, Fukuda S and Kai S. 2008. Impact of market liberalization on international pulses trade of Myanmar and India. Journal of the Faculty of Agriculture, Kyushu University 53: 553561. REFERENCES Redddy AA. 2007. Commodity Market Integration: Case of Asian Rice Markets. The IUP Journal of Applied Economics, VI: 21-44. Ajjan N, Shivkumar KM, Murugananthi D. and Padmavathi P. 2009. Red Chillies. CARDS Commodity Series - 2 (TNAU, Coimbatore). Reddy AA and Reddy GP. 2011. Integration of Wholesale Prices of Groundnut Complex. Indian Journal of Agricultural Marketing 25: 89-108. Bollerslev T. 1986. Generalized Autoregressive Conditional Heteroscedasticity. Journal of Econometrics 31:307–27. Chaudhari DJ and Pawar ND. 2010. Growth, instability and price analysis of pigeon pea (Cajanus Cajan Lin.) in Marathwada region. Sekhar CSC. 2003. Volatility of agricultural prices–an analysis of major international and domestic markets. Working Paper No. 103 (Indian Council for Research on International Economic Relations). Journal of Food Legumes 25(4): 348-350, 2012 Short Communication Genetic variability, character association and path coefficient analyses in faba bean B.K. CHAUBEY, C.B. YADAV, K. KUMAR and R.K. SRIVASTAVA Department of Genetics & Plant Breeding, N. D. University of Agriculture and Technology Kumarganj, Faizabad224 229 (U.P.) India; E-mail: [email protected] (Received: March 31, 2012; Accepted: November 6, 2012) ABSTRACT Present investigation was carried out involving seventy diverse germplasm lines of Faba bean. Seed yield/plant showed highly significant and positive correlation with number of pods/plant, biological yield/plant, number of branches/plant, number of seeds/pod, 100-seed weight and harvest index. Path analysis identified biological yield/plant, harvest index, number of pods/ plant, 100-seed weight, number of seeds/pod and plant height as important components having high order direct effects, while number of pods/plant, number of branches/plant, plant height and number of seeds/pod via biological yield/plant showed maximum indirect effect on seed yield. Key words: Character association, Faba bean, Genetic variability, Path coefficient analysis. Faba bean (Vicia faba L.) is an important pulse crop of the world cultivated under both irrigated and rainfed conditions. It is grown as Rabi crop in diverse agro-ecological si tuat ions fro m hi lls to plai ns and even under po or management. In India, it is grown in a sizeable acreage in Bihar, Madhya Pradesh and some parts of Uttar Pradesh. Its green pods are used as vegetable and dry seeds are used as split dal and in the preparation of besan. It has great production potential which has not been realized so far. Accordingly, the present study was carried out to evaluate the available germplasms to work out the character association and, direct and indirect effects of different attributes with respect to yield. The experimental material for the present investigation consisted of 70 germplasm lines of Faba bean along with check varieties e.g.,’PRT-7’, ‘PRT-12’ and ‘Vikrant’, grown in Augmented Block Design with three checks repeated after every 10 lines of the test entries. The experiment was carried out at the Student’s Instructional Farm, Narendra Dev University of Agriculture and Technology (NDUAT), Faizabad (U.P.). Each accession was grown in double row of 4 m length with inter row spacing of 30 cm. All the recommended cultural practices were adopted to raise a good crop. The observations on plant height, number of branches/plant, number of pods/ plant, number of seeds/pod, 100-seed weight (g), biological yield/plant (g), seed yield/plant (g), harvest index (%), and protein content, were recorded on five randomly selected plants, while days to 50% flowering and days to maturity on plot basis. The mean data were subjected to analysis of variance following Federer (1956). Estimation of correlation coefficient was done following Searle (1961). Path coefficient analysis was done as suggested by Dewey and Lu (1959). Analysis of variance exhibited significant differences for all the accessions indicating presence of sufficient genetic variability among the accessions. Correlation analysis (Table 1) revealed that seed yield/plant was significantly and positively correlated with number of pods/ plant, biological yield/plant, number of branches/plant, number of seeds/pod, 100-seed weight and harvest index indicating that selection based on these characters may result in higher yield, which was in close agreement with earlier findings of Vandana and Dubey (1993), Abo-Elwafa and Bakheit (1999) and Patel and Acharya (2011). Biological yield/plant was highly significant and positively correlated with number of pods/plant, number of branches/plant, plant height and number of seeds/pod. Interestingly, there were significant correlations existing among the above characters as well as seed yield/plant which, suggested that these characters may be considered for improvement of seed yield. Furt her, based o n these relationships, it can be presumed that for improving yield in faba bean, a model plant type would be that with high biological yield, higher number of pods/plant, increased number of branches/plant and higher seeds/per pod. Harvest index also showed positive correlation with seed yield/plant while negatively correlated with plant height, biological yield/plant and days to 50% flowering. Association of these characters with grain yield/plant elucidates the importance of proper source to sink relationship. It would be rational to expect that a genotype which has smaller vegetative period as in present case i.e., early flowering will have greater ability to give more yield than a genotype with delayed flowering. Similarly a genotype which has more number of branches, more number of pods, more number of seeds, higher 100-seed weight and high biological yield is expected to fill the sink to larger extent (Pace et al., 1979; Huang, 1983 and Ramgiry and Bansal 1997). Path analyses showed highest positive direct effects on the grain yield through biological yield / plant followed by harvest index and number of pods per plant (Table 2). Considering both correlation and path coefficient, it is crystal clear that these characters are the most important for realizing maximum genetic gain through selection in faba bean. Chaubey et al.: Genetic variability, character association and path coefficient analyses in faba bean 349 Table 1. Estimates of simple correlation coefficients between different pairs of characters in faba bean Characters Days to 50% flowering Days to maturity Plant height Branches/ plant Pods/ plant Seeds/ pod 100seed weight Biological yield/ plant Seed yield/ plant Harvest index Protein content Days to 50% flowering (no.) 1.00 -0.046 0.068 -0.167 -0.130 0.062 0.007 -0.065 -0.066 -0.467** -0.090 1.00 0.217 -0.071 -0.096 0.004 -0.043 0.110 -0.030 -0.210 0.355** 1.00 0.150 0.167 0.120 -0.063 0.442** 0.192 -0.406** 0.066 1.00 0.655** 0.253* 0.226 0.607** 0.633** -0.026 -0.104 1.00 0.369** 0.208 0.663** 0.855** 0.141 -0.088 1.00 0.041 0.329** 0.429** 0.059 0.035 1.00 0.170 0.405** 0.286* 0.085 1.00 0.700** -0.441** 0.081 1.00 0.281** -0.017 1.00 -0.117 Days to maturity (no.) Plant height (cm) Branches/ plant (no.) Pods/plant (no.) Seeds/pod (no.) 100-seed weight (g) Biological yield/ plant (g) Seed yield/ plant (g) Harvest index (%) Protein content (%) 1.00 *, **: Significant at P=0.05 & 0.01, respectively Table 2. Direct and indirect effects of different characters on seed yield/plant in faba bean Characters Days to 50% flowering Days to maturity Plant height Branches/ plant Pods/plant Seeds/pod 100seed weight Biological yield/ plant Harvest index Protein content Correlation with seed yield Days to 50% flowering 0.0378 -0.0015 0.0032 -0.0029 -0.0322 0.0031 0.0004 -0.0471 -0.0268 0.0003 -0.0655 Days to maturity -0.0017 0.0314 0.0103 -0.0012 -0.0238 -0.0002 -.0028 0.0800 -0.1202 -0.0013 -0.0295 Plant height 0.0026 0.0068 0.0477 0.0026 0.0414 0.0059 -.0041 0.3219 -0.2329 -0.0002 0.1917 Branches/ plant -0.0063 -0.0022 0.0071 0.0173 0.1626 0.0125 0.0145 0.4421 -0.0149 0.0004 0.6331 Pods/plant -0.0049 -0.0030 0.0080 0.0113 0.2483 0.0182 0.0133 0.4828 0.0806 0.0003 0.8551 Seeds/pod 0.0023 -0.0001 0.0057 0.0044 0.0915 0.0495 0.0026 0.2394 0.0338 -0.0001 0.4290 100-seed weight 0.0003 -0.0013 -.0030 0.0039 0.0516 0.0020 0.0642 0.1238 0.1640 -0.0003 0.4051 Biological yield/ plant -0.0024 0.0035 0.0211 0.0105 0.1646 0.0163 0.0109 0.7284 -0.2526 -0.0003 0.6999 Harvest index -0.0018 -0.0066 -.0194 -0.0005 0.0349 0.0029 0.0184 -0.3210 0.5732 0.0004 0.2807 Protein content -0.0034 0.0112 0.0031 -0.0018 -0.0217 0.0017 0.0055 0.0593 -0.0670 -0.0037 -0.0169 Residual effect= 0.0529, Direct effect: diagonal (bold) 350 Journal of Food Legumes 25(4), 2012 Simultaneously, low value of direct effects recorded in positive direction for 100-seed weight, number of seeds/pod, plant height, days to 50% flowering, days to maturity and number of branches/plant indicating that direct contribution of these traits might lead to increase in grain yield if other variables remain constant, which was in conformity with earlier findings of Salem (1982) and Bora et al. (1998). The indirect contribution of biological yield/plant via number of pods/plant, 100-seed weight via harvest index and number of branches/plant via number of pods/plant also showed high order positive effect indicating that these characters are important contributors of grain yield. Biological yield/plant and plant height via harvest index had considerable negative indirect effect on seed yield. Some of earlier reports have also identified these characters as important indirect contributors in the expression of seed yield in faba bean (Reddy et al. 2002). Thus, harvest index and number of seeds/pod emerged as most important traits to be considered in the development of high yielding genotypes of faba bean. 518. Federer WT. 1956. Augmented design, “Hawain Planters” Record. 55: 191-208. Habetinek J, Ruzickova M and Soucek J.1982. Variability and correlations in some quantitative characters in a collection of broad bean varieties (Faba vulgaris Moench). Sbornik Vysoke, Skolly Zemadelske V Praze, A. 36:79-92. Huang WT, Li FQ, Jiang XY and Li HY.1983. Correlation and path coefficient analyses of characters in Vicia faba. Hereditas 5:21-23. Pace C-de and Bond DA, Scarascia-Mugnozza GT and Poulsen MH. 1979. Chracteristics with significant correlations to seed yield and path analysis in broad bean populations grown in southern Italy. In: Proceedings of seminar in the EEC programme of coordination of Research on Plant Proteins, held at Bari, Italy. Pp 144-167. Patel JB and Acharya S. 2011. Genetic divergence and character association in Indo-African derivatives of pigeonpea [Cajanus cajan (L.) Millsp.]. Journal of Food Legumes 24:198-201. Ramgiry SR and Bansal YK. 1997. Correlation and path coefficient studies for yield and nodule characters in broad bean (Vicia faba L.). Advances in Plant Sciences 10:207-211. REFERENCES Reddy SRR, Gupta SN, Verma PK and Rai L. 2002. Genetic variation, correlation and path coefficient analysis under normal sown conditions in Vicia faba L. Forage Research 28: 63-66. Abo-Elwafa A A and Bakheit BR.1999. Performance, correlation and path coefficient analysis in Faba bean. Asian Journal of Agricultural Sciences 30: 77-92. Salem SA. 1982. Variation and correlations among agronomic characters in a collection of beans (Vicia faba L.). Journal of Agricultural Sciences 99: 541-545. Bora GC, Gupta SN, Tomar YS and Singh S. 1998. Genetic variability, correlation and path analysis in Faba bean (Vicia faba). Indian Journal of Agricultural Sciences 48:212-214. Searle SR. 1961. Phenotypic, genotypic and environmental correlation. Biometrics 17:474-480. Dewey DR and Lu KH. 1959. Correlation and path coefficient analysis of crested wheat grass seed production. Agronomy Journal 51: 515- Vandana and Dubey DK. 1993. Path analysis in Faba bean. FABIS Newsletter 32:23-24. Journal of Food Legumes 25(4): 351-354, 2012 Short Communication A comparative study of hybrid and inbred cultivars for germination and other related traits of pigeonpea M. BHARATHI and K.B. SAXENA International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Patancheru - 502 324(Andhra Pradesh), India; E-mail: [email protected] (Received: August 10, 2012; Accepted: November 27, 2012) ABSTRACT Eleven pigeonpea hybrids and 4 inbred cultivars were evaluated to study seed germination and seedling growth parameters using ‘Between Paper Towel’ (BPT) method. The test was conducted over a period of 10 days at room temperature in a seed germinator. Data were rec orded on test we ight, germination, root length, shoot length, shoot to root ratio, seedling dry weight and seedling vigour index. Significant differences were observed among entries for all the traits studied. Significantly greater values for germination (99.5%), root length (11.6 cm), seedling vigour index (1832), seedling dry weight (4.7 g) and test weight (11.4 g) were recorded for hybrids compared to inbred lines. The presence of longer root in the hybrids resulted in producing lengthier roots and greater tolerance to drought during early growth stages. Thus, greater seedling vigour of the hybrids would also contribute to their higher plant vigour and seed yields. Key words: Germination, Hybrids, Inbred cultivars, Pigeonpea, Seedling vigour index Pigeonpea[Cajanus cajan (L.) Millsp.] is an important pulse crop predominantly grown in the tropics and subtropics for their high protein grains. It has been observed that despite increase in its area and production, the national mean yield of pigeonpea (774 kg/ha) has remained consistently low over the last five decades. To break this yield barrier, a hybrid breeding technology based on cytoplasmic nuclear malesterility (Saxena et al. 2005) and partial natural out-crossing systems was developed (Saxena et al. 2009) at ICRISAT. In comparison to other grain legumes, the growth rate of pigeonpea seedling is slow (Brakke and Gardner 1987) and this condition normally persists for about 30-40 days from sowing and, thereby, makes pigeonpea less competitive to weeds and cereal inter-crops. The cultivation of high yielding pigeonpea hybrids could contribute in overcoming the productivity constraints to some extent due to their better germination, establishment of uniform plant stand, faster seedling growth and a high yield potential. Therefore, the present study was undertaken to compare germination and related traits in pigeonpea hybrid and inbred cultivars. Eleven pigeonpea hybrids and 4 inbred cultivars were identified for this study. The experiment was conducted in 3 replications in a randomized complete block design (RCBD) with a sample size of 100 seeds. The germination test was conducted as per International Seed Testing Association (ISTA) rules using ‘Between Paper Towel’ (BPT) method (ISTA 200 7). The seeds were pre-t reat ed with tetramethylthiuramdisulphide @ 2.5 g/kg and placed in between layers of wet germination papers. The rolled paper towels were placed in a tray with 1-2 cm of distilled water for maintaining sufficient moisture. These trays were kept inside the Percival Scientific Seed Germinator (Model I-35-LLVL) for 10 days. Data on test weight (g) was recorded as per ISTA (2007). The data on germination (%), root length (cm) and shoot length (cm) were recorded on 10th day after incubation. Subsequently, the seedlings were dried inside craft paper bags at 60±2°C temperature for 48 h and the average dry weight (mg) of seedlings per sample was calculated. Seedling vigour indices were estimated using the following formula (AbdulBaki and Anderson 1973): I. Seedling Vigour Index = Germination % × (Total seedling length) Where, Total seedling length = Root length + Shoot length II. Seedling Vigour Index = Germination % × Seedling dry weight Statistical analyses of the data were performed using SAS version 9.2, Anon (2008). The analysis of variance revealed highly significant differences among the entries for all the characters studied. The partitioning of t reatment mean squ ares revealed significant differences among hybrids for germination per cent, seedling dry weight, shoot length, root length, shoot: root ratio, and test weight (Table 1). Among inbred cultivars, significant differences were observed for germination per cent, seedling dry weight, shoot length, and test weight. On average, the hybrids exhibited significantly higher germination (99.5±0.26%) over inbred cultivars (96.8±0.26%). Mercer et al. (2006) also reported that hybrids exhibited increased seed germination and decreased dormancy over their parents in 352 Journal of Food Legumes 25(4), 2012 Table 1. Statistical significance of treatments on various physiological parameters Source df Germination % Root length (cm) Shoot length (cm) Shoot: root ratio Seedling dry weight (mg) 1.38** 0.74** 0.18** 11.40** Test weight (g) 2.60** 1.91* 1.67 11.71** Seedling vigour index (I) 80054** 53912 31074 488419** Treatments Hybrids Inbred cultivars Hybrid vs. Inbred cultivars Error 14 10 3 1 0.069** 0.006 0.074* 0.684** 153.22** 85.05** 6.30 1277.24** 123.77** 143.90** 71.10** 70.86** 30 0.02 13.77 10.64 0.93 25477 0.03 0.17 5.68** 3.04** 2.21** 42.47** *, **: significant at P = 0.01 and 0.05, respectively Table 2. Variation for different traits observed among hybrids and inbred cultivars ten days after germination Genotype Hybrids ICPH 4438 ICPH 4430 ICPH 2441 ICPH 2363 ICPH 3310 ICPH 2671 ICPH 4022 ICPH 4013 ICPH 2740 ICPH 3762 ICPH 2364 Mean Inbred cultivars ICPL 88039 Asha UPAS 120 Maruti Mean CV (%) SEm (+) LSD (5%) Germination (%) Root length (cm) Shoot length (cm) Shoot: root length Seedling vigour index Seedling dry weight (g) Test weight (g) 99.3 99.0 100.0 100.0 99.7 100.0 99.7 98.7 99.3 99.3 100.0 99.5 12.38 12.04 12.34 11.61 12.31 11.64 11.07 11.85 10.39 11.41 10.16 11.56 8.62 8.47 6.59 7.06 6.36 5.46 7.44 6.23 6.45 6.14 6.12 6.81 0.73 0.81 0.58 0.65 0.54 0.49 0.71 0.61 0.67 0.89 0.65 0.67 2072 2006 1895 1866 1855 1840 1838 1758 1728 1663 1629 1832 5.6 5.2 4.1 4.5 4.5 5.2 4.0 4.8 4.1 4.8 4.6 4.7 11.0 12.5 12.5 11.5 11.5 10.0 10.0 11.5 13.0 10.5 11.5 11.4 98.3 97.7 94.8 96.2 96.8 1.6 0.13 0.26 9.63 9.87 10.09 9.65 9.81 33.4 0.44 0.86 7.95 7.48 7.24 6.24 7.23 47.2 0.38 0.75 0.91 0.95 0.75 0.73 0.84 135.3 0.11 0.22 1700 1655 1558 1472 1596 9.0 130.47 266.16 3.9 3.5 3.3 3.4 3.5 3.8 0.13 0.28 9.5 10.0 8.0 9.0 9.13 3.8 0.33 0.68 sunflower. Mean root length in the hybrids (11.56±0.86 cm) was significantly greater than that of inbred cultivars (9.81±0.86 cm) indicating expression of hybrid vigour for this trait (Table 2). In contrast, the shoots were longer in the inbred cultivars than that in hybrids. This resulted in low shoot: root ratio in the hybrids. According to Saxena et al. (1992), one month old seedlings of pigeonpea hybrids produced 44% more shoot mass and 43% more root mass as compared to the inbred cultivars. This data set indicated that the food reserves in the hybrid seed contri buted predominantly towards root development. The root and shoot or total seedling length is known to influence the vigour of seedling which can be used for comparing the genotypes by a parameter called ‘seedling vigour index’ that takes into account the germination per cent of the individual genotype, giving an overall understanding of their ability to express potential growth at the seedling stage itself. The present study also indicated that the hybrids were significantlyvigorous (with the index, 1832±266.2) than the inbred cultivars (1596±266.2). Among the hybrids, ‘ICPH 4438’ had the highest seedling vigour index (2072) and ‘ICPH 2364’ had the lowest one (1629). Seedling vigour index was also found to be highly correlated with germination per cent, root length, seedling dry weight and test weight suggesting the important role of these traits in the manifestation of vigour in hybrid seedlings (Table 3). In pigeonpea, variation for seed yield is primarily attributed to the differences in crop growth rates. Chauhan et al. (1995) reported that the yield in pigeonpea hybrid was greater than inbred cultivars; and was primarily associated with higher crop growth rates and the early vigour. They also reported that the differences in plant vigour between hybrids and inbred cultivars began to appear during early seedling stage which became more pronounced with time enabling them suitable for competitive situations. In the present set of materials, test weight was highest in hybrids (11.4±0.33 g) as compared to inbred cultivars (9.1±0.33 g). During the first 10 Bharathi & Saxena: A comparative study of hybrid and inbred cultivars for germination and other related traits of pigeonpea 353 Table 3. Correlation coefficients observed among different seedling traits Characters Seedling dry weight (mg) 0.667** Germination (%) Seedling dry weight Shoot length Root length Shoot: root ratio Test weight *, **: significant at P = 0.01 and 0.05, respectively Shoot length (cm) -0.153 0.091 days of germination, relatively more quantity of reserved carbohydrates was mobilized for the growth of root in the hybrids. On the contrary, in the inbred cultivars, more carbohydrates were translocated to the development of shoot. Narayanan et al. (1981) reported that the seed reserves rather than size of the shoot and root length were important factors in determining seedling size. It was also evident from the present study that the seed size was positively related to germination % (r = 0.708**) and seedling vigour (r = 0.766**). The data also suggested that large seeded hybrids with their relatively vigorous roots are likely to encounter early season drought better than the small seeded hybrids or inbred cultivars. Olisa et al. (2010) also suggested that the germination in pigeonpea could be enhanced through the choice of seed size as the cultivars with higher seed weight were found to have higher germination. The mean dry weight of the hybrid seedlings (4.7±0.28 mg) was significantly more in comparison to inbred cultivars (3.5±0.28 mg) indicating the fact that larger seeds had produced greater seedling biomass and thus, higher dry matter. Narayanan and Sheldrake (1974) demonstrated that the seedling dry weight in pigeonpea inbred cultivars was directly proportional to the test weight confirming again that seedling growth was a consequence of reserve assimilates in seeds. Seedling dry weight was found to be positively associated with seed size, root length and seedling vigour index. The seedling dry matter also indicated that vigour of the pigeonpea seedling in terms of biomass was highly correlated with their longer root and shoot (Kumar et al. 2011). Identification of genotypes with high germination and fast initial seedling growth is likely to result in rapid canopy develo pment fo r greater li ght int erceptio n, great er suppression of weeds and better seedling establishment. Pigeonpea hybrids are more beneficial than inbred cultivars with respect to enhanced seedling vigour, greater drought tolerance, greater disease resistance and increased grain yield (Saxena et al. 1992). Physiological studies have also confirmed that pigeonpea hybrids were superior to inbred cultivars with respect to seedling vigour, growth rate, biomass production, drought resistance and seed yield (Saxena et al. 1996). The present study also showed that large seeded hybrids had better germination and more vigorous root system than those in inbred cultivars. Significant and positive correlation between seed size and yield was also reported by Patel and Root length (cm) 0.598* 0.671** 0.045 Shoot: root ratio -0.409 -0.429 0.552* 0.505 Test weight (g) 0.708** 0.893** 0.063 0.718** -0.279 Seedling vigour index (I) 0.639** 0.738** 0.445 0.858** 0.229 0.766** Acharya (2011). Thus, pigeonpea hybrids by virtue of their greater root mass and depth have better abilityto mine receding soil water and tolerate drought during its early phases of growth. The superiority of hybrids in terms of vigour can be identified at early growth stage which may be related to vigour at later developmental stages associated with seed yield. Correlation studies also indicated the association of germination and related traits with vigour of the hybrid seedlings. The present study also indicated the benefits of breeding large seeded hybrids which would give better germination and longer root with better drought tolerance at early growth stages. ACKNOWLEDGEMENTS The authors are thankful for the financial support from National Food SecurityMission (NFSM) of DAC, MOA, India. Thanks are also due to Dr. Abhishek Rathore, Scientist (Biometrics) for his support in statistical analysis, and to Dr. L. Krishnamurthy of ICRISAT for his valuable comments. REFERENCES Abdul-Baki AA and Anderson JD. 1973. Relationship between decarboxylation of glutamic acid and vigour in soybean seed. Crop Science 13: 222-226. Brakke MP and Gardner FP. 1987. Juvenile growth in pigeonpea, soybean, and cowpea in relation to seed and seedling characteristics. Crop Science 27: 311-316. Chauhan YS, Johansen C and Saxena KB. 1995. Physiological basis of yield variation in short-duration pigeonpea grown in different environments of the semi-arid tropics. Journal of Agronomy and Crop Science 174: 163-171. ISTA. 2007. International Rules for Seed Testing International Seed Testing Association (ISTA), Switzerland. Kumar RR, Krishna K and Naik GR. 2011. Variation of sensitivity to drought stress in pigeonpea (Cajanus cajan (L.) Millsp.) cultivars during seed germination and early seedling growth. World Journal of Science and Technology 1: 11-18. Mercer KL, Shaw RG and Wyse DL. 2006. Increased germination of diverse crop-wild hybrid sunflower seeds. Ecological Applications 16: 845-854. Narayanan A and Sheldrake AR. 1974. Effect of seed size on growth of seedlings in pigeonpea. In: Annual Report Pigeonpea Physiology, International Crops Research Institute on Semi-Arid Tropics, Patancheru, India. Pp. 39-43. Narayanan A, Saxena NP and Sheldrake AK. 1981. Varietal differences 354 Journal of Food Legumes 25(4), 2012 in seed size and seedling growth of pigeonpea and chickpea. Indian Journal of Agricultural Sciences 51: 289-393. Workshop on ‘New Frontiers in Pulses Research and Development’, 10-12 November 1989, Kanpur, India. Pp. 58-69. Olisa BS, Ajayi SA and Akande SR. 2010. Physiological quality of seeds of promising African yam bean (Sphenostylisstenocarpa (Hochst, ExA, Rich) Harms) and pigeonpea (Cajanus cajan (L.) Millsp.) landraces. Research Journal of Seed Science 3: 93-101. Saxena KB, Chauhan YS, Laxman Singh, Kumar RV and Johansen C. 1996. Research and development of hybrid pigeonpea. Research Bulletin 19. International Crops Research Institute for the SemiArid Tropics, Patancheru 502324, Andhra Pradesh, India. Pp.20. Patel JB and Acharya S. 2011. Genetic divergence and charcter association in Indo-African derivatives of pigeonpea [Cajanus cajan (L.) Millsp.]. Journal of Food Legumes 24: 198-201. Saxena KB, Kumar RV, Srivastava N and Shiying B. 2005.A cytoplasmicnuclear male-sterility system derived from a cross between Cajanus cajanifolius and Cajanus cajan. Euphytica 145: 289-294. SAS. 2008. SAS/STAT ® 9.2 Users guide, SAS Institute Inc., Cary, NC, USA. Saxena KB. 2009. Evolution of hybrid breeding technology in pigeonpea. In: M Ali and Shiv Kumar (Eds), Milestones in Food Legume Research. Indian Institute of Pulses Research, Kanpur, India. Pp. 82-114. Saxena KB, Chauhan YS, Johansen C and Singh L. 1992. Recent developments in hybrid pigeonpea research. In: Proceedings of Journal of Food Legumes 25(4): 355-357, 2012 Short communication Screening of chickpea (Cicer arientinum L.) genotypes for identification of source of resistance to Botrytis grey mould LAJJA VATI, K.P.S. KUSHWAHA and ABHIJEET GHATA Department of Plant Pathology, College of Agriculture, GBPUA&T, Pantnagar - 263 145, Uttarakhand, India; E-mail: [email protected] (Received: April 04, 2012; Accepted: November 10, 2012) ABSTRACT In order to identify the source of genetic resistance to Botrytis grey mould (BGM), thirty genotypes of chickpea were screened under field condition during 2009-10 and 2010-2011. None of the genotypes was found to be immune and highly resistant. However, 15 genotypes namely ‘ICCV 05502’, ‘ICCV 05506’, ‘ICCV 05509’, ‘ICCV 05522’, ‘ICCV 05528’, ‘ICCV 05529’, ‘ICCV 05553’, ‘EC 516891’, ‘EC 516696’, ‘EC 516720’, ‘EC 516806’, ‘EC 516670’, ‘EC 516878’, ‘ICC 4954’ and ‘ICC 4951’ were found resistant to BGM. These genotypes can be used in breeding programmes to develop varieties resistant to BGM. Key words: Breeding, Chickpea, Mould, Resistance Chickpea (Cicer arientinum L.) is a self-pollinated diploid annual with genome size of approximately 750 Mbp (Arumuganathan and Earle 1991). It is the third most important grain legume crop in the world (Bakr et al., 2002; Pande et al., 2006). Chickpea is valued for its nutritive seeds with high protein content ranging from 5.3 to 28.9% (Hulse 1991). It is grown in the Indian sub-continent, West Asia, North Africa (WANA), the Mediteterranean basin, the Americas and the Australia (Croser et al. 2003). However, the vulnerability of this crop to biotic stresses such as BGM, Ascochyta blight, Fusarium wilt, nematodes and pests and abiotic stresses (drought and cold) reduce its yield substantially. Among various fungal diseases, BGM caused by Botrytis cinerea Pers. ex. Fr. causes considerable yield loss by reducing plant population in the field. The wide variety of symptoms on different plant parts may suggest that B. cinerea has a large ‘arsenal of weapons’ to attack its host plants. Among the necrotrophic and polyphagous fungi, the grey mould agent is one of the most – studied models (Van Kan et al., 2006). Heavy mortality of flowers results in poor pod formation due to BGM infection. Drooping of the infected tender terminal branches is a common field symptom (Pande et al., 2005). It causes extensive crop losses in most regions of the world due to the fact that environmental condition favourable to chickpea crop (>350 mm annual rainfall, 23-25ºC) also favour the disease. World wide losses from this fungus account for 20% of the harvest of the affected crops, and their cost is estimated at 101000 billion euros per year and the market size for anti botrytis products has been 15-25 million US dollars in recent years (Genoscope 2008). Therefore, controlling this disease is essential to ensure stable chickpea production. Eradication of this soil-borne pathogen is difficult because of its polyphagous nature and its survival in the soil through its resting structures. As fu ngicides are costly, and unfriendly t o our ecosystem, it is imperative to identify the source of its resistance and exploit it to develop resistant varieties of chickpea through breeding approaches. Therefore, in this study efforts have been made to screen thirty genotypes against BGM in chickpea. Beside this, such screenings were also conducted previously. But the present study was carried out to observe whether new races of the pathogen evolved to cause severe damage in the locality and the screened genotypes showing resistance in both years can exploit for further breeding programme. Thirty genotypes of chickpea (Table 1) were used for screening purpose. These genotypes were obtained from the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad. Direct seeding was performed in November 2009 and 2010 to record observations on the disease. A basal dose of N, P and K was supplied @ 20, 60 and 50 kg/ha, respectively (Suyal 2010). Hand weeding was performed at 2-week interval during the crop season. Irrigation was given at 5 weeks after sowing. Plots (1.0 × 4.0 m) were established in a randomized complete block design (RCBD) with three replications (Gomez and Gomez 1984). Plant-to-plant distance within the row was kept at 10 cm and row to row distance was maintained at 30 cm. A plot with three rows (4 m length) was kept for each genotype. Methodology for inoculation was followed in the present investigation as suggested by Pande (2010). During both years, field screening was performed in the Pulse Pathology Block, N.E. Borlaug Crop Research Center, GBPUAT, Pantnagar. Disease severity (disease index) on each genotype in each replication was recorded three times at the interval of 15 days and first observation was recorded 10 days after inoculation using the following formula: Disease index (DI) = of numerical ratings × 100/ No. of plants observed × highest degree of rating Journal of Food Legumes 25(4), 2012 Table1. Botrytis grey mould reaction of chickpea genotypes during 2009-11 S.N. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 Genotype ICCV04509 ICCV04512 ICCV04513 ICCV04514 ICCV05501 ICCV05502 ICCV05506 ICCV05509 ICCV05522 ICCV05526 ICCV05527 ICCV05528 ICCV05529 ICCV05530 ICCV05531 ICCV05532 ICCV05533 ICCV05535 ICCV05553 EC 516891 EC 516671 EC516696 EC 516720 EC516806 EC 516670 EC516878 EC516012 EC516755 ICC 4954 ICC4951 Cd at 5 % (%)a Disease severity 2009-10 2010-11 23.0 (0.496) Ib 28.0 (0.553) MS 18.0 (0.438) I 23.0 (0.496) I 18.0 (0.438) I 18.0 (0.438) I 28.0 (0.553) MS 33.0 (0.611) MS 23.0 (0.496) I 23.0 (0.496) I 3.0 (0.174) R 3.0 (0.174) R 4.7 (0.211) R 3.0 (0.174) R 3.0 (0.174) R 4.7 (0.211) R 4.7 (0.211) R 4.7 (0.211) R 23.0 (0.496) I 23.0 (0.496) I 18.0 (0.438) I 23.0 (0.496) I 3.0 (0.174) R 3.0 (0.174) R 4.7 (0.211) R 3.0 (0.174) R 23.0 (0.496) I 28.0 (0.553) MS 23.0 (0.496) I 19.7 (0.445) I 28.0 (0.553) MS 23.0 (0.496) I 23.0 (0.496) I 23.0 (0.496) I 23.0 (0.496) I 28.0 (0.553) MS 3.0 (0.174) R 4.7 (0.211) R 3.0 (0.174) R 3.0 (0.174) R 23.0 (0.496) I 19.7 (0.445) I 4.7 (0.211) R 3.0 (0.174) R 4.7 (0.211) R 4.7 (0.211) R 3.0 (0.174) R 3.0 (0.174) R 4.7 (0.211) R 4.7 (0.211) R 3.0 (0.174) R 3.0 (0.174) R 19.7 (0.445) I 28.0 (0.553) MS 28.0 (0.553) MS 23.0 (0.496) I 3.3 (0.183) R 3.0 (0.174) R 3.0 (0.174) R 4.7 (0.211) R 0.12 0.12 a Figures in parenthesis are arcsine transformed values. Averaged over three replications b Level of resistant reaction; I = intermediate, MS = moderately susceptible, R = resistant Plants were considered diseased when drooping occur of the infected tender terminal branches, is a common field symptom. The data obtained on disease severity were analyzed to determine the level of pathogenicity reaction among the tested chickpea genotypes. The disease reaction of the genotypes was quantified separately for two different years. A final ANOVA was developed over the pooled value of two years on the basis of their mean disease severity as per 1 to 9 disease rating scale suggested by Hawthorne et al., (2006). Before analysis, values were transformed using arcsine transformation. Statistical analyses were made using the Statistical Package for Social Sciences Version 16.0 (SPSS 16.0, SPSS Inc.) programme. The two -year trial data for screeni ng chickpea genotypes against BGM is presented in Table 1. The observations indicated that none of the genotype was observed without BGM infection. Similar screening of chickpea genot ypes has been do ne at the ICRISAT, Hyderabad, and some other State Agricultural Universities by former workers (Verma et al. 1981, Verma and Gill 1981, Haware and Nene 1982, Pandey et al. 1982, Chaube et al. 1983). Keeping in view of development of new races, the present screeni ng study was necessary as previo us screenings were done much earlier. Disease severityranged from 3.0 to 33.0%. However, 15 genotypes (ICCV05502, ICCV05506, ICCV05509, ICCV05522, ICCV05528, ICCV05529, ICCV05553, EC516891, EC516696, EC516720, EC516806, EC516670, EC516878, ICC 4954 and ICC4951) showed minimum disease severity (<5%), and thus classified as resistant (R). In both years, genotypes under R category showed similar reaction (Table 2). The present findings when compared with the results of other two locations (ICRISAT and Ludhiana), it was observed that germplasm lines ‘ICCV 05502’, ‘ICCV 05506’, ‘ICCV 05509’, ‘ICCV 05522’, ‘ICCV 05529’, ‘ICCV 05553’, ‘EC 516891’, ‘EC 516720’, ‘EC 516806’ and ‘EC 516878’ showed resistant reaction at both Pantnagar and ICRISAT locations (Anonymous 2010). Five genotypes (ICCV04509, ICCV04514, ICCV05530, ICCV05535 and EC516012) were classified as moderately susceptible (MS). Different reactions of the same genotype over years were obtained, particularly for the intermediate reaction (I). Therefore, the genotypes were categorised under a specific reaction level on the basis of the pooled value (Fig. 1). The genotypes that showed intermediate disease reaction had possibly little scope of their use for further study. This result was consistent during both the year of study. Similar results with analogous methodology were reported by Singh et al. (1982). Disease incidence (%) 356 35 28 21 14 7 0 1 4 14 16 18 28 2 3 5 10 11 15 17 21 27 6 7 8 9 12 13 19 20 22 23 24 25 26 29 30 MS I R Reaction level Fig 1. Reaction of chickpea genotypes against B. cinerea (pooled value over two years). Genotypes numbers under a specific reaction level are referred from Table 1. Error bars are standard error of the mean Table 2. Analysis of variance for the effect of genotype on disease severity tested in two years (2009-11) Variable Genotype Reaction levela MS I R a Year 2009-10 2010-11 2009-10 2010-11 2009-10 2010-11 SS MS F-value 0.2E-06b 0.1E-06 0.0000 0.0080 0.0020 0.2499 0.2526 0.0022 0.2575 0.0177 0.0019 0.1599 0.0146 0.0010 0.6120 0.0136 0.0009 0.6834 Level of resistant reaction; I = intermediate, MS = moderately susceptible, R = resistant b Data transformed in arcsine prior to ANOVA. Vati et al.: Screening of chickpea genotypes for identification of source of resistance The genotypes grouped under MS can be eliminated for future breeding programme. Similar type of screening for disease severity of BGM in chickpea has been reported (Tripathi and Rathi 1992). An analysis of variance (ANOVA) was made including two experiments, where genotypes had strong significant effect on disease severity over two-years estimation. However, experiment had a poor significant effect on disease severity. Furthermore, disease severity was significantly affected by the interaction of genotypes × experiment, meaning experiments have effect on disease severity of BGM of chickpea. Therefore, separate analyses were performed for different reaction levels in each year (Table 2). In all reaction levels of each year showed non significant effect. However, a strong effect of genotype reaction against the BGM disease was detected in both years for resistant genotypes. Hence, the pattern showed that the genotypes under resistance level could be cultivated over period. Resistance genotypes showing susceptibility to BGM were often recognised as breakdown o f gene by the pathogen or development of a new pathogenic race of a certain region. The present study revealed the absence of complete resistance to B. cinerea in the used set of chickpea genotypes. These genotypes may be utilized for improving BGM resistance in chickpea. REFERENCES Anonymous. 2010. Annual Report. All India Coordinated Research Project (AICRP) on chickpea. Pp.176. Bakr MA, Rahman MM and Ahmed AU. 2002. Manifestation of Botrytis grey mould of chickpea in Bangladesh. In: MA Bakr, KHM Siddique and C Johansen (Eds), Integrated Management of Botrytis Grey Mould of Chickpea in Bangladesh and Australia, International Crops Research Institute for the Semi-Arid Tropics, India. Pp. 63-69. 357 Genoscope. 2008. Botrytis cinerea estimated losses for vineyards in France amount to 15-40 % of the harvest, depending on climatic conditions. In: Sequencing Projects of Botrytis cinerea. Gomez KA and Gomez AA. 1984. Statistical procedures for agricultural research. John Wiley & Sons, Singapore. Haware MP and Nene YL. 1982. Screening of chickpea for resistance to Botrytis grey mould. International Chickpea Newsletter 6: 18. Hawthorne W, Davidson J, McMurray L, Lindbeck K and Brand J. 2006. Chickpea disease management strategy southern region. Pp. 4-6. Hulse JH. 1991. Nature, composition and utilization grain legumes uses of tropical legumes. In: Proceedings of the Consultants Meeting (CM91), ICRISAT, Patancheru, India. Pp. 11-27. Pande S, Krishna KG and Rao JN. 2005. Marigold: A diagnostic tool for BGM forcasting and management in chickpea. E journal (WWW.ICRISAT.Org) 1: 1. Pande S, Galloway PM, Gaur, Siddique KHM and Tripathi HS. 2006. Botrytis grey mould of chickpea: A review of biology, epidemiology and disease management. Australian Journal of Agricultural Research 57: 1137-1150. Pandey MP, Pandya BP, Chaubey HS, Tewari SK and Beniwal SPS. 1982. Screening cultivars and genetic stocks of chickpea for resistance to Ascochyta blight. ICN 7: 15-16. Pande S. 2010. Guidelines: Method of inoculation. In: International chickpea Botrytis Gray Mold Nursery Pp. 3. Singh G, Kapoor S and Singh K. 1982. Screening chickpea for grey mould resistance. International Chickpea Newsletter 7: 13. Suyal U. 2010. Studies on histopathology, molecualar charactrization and management of Botrytis cinerea Pers. Ex. Fr., the incitant of grey mould of chickpea. Ph.D. Thesis, GBPUA&T, Pantnagar, India. Tripathi, HS and Rathi YPS. 1992. Resistance to Botrytis grey mould in chickpea: Screening technique and identification of resistance source. Journal of Mycology and Plant Pathology 30: 231-232. Van Kan JAL. 2006. Licensed to kill: the lifestyle of a necrotrophic plant pathogen. Trends in Plant Science 11: 247-253. Chaube HS, Beniwal SPS, Tripathi HS and Nene YL. 1980. Field Screening of Chickpea for resistance to Botrytis grey mould. International Chickpea Newsletter 2: 16. Verma NN, Singh G, Sandhu TS, Singh H, Sandhu SS, Singh K and Bhullar BS. 1981 Sources of resistance to gram blight and grey mould. International Chickpea Newsletter 4: 14. Croser JS, Clarke HJ and Siddique KHM. 2003. Utilization of wild Cicer in chickpea improvement progress, constraints and prospects. Australian Journal of Agricultural Research 54: 429-444. Verma MM and Gill AS. 1981. Field screening of chickpea segregating material in multiple disease sick plot. ICN 4:14. Journal of Food Legumes 25(4): 358-360, 2012 Short Communication Management of Fusarium wilt of lentil using antagonistic microorganisms in Tarai region of Uttarakhand ANKITA GARKOTI and H.S. TRIPATHI Department of Plant Pathology, G.B. Pant University of Agriculture and Technology, Pantnagar - 263 145, Uttarakhand, India; E-mail: [email protected] (Received: May 26, 2012; Accepted: October 23, 2012) ABSTRACT Two bio-agents, Trichoderma harzianum and Pseudomonas fluorescence were screened in vitro against Fusarium wilt pathogen of lentil by using dual culture technique. Observations revealed that T. harzianum grew faster than P. fluorescens. All the isolates of T. harzianum significantly checked the growth of the test pathogen. Maximum inhibition (71.8%) after 96 hrs of incubation was recorded with isolate of Th 5 followed by Th 39 (70.8%) and Th 14 (68.9%). Isolates of P. fluorescence were found less effective in comparison to T. harzianum in inhibiting the growth of the test pathogen (Fusarium oxysporum f.sp. lentis). Key words: Bio-agents, Fusarium wilt, Lentil Lentil (Lens culinaris Medic.) is a protein-rich food legume grown throughout northern and central India having total area of 1.59 million ha with a total production of 0.94 million tons and productivity 591 kg/ha (AICRP on MULLaRP 2011-2012). Among the soil-borne diseases, Fusarium wilt caused by Fusarium oxysporum f.sp. lentis (Fol) is the most important biotic constraint to productivity of lentil worldwide (Bhalla et al. 1992). Lentil wilt caused by Fol, is one of the most widespread and destructive diseases. Symptoms include wilting of top leaves followed by entire plant (Khare 1980). The disease may cause complete crop failure under favorable conditions for disease development, and can be the major limiting factor for lentil cultivation in certain areas (Chaudhary and Amarjit 2002). Biological control using antagonistic microorganisms is an alternative method to the fungicides and provides an opportunity for ecological based approach to integrated pest management in sustainable agriculture in crop production systems (Cook and Granados 1991, Singh et al. 1999, Sutton and Peng 1993). Antagonistic Trichoderma species are considered as promising biological control agents against numerous phytopathogenic fungi including F. oxysporum (Sarhan et al. 1999). Hence, in the present study fungal and bacterial bio-agents were tried as alternative method under organic farming for lentil wilt disease management particularly in Uttarakhand state. Fusarium oxysporum f.sp. lentis was isolated from naturally infected lentil roots and thereafter same was purified by single spore method and maintained on PDA in culture tubes at 28+ 1°C in an incubator for further studies. Different strains of bio-control agents viz., Trichoderma harzianum and Pseudomonas fluorescence were procured from Biocontrol laboratory of the Department of Plant Pathology, GBNPUA&T, Pantnagar and initially screened by following dual culture technique. For fungal antagonist 20 ml of sterilized melted PDA was poured in 90 mm diameter Petri plates. After solidification of medium, 5 mm disc of the antagonist and test pathogen were cut with the help of a sterilized cork borer from the edge of 5 day old culture and placed in straight line at distance of 5 mm from the edge. Three replications were maintained for each treatment and petri plates without antagonist served as control. For bacterial antagonist (same amount of melted PDA + King’s B medium in 1:1 ratio) was used. Five mm sterilized paper disc were dipped in bacterial suspension and were placed at the opposite corner of the petri plate containing Fol disc on solidified medium. The pathogen without antagonist served as check. The inoculated petri plates were incubated at 28±10C and linear growth of the bio-agent was observed consequently for four days to record different stages of antagonism. The per cent inhibition of both antagonists was determined with the help of mean colony diameter and calculated by using formula described earlier (Mckinney 1923). I C-T C Where, I = % inhibition C = colony diameter in control T = colony diameter in treated medium The effect of bi o-agents (T. harzianu m and P. fluorescence) for their antagonistic potential against the test pathogen (F. oxysporum f.sp. lentis) was assessed by dual culture technique. Observations on the colonization of the test pathogen by the selected bio-agents indicated their varied antagonistic potentiality (Table 1 and 2). The strains of T. harzianum grew at a faster rate than the test pathogen F. oxysporum f.sp. lentis. All the strains of Trichoderma significantly checked the growth of the pathogen. After 24 h of inoculation of antagonist and test Garkoti & Tripathi: Management of Fusarium wilt of lentil using antagonistic microorganisms pathogen, colony diameter of each of them developing on PDA, individually and after 48 h of inoculation, both the cultures came in contact of each other. The growth of antagonist overlapped the mycelia of the test pathogen after 72 h of incubation. After 96 h of incubation, T. harzianum completely covered the growth of the test pathogen. All the ten isolates of T. harzianum significantly controlled the growth of the test pathogen. Maximum inhibition 71.8% after 96 h of incubation was recorded in Th 5 followed by Th 39, where inhibition was 70.8%. Minimum inhibition (49.3%) was recorded in the culture treated with Th3. A clear zone of inhibition between F. oxysporum and P. fluorescence was observed. All the strains of P. fluorescence were found effective in inhibiting the growth of test pathogen. After 96 h of incubation, maximum inhibition was observed in strain 12, in which per cent inhibition was 64.2 followed by strain 4 where it was 61.4. Minimum per cent inhibition was 35.8 observed in strain 7. Biological control is the best alternative, especially 359 against soil-borne pathogens such as Fusarium spp. (Akrami et al. 2011). Trichoderma species differentially limited the colony growth of the pathogen, overgrew the pathogen colony and produced yellow pigment (Dolatabadi et al. 2011). The majority of Trichoderma species is antagonist of phytopathogenic fungi and has been broadly used as the most important biocontrol agent (Tjamos et al. 1992, Akrami et al. 2011). From several studies, it has been confirmed that Trichoderma spp. have antagonistic effects against diversity of soil-borne pathogens (Grondona et al. 1997, Bajwa et al. 2004). Results revealed that all strains of T. harzianum were effective as it grew over and parasitized F. oxysporum f.sp. lentis and checked the growth in dual culture technique. The strains of T. harzianum were fast growing than pathogen. The suppression in growth may be due to the lack of nutrition for growth of pathogen and production of certain inhibitory chemicals by antagonists in culture. The growth inhibition may be due to the hyphal parasitization and production of wall degrading enzymes by the test antagonists. However, this needs further investigation. Table 1. In vitro efficacy of T. harzianum isolates on growth of Fusarium oxysporum f.sp. lentis T. harzianum isolates Th-1 Th-3 Th-5 Th-9 Th-14 Th-39 Th-56 Th-60 Th-66 Th-70 Check SEm (±) CD (P=0.05) CV (%) *Mean of three replications 48 h 8.3 11.4 5.3 10.5 6.6 6.1 8.8 8.8 10.2 9.6 13.8 0.47 1.39 9.08 Radial growth (mm)* 72 h 11.5 16.8 7.3 16.0 10.8 9.0 12.1 13.2 15.3 14.0 26.0 0.59 1.73 7.40 96 h 13.1 20.7 11.5 20.1 12.7 11.9 14.4 15.2 18.8 16.6 40.8 0.56 1.65 5.48 48 h 39.9 17.4 61.6 23.9 52.2 55.8 36.2 36.2 26.1 30.4 Per cent inhibition 72 h 55.8 35.4 71.9 38.5 58.5 65.4 53.5 49.2 41.2 46.2 96 h 67.9 49.3 71.8 50.8 68.9 70.8 64.7 62.7 53.9 59.31 Table 2. In in vitro efficacy of P.fluorescens isolates on growth of Fusarium oxysporum f.sp. lentis P. fluorescens isolates FL P-2 FL P-4 FL P-6 FL P-7 FL P-12 FL P-18 FL P-25 FL P-27 FL P-28 FL P-31 Check SEm (±) CD (P=0.05) CV (%) *Mean of three replications 48 h 7.0 6.3 7.6 8.0 6.0 6.3 6.6 7.6 6.6 6.3 13.0 0.36 1.06 8.45 Radial growth (mm)* 72 h 11.3 10.3 12.0 13.0 9.3 10.6 11.6 11.6 11.0 11.6 21.3 0.68 2.02 9.79 96 h 19.3 14.0 20.6 23.3 13.0 14.6 19.6 21.0 17.3 17.0 36.3 0.97 2.85 8.58 48 h 46.2 51.5 41.5 38.5 53.8 51.5 49.2 41.5 49.2 51.5 Per cent inhibition 72 h 46.9 51.6 43.7 38.9 56.3 50.2 45.5 45.5 48.4 45.5 96 h 46.8 61.4 43.3 35.8 64.2 59.8 46.0 42.1 52.3 53.2 360 Journal of Food Legumes 25(4), 2012 REFERENCES Akrami M, Golzary H and Ahmadzadeh M. 2011. Evaluation of different combinations of Trichoderma species for controlling Fusarium rot of lentil. African Journal of Biotechnology 10: 2653-2658. All India Coordinated Research Project on MULLaRP. 2011-2012. Project Coordinator’s Report (Rabi Crops), Indian Institute of Pulses Research, Kanpur. Bajwa R, Mukhtar I and Anjum T. 2004. In vitro biological control of Fusarium solani- cause of wilt in Dalbergia sissoo Roxb. Mycopathology 2: 11-14. Bhalla MK, Nozzolillo C and Schneider E.1992. Observation on the responses of lentil root cells to hypha of Fusarium oxysporum. Journal of Phytopathology 135: 335-341. Chaudhary RG and Amarjit K. 2002. Wilt disease as a cause of shift from lentis cultivation in Sangod Tehsil of Kota, Rajasthan. Indian Journal of Pulses Research 15: 193–194. Cook RJ and Granados RR. 1991. Biological control: making it work. In: MJF MacDonald (ed). Agricultural Biotechnology At The Cross Roads. National Agricultural Biotechnology Council, Ithaca. Pp. 213-227. Dolatabadi HK, Goltapeh EM, Mohammadi N, Rabiey M, Rohani N and Varma A. 2011. Evaluation of different combinations of Trichoderma species for controlling Fusarium rot of lentil. African Journal of Biotechnology 10: 2653-2658. Grondona I, Hermosa R, Tejada M, Gomis MD, Mateos PF, Bridge PD, Monte E, and Garcia Acha I. 1997. Physiological and biochemical characterization of Trichoderma harzianum, a biological control agent against soilborne fungal plant pathogens. Applied and Environmental Microbiology 63: 3189–3198. Khare MN. 1980. Wilt of Lentis. Jawaharlal Nehru Krishi Vishwa Vidyalaya, Jabalpur, M.P., India. Pp.155. Sarhan MM , Ezzat SM and Al- Tohamy MR. 1999. Application of Trichoderma hamatum as a Biocontroller against Tomato Wilts Disease Caused by Fusarium oxysporum f. lycopersici. Egyptian Journal of Microbiology 34: 347-376. Singh PP, Shin YC, Park CS and Chung YR. 1999. Biological control of Fusarium wilt of cucumber by chitinolytic bacteria. Phytopathology 89: 92-99. Tjamos EC, Papavizas GC and Cook RJ. 1992. Biological control of plant diseases. Progress and challenges for the future. Plenum Press, New York. USA. Journal of Food Legumes 25(4): 361-363, 2012 Short communications Effect of phosphorus and zinc on yield and economics of mothbean under semiarid conditions L.R. YADAV, POONAM CHOUDHARY, SANTOSH, O.P. SHARMA and MEENU CHOUDHARY S.K.N. College of Agriculture (Swami Keshwanand Rajasthan Agriculture University, Bikaner), Jobner, (Rajasthan) 303329, India; E-mail: [email protected] (Received: March 17, 2012; Accepted: December 06, 2012) ABSTRACT A field experiment was conducted during rainy (kharif) season of 2007 at Agronomy farm, S.K.N. College of Agriculture, Jobner (Rajasthan) to study the effect of phosphorus and zinc on mothbean under semi arid conditions on loamy sand soil. Twenty treatment combinations comprising five levels of phosphorus (0, 10, 20, 30 and 40 kg P2O5/ha) and four levels of zinc (0, 2, 4 and 6 kg Zn/ha) were tried in a RBD with three replications. Application of 30 kg P 2 O 5/ha significantly enhanced plant height, dry matter accumulation/plant, effective nodules and dry weight of nodules, pods/plant, seeds/pod, seed and straw yields, net returns, nutrient (N, P, K and Zn) content in seed and straw and their total uptake. Zinc applied at 4 kg/ha (on par with 6 kg/ha) was optimum as it significantly increased plant height, dry matter accumulation/plant, chlorophyll content in plant leaves at 40 days after sowing (DAS), per plant effective nodules and its dry weight, number of pods, seeds/pod, seed and straw yields, net returns, nutrient (N, P, K and Zn) content in seed and straw and their total uptake over the lower doses (both 2 kg Zn/ha and control). Key words: Mothbean, Net returns, Phosphorus levels, Zinc levels Mothbean (Vigna aconitifolia [Jacq.] Marechal) is a hardy and drought tolerant crop among kharif pulses and largely grown in arid and semi-arid regions. It is also one of the most assured and remunerative crops in arid regions due to i ts ext reme t olerance to moisture stress and high temperature. Because of its very short maturity period, it is highly suitable for low rainfall areas of western Rajasthan (Yadav et al. 2004) under Indian subcontinent. Nutritionally it is also good as it contains 20.5% easily digestible protein rich in lysine and tryptophan and other essential amino acids. It has the inherent capability for biological nitrogen fixation for meeting its own N need and thus, helps in enriching soil nitrogen status. The state of Rajasthan is the largest producer of mothbean and contributes 80 % share of country’s production. However, its productivity is very low. Thus, harnessing its productivity potential through use of improved agronomy like, balanced fertilization is a viable option in the arid and semi arid regions of the country. Amongst nutrients, phosphorus is deficient particularly in light textured soils and its deficiency has been recognized as a major bottleneck in realizing the potential yield of mothbean (Patel et al. 2004). P is also an important mineral element for grain legumes as it helps in root development, synthesis of phosphates and phospho-proteins, energy fixing and releasing process in plants and improving seed quality (Singh and Yadav 2008). Besides major and secondary nutrients, pulses require zinc for completion of its life cycle. It is reported that majority of pulse growing regions of India are low in zinc availability (Singh et al. 2011). Moreover, in light textured semi arid and arid soils of western Rajasthan, zinc deficiency is widespread because of low soil organic carbon (SOC) and alkaline soil reaction (pH). Zi nc also pl ays an i mpo rtant role in photosynthesis, sugar transformation, protein synthesis, water absorption, flowering and seed setting. Hence, the present invest igat ion was undertaken to harness t he productivity of mothbean by P and zinc application especially under semi-arid conditions. A field experiment was conducted during kharif 2007 at Agronomy farm, S.K.N. College of Agriculture, Jobner (Rajasthan), India. The loamy sand soil was alkaline (pH 8.2) in reaction, low in SOC (0.14%), available N (133 kg/ha), P (16.3 kg P2O5 /ha) and available Zn (0.4 ppm) and medium in available K(150 kg/ha). Twenty treatments involving five levels of P (0, 10, 20, 30 and 40 kg P2O5/ha) and four levels of Zn (0, 2, 4 and 6 kg Zn/ha) were tried in a randomized complete block design (RCBD) with three replications. The total rainfall received during the crop period was 114.5 mm. Mothbean ‘RMO-40’ was sown during second week of July at 30 x 5 cm spacing using 15 kg seed/ha and harvested in the second week of September. A uniform basal dose of 15 kg urea-N/ha was applied to all the plots and was adjusted with N supplied through diammonium phosphate (DAP) as per treatment. Whole of P through DAP and Zn through zinc sulphate were drilled in earmarked plots as per treatment before sowing. The amount of sulphur supplied through zinc sulphate was also adjusted with elemental sulphur to make it uniform to all plots. Thus, the treatments were compared with respect to P and Zn only keeping all other factors constant. Standard methods were used for determination of P and Zn content in plant and their uptake were also calculated. The net monitory returns 362 Journal of Food Legumes 25(4), 2012 were calculated on the basis of prevailing market rates. weight of nodules over both 2 kg Zn/ha and control (Table 1). Increasing levels of zinc up to 4 kg/ha also significantly increased yields (seed and straw yield), yield attributes (pods/ plant and seeds/pod) and net return of mothbean. In relative terms, application of 4 kg Zn/ha recorded 42.6, 30.4, 23.9 and 54.6 % in respect of pods/plant, seed and straw yields and net return over control, respectively. However, test weight of mothbean was significantly increased up to 2 kg Zn/ha (5.9 % over control) as higher doses of Zn beyond 2 kg/ha (4 and 6 kg/ha) were found statistically at par with each other (Table 1). This might be due to increased yield attributes and seed yield because of its pertinent role in enhanced nitrogen metabolism thereby increasing its availability to the plants for efficient growth and development by way of enhanced partitioning of photosynthates towards newly formed sink i.e. pods and seeds. These results corroborate the findings of Singh and Sharma (2005) in mothbean. Phosphorus application up to 30 kg P2O5/ha significantly enhanced plant height, dry matter accumulation/plant, effective nodules and dry weight of nodules (Table 1). Application of higher P levels viz., 30 and 40 kg P2O5/ha (both on par) also recorded significantly higher dry matter accumulation and registered an increase of 50.6, and 56.1 % over the control, respectively. However, leaf chlorophyll content in mothbean recorded at 40 DAS was significantly increased up to 20 kg P2O5/ha (on par with 30 and 40 kg P2O5/ ha) over control (16.2%) and 10 kg P2O5/ha (6.1%), respectively (Table 1). These results are in close agreement with Luikham et al. (2005). Similar to dry matter, yield attributes viz., pods/ plant and seeds/pod were significantly improved due to application of 30 kg P2O5/ha (on par with 40 kg P2O5/ha) over the preceding levels of phosphorus including the control. Test weight however, increased significantly up to 20 kg P2O5/ ha over the control and 10 kg P2O5/ha as higher doses of phosphorus failed to bring out significant variations in test weight. Seed and straw yield of mothbean also increased significantly with increasing levels of P up to 30 kg P2O5/ha and per cent increases in the above yields were to the tune of 39.5, 18.9, 6.2 and 28.1, 12.0, 5.2 over control, 10 and 20 kg P2O5/ha, respectively. Similar to yields, maximum net return was obtained following application of 30 kg P2O5/ha (on par with 40 kg P2O5/ha) and was superior (by ` 882/ha) over control and other lower doses of P. Application of P might have resulted in increased carbohydrate accumulation (higher biomass) and their remobilization to reproductive parts of the plants, being the closest to the sink and hence, resulted in increased flowering, fruiting and seed formation (Nadeem et al. 2004). Phosphorus fertilization @ 30 kg P2O5/ha (on par with 40 kg P2O5/ha) to mothbean also significantly increased protein and N content in seed, N content in straw and their total uptake (12.4, 12.3, 24.5 and 59.7 % respectively, over control). The improvement in protein content was due to higher uptake of N in plant (Gupta et al. 2006). Application of P might have improved nodules number and root growth which ultimately increased nutritional environment in rhizosphere as well as in pl ant syst em l eadi ng i n increased translo cati on and consequently uptake of nutrients. Since protein content is dependent on N content in seed, thus, increased N content in seed has increased protein content in seed which reflects the better nutritional environment in rhizosphere. These results are in close conformity with the findings of Gupta et al. (2006) in urdbean. Application of 4 kg Zn/ha also significantly increased plant height, dry matter accumulation (34.6 % over control), leaf chlorophyll content at 40 DAS, effective nodules and dry Zinc fertilization to mothbean also significantly increased protein and N content in seed and straw, and its total uptake (Table 2). Application of 4 kg Zn/ha significantly Table 1. Effect of P and Zn fertilization on growth, seed yield and its attributes and economics of mothbean* Treatments Plant height at harvest (cm) P levels (kg P2O5 /ha) 0 22.1 10 27.8 20 30.5 30 32.7 40 33.9 CD(P=0.05) 2.13 Zn levels (kg Zn/ha) 0 24.5 2 28.7 4 31.2 6 33.1 CD(P=0.05) 1.91 *Interaction not significant Dry matter accumulation at harvest (g/plant) Chlorophyll content at 40 DAS (mg/g) Effective nodules/ plant Dry weight of Pods/ Seeds/ Test Seed nodules/ plant pod weight (g) yield (kg/ha) plant (mg) 7.3 8.8 10.1 11.0 11.4 0.8 2.84 3.11 3.30 3.32 3.32 0.18 8.39 10.21 10.61 11.93 12.68 0.76 52.7 61.1 67.3 72.4 76.8 4.6 17.4 22.4 25.2 27.4 28.1 1.82 4.2 4.8 5.2 5.5 5.7 0.34 26.5 28.0 29.4 29.5 29.6 1.34 7.8 9.6 10.5 10.9 0.7 2.75 3.15 3.42 3.49 0.16 8.59 10.48 11.79 12.42 0.68 53.6 64.1 71.8 74.6 4.15 19.0 23.9 26.4 27.1 1.63 4.3 4.9 5.4 5.7 0.31 27.2 28.8 29.1 29.3 1.20 Straw yield (kg/ha) Net return (`/ha) 668 784 878 932 938 33 1665 1903 2026 2132 2174 110 7632 9886 11573 12514 12495 755 697 830 909 924 30 1686 1977 2090 2167 98 7895 10666 12204 12515 960 Yadav et al.: Effect of P and Zn in mothbean under semiarid conditions 363 Table 2. Effect of P and Zn fertilization on protein, nutrient content and total uptake by mothbean* Treatments Protein content in seed (%) P levels (kg P2O5 /ha) 0 10 20 30 40 CD (P=0.05) Zn levels (kg Zn/ha) 0 2 4 6 CD (P=0.05) N content (%) Seed Straw Total N uptake (kg/ha) P content (%) Seed Straw Total P uptake (kg/ha) Zn content (ppm) Seed Straw Total Zn uptake (g/ha) 20.2 21.5 22.2 22.7 22.9 0.7 3.23 3.44 3.55 3.63 3.67 0.11 1.65 1.82 1.92 2.06 2.08 0.07 48.6 61.8 70.2 77.6 79.4 5.0 0.38 0.43 0.48 0.50 0.50 0.01 0.23 0.25 0.27 0.28 0.28 0.01 6.28 8.10 9.66 10.57 10.88 0.51 25.1 28.3 30.7 31.8 32.8 1.3 20.0 21.0 21.7 22.2 22.6 0.9 49.5 62.5 71.2 76.9 79.9 4.0 21.2 21.8 22.3 22.3 0.6 3.39 3.49 3.57 3.57 0.10 1.84 1.90 1.93 1.94 0.06 54.9 66.8 72.7 75.0 4.5 0.41 0.46 0.48 0.48 0.01 0.23 0.27 0.27 0.28 0.01 6.74 9.10 10.07 10.45 0.46 26.8 28.0 31.7 32.6 1.2 19.5 20.7 22.5 23.0 0.8 51.6 64.3 75.6 80.0 3.6 *Interaction not significant increased N content in seed and straw and total uptake to the tune of 5.1, 5.0, 4.9 and 32.4 per cent over control, respectively. However, higher dose of Zn (6 kg/ha) could not bring in significant variation in N, P and Zn content and uptake. This might be due to positive response of applied Zn in deficient soils of arid and semi arid regions thereby, increased total uptake of Zn by mothbean. Similar, finding was reported by Saini (2003). It could be inferred from the above that application of 30 kg P2O5/ha and 4 kg Zn/ha was found effective for higher productivity and monetary returns of mothbean under the existing condition of semi arid regions of Rajasthan. REFERENCES Gupta A, Sharma GD and Chopra P. 2006. Effect of biofertilizer and phosphorus levels on yield attributes, yield and quality of urdbean (Vigna mungo). Indian Journal of Agronomy 51: 142-144. Luikham E, Lhungdim J and Singh A. 2005. Influence of sources and levels of phosphorus on growth and yield of greengram [Vigna radiata (L.) Wilzek]. Legume Research 28: 59-61. Nadeem MA, Ahmad R and Ahmad MS. 2004. Effect of seed inoculation and different fertilizer levels on growth and yield of mungbean (Vigna radiata).Journal of Agronomy 3: 40-42. Patel MM, Patel IC, Patel RM, Tikka SBS and Patel BS. 2004. Response of mothbean to row spacing and phosphorus under rainfed conditions. Indian Journal of Pulses Research 17: 91-92. Saini RK. 2003. Effect of zinc and iron on growth and yield of mothbean [Vigna acconitifolia (Jacq.) Marechal] in arid western Rajasthan. M.Sc. (Ag.) Thesis, Rajasthan Agricultural University, Bikaner. Singh KK, Praharaj CS, Choudhary AK, Kumar N and Venkatesh MS. 2011. Zinc response to pulses. Indian Journal of Fertilizers 7: 118126. Singh RS and Yadav MK. 2008. Effect of phosphorus and biofertilizers on growth, yield and nutrient uptake of long duration pigeonpea under rainfed condition. Journal of Food Legumes 21: 46-48. Singh V and Sharma SK. 2005. Response of mothbean ( Vigna acconitifolia L.) to zinc and thiourea under dryland condition. Annals of Agricultural Research, New Series 26: 313-314. Yadav GL, Kumawat PD and Babel AL. 2004. Effect of nitrogen, phosphorus and Rhizobium inoculation on mothbean. Indian Journal of Pulses Research 17: 95-96. Journal of Food Legumes 25(4), 2012 i List of Referees for Vol. 25 (4) The Editorial Board gratefully acknowledges the help rendered by following referees in reviewing manuscripts for the Vol. 25 (4), December 2012. Akram Dr M Kumari Dr (Ms) Jayoti Singh Dr NP IIPR, Kanpur NBPGR, New Delhi IIPR, Kanpur Basu Dr PS Lakhmi Dr Swarn Singh Dr ON IIPR, Kanpur IARI, New Delhi BHU, Varanasi Chaudhary Dr AK Malathi Dr (Ms) VG Singh Dr Rakesh IIPR, Kanpur IARI, New Delhi NBPGR, New Delhi Gupta Dr (Ms) Om Panwar Dr JDS Singh Dr Sarvjeet JNKVV, Jabalpur IARI New Delhi PAU, Ludhiana Gupta Dr SC Praharaj Dr CS Tripathi Dr AK ARS, Durgapura, Rajasthan IIPR, Kanpur CSAUAT, Kanpur Iquabal Dr Asif Pratap Dr Aditya Verma Dr Prasoon IASRI, New Delhi IIPR, Kanpur IIPR, Kanpur Jha Dr SK Raje Dr Ranjeet Vishwanath Dr KP IARI, New Delhi IARI, New Delhi UAS, Raichur Katiyar Dr PK Ram Dr Baldev Wadaskar Dr R IIPR, Kanpur ZARS, Kota Akola Kaur Dr (Mrs) Lavinder Reddy Dr AR Yadav Dr ND PAU, Ludhiana ICARISAT, Patencheru CAZARI, Bikaner Kumar Dr D Singh Dr Archana Yadav Dr SK CAZRI, Jodhpur IGFRI, Jhansi CRIDA, Hyderabad Kumar Dr Rajendra Singh Dr IP DSR, Mau IIPR, Kanpur Kumar Dr Rajesh Singh Dr Jagdish IIPR, Kanpur IIPR, Kanpur ii Journal of Food Legumes 25(4), 2012 REVIEWER INDEX (2012) The Editorial Board gratefully acknowledges the help rendered by following referees in reviewing manuscripts for the Vol. 25 (2012) Akram Dr M IIPR, Kanpur 208024 (U.P.) Gabhane Dr VV PDKV, Akola 444104 Basu Dr PS IIPR, Kanpur 208024 Gill Dr BS PAU,Ludhiana141004 (PB) Bhatia Dr VS DSR, Khandwa Road, Indore 452 001 (M.P.) Gupta Dr (Mrs) Om JNKVV,Jabalpur 482004(MP) Bhatnagar Dr Pooja ICRISAT, Patancheru, Hyderabad Choudhary Dr AK IIPR Regional Station, Dharwad 580 005 (Karnataka) Gupta Dr Sanjeev IIPR, Kanpur 208024 Gupta Dr SC ARS, Durgapura, Rajasthan Kumar Dr D CAZRI Jodhpur 342 003 (Rajasthan) Kumari Dr (Ms) Jayoti NBPGR, New Delhi 110012 Kumar Dr. Jitendra IIPR, Kanpur 208024 (U.P.) Kumar Dr. Narendra IIPR,Kanpur 208024 (U.P.) Kumar Dr Rajendra DSR, Mau Harer Dr PN MPKV Akola 444 104 (M.H.) Kumar Dr Rajesh IIPR, Kanpur 208024 Hussain Dr Aftab UAS, Bangalore Kumar Dr Ram Vinod IGFRI, Jhansi (U.P.) Choudhary Dr VP PDFSR, Modipuram Meerut 250110 Iquabal Dr Asif IASRI, New Delhi-110012 Kumar Dr Sanjeev ICER, Patna Das Dr Anchal IARI, New Delhi-110012 Jha Dr SK IARI, New Delhi-110012 Mahapatra Dr SD CRRI, Cuttack (Odisha) Das Dr Manish DMAPR Anand 387 310 (Gujarat) Kalaimagal Dr (Mrs) T TNAU Coimbatore 641003 (T.N.) Malathi Dr (Ms) VG IARI, New Delhi 110012 Datta Dr S IIPR, Kanpur 208024 Kandan DrA NBPGR, New Delhi 110012 Mallikarjuna Dr N ICRISAT Patancheru, Hyderabad Dillon Dr MK IARI, New Delhi 110012 Katiyar Dr PK IIPR, Kanpur 208024 Mandal Dr Asit B CRIJAF, Barrackpore (WB) Dixit Dr GP IIPR, Kanpur 208024 (UP) Kaur Dr (Mrs) Jagmeet PAU, Ludhiana 141 004 Mudalagiriyappa Dr UAS,GKVK, Bangalore 560065 Dixit Dr Harsh IARI, New Delhi 110 012 Kaur Dr (Mrs) Lavinder PAU, Ludhiana 141 004 Naimuddin Dr IIPR, Kanpur 208024 Dudeja Dr SS CCSHAU Hisar 125 004 (Haryana) Khan Dr MR AMU, Aligarh Nath Dr Rajiv BCKVV, Kalyani (WB) Choudhary Dr RG IIPR, Kanpur 208024 Journal of Food Legumes 25(4), 2012 Panda Dr PK WTC, Bhubaneswar (Odisha) Pandaya Dr M IIVR, Varanasi 221 305 (U.P.) Panwar Dr JDS IARI New Delhi 110012 Patil Mr Prakash G IIPR, Kanpur 208 024 (UP) Praharaj Dr Chandra Sekhar IIPR, Kanpur 208024 (UP) Prakash Dr Vijay ARS, Sriganganagar 335 001 (Rajasthan) Pratap Dr Aditya IIPR, Kanpur 208024 Rai Dr AB IIVR, Varanasi 221 305 (U.P.) Raghvani Dr B R JAU, Junagadh 362001 Raje Dr RS IARI, New Delhi 110012 Ram Dr Baldev ZARS, Kota (Rajasthan) Reddy DrA Amarender ICRISAT Patancheru 502 324 (AP) Reddy Dr AR ICARISAT Patancheru 502 324 (AP) Sah Dr (Mrs) Uma IIPR, Kanpur 208024 (UP) Samad Dr A CIMAP, Lucknow Saxena Dr KB ICRISAT, Patancheru, Hyderabad Sengupta Dr Kajal BCKV, Nadia, 741 252 West Bengal Singh Dr (Mrs) Archana IGFRI, Jhansi 284128 (U.P.) iii Tripathi Dr AK CSAUAT, Kanpur 208002 (U.P.) Upadhyay Dr JP RAU, Pusa, Samastipur (Bihar) Singh Dr Awnindra CARI, Portblair Varshney Dr Rajeev K ICRISAT Patancheru 502 324 (AP) Singh Dr I P IIPR, Kanpur 208024 (UP) Venkatesh DR MS IIPR, Kanpur 208024 (U.P.) Singh Dr Jagdish IIPR, Kanpur 208024 (UP) Verma Dr DK IARI Regional Station, Indore (MP) Singh Dr NP IIPR, Kanpur 208024 (UP) Singh Dr ON BHU, Varanasi Singh Dr PK IAS, BHU, Varanasi Singh Dr Rakesh NBPGR, New Delhi Singh Dr RC CIAE, Bhopal 462 038 Singh Dr Sarvjeet PAU.,Ludhiana 14100 Sohrab, Dr SS King Abdul Aziz University Saudi Arabia Solanki Dr IS IARI Regional Research Station Pusa, Samastipur 848125 Subramanian Dr S IARI, New Delhi -110012 Swarnlakshami Dr IARI, New Delhi 110012 Verma Mr Prasoon IIPR, Kanpur 208024 (U.P.) Vijaylaxmi Dr (Mrs.) IIPR, Kanpur 208024 (UP) Vishwanath Dr KP UAS, Raichur (KTK) Wadaskar Dr R Akola (M.H.) Waldia Dr RS CCSHAU, Hisar 125 004 (Haryana) Yadav Dr ND CAZARI, Bikaner (Rajasthan) Yadav Dr Rasmi NBPGR, New Delhi-110012 Yadav Dr RC CCS HAU, Hisar 125004 Yadav Dr SK CRIDA, Hyderabad iv Journal of Food Legumes 25(4), 2012 AUTHOR INDEX A Ahmad Shahid (175) Akhtar J. (81) Akken M.K. (314) Akram Mohd (54,286) Alipatra A. (37) Andhalkar A.S. (128) B Babbar Anita (70,147) Bairwa R.K. (211) Balai O.P. (109) Bandopadhyay P. (37) Banerjee H. (37) Bansal Ravindra (18) Bansal Ravindra (273) Bansode V.V. (321) Barkhade U.P. (162) Barpete Surendra (14) Bhalkare S.K. (215) Bharathi M. (96) Bharathi M. (351) Bhardwaj R. (234) Bhardwaj S.K. (246) Bhareti Priyanka (89) Bhatia Ranjana (294) Bhattacharya A (50) C Chandra Subhash (326) Chandra Subhash (71) Changkija S.A.P.U.(282) Chaubey B.K. (93) Chaubey B.K. (348) Chaudhari D.J. (89,344) Chaudhary R.G (61) Chauhan Richa (187) Chauhan V.B. (76) Chimote, V.P. (9) Choudhary Meenu (361) Choudhary Poonam (361) D Dahiya S.S. (153) Deb D. (112) Dhanasekar P. (25) Dhiman Sushil (246) Dixit S.P. (246) Durairaj C. (83) G Ganeshmurthy A.N. (116) Gangwar S. (45) Garg S.K. (131) Garkoti Ankita (103) Garkoti Ankita (358) Gera Rajesh (294) Gera Rajesh (39) Ghai Navita (206) Ghatak Abhijeet (355) Gill B.S. (314) Gill K.K. (125) Gill R.K. (159) Gill. B.S. (206) Gopalakrishna (273) Goud V.V. (128,243) Gowda M. Byre (194) Goyal Meenakshi (206) Goyal Reeti (59) Goyal Reeti (314) Gupta Om (139) Gupta S.C. (45) Gupta S.K. (234) Gupta Sudhir Kumar (273) H Holmesheoran M.E. (334) I Imtiaz M. (79) Iquebal M.A. (31,147) J Jadhao V.P. (215) Jadhav A.S. (9) Jat H.L. (227) Jat Shankar Lal (239) Jayalakshmi V. (94) Jayamani P. (279) Jayaram Neetha (135) Jyothirmayi G. (94) K Kale H.B. (243) Kale A.A. (9) Kamaluddin (175) Kandalker V.S. (231) Kant Rama (1,102) Kaur Gurpreet (234) Kaur Jagmeet (206) Kaur Jasdeep (206) Kaur Livinder (79) Kaur Prabhjit (234) Kaur Ramndeep (234) Keval Ram (249) Khan M.N. (175) Khanna Veena (125) Khare D. (200) Khode N.M. (243) Khulbe R.K. (89,183) Kotecha P.M. (321) Krishna Bal (61) Krishna K. Ram (109) Kumar Ajay (234) Kumar C.V.S. (334) Kumar D. (1) Kumar D. (255) Kumar Deepak (18) Kumar Dhirendra (66) Kumar J. (165) Kumar K. (66,348) Kumar Narendra (41,116,131) Kumar Rajesh (61,85,340) Kumar Rakesh (121) Kumar Sanjeev (179) Kumar Shiv (14) Kumar Varun (294) Kumar Y. (81) Kumari Anupma (121) Kumawat S.R. (156) Kushwaha K.P.S. (355) L Lajjavati (100) Lakhera J.P. (227,326) Laxmi Vijay(300) M Malhotra R.S. (79) Manjunath B. (135) Meena D.S. (306) Meena H.N. (239) Meena H.P. (165) Mishra J.P. (41,55,310) Mishra Madhuri (139) Mondal C. (37) Mula M.G. (334) Mula R.P. (334) Journal of Food Legumes 25(4), 2012 Muniyappa V. (135) Murmu H. (81) Muthaiah A.R. (171) N Naik Satheesh S.J. (194) Naimuddin (31,54,286) Naphade S.A. (344) Narasimhamurthy G.M. (249) P Pannu R.K. (153) Panwar R.K. (183) Parihar C.M. (239) Parmar Dinesh (14) Parmar R.K. (231) Parmila C.K. (194) Patil A.N. (142,215) Pawar V.D. (9,321) Pawar S.V. (9) Praharaj C.S. (41,61,310,330) Prakash V. (147) Prameela H.A. (135) Punia S.S. (306) Purushottam (61,330,340) R Rai V.P. (179) Rajesh Kumar J. (83) Ram Baldev (51,306) Ram Hari (125) Ramappa H.K. (194) Ramesh S. (194) Rathi Manisha (139) Rathod K.S. (153) Ravikumar R.L. (18) Reddy C.K.K. (94) Reedy K.S. (25) Reena G.A. Mary (194) Rodge A.B. (255) Roy A.K. (112) Roy Rina (236) Rungsung S Sah Uma (340) Sahu Pooja (200) Saini N. (200) Sandhu J.S. (79,150,234) Santosh (361) Sarika (31) Sathya M. (279) Satpute N.S. (162) Savitha B.N. (18) Saxena K.B. (334,351) Sekhon H.S. (125) Sepehya Swapana (246) Sewak Shiv (31) Sharma O.P. (361) Sharma S.K. (227) Sharma Sucheta (314) Sharma N.C. (14) Shende N.V. (222) Shinde G. (18) Shivay Yashbir Singh (239) Shukla R.K. (291) Shukla S.S. (70) Singh A.K. (179) Singh Amitesh Kumar (73) Singh Anupma (183) Singh Archana (112,187) Singh Avtar (236) Singh Bansa (58) Singh D.P. (89) Singh Guriqbal (125) Singh Inderjit (150) Singh J.P. (76) Singh Johar (159) Singh K.K. (41,58,116,310) Singh Lakhan (330) Singh M.K. (131) Singh Manpreet (159) Singh N.P. (31,97,187) Singh O.N. (121) Singh P. (227, 326) Singh P.K. (81) Singh P.S. (36, 291) Singh Prakash (66) Singh R.B. (76) Singh R.S. (73) Singh Rupinderpal (150) Singh S.K. (61,330,340) v Singh Sarvjeet (150,159) Singh U.P. (112) Singhn Ranjeet (66) Sirari A. (79) Sirvastava R.K. (1) Solanki, R.K. (31) Srimathy M. (279) Srinivasarao C.H. (116) Srivasta A.N. (200) Srivastava R.K. (348) Srivastava A. (147) Surinder Kaur (76) Swarnalakshmi K. (116) Srivastav R.K. (1,102) Srivastava S.P. (70) Srivastava Arpita (70) Srivastava C.P. (179) T Tetarwal J.P. (306) Thakare S.S. (222) Thiruvengadam V. (171) Tyagi N. (179) Thorat S.S. (321) Tikle A.N. (231) Tiwari Archna (187) Tripathi H.S. (358) Tripathi N. (147,200) U Ughade Jayashri (142) V Vati Lajja (355) Venkatesha S.C. (194) Verma Prasoon (131) Vijyalakshmi (50,45) W Wadaskar R.M. (142,215) Wungsem (282) Y Yadav L.R. (106,361) Yadav B.L. (156) Yadav C.B. (348) Yadav Indu Singh (97) Yadav N.K. (291) vi Journal of Food Legumes 25(4), 2012 SUBJECT INDEX A Achievement motivation (227) ADF test (344) Amino acids (286) Anti-nutrients (321) ARCH- GARCH (344) Arid legumes (255,273,326) Ascochyta blight (79) Ascochyta rabiei (79) Association (227) Azuki bean (273) B B-biotype (135) Bed planting (236) Bio-agents (358) Bio-efficacy (291) Bio-fertilizer (121) Bioinculants (73) Blackgram (24,89) Boron (37) Breeding (355) Broadcasting (131) Bulk Method (165) Bundelkhand region (61,330) C Calendar based application (142,215) CGMS lines (231) Character Association (93,348) Chickpea (27,31,37,41,70,94,97,100, 139, 142,147, 150,165, 187, 234, 236, 291, 310, 355) Chickpea regeneration (9) Chlorophyll content (45) Cluster analysis (24,31, 70,109,147) Coat protein (286) Co-integration (344) Combining ability (231) Communication behavior (326) Compact dwarf (25) Competitive incides (128) Constraint analysis (340) Conventional tillage (236) Cooking characteristics (321) Correlation (66, 70,179) Cowpea (273,255) Credit behavior (227) Crop geometry (243) Cropping Sequence (58) D D2 analysis (279) Demonstration (326) Dibbling (131) Diseases (200) Dolichos Bean (18) Drought (94) Dry matter accumulation (50) Dry matter production (73) Dry root rot (139) Durable resistance (79) E Economic (236,239,243) Economic motivation (326) Embryonic axis (9) Epistasis (1) ET (41) F Faba bean (348) Farmers participation (330) Fenugreek (156) Fertilizer level (159) Field bean (18) Fieldpea (121,310) Fodder yield (109) FPARP (330) French bean (54) Functional properties (321) Fusarium wilt (81,358) G G × E interaction (282) Garden pea (246) Gene action (1) General combining ability (171) Generation mean analysis (1) Genetic divergence (150,279) Genetic diversity (18,89,147,194,200) Genetic male sterility (171) Genetic variability (70,348) Genetics variation (109) Genotypes (125) Genotypic correlation (31) Germination (351) Germplasm (18,31) Gibberellic acid (25) Grain yield (73,109) Gram pod borer (142) Granger Causality Test (344) Grass pea (109) Groundnut bud necrosis virus (54) Growth and reproductive stages (187) Growth (37,222) Guar (255) H Harvest index (159) Helicoverpa armigera (83,291) Heritability (66) Heterobeltiosis (102) Horse gram (255) Hybrids (351) I Imazethapyr (306) Inbred cultivars (351) Inbreeding depression (102) Induced variability (109) Indigenous pheromone lures (83) Inheritance (94) Inorganic nutrient (121) Interspecific derivatives (150) Insecticides (36,249) Integrated nutrient management (246) Iron (45) Irrigation (45) Irrigation (300) ISSR markers (89) K Kabuli chickpea (79) L LAI (73) Land equivalent ratio (128) Lathyrus sativus (109) Leaf angle (50) Leaf area (50) Leaf chlorophyll (94) Legumes – wheat rotation (116) Lentil (66,81,300,358) Lentil genotypes (159) Lodging (179) M Mahalanobis D2 statistics (150) Melanagromyza obtusa (249) Meloidogyne javanica (58) Micrografting (97) Mid parent heterosis (102) Milling quality (70) Molecular marker (200) Molecular markers (147) Molybdenum (45) Morpho-physiological traits (206) Mothbean (255,361) Mould (355) Mulch (310) Mulching (41) Mungbean (37,50,89,109,135, 211,) Mutant progenies (109) MYMIV (286) MYMV (135) Journal of Food Legumes 25(4), 2012 N N Management (153) NaCl (187) Nematode (58) Net returns (361) Newer insecticides (142,215) Nod C (294) Nodulation (125) NPK uptake (153,246) NPKSZn (243) NSm gene (54) Nucleotides (286) Nutrient accumulation (239) Nutrient availability (239) Nutrient uptake (128) O Oil (314) Organic nutrient (121) P Parity Index (222) Path analysis (70,348) Path and cluster analysis (179) Path coefficient analysis (348) Path coefficient (66) PCR (286) Pea (179) Pedigree Method (165) Pendal type (18) Phaseolus vulgaris (54) Phenological days (50) Phenotypic correlation (31) Phosphorus levels (361) Phosphorus levels (73,211,361) Photoperiod (25) Photosynthetic rate (50) Phylogeny (294) Phytic acid (314) Pigeonpea (25,76,162,171, 194,215, 231,334,344) Pigeonpea hybrid (243) Pigeonpea Podfly (249) Pigeonpea Transplanting (128) Plant growth regulators (206) Planter (131) Planting time (125) Pod borer complex (162,215) Polymorphism (89,194) Pra-harvest sprouting (183) Presoak treatment (321) Price movement (344) Principal Component Analysis (31,109) Progressiveness (227) Protein (314) PSB (45) Pulse production technologies (340) Pulse treatment (9) Pulses (61,330) Purple seed stain (200) Q QTLs Soybean (200) Qualitative short day (25) Quality seed production (75) Quality traits (66) Quizalofop ethyl (306) R Rabi (25) Rainfed (61) 16S Rdna (294) Regeneration (97) Residues incorporation (116) Resistance (139,355) RFLP (294) Rhizobia (294) Rhizobium (45) Rhizoctonia bataticola (139) Rice bean (282,321) Rice fallow (25) Rice-wheat rotation (239) Root-knot nematode (58) RSC water (156) Rynaxypyr (162) S Salinity tolerance (187) Seasonal variation (344) Seed bed configurations (310) Seed drill (131) Seed inoculation (45) Seed size (234) Seed village system (334) Seed village system (334) Seed Yield (41,116,125,156,206,211,234, 282,300,310) Seedling vigour index (351) Selected Bulk Method (165) Single Seed Descrent Method (165) Socio economic status (326) Socio-economic status (227) Soil application (45) Soil fertility (128,153) Soil Properties (116) Sowing dates (159) Sowing equipment (131) vii Soybean genotypes (314) Soybean (128,153,175,206,222) SPAD chlorophyll meter (94) Specific combining ability (171) Spinosad (249) SSR markers (194,273) SSR (200) Stability analysis (175,282) Standard heterosis (102) Stem canker (76) Sterility Mosaic Disease (194) Sulphur levels (211) Summer legumes (239) T Technological input assessment (61) Technology adoption (340) Technology dissemination process (340) Temporel variation (222) Thidiazuron (9) Tillage (41) Total soluble sugars (314) Transferability (273) Transgenics (97) Transmission (135) Trap catches (83) Tropical legumes 2 (334) Truthfully labeled seed (334) Trypsin inhibitor (314) U Urdbean (1,58,102,125) V Variation (286) Varietal improvement (81) Vermicompost (121) Vermicompost Zn mobility ratio (156) Vigna (89) Vigna mungo (1,279) Vigna species (273) W Water use efficiency (41,310) Weather parameters (83) Weed control efficiency (306) Weed index (306) Whitefly (135) Y Yield (37,66,175,179) Yield attributes (211,206,246) Yield components (175,300) Z Zero tillage (236) Zinc levels (361) viii Journal of Food Legumes 25(4), 2012 Indian Society of Pulses Research and Development Indian Institute of Pulses Research, Kanpur – 208 024 ISPRD Fellowship Awards 2012 To encourage pulses research and development, ISPRD admits its members as Fellows. Applications in the prescribed proforma are invited from eligible ISPRD members for the award of ISPRD Fellowship for the year 2012. Any member is eligible if he/she has been the member of the Society continuously preceding last 5 years and has at least 3 research papers related to food legumes (out of which, one must have been published in the Journal of Food Legumes). Only those 2 research papers, which were published in other Journals having NAAS rating at or above par with Journal of Food Legumes, will be considered. Filledin applications along with necessary enclosures should be submitted to the Secretary, Indian Society of Pulses Research and Development, IIPR, Kanpur 208 024 (U.P.) by February 28, 2013. Those who are already Fellows of the Society need not apply. -sdA K Choudhary Secretary, ISPRD [email protected] Journal of Food Legumes 25(4), 2012 ix PROFORMA Indian Society of Pulses Research and Development Indian Institute of Pulses Research, Kanpur 208 024 ISPRD Fellowship Awards 2012 1. Name in Full : 2. Father’s Name : 3. Date of Birth : 4. Designation : 5. Field of specialization : 6. Address (with Telephone No., E-mail and Fax) Passport Size Photo Office : ________________________________________________________________________________________ Residence : ____________________________________________________________________________________ 7. Academic career Degree 8. University/Institution Year Distinction, if any Employment Record and Experience Designation Organization Period 9. Enlist only three best publications indicating (a) Name of author(s), (b) year, (c) title, (d) name of journal, volume no. and page nos. 10. Date/year of life/ordinary membership of ISPRD. 11. Special services rendered to ISPRD. Signature of applicant with date Head of Institute/Organization (Optional) x Journal of Food Legumes 25(4), 2012 Obituary Dr Laxman Singh: Former Project Director, Directorate of Pulses Research, Kanpur Dr Laxman Singh (78), an eminent scientist and research manager passed away on 19, December 2012 at his residence in Indore (Madhya Pradesh). He was born on 4 April, 1935 in Naulana, Gautampura, Indore (MP). Dr Singh is survived by a son and a daughter. He completed his B Sc from Gwalior and M Sc (Ag) from Agriculture College, Kanpur. He also received PhD from a US university. Dr Singh worked as Professor in JNKVV, Jabalpur before joining as Project Director, Directorate of Pulses Research, (Presently IIPR), Kanpur. He was life member of ISPRD. He also served as consultant in the Caribbean Agricultural Research & Development Institute (CARDI), Trinidad and later on served ICRISAT, Hyderabad as Principal Pigeonpea Breeder (1986 - 1997). He also lived in the US but surrendered his green card and returned to India and started welfare work for the society. Dr Singh had a great sense of humor and was compassionate person. As per his last wish, his eyes and body were donated for medical research to MGM Medical College, Indore (MP). May almighty give the bereaved family the strength and courage to face this loss. Journal of Food Legumes 25(4), 2012 Instructions to Authors Journal of Food Legumes (formerly Indian Journal of Pulses Research) publishes original papers, short communications and review articles by renowned scientists, covering all areas of food legumes research. The paper should not have been published or communicated elsewhere. Authors will be solely responsible for the factual accuracy of their contribution. Language of publication is English (British). Please send your manuscript to following address: Secre tary ISPRD Indian Institute of Pulses Research Kalyanpur, Kanpur 208 024, India Email: [email protected] Manuscript must be submitted through e-mail. You should also submit a hard copy of your manuscript for our official record. Besides author(s) is required to submit a certificate that the paper is exclusive for Journal of Food Legumes. Manuscripts must conform to the Journal style (see the latest issue). Correct language is the responsibility of the author. After having received your contribution (date of submission), there will be a review process before the editorial board takes decision regarding acceptance for publication. One copy of the revision together with the original manuscript must be returned to the subject editor or Secretary. The submitted paper must be one complete word document file comprising a title page, abstract, text, references, tables, figure legends and figures. When preparing your text file, please use only Times New Roman for text (12 point, double spacing) and Symbol font for Gr eek letters to a void ina dver tent cha racter substitutions. Format Every original paper should be divided into the following five sec tions: ABSTRACT, Key words, INTRODUCTION, MATERIALS AND METHODS, RESULTS AND DISCUSSION, and REFERENCES. The manuscript should be typed on one side of the paper only, double spaced, and with 4-cm margins with page and line numbers. The main title must be capital bold. Subheading must be bold italic and Sub-sub heading normal italic. At the head of the manuscript, the following information should be given: the title of the paper, the name(s) of the author(s), the institute where the research was carried out, the present addresses of the authors (foot note) and of the corresponding author (if different from above Institute). Authors are required to provide running title of the paper. You must supply an E-mail address for the corresponding author. The abstract should contain at least one sentence on each of the following: objective of investigation (hypothesis, purpose, aim), experimental material, method of investigation, data collection, result and conclusions. Maximum length of abstract is 175 words. Up to 10 key words should be added at the end of the abstract and separated by comma. Key words must be arranged alphabatically (e.g., EMS, Gamma ray, Mungbean, Mutations, Path coefficient, ......). Each figure, table, and bibliographic entry must have a reference in the text. Any correction requested by the reviewer should also be integrated into the file. Manuscript file including tables must be in MS Word and Windows-compatible and must not contain any files other than those for the current manuscript. Please do not import the figures into the text file. The text should be prepared using standard software (Microsoft Word); do not use automated or manual hyphenation. xi Length Manuscripts should not exceed a final length of 15 printed pages, i.e., 5,000 words, including spaces required for figures, ta bles and lis t of references. Manuscr ipts for short communications should not exceed 3000 words (3 printed pages, with not more than a total of 2 figures or tables). Units, abbreviations and nomenclature For physical units, unit names and symbols, the SI-system should be employed. Biological names should be given according to the latest international nomenclature. Botanical and zoological names, gene designations and gene symbols are italicised. Yield data should be reported in kg/ha. The name of varieties or genotypes must start and end with single inverted comma (e.g., ‘Priya’, ‘IPA 204’, ......). Tables and Figures Tables and figures should be limited to the necessary minimum. Please submit reproducible artwork. For printing of coloured photogr aph, aut hors wil l be charged Rs. 4000/- per photograph. It is essential that figures are submitted as highresolution scans. References The list of references should only include publications cited in the text. They should be cited in alphabetical order under the first author ’s name, listing all author s, the ye ar of publication and the complete title, according to the following examples: Becker HC, Lin SC and Leon J. 1988. Stability analysis in plant breeding. Plant Breeding 101: 1-23. Sokal RR and Rholf FJ. 1981. Biometry, 2nd Ed. Freeman, San Francisco. Tandon HLS. 1993. Methods of Analysis of Soils, Plants, Water and Fertilizers (ed). Fertilizer Development and Consultation Organization, New Delhi, India. 143 pp. Singh DP. 1989. Mutation breeding in blackgram. In: SA Farook and IA Khan (Eds ), Breedi ng Food Legumes. Premier Publishing House, Hyderabad, India. Pp 103-109. Takkar PN and Randhawa NS. 1980. Zinc deficiency in Indian soils and plants. In: Proceedings of Seminar on Zinc Wastes and their Utilization, 15-16 October 1980, Indian Lead-Zinc Information Centre, Fertilizer Association of India, New Delhi, India. Pp 13-15. Satyanarayan Y. 1953. Photosociological studies on calcarious plants of Bombay. Ph.D. Thesis, Bombay University, Mumbai, India. In the text, the bibliographical reference is made by giving the name of the author(s) with the year of publication. If there are two references, then it should be separated by placing ‘comma’ (e.g., Becker et al. 1988, Tandon 1993). If references are of the same year, arrange them in alphabatic order, otherwise arrange them in ascending order of the years. While preparing manuscripts, authors are requested to go through the late st issue of the journal. Authors are also required to send the names & E-mail address of at least 3-4 reviewers appropriate to their articles. 16. Dissemination of pulse production technologies in Uttar Pradesh : A micro-level analysis Rajesh Kumar, S.K. Singh, Purushottam and Uma Sah 17. Pigeonpea (Cajanus cajan L.) price movement across major markets of India D.J. Chaudhari and A.S. Tingre SHORT COMMUNICATIONS 18. Genetic variability, character association and path coefficient analysis in faba bean B.K. Chaubey, C.B. Yadav, K. Kumar and R.K. Srivastava 19. A comparative study of hybrid and inbred cultivars for germination and other related traits of pigeonpea M. Bharathi and K.B. Saxena 20. Screening of chickpea (Cicer arientinum L.) genotypes for identification of source of resistance to Botrytis grey mould Lajja Vati, K.P.S. Kushwaha and Abhijeet Ghata 21. Management of Fusarium wilt of lentil using antagonistic microorganisms in Tarai region of Uttarakhand Ankita Garkoti and H.S. Tripathi 22. Effect of phosphorus and zinc on yield and economics of mothbean under semiarid conditions L.R. Yadav, Poonam Choudhary, Santosh, O.P. Sharma and Meenu Choudhary List of Referees for Vol. 25 (4) Reviewer Index (2012) Author Index (2012) Subject Index (2012) ISPRD Fellowship awards, 2012 Obituary 340 344 348 351 355 358 361 i ii iv vi viii x ISSN 0970-6380 Online ISSN 0976-2434 Volume 25 Journal of Food Legumes I SPR D 1987 Number 4 December 2012 Contents RESEARCH PAPERS 1 Status, scope and strategies of arid legumes research in India- A review D. Kumar and A.B. Rodge 2. Transferability of cowpea and azuki bean derived SSR markers to other Vigna species Ravindra Bansal, Sudhir Kumar Gupta and T. Gopalakrishna 3. Genetic diversity studies in blackgram (Vigna mungo L. Hepper) M. Srimathy, M. Sathya and P. Jayamani 4. Stability analysis for seed yield and its component traits in rice bean (Vigna umbellata) Wungsem Rungsung and S.A.P.U. Changkija 5. Sequence comparison of coat protein gene of Mungbean yellow mosaic India virus isolates infecting mungbean and urdbean crops Naimuddin and M. Akram 6. Bio-efficacy of some insecticides against H. armigera (Hubner) on chickpea (Cicer arietinum L.) P.S. Singh, R.K. Shukla and N.K. Yadav 7. Comparison of nodC and 16S rDNA gene analysis of rhizobia associated with legumes of arid and semi-arid regions of Haryana Rajesh Gera, Ranjana Bhatia and Varun Kumar 8. Phenology, dry matter distribution and yield attributes under normal and drought stress conditions in Lentil (Lens culinaris Medik.) Vijay Laxmi 9. Efficacy of post emergence herbicides on weed control and seed yield of rajmash (Phaseolus vulgaris L.) Baldev Ram, S.S. Punia, D.S. Meena and J.P. Tetarwal 10. Enhancing water use efficiency and production potential of chickpea and fieldpea through seed bed configurations and irrigation regimes in North Indian Plains J.P. Mishra, C.S. Praharaj and K.K. Singh 11. Variability in the nutrients, antinutrients and other bioactive compounds in soybean (Glycine max (L.) Merrill) genotypes Reeti Goyal, Sucheta Sharma and B.S. Gill 12. Effect of presoak treatment on cooking characteristics and nutritional functionality of rice bean V.D. Pawar , M.K. Akkena, P.M. Kotecha, S.S. Thorat and V.V. Bansode 13. Factors associated with economic motivation of legume growers in desert area of Rajasthan Subhash Chandra, P.Singh and J.P. Lakhera 14. Farmers participatory approach in seed multiplication of pulses in Bundelkhand region - A case study Purushottam, S.K. Singh, C.S. Praharaj and Lakhan Singh 15. Tropical Legumes 2 pigeonpea seed system in India: An analysis M.E. Holmesheoran, M.G. Mula, C.V.S. Kumar, R.P. Mula and K.B. Saxena Published by Dr AK Choudhary, Secretary on behalf of The Indian Society of Pulses Research and Development (www.isprd.in) from Indian Institute of Pulses Research, Kanpur-208 024 Phone : 09019870914 E-mail: [email protected], [email protected] at Army Printing Press, 33, Nehru Road, Sadar Cantt. Lucknow-2 Ph.: 0522-2481164 For free download of JFL articles, please also visit: www.indianjournals.com 255 273 279 282 286 291 294 300 306 310 314 321 326 330 334