October 2-7, 2012 Hyderabad Marriott Hotel & Convention Centre Hyderabad, India Patrons: William D Dar, Director General International Crops Research Insititute for the Semi-Arid Tropics, India S Ayyappan, Director General, Indian Council of Agricultural Research, India Chair: Rajeev K Varshney, International Crops Research Insititute for the Semi-Arid Tropics (ICRISAT) Organizing Committee: S K Datta, Indian Council of Agricultural Research, India C L L Gowda, International Crops Research Insititute for the Semi-Arid Tropics, India H S Gupta, Indian Agricultural Research Institute, India D A Hoisington, International Crops Research Insititute for the Semi-Arid Tropics, India J B Misra, Directorate of Groundnut Research, India N Nadarajan, Indian Institute of Pulses Research, India Amita Pal, Bose Institute, India N K Singh, National Research Centre on Plant Biotechnology, India S K Srivastava, Directorate of Soybean Research, India A K Tyagi, National Institute of Plant Genome Research, India International Advisory Board: David Bertioli,University of Brasilia, Brazil Paula Bramel, International Institute of Tropical Agriculture, Nigeria Doug Cook, Univeristy of California-Davis, USA Martin Crespi, ISV-CNRS, France Noel Ellis, Aberystwyth University, UK Peter Gresshoff, University of Queensland, Australia Maria Harrison, BT Institute, Cornell University, USA Georgina Hernández, UNAM, Mexico Sachiko Isobe, Kazusa DNA Research Institute, Japan Scott Jackson, University of Georgia, USA Günter Kahl, University of Frankfurt am Main, Germany Suk-Ha Lee, Seoul National University, Korea Da Luo, Sun Yat-sen University, China Teresa Milan, Universidad de Córdoba, Spain Maria Monteros, Noble Foundation, USA Lijuan Qiu, NFCRI/CAAS, China Karam Singh, CSIRO Plant Industry, Australia Richard Thompson, INRA-Dijon, France Bert Vandenberg, University of Saskatchewan, Canada Programme VI International Conference on Legume Genetics and Genomics (ICLGG) (October 2-7, 2012) Hyderabad Marriott Hotel & Convention Centre Hyderabad, India Tuesday, October 2, 2012 09:00–17.00 Registration & Parallel Workshops Time Speaker Inaugural Session Title 17.00-17.10 17.10-17.30 17.30-17.50 17.50-18.30 Welcome Inaugural address Inaugural address Legume genomics in agriculture: An example of harnessing Medicago genomics for forage improvement 19.00 Rajeev Varshney, ICRISAT William Dar, DG, ICRISAT Swapan Datta, DDG-CS, ICAR Richard Dixon, The Noble Foundation Welcome dinner Wednesday, October 3, 2012 Theme: Next Generation Genomics Co-chairs: David Bergvinson & Tim Close 8.30-9.00 Scott Jackson (USA) 9.00-9.30 Suk-Ha Lee (Korea) Genome sequence for common bean and analysis of wild relatives Toward completion of genome sequence of mungbean 9.30-9.45 9.45-10.00 10.00-10.30 Noel Ellis (UK) Robert Stupar (USA) RAD tags for the characterization of fast neutron mutants in pea Exploring structural variation in the soybean genome Coffee Break Theme: Symbiosis and Development (I) 10.30-11.00 11.00-11.30 11.30-12.00 12.00-12.15 12.15-12.30 Giles Oldroyd (UK) Jens Stougaard (Denmark) Peter Gresshoff / Paul Scott (Australia) Julia Frugoli (USA) Andreas Niebel (France) 12.30-13.30 Lunch Theme: Symbiosis and Development (II) Co-chairs: Georgina Hernandez & Karam Singh Decoding calcium oscillations during symbiotic signaling The role of LysM type receptors in Nod Factor perception Sensing, signals, suppression, symbiosis, soybean The autoregulation of nodulation in Medicago truncatula The CCAAT-box binding factors MtNF-YA1&2 are transcriptional regulators required for Nod factor signaling, rhizobial infection and nodule development Co-chairs: Richard Thompson & Jens Stougaard Functional genomics of symbiotic nitrogen fixation in Medicago truncatula Transcriptome of common bean (Phaseolus vulgaris) through RNA-seq: nodulation, symbiotic nitrogen fixation, transcription factors Antimicrobial peptides in symbiosis Multi-locus molecular phylogeny and allelic variation in a transcription factor gene suggest the multiple independent origins of kabuli chickpea The same or different? Flowering on time in the model legume Medicago 13.30-14.00 Michael Udvardi (USA) 14.00-14.30 Georgina Hernandez (Mexico) 14.30-15.00 15.00-15.15 Attilla Kereszt (Hungary) R Varma Penmetsa (USA) 15.15-15.30 Jo Putterill (New Zealand) 15.30-16.00 16.00-19.00 Coffee Break Poster Session: One minute oral presentation and poster presentation (snacks and drinks will be available during poster presentation) i VI ICLGG - Programme Thursday, October 4, 2012 Theme: Evolution and Diversity Time Speaker 8.30-9.00 David Bertioli (Brazil) 9.00-9.30 Jeff Doyle (USA) 9.30-09.45 Steven Cannon (USA) 09.45-10.00 10.00-10.30 Eric von Wettberg (USA) Coffee Break Co-chairs: Lijuan Qiu & Rajeev Varshney Title The repetitive component of the A genome of peanut (Arachis hypogaea L.) and its role in the erosion of genome sequence similarity over time Diversity and evolution of the perennial Glycine allopolyploid complex Using high-throughput sequencing data to speed the domestication of Apios americana - a potential new legume crop Legume and symbiont phylogeography of South Florida Theme: Harnessing Germplasm Resources 10.30-11.00 CLL Gowda (India) 11.00-11.30 11.30-11.45 Bert Vandenberg (Canada) HD Upadhyaya (India) 11.45-12.00 Timothy Close (USA) 12.00-12.15 Aditya Pratap (India) 12.15-12.30 MC Millawithanachi (Sri Lanka) 12.30-13.30 Lunch Theme: Abiotic Stress Co-Chairs: JB Misra & Clarice Coyne Genetic resources and genomics for enhancing the efficiency of crop improvement in grain legumes Interspecific convergence of lentil genetics and genomics Legumes germplasm subsets to select diverse trait-specific lines for use in improvement programs Coupling genome resources to biological traits and breeding objectives in cowpea (Vigna unguiculata (L.) Walp) Development of extra short duration genotypes in greengram (Vigna radiata L. Wilczek) Present status of cultivation, constraints, level of on-farm improvement and breeding requirements of cowpea (Vigna unguiculata L. Walp) based on survey in Monaragala district of Sri Lanka Co-chairs: Kailash Bansal & M Udayakumar 13.30-14.00 Henry Nguyen (USA) Exploitation of root system architecture and plasticity for improving drought resistance in grain legumes Transcriptional networks regulating legume root architecture adaptation to different environmental conditions Salinity-induced expression landscape: Fast reply transcripts and transcript variants for oxygen burst and signal transduction in chickpea An integrated framework combining crop physiology, genetics, and crop simulation modeling to improve drought resistance across legumes in the semi-arid tropics Identification of drought-induced transcription factors in peanut (Arachis hypogaea L.) 14.00-14.30 Martin Crespi (France) 14.30-15.00 Günter Kahl (Germany) 15.00-15.15 Vincent Vadez (India) 15.15-15.30 Charles Chen (USA) 15.30-16.00 Coffee Break 16.00-19.00 Poster Session: One minute oral presentation and poster presentation (snacks and drinks will be available during poster presentation) Friday, October 5, 2012 8.00-17.00 City tour & ICRISAT visit 18.30- 21.00 Cultural Program & Cocktail dinner @ ICRISAT ii VI ICLGG - Programme Saturday, October 6, 2012 Theme: Pathogenesis and Disease Resistance Time Speaker Title 8.30-9.00 Karam Singh (Australia) 9.00-9.30 Valerie Geffroy (France) 9.30-9.45 Sampa Das (India) 9.45-10.00 Patricia Guimaraes (Brazil) 10.00-10.30 Coffee Break Resistance to sap-sucking insect pests and soil borne necrotrophic fungal pathogens in a model legume, Medicago truncatula Evolution of subtelomeric disease resistance gene clusters in common bean (Phaseolus vulgaris) Molecular insight into the early defense signal modulation in chickpea upon Fusarium oxysporum f. sp. Ciceris attack Gene expression revealed during the interaction of resistant Arachis stenosperma and Meloidogyne arenaria Theme: Genomic Resources and Trait Mapping 10.30-11.00 Sachiko Isobe (Japan) 11.00-11.30 Ana Maria Torres (Spain) 11.30-11.45 Lutz Froenicke (USA) 11.45-12.00 Rohini Garg (India) 12.00-12.15 12.15-12.30 Kyujung Van (Korea) Eigo Fukai (Japan) 12.30-13.30 Lunch Theme: Genomics- Assisted Breeding 13.30-14.00 Jean-Marcel Ribaut (Mexico) 14.00-14.30 14.30-14.45 N Nadarajan (India) Bunyamin Taran (Canada) 14.45-15.00 15.00-15.15 Pooran Gaur (India) Donal O'Sullivan (UK) 15.15-15.30 Daniel Fonceka (France) 15.30-16.00 Coffee Break Co-chairs: Madan Bhattacharya & Amita Pal Co-chairs: Vidya Gupta & Scott Jackson Identification of optimum DNA markers sets for MAS in red clover with a novel approach based on ‘Adaboost’ algorithm Genomics research for faba bean (Vicia faba L.) breeding applications Generation of ultra-dense genetic maps for peanut A and B genome RIL populations and a tetraploid mapping population Deep sequencing of chickpea transcriptome for gene discovery, marker development and gene expression studies Mungbean genome-wide SNP detection by Illumina HiSeq Establishment of a Lotus japonicus gene tagging population using the endogenous retrotransposon LORE1 Co-chairs: Noel Ellis & Teresa Millan Innovative approaches to increase tropical legume productivity in Africa and South Asia Genomics enabled molecular breeding of pulse crops Molecular marker platforms for chickpea, lentil, pea and common bean breeding Marker-assisted breeding for drought tolerance traits in chickpea SNP discovery and validation for genomic-assisted breeding of faba bean (Vicia faba L.) QTL mapping in AB-QTL and CSSL populations derived from the cross between a cultivated peanut (Arachis hypogaea L.) variety and a synthetic wild amphidiploid Special Session: Young Scientists in Legume Genetics and Genomics (co-sponsors: NSF, GRDC, GCP and SPG) 16.00-16.15 Wendy Vu (USA) 16.15-16.30 Vikas Belamkar (USA) 16.30-16.45 16.45-17.00 17.00-17.15 Aiko Iwata (USA) Jerome Verdier (USA) Himabindu K (India) 17.15-17.30 Matthew Nelson (Australia) iii Co-chairs: Doug Cook, Omid Ansari, Ndeye Ndack & Jeff Parker Genetic variation of parental environmental effects on seed transcriptome and transgenerational epigenetic inheritance of offspring response to salinity in M. truncatula Genome-wide sequence analysis and expression profiling using RNA-Seq implicates soybean (Glycine max) HD-Zip gene family members in dehydration and salt stress responses Genome structure, evolution and diversity in the genus Phaseolus Functional genomics of M. truncatula seed development Deciphering the role of stress responsive genes in chickpea (Cicer arietinum L.) Gene diversity and marker development in narrow-leafed lupin (Lupinus angustifolius L.) VI ICLGG - Programme 17.30-17.45 Alice Kosgei (Kenya) 17.45-18.00 Michael Fedoruk (Canada) 19.00 Gala dinner @ Hotel Marriott Sunday, October 7, 2012 Theme: Nutrition and Quality 8.30-9.00 Richard Thompson (France) 9.00-9.30 Claire Domoney (UK) 9.30-9.45 Ravindra Chibbar (Canada) 9.45-10.00 Sabhyata Bhatia (India) 10.00-10.30 Coffee Break Application of marker assisted selection to improve drought tolerance in adapted Kenyan chickpea varieties QTL analysis and heritability of seed size and shape In lentil (Lens Culinaris Medic.) Co-chairs: Irvin Widders & Tom Warkentin Dissecting the role of the endosperm in regulating Medicago truncatula seed filling Exploiting novel mutations affecting seed composition and visual traits in Pisum sativum L. (pea) for improved food and feed uses Genetic analysis of seed carbohydrates in chickpea (Cicer arientinum L.) Utilizing molecular mapping and transcriptome analysis to elucidate development of seed and nodule for enhancing yield and nutrition in chickpea Theme: Translational Genomics 10.30-11.00 11.00-11.30 Doug Cook (USA) T J Higgins (Australia) 11.30-12.00 Gengyun Zhang (China) 12.00-12.15 P Janila (India) 12.15-12.30 SR Uppalapati (USA) 12.30-13.30 Lunch Co-chairs: Jeff Ehlers & Howard Valentine Harvesting legume genomes for agronomic traits Protecting cowpeas from field and storage pests using gene technology Whole genome molecular marker assisted crop breeding based translational genomics approach in BGI Groundnut breeding at ICRISAT: From tools of genetics to genomics A search for novel sources of resistance in Medicago truncatula to improve fungal resistance in crop legumes Closing Ceremony 13.30-13.45 13.45-14.00 14.00–14.15 14.15–14.30 iv Summary- CLL Gowda Closing remarks- Dave Hoisington ICLGG Business Vote of thanks- Rajeev Varshney VI ICLGG - Programme Abstracts Inaugural Lecture RICHARD A. DIXON Director Plant Biology Division The Samuel Roberts Noble Foundation USA radixon@noble.org Richard A. Dixon is Distinguished Professor and Samuel Roberts Noble Research Chair, Senior Vice President, and Director of the Plant Biology Division at the Samuel Roberts Noble Foundation. He holds Adjunct Professorships at Rice University, the University of Texas at Austin, and the University of Oklahoma. He received his Bachelor’s and Doctoral degrees in Biochemistry and Botany from Oxford University (UK), and postdoctoral training in Plant Biochemistry at Cambridge University (UK). He was awarded the Doctor of Science degree for his research achievements by Oxford University in 2004. His research interests center on molecular biology and metabolic engineering of plant natural product pathways and cell walls in legumes and bioenergy crops. He has published over 400 papers on these and related topics in international journals, and has been named by the Institute for Scientific Information as one of the 10 most cited authors in the plant and animal sciences. Professor Dixon is Co-Editor-in-Chief of the journal BioEnergy Research, and a member of the Editorial Boards of four other international journals. He is Fellow of the American Association for the Advancement of Science, and was elected to membership of the US National Academy of Sciences in 2007. Selected Publications: 1. Chen F, Dixon RA. (2007). Lignin modification improves fermentable sugar yields for biofuel production. Nature Biotechnology 25: 759-761. 2. Pang Y, Peel GJ, Sharma SB, Tang Y, Dixon RA. (2008). A transcript profiling approach reveals an epicatechin-specific glucosyltransferase expressed in the seed coat of Medicago truncatula. Proceedings of the National Academy of Sciences USA 105: 14210-14215. 3. Naoumkina MA, Modolo LV, Huhman DV, Urbanczyk-Wochniak E, Tang Y, Sumner LW, Dixon RA. (2010). Genomic and co-expression analyses predict multiple genes involved in triterpene saponin biosynthesis in Medicago truncatula. Plant Cell 22: 850-866. 4. Wang H, Avci U, Nakashima J, Hahn MG, Chen F, Dixon RA. (2010). Mutation of WRKY transcription factors initiates pith secondary wall formation and increases stem biomass in dicotyledonous plants. Proceedings of the National Academy of Sciences USA 107, 22338-22343. 5. Gallego-Giraldo L, Escamilla-Trevino L, Jackson LA, Dixon RA. (2011). Salicylic acid mediates the reduced growth of lignin down-regulated plants. Proceedings of the National Academy of Sciences USA 108, 20814-20819. VI ICLGG - Abstracts Legume genomics in agriculture: An example of harnessing Medicago genomics for forage improvement L0 Dixon RA*, Chen F, Gallego-Giraldo L, Liu C, Zhao Q Plant Biology Division, Samuel Roberts Noble Foundation, Oklahoma, USA *E-mail: radixon@noble.org Abstract The available genetic and genomic resources in the model legume Medicago truncatula provide an excellent platform for trait development in the closely related forage species alfalfa (Medicago sativa). Two traits are of particular importance in this species; forage digestibility and protein quality (high by-pass protein and lower bloat potential). Lignocellulosic biomass such as alfalfa forage becomes more recalcitrant to digestion as it matures, and this is, at least in part, due to the presence of the cell wall polymer lignin. We have utilized the Medicago genomic and genetic resources to understand the pathways of lignin biosynthesis in this species and to modify lignin content and composition, leading to development of lines with good agronomic properties and commercially useful improvements in forage quality; these are currently undergoing regulatory assessment. However, in some low lignin alfalfa lines, the gains in cell wall digestibility are partially offset by reductions in plant yield. Targeting transcriptional regulators of the lignin or whole secondary cell wall pathways provides an alternative strategy for lignin down-regulation that potentially avoids metabolic spillover effects. Forward genetic screening of a transposon mutagenized population of M. truncatula has led to the identification of NAC and WRKY master switches that work as positive and negative regulators of lignification, respectively, and manipulation of lignin biosynthetic and regulatory genes has potential to deliver bioenergy feedstocks or forages combining increased cell wall density with improved digestibility. Finally, we will discuss attempts to engineer condensed tannins into alfalfa foliage in order to develop lines with better protein utilization efficiency; again, these studies are largely facilitated by the availability of resources for forward and reverse genetics, as well as transcriptome profiling, in M. truncatula. VI ICLGG - Abstracts Abstracts (and biography of invited speakers) Theme: Next Generation Genomics SCOTT A. JACKSON Director Center for Applied Genetic Technologies University of Georgia USA sjackson@uga.edu Scott Jackson is a Professor of Plant Genomics at the University of Georgia where he is the Director of the Center for Applied Genetic Technologies. His research focuses on the application of plant genomics to solve relevant agricultural problems, primarily on legumes and rice. Currently his lab is working on the discovery and implementation of useful genetic diversity for soybean and the leveraging genomics for related legumes (common bean, pigeonpea and cowpea). In rice, he has been part of a team that developed genomics infrastructure spanning the entire Oryza genus that is being used to clone genes, track introgressions and understand genome evolution and domestication. Selected Publications: 1. Jackson SA, Iwata A, Lee S-H, Schmutz J, Shoemaker RC. (2011). Sequencing crop genomes: approaches and applications. New Phyt. doi: 10.1111/j.1469-8137.2011.03804.x. 2. Sanyal A, Jetty A, Lu F, Yu Y, Rambo T, Currie J, Kollura K, Kim HR, Ma J, Wing R, Jackson SA. A genus-wide comparison of the Hd1 region reveals gene lability within the genus Oryza and disruptions to microsynteny in sorghum. Biol. and Mol. Evol. doi:10.1093/molbev/msq133. 3. Schmutz G, Stacey J, Specht D, Rokhsar RC, Shoemaker SA, Jackson. (2009). Genome sequence of the paleopolyploid soybean (Glycine max (L.) Merr.). Nature 463: 178-183. 4. Cannon SB, May GD, Jackson SA. (2009). Update on comparative genomics of legumes. Three sequenced legume genomes and many crop species: rich opportunities for translational genomics. Plant Physiol. doi: 10.1104/pp.109.144659. 5. Gill N, Findley S, Walling JG, Ma J, Stacey G, Doyle J, Jackson SA. (2009). Molecular and chromosomal evidence for allopolyploidy in soybean, Glycine max (L.) Merr. Plant Physiol. 0.1104/pp.109.137935. VI ICLGG - Abstracts Genome sequence for common bean and analysis of wild relatives 1* 1 2 5 3 Jackson SA , Iwata A , McClean P , Cregan P , Schmutz J , Rokhsar D I-NGG01 4 1 University of Georgia, Athens North Dakota State University, Fargo, USA 3 HudsonAlpha Institute, Huntsville, USA 4 Joint Genome Institute, Walnut Creek,USA 5 USDA-ARS, Beltsville, USA 2 *E-mail: sjackson@uga.edu Abstract Common bean (Phaseolus vulgaris) is an important food crop in much of the developing world. We report a genome sequence for common bean composed of both next generation and Sanger sequences to result in a high quality, genetically anchored sequence. A majority of the sequences were 454derived but fosmid and BAC end sequences were used to provide better scaffolding and contiguity resulting in a genome where the contig N50 and scaffold N50 were 39 kb and 38.2 Mb, respectively. Over 9000 SNP markers were developed to anchor the contigs and scaffolds providing eleven chromosome-scale pseudomolecules. To increase the value of the genome sequence a variety of additional studies were undertaken including resequencing of 167 diverse lines from the two domestication centers to get a better understanding of the genetics of the double domestication that occurred in common bean. Cytogenetics were undertaken to understand chromosome structure and evolution within the genus Phaseolus with particular focus on highly repetitive DNA elements and centromere structure. VI ICLGG - Abstracts SUK-HA LEE Professor Department of Plant Science College of agriculture and Life Sciences Seoul National University Korea sukhalee@snu.ac.kr Lee’s main research area is working on breeding, genetics, and genomics of legume crops with a specific emphasis on soybean crop including the development and application of SNP marker. His group identified QTLs for some agronomic traits such as disease resistance genes and seed protein content, and used fine mapping and RNA-seq technology to identify a BLP resistance gene. Using NGS technology, his group has re-sequenced a G. soja genome, wild relative of cultivated soybean (Glycine max) to understand the soybean crop domestication history. Recently, Lee is sequencing the whole genome of mungbean as well as its relative vigna species. Currently, he is serving as one of editorial board member of Molecular Breeding and Euphytica. Selected Publications: 1. Shin JH, Van K, Kim KD, Lee Y-H, Jun T-H, Lee S-H. (2012). Molecular sequence variations of the lipoxygenase-2 gene in soybean. Theor. Appl. Genet. 124: 613-622. 2. Kim MY, Van K, Kang YJ, Kim KH, Lee S-H. (2011). Tracing soybean domestication history: From nucleotide to genome. Breed Sci. 61: 445-452. 3. Kim KH, Kang YJ, Kim DH, Yoon MY, Moon J-K, Kim MY, Van K, Lee S-H. (2011). RNA-Seq analysis of a soybean near-isogenic line carrying bacterial leaf pustule-resistant and –susceptible alleles. DNA Res. 18: 483-497. 4. Liu W, Kim MY, Kang YJ, Van K, Lee Y-H, Srinives P, Yuan DL, Lee S-H. (2011) QTL identification of flowering time at three different latitudes reveals homeologous genomic regions that control flowering in soybean. Theor. Appl. Genet. 123: 545-553. 5. Van K, Kim DH, Shin JH, Lee S-H. (2011) Genomics of plant genetic resources: past, present, and future. Plant Genetics Resources 9: 155-158. VI ICLGG - Abstracts Toward completion of genome sequence of mungbean Lee S-H 1, 2 I-NGG02 * 1 Department of Plant Science and Research Institute for Agriculture and Life Sciences, Seoul National University, Seoul, Korea 2 Plant Genomics and Breeding Research Institute, Seoul National University, Seoul, Korea *E-mail: sukhalee@snu.ac.kr Abstract Mungbean (Vigna radiata (L.) Wilczek) belongs to the genus Vigna savi, subgenus Ceratotropis (also as known as Asian Vigna), section Ceratotropis. It is diploid in nature and has 11 chromosomes (2n=2x=22). Genome size of mungbean is relatively small and it was estimated to be approximate 635 Mb (1.3 pg/2C) by flow cytometry or approximate 548 Mb by K-mer frequency analysis. To construct the mungbean reference genome sequence, Korean mungbean cultivar, Sunwhanokdu, was sequenced using Illumina HiSeq 2000 with one fragment library and two jumping libraries. Produced Illumina short reads were de novo assembled by Allpath-LG software, constructing 93 scaffolds over N50 (=1.2 Mb) with 6 Mb of the maximum length. The total length of 2,927 scaffolds of > 897 bp was approximately 431 Mb, covering 68% of genome size. To validate the assembled scaffolds, we mapped sequence reads from 500 bp insert library back to scaffolds. The 84.93 % of total reads were safely mapped and 75.81% were properly mapped. Additionally, we enabled to obtain 6.5 million GS-FLX reads, corresponding to 3.06 Gb. Assembled contigs were used to validate scaffold by alignment.. Meanwhile, to construct high density genetic linkage map, DNA markers, such as simple sequence repeats and single nucleotide polymorphisms, were developed between genome sequences of the mapping parents, Sunhwadokdu and Gyeonggijaerae 5. VI ICLGG - Abstracts S-NGG03 RAD tags for the characterisation of fast neutron mutants in pea Hofer JMI, Hegarty M, Blackmore T, Camargo-Rodriguez A, Ellis THN *1 IBERS, Aberystwyth University, Aberystwyth, UK *E-mail: noe2@aber.ac.uk Abstract Many mutants altered in leaf form have been described in pea. These provide a resource for understanding the determination of compound leaf form if we can isolate and characterise the corresponding genes. In the absence of a transposon or t-DNA tagging system, we developed a Fast Neutron (FN) deletion mutant population (in the line JI2822) to see whether this could be used for the systematic isolation of genes. We have used a variety of approaches for the identification of genes deleted in this mutant population. Of these only one, was a genome wide survey and it used AFLP analysis. Second generation DNA sequencing methods offer a more direct approach than AFLP. Here we report on progress using RAD tags to facilitate the identification of deleted genes that are candidates for causative mutations. VI ICLGG - Abstracts Exploring structural variation in the soybean genome 1 2 1 1 S-NGG04 1 1 Stupar RM *, McHale LK , Stec A , Anderson JE , Michno JM , Xu WW , 1 1 1,3 1 4 1,3 Haun WJ , Bhaskar PB , Bolon YT , Roessler J , Jeddeloh JA , Vance CP 1 University of Minnesota, Minnesota, USA Ohio State University, Ohio, USA 3 United States Department of Agriculture, Minnesota, USA 4 Roche NimbleGen Inc., Madison, USA 2 *Email : rstupar@umn.edu Abstract Genome-wide structural variation, such as large deletions and duplications, are hypothesized to drive important phenotypic variation. Recent advances involving array hybridization and targeted resequencing now allow for the genome-wide assessment of structural and gene content variation within a species. We have used these methods to assess natural and induced genome variation in soybean cultivars and mutant lines. High rates of natural structural variation have been observed within gene-rich regions that harbor clustered multi-gene families, particularly the Nucleotide Binding (NB) and Receptor-Like Protein (RLP) classes, both of which are important for plant biotic defense responses. The co-localization of structural variants with elements of the plant defense response signal transduction pathways provides insight into the mechanisms and frequencies of soybean resistance gene evolution. Emerging technologies may soon allow for the establishment of genetic stocks with targeted structural variants that can address new hypotheses about the relationship between structural variation and disease gene evolution in soybean. VI ICLGG - Abstracts Theme: Symbiosis and Development GILES OLDROYD John Innes Centre (JIC) Norwich UK giles.oldroyd@jic.ac.uk Western agricultural systems are reliant on the application of inorganic nitrogen fertilisers that greatly enhance yield. However, production and application of nitrogen fertilisers account for a significant proportion of fossil fuel usage in food production and the major global source of nitrous oxide emissions, a very potent greenhouse gas. Prof Giles Oldroyd studies the mechanisms by which some species of plants are capable of forming beneficial interactions with nitrogen fixing bacteria that provide a natural source of nitrogen for plant growth and with mycorrhizal fungi that facilitate nutrient capture from the soil. A long term aim of this research is to reduce agricultural reliance on nitrogen fertilisers. Giles completed his PhD in 1998 at the University of California, Berkeley, studying plant/pathogen interactions and then moved to Stanford University to complete a postdoc on legume/rhizobial interactions in the laboratory of Prof. Sharon Long. He has been an independent researcher at the John Innes Centre since 2002. From 2002 to 2007 he held a BBSRC David Philips Fellowship and since 2007 has been a tenured member of the JIC faculty. In 2008 he was made the head of the Plant Biotic Interactions group at the John Innes Centre and during 2010-2011 he was the Deputy Director. He has been recognised by a number of awards for his research: EMBO young investigator; European Research Council young investigator; Society of Experimental Biology Presidents medal; Royal Society Wolfson Research Merit award and the BBSRC David Phillips Fellowship. Selected Publications: 1. Young N, Debelle F, Oldroyd G. (joint first authors) et al. (2011). The Medicago genome provides insights into the evolution of rhizobial symbioses. Nature 480 520-524. 2. Capoen W, Sun J, Wysham D, Otegui M, Venkateshwaran M, Hirsch S, Miwa H, Downie A, Morris R, Ane J-M, Oldroyd G. (2011). The nuclear membranes control symbiotic calcium signalling of legumes. Proc. Natl. Acad. Sci. USA 108: 14348-14353. 3. Kosuta K, Hazledine S, Sun J, Miwa H, Morris R, Downie JA, Oldroyd G. (2008). Differential and chaotic calcium signatures in the symbiosis signaling pathway of legumes. Proc. Natl. Acad. Sci. USA 105: 9823-9828. 4. Gleason C, Chaudhuri S, Yang T, Munoz A, Poovaiah B, Oldroyd G. (2006). Legume nodulation independent of rhizobia is induced by a calcium activated kinase lacking autoinhibition. Nature 441: 1149-1152. 5. Kaló P, Gleason C, Edwards A, Marsh J, Mitra R, Hirsch S, Jakab J, Sims S, Long S, Rogers J, Kiss G, Downie JA, Oldroyd G. (2005). Nodulation signaling in legumes requires NSP2, a member of the GRAS family of transcriptional regulators. Science 308:1786-1789. VI ICLGG - Abstracts Decoding calcium oscillations during symbiotic signalling I-SAD01 Oldroyd G*, Miller B, Miyahara A, Pratap A, Swainsbury D, Zhou L, Bornemann S, Morris R John Innes Centre, Norwich, UK *E-mail: giles.oldroyd@jic.ac.uk Abstract The establishment of rhizobial and mycorrhizal symbioses requires the common symbiotic signalling pathway that utilises oscillations in calcium as a secondary messenger. Decoding the calcium oscillations is the function of a calcium and calmodulin dependent protein kinase, CCaMK. This protein is unique among calcium-activated kinases in its dual ability to bind free calcium via EF-hand domains and calcium in a complex with calmodulin. Calcium binding to the EF-hand domains activates autophosphorylation on threonine 271 and mutation of this residue autoactivates the protein. This implies that threonine 271 phosphorylation negatively regulates the protein and the dephosphorylation of threonine 271 is sufficient to activate CCaMK. Considering the importance of threonine 271 phosphorylation we generated state specific antibodies that can measure the phosphorylation status of this residue. Whereas calcium binding to the EF-hands activates threonine 271 phosphorylation, calmodulin binding protects this residue from phosphorylation. Thus we propose that calcium binding supresses CCaMK, while calmodulin binding activates CCaMK. We have ascertained the binding kinetics for calcium and calmodulin binding and conclude that CCaMK measures the basal calcium concentration via the EF-hand domains, which maintains the protein in a stably inactive state, while calmodulin binding will only occur during symbiotic calcium oscillations. Mathematical modelling predicts that these calcium binding behaviours creates a robust molecular switch that is reversably activated by calcium oscillations. VI ICLGG - Abstracts JENS STOUGAARD Department of Molecular Biology and Genetics Aarhus University Denmark stougaard@mb.au.dk Jens Stougaard is Professor of Molecular Biology and Genetics at Aarhus University and Director of the Centre for Carbohydrate Recognition and Signalling (CARB). Jens Stougaard leads a group studying genes regulating development of nitrogen fixing root nodules and mycorrhiza formation in legumes. Currently the mechanisms of Nod-factor perception, the function of receptors involved and the downstream signal transduction cascades are in focus. The plant model system used for this research is Lotus japonicus that is also used for investigating the long range signalling integrating root nodule development into the general developmental program of the plant. Genetics, genomics and biochemical methods are used to identify and characterise components of regulatory circuits. In order to improve the genetic analysis and to establish a system for reverse genetics, a large-scale insertion population based on the germ-line specific activity of the LORE1 retroelement is being established and made available to the community. Selected Publications: 1. Broghammer A, Krusell L, Blaise M, Sauer J, Sullivan JT, Maolanon N, Vinther M, Lorentzen A, Madsen EB, Jensen KJ, Roepstorff P, Thirup S, Ronson CW, Thygesen MB, Stougaard J. (2012). Legume receptors perceive the rhizobial lipochito-oligosaccharide signal molecules by direct binding. PNAS (accepted). 2. Urbański DF, Małolepszy A, Stougaard J, Andersen SU. (2012). Genome-wide LORE1 retrotransposon mutagenesis and high-throughput insertion detection in Lotus japonicus. Plant J 69, 731-741. 3. Krusell L, Sato N, Fukuhara I, Koch BEV, Grossmann C, Okamoto S, Oka-Kira E, Otsubo Y, Aubert G, Nakagawa T, Sato S, Tabata S, Duc G, Parniske M, Wang T, Kawaguchi M, Stougaard J. (2011). The Clavata2 genes of pea and Lotus japonicus affect autoregulation of nodulation and shoot architecture. Plant J. 65, 861-871. 4. Madsen E, Antolín-Llovera M, Grossmann C, Ye J, Vieweg S, Broghammer A, Krusell L, Radutoiu S, Jensen O, Stougaard J, Parniske M. (2011). Autophosphorylation is essential for in vivo function of the Lotus japonicus Nod Factor Receptor 1 and receptor mediated signalling in cooperation with Nod Factor Receptor 5. Plant J. 65, 404-417. 5. Madsen LH, Tirichine L, Jurkiewicz A , Sullivan JT, Heckmann AB, Bek AS, Ronson CW, James EK, Stougaard J. (2010). The molecular network governing nodule organogenesis and infection in the model legume Lotus japonicus. Nature Communications DOI:10.1038/ncomms1009. VI ICLGG - Abstracts The role of LysM type receptors in Nod Factor perception I-SAD02 Stougaard J Aarhus University, Gustav Wieds Vej 10, Denmark E-mail:stougaard@mb.au.dk Abstract Development of root nodules in legumes in response to signals secreted from rhizobia is an example of inducible organ formation. Lipochito-oligosaccharides (Nod-factors), consisting of substituted ,1-4 N-acetylglucosamine (chitin) backbones are the rhizobial morphogenic signal inducing root hair deformation and cell division leading to formation of nodule primordia. An important determinant of bacterial host specificity is the structure of the Nod-factor suggesting that a plant receptor is involved in signal perception and signal transduction initiating the plant developmental response. The role of the two Lotus japonicus LysM type serine/threonine receptor kinases, NFR1 and NFR5, in perception of Nod-factor signals from bacterial microsymbionts will be discussed. The extracellular domains of the two trans-membrane kinases carries LysM domains suggesting that they may be directly involved in binding of the rhizobial lipochitin-oligosaccharide signals and in deciphering the structure of lipochitin-oligosaccharides. Biochemical experiments addressing this question will be presented and the involvement of NFR1 and NFR5 receptor kinases in the earliest physiological and cellular responses will be illustrated. In addition to the NFR1 and NFR5 receptors, the Lotus japonicus genome encodes fifteen hitherto uncharacterised LysM type receptor kinases. The functional role of these receptors is currently investigated using a number or approaches including TILLING mutants and expression studies. VI ICLGG - Abstracts PETER GRESSHOFF Director ARC Centre of Excellence for Integrative legume Research (CILR) The University of Queensland Australia director.cilr@uq.edu.au Professor Peter Gresshoff, Director, ARC Centre of Excellence for Integrative Legume Research (CILR), is a plant developmental geneticist, who uses molecular and genetic tools to understand (a) lateral organ differentiation and the genetic control of Bradyrhizobium induced-nodule formation in soybean and Lotus japonicus roots, and (b) the functional genomics of sustainable biofuel production in the legume tree Pongamia pinnata. He pioneered the use of induced mutagenesis to isolate non-nodulation and supernodulation mutants; he laid the basis to the concept of ‘Autoregulation of Nodulation’ (AON) by publishing the first supernodulation mutants, the recognition of shoot control, the positional cloning of the causative LRR-RK GmNARK gene, and systemic peptide and metabolite signals in nodulation. He is a Fellow of AAAS, the Indian National Academy of Agricultural Sciences, and the Russian Academy of Agricultural Sciences, and received the prestigious Chinese Academy of Science (CAS) International Professorial Fellowship (2010). Professor Gresshoff has published over 300 peer-reviewed journal papers, including Nature, Science and PNAS, edited 10 books and holds 12 biotechnology-related patents. His hindex is 42, with about 9,200 citations, 12 cited more than 100 times (one 1,900 times). In 2006, he chaired the ICLGG3 meeting in Brisbane, Australia. Selected Publications: 1. Carroll BJ, McNeil DL, Gresshoff PM. (1985). Isolation and properties of soybean mutants which nodulate in the presence of high nitrate concentrations. Proc. Natl. Acad. Sci. USA 82: 4162-4166. 2. Delves AC, Mathews A, Day DA, Carter AS, Carroll BJ, Gresshoff PM. (1986). Regulation of the soybean-Rhizobium symbiosis by shoots and root factors. Plant Physiol. 82: 588-590. 3. Searle IR, Men AM, Laniya TS, Buzas DM, Iturbe-Ormaetxe I, Carroll BJ, Gresshoff PM. (2003) Long distance signalling for nodulation control in legumes requires a CLAVATA1-like receptor kinase. Science 299: 109-112. 4. Lin Y-H, Ferguson BJ, Kereszt A, Gresshoff PM. (2010) Suppression of hypernodulation in soybean by a leaf-extracted, NARK- and Nod-factor-dependent small molecular fraction. New Phytologist 185: 1074-1086. 5. Reid DE, Ferguson BJ, Gresshoff PM. (2011). Inoculation- and nitrate-induced CLE peptides of soybean control NARK-dependent nodule formation. Mol. Plant Microbe Interactions 24: 606618. VI ICLGG - Abstracts Sensing, signals, suppression, symbiosis, soybean I-SAD03 Gresshoff PM, Reid D, Lin Y-H, Hayashi S, Indrasumunar A, Mirzaei S, Batley J, Ferguson BJ The University of Queensland, Brisbane, Australia *E-mail:p.gresshoff@uq.edu.au Abstract Plants regulate the number of lateral organs through sensing of environmental and internal developmental signals. For example, legumes regulate the number of nitrogen fixing nodules through systemic signals. Prior nodulation events (AON), acid soils and nitrate are major controls acting in either local or systemic manner. AON in all legumes functions through an ubiquitously phloem-expressed LRR receptor kinase gene (GmNARK, MtSUNN, LjHAR1; all Arabidopsis CLV1-related), that requires the sensing of nodule primoridia. Inoculation-related AON depends on leaf phloem parenchyma where GmNARK and interacting proteins result in long-distance negative regulation of further cell divisions in the root cortex/pericycle. We have characterised candidate molecules for these long distance signals moving between root and shoot, and reverse. CLE peptide genes, related to Arabidopsis CLV3, induced by rhizobia or nitrate in soybean, when overexpressed in soybean roots suppress nodulation, in a NARK-dependent fashion (1,2). Site-directed mutagenesis of CLE peptide residues, as well as domain swaps between locally acting NIC1 and systemically acting RIC1 peptides, monitored for loss of suppression after overexpression will be discussed. A small (<1 kDa) metabolite produced in leaf in a NARK-dependent manner if fed by a novel petiole feeding assembly (3,4), suppresses nodulation. The presentation will summarise our combined genetic, functional genomics and biochemical understanding of the nodulation-control network. VI ICLGG - Abstracts S-SAD04 The autoregulation of nodulation in Medicago truncatula Schnabel EL, Kassaw TK, Crook A, Frugoli JA* Clemson University, Clemson, USA *E-mail: jfrugol@clemson.edu Abstract The formation of nitrogen-fixing nodules is tightly controlled by a long-distance signaling system in which nodulating roots signal to shoot tissues to suppress further nodulation. We have previously identified several mutants in Medicago truncatula that lack the ability to regulate nodule number from the shoot (sunn and lss) or from the root (rdn1). Split-root and inverted Y-graft experiments exploring autoregulation over time in these mutants and wild type plants identified both an early and later time point at which an autoregulatory signal is perceived. All autoregulatory mutants are defective in the early signal, while mutant response toward the second signal varies. We also report a suppressor of the sunn-1 allele and the lss lesion, a mutation in the MtCRE1 cytokinin receptor. Unlike Mtcre1 alleles identified by others, this suppressor allele has almost no effect on nodule number alone; rather its effect on nodulation is observed only in plants with disruptions of the SUNN pathway. Plants carrying the allele have shorter roots and reduced lateral root density, but the effect on cytokinin associated phenotypes such as growth on BAP are weak, suggesting the identified mutation is important to a specific signal transduction pathway involving cytokinin. The combination of these findings, our work on interacting partners of the SUNN kinase, and data from other groups working in M. truncatula allows us to postulate a general signal transduction pathway for nodule number regulation that includes SUNN, LSS, RDN1 and CLE peptides. VI ICLGG - Abstracts The CCAAT-box binding factors MtNF-YA1&2 are transcriptional regulators required for Nod factor signaling, rhizobial infection and nodule development 1 1 1 1 2 S-SAD05 1 Laporte P , Laloum T , Baudin M , Frances L , Breakspear A , Cerri MR , 1 1 1 1 2 1 Lepage A , Fournier J , Catrice O , Jardinaud F , Murray J , Gamas P , 1 1 Fernanda de Carvalho-Niebel , Niebel A * 1 LIPM, INRA-CNRS, Castanet-Tolosan, France John Innes Center, Norwich, UK 2 *Email : aniebel@toulouse.inra.fr Abstract MtNF-YA1& 2 encode transcription factors (TFs) of the CCAAT-box binding factor family which are strongly expressed in Medicago truncatula during early phases of the symbiotic interaction with Sinorhizobium meliloti. In animal systems, NF-YA proteins are known to associate with NF-YB and NF-YC subunits to form heterotrimeric hubs which interact with other TFs, and promote specific processes, in particular cell cycle progression. Whereas in animals each NF-Y subunit is encoded by a single gene, structural diversification in plants has lead to the emergence of gene families comprising between 8 and 39 members depending on the subunit and the plant species. This diversification is also apparent at the functional level where NF-Y encoding genes are involved in processes as diverse as embryo development, flowering time control, and drought or ER-stress. We have analysed in detail the symbiotic expression patterns of the two closely related MtNF-YA1& MtNFYA2 genes by q-RTPCR, mRNA in situ hybridization and promoter-GUS analysis. These analyses together with functional RNAi and mutant characterization have shown that these two TFs play partially overlapping roles during Nod factor signalling, as well as during subsequent rhizobial infection and nodule development. Furthermore, in order to better understand their functions and the symbiotic pathways regulated by these TFs we have recently started to characterise their symbiotic NF-YB and NF-YC partners as well as their potential targets during different steps of the symbiotic legume-Rhizobium interaction. VI ICLGG - Abstracts MICHAEL UDVARDI Professor The Samuel Roberts Noble Foundation USA mudvardi@noble.org Dr. Udvardi earned his Ph.D. from the Australian National University (ANU) in Canberra in 1989 and received postdoctoral training in the USA and Australia from 1989-1994. He worked as a Lecturer then Senior Lecturer at the ANU from 1994-1998, as an Associate Professor at the Max Planck Institute of Molecular Plant Physiology in Golm, Germany, from 1998-2006, and as a Full Professor at the Samuel Roberts Noble Foundation in Ardmore, USA, from 2006 until the present. Dr. Udvardi has made seminal contributions to our understanding of symbiotic nitrogen fixation in legumes, using biochemical, molecular, genetic, and genomic methods. His group has also contributed to the understanding of plant acclimation and adaptation to abiotic stress, and to the genetic regulation of seed development and metabolism in legumes. He was part of a large team that sequenced and analyzed the Medicago truncatula genome and he currently leads efforts to sequence the alfalfa (Medicago sativa) genome. Selected Publications: 1. Benedito VA, Torres-Jerez I, Murray JD, Andriankaja A, Allen S, Kakar K, Wandrey M, Verdier J, Zuber H, Ott T, Moreau S, Niebel A, Frickey T,Weiller G, He J, Dai X, Zhao PX, Tang Y, Udvard MK. (2008). A gene expression atlas of the model legume Medicago truncatula. Plant J. 55: 50413. 2. Kang Y, Han Y, Torres-Jerez I, Wang M, Tang Y, Monteros M, Udvardi M. (2011). System responses to long-term drought and re-watering of two contrasting alfalfa varieties. Plant Journal 68: 871-889. 3. Young ND et al. (2011). The Medicago genome provides insight into the evolution of rhizobial symbioses. Nature 480: 520-4. 4. Verdier J, Zhao J, Torres-Jerez I, Ge S, Lui C, He X, Mysore KS, Dixon RA , Udvardi MK. (2012). MtPAR MYB transcription factor acts as an on-switch for proanthocyanidin biosynthesis in Medicago truncatula. Proc. Natl. Acad. Sci. U S A. 109:1766-1771. 5. Pislariu CI, Murray J, Wen J, Cosson V, Duvvuru Muni RR, Wang M, Benedito V, Andriankaja A, Cheng X, Torres-Jerez I, Mondy S, Zhang S, Taylor M, Tadege M, Ratet P, Mysore KS, Chen R, Udvardi MK. (2012). A Medicago truncatula tobacco-retrotransposon (Tnt1)-insertion mutant collection with defects in nodule development and symbiotic nitrogen fixation. Plant Physiol. (in press). VI ICLGG - Abstracts Functional genomics of symbiotic nitrogen fixation in Medicago truncatula 1* 1 1 1 1 I-SAD06 1 Udvardi M , Pislariu C , Kryvoruchko I , Sinharoy S , Shibasaki K , Wang M , 1 1 1 1 1 1 1 Torres-Jerez I , Taylor M , Zhang S , Cheng X , Wen J , He J , Tang Y , 1 1 2 3 Chen , Kirankumar Mysore , Pascal Ratet , Vagner A Benedito , Giles Oldroyd 4 4 R , Murray JD 1 Samuel Roberts Noble Foundation, Ardmore, USA ISV-CNRS, Gif sur Yvette, France 3 West Virginia University, USA 4 John Innes Centre, Norwich, UK 2 * E-mail: mudvardi@noble.org Abstract The past few years have seen the development of tremendous tools and resources for functional genomics in Medicago truncatula, including the M. truncatula Gene Expression Atlas (MtGEA), Tnt1-insertion and fast-neutron deletion mutant populations, and, most recently, a complete genome sequence. These resources are proving to be instrumental in systematic studies of gene function in this species. Symbiotic nitrogen fixation (SNF) in legumes involves thousands of plant (and bacterial) genes that are expressed during nodule development and cellular differentiation. Many of these genes are up-regulated during nodule development and a substantial number are essential for SNF. We have carried out systematic forward- and reverse-genetic screens to identify novel genes required for SNF in Medicago. Forward-screening of 9,300 Tnt1 mutant lines yielded 156 lines defective in SNF. Mutants were sorted into six distinct phenotypic categories: 72 non-nodulating mutants (Nod-), 51 mutants with totally-ineffective nodules (Nod+ Fix-), 17 mutants with partially-ineffective nodules (Nod+ Fix+/-), 27 mutants defective in nodule emergence, elongation, and nitrogen fixation (Nod+/- Fix-); 1 mutant with delayed and reduced nodulation, but effective in nitrogen fixation (dNod+/- Fix+), and 11 supernodulating mutants (Nod++Fix+/-). Analysis of flanking sequence tags (FSTs) and gene-specific PCR identified insertion alleles of the following known symbiotic genes: NIN, DMI1, DMI2, DMI3/CCaMK, NSP1, NSP2, SKL, ERN1, and SUNN. However, mutants containing these defective alleles account for a minority of the Tnt1 SNF mutants, indicating that many novel symbiotic genes have been tagged and remain to be discovered amongst these 156 lines. Morphological and developmental features of several new symbiotic mutants will be presented. Reverse-genetics, using the Tnt1-insertion population is being used to determine the functions of selected transcription factors (TFs) and transporters that are induced during nodule development. One of the TFs, a C2H2 family member is nodule-specific, expressed mainly in the invasion zone, controls symbiosome maturation after bacterial endocytosis from the infection threads, and is essential for SNF. Sixteen transporter genes are being investigated using reverse-genetics and mutants in several of these are defective in SNF. For example, defects in MtMATE5, a transporter of the multidrug and toxin efflux family, result in smaller nodules with less leghemoglobin, reduced nitrogen fixation ability, and stunted growth of shoots. Based on sequence conservation, we hypothesize that MtMATE5 might transport citrate or citratechelated iron, possibly to bacteroids. The phenotypes of these mutants will be presented. This work was supported by the National Science Foundation, the USDA CSREES-NRI, and the Samuel Roberts Noble Foundation VI ICLGG - Abstracts GEORGINA HERNANDEZ Professor Center for Genomic Sciences (CCG) National University of Mexico (UNAM) Mexico gina@ccg.unam.mx Georgina Hernández is a Professor from the Center for Genomic Sciences (CCG) -previously Nitrogen Fixation Research Center (CIFN)- from the National University of Mexico (UNAM) in Cuernavaca, Mexico. She served as the Director of the Center from 1997 to 2005. Currently she serves as the leader of the Eukaryotic Functional Genomics Research Program from CCG. She received her undergraduate and PhD degrees from UNAM. She did a postdoctoral training in BioTechnica International, Inc. in Cambridge, MA, and a sabbatical leave at the University of Minnesota – USDA in Carroll Vance's laboratory and at the Max Planck Institute for Plant Molecular Physiology (Potsdam, Germany) in Michael Udvardi's group. She is member of the Mexican Academy of Sciences and of the National Researchers System from the Mexican government. She has organized several international and national meetings including ICLGG V (2008). She initiated and has co-coordinated the international consortium for Phaseolus genomics: "Phaseomics". The research area from her group is the functional genomics of common bean (Phaseolus vulgaris): symbiosis with rhizobia and response to nutrient-stress. Her current projects include Phaseolus transcriptomics through RNA-seq and the functional analysis of transcription factors and miRNAs as global regulators in common bean. Selected Publications: 1. Mendoza-Soto AB, Sánchez F, Hernández G (2012). MicroRNAs as regulators in plant metal toxicity response. Frontiers in Plant Science 3:105. doi: 10.3389/fpls.2012.00105. 2. Valdés-López O, Yang SS, Aparicio-Fabre R, Graham PH, Reyes JL, Vance CP, Hernández G (2010). MicroRNAs expression profile in common bean (Phaseolus vulgaris) during nutrient deficiency stresses and manganese toxicity. New Phytologist. 187: 805-818. 3. Hernández G, Valdés-López O, Ramírez M, Goffard N, Weiller G, Aparicio-Fabre R, Fuentes SI, Erban A, Kopka J, Udvardi MK, Vance CP (2009). Global changes in the transcript and metabolic profiles during symbiotic nitrogen fixation in phosphorus-stressed common bean plants. Plant Phsyiology. 151: 1221-1238. 4. Valdés-López O, Arenas-Huertero C, Ramírez M, Girard L, Sánchez F, Vance CP, Reyes JL, Hernández G (2008). Essential role of MYB transcription factor: PvPHR1 and microRNA: PvmiR399 in the phosphorus deficiency signaling in common bean roots. Plant Cell and Environment 31: 1834-1843. 5. Gepts P, Aragao F, de Barros E, Blair MW, Brodani R, Broughton WJ, Galasso I, Hernández G, Kami J, Lariguet P, McClean P, Melotto M, Miklas P, Pauls P, Pedrosa-Harand A, Porch T, Sánchez F, Sparvoli F, Yu K (2008). Genomics of Phaseolus beans, a major source of dietary protein and micronutrients in the tropics. In: “Genomics of tropical crop plants” Moore PH, and Ming R (eds). ISBN: 978-0-387-71218-5. Springer New York pp. VI ICLGG - Abstracts Transcriptome of common bean (Phaseolus vulgaris) through RNA-seq: nodulation, symbiotic nitrogen fixation, transcription factors 2 1 1 I-SAD07 3 Hernández G , O’Rourke JA , Íñiguez LP , Nova-Franco B , Reyes JL , 4 5 6 2 McClean P , Shoemaker RC , Jackson SA , Vance CP 1* 1 Universidad Nacional Autónoma de México (UNAM),Morelos, México USDA-ARS, University of Minnesota, USA 3 Instituto de Biotecnología - UNAM, Morelos, México 4 North Dakota State University, Fargo, USA 5 USDA-ARS, Iowa State University, Ames, USA 6 University of Georgia, Athens, USA 2 *E-mail: gina@ccg.unam.mx Abstract The advent of next generation sequencing (RNA-seq) allows identifying all expressed transcripts in tissues of interest. The whole transcriptome study of common bean (Phaseolus vulgaris), the most important legume for human consumption, was performed through RNA-seq with Illumina-Sollexa platform. This included samples from five tissues (seeds, pods, leaves, roots, and nodules) of P. vulgaris cv. Jamapa at three stages of development that generated ca. 25 million 36-bp reads per sample. From these, 80, 462 transcripts were assembled and ca. 50,000 were annotated. The study identified 43,708 transcript assemblies (>100bp) that were expressed in seeds, 53,447 in pods, 54,665 in leaves, 59,337 in roots, and 53,099 in nodules. The libraries included: nitrogen-fixing nodules, non-fixing nodules –elicited by an ineffective rhizobia- and roots associated with the nodules. We are performing a comprehensive transcriptome study of nodules vs. roots (inoculated and non-inoculated as control) to identify the transcripts involved in relevant processes of nodulation and symbiotic nitrogen fixation in the common bean Rhizobium tropici association. Derived from transcriptomic data, our current projects include the functional analysis of the global regulators: transcription factors (TF) and microRNAs. The common bean transcriptome revealed 3,567 transcripts annotated as TF belonging to 52 gene families. We are analyzing the TF differential expression in roots vs. nodules aiming to identify key TF for nodulation and SNF. TFs that are targets of miRNAs highly expressed in common bean nodules have been identified, these may play relevant roles in signaling / regulatory pathways of the rhizobia-legume symbiosis. VI ICLGG - Abstracts ATTILA KERESZT Institute of Biochemistry Biological Research Centre of the Hungarian Academy of Sciences (HAS) Szeged Hungary kereszta@baygen.hu I graduated and then obtained my PhD from the Szeged University working at BRC HAS under the supervision of Adam Kondorosi to identify bacterial genes involved in cytochrome c biogenesis and surface polysaccharide production (1). During my first post-doc period, I joined the group of Thierry Huguet at LIPM (Castanet-Tolosan, France) where we constructed the first genetic map of the model legume Medicago truncatula (2). After returning to Szeged, I participated in the map-based cloning of NORK/SYMRK required for rhizobial and mycorrhizal symbioses (3). I spent three years at the ARC CILR (Brisbane, Australia) where I was involved in the cloning of soybean nodulation genes and in the development of bioassays to identify long-range signaling molecules of autoregulation of nodulation (4). Since affiliating with the former BayGen Institute I am working on plant peptides governing bacteroid differentiation (5). Selected Publications: 1. Kereszt A, Kiss E, Reuhs B L, Carlson R W, Kondorosi A, Putnoky P (1998). Novel rkp gene clusters of Sinorhizobium meliloti involved in capsular polysaccharide production and the invasion of the symbiotic nodule: rkpK gene encodes a UDP-glucose dehydrogenase. J Bacteriol 180:5426–5431. 2. Thoquet P, Ghérardi M, Journet EP, Kereszt A, Ané JM, Prosperi JM, Huguet T (2002). The molecular genetic linkage map of the model legume Medicago truncatula: an essential tool for comparative legume genomics and the isolation of agronomically important genes. BMC Plant Biology 2:1. 3. Endre G, Kereszt A, Kevei Z, Mihacea S, Kaló P, György B. Kiss (2002). A receptor kinase gene regulating symbiotic nodule development. Nature 417:962 - 966. 4. Lin Y-H, Ferguson B-J, Kereszt A, Gresshoff PM (2010). Suppression of hypernodulation in soybean by a leaf-extracted, NARK and Nod factor-dependent, low molecular mass fraction. New Phytol 185: 1074-1086. 5. Van de Velde W, Zehirov G, Szatmari A, Debreczeny M, Ishihara H, Kevei Z, Farkas A, Mikulass K, Nagy A, Tiricz H et al (2010). Plant peptides govern terminal differentiation of bacteria in symbiosis. Science 327:1122-1126. VI ICLGG - Abstracts Antimicrobial peptides in symbiosis 1,3* 2 I-SAD08 1 1 3 1 Kereszt A , Megaert P , Abraham E , Ordogh L , Tiricz H , R Mikulass K , 1 3 1 1 1,3 4 4 Szucs A , Farkas A , Klement E , Durgo H , Maroti G , Horvath B , Kalo P , 1,2 Kondorosi E 1 Hungarian Academy of Sciences, Szeged, Hungary Institut des Sciences du Vegetal – CNRS, Gif-sur-Yvette, France 3 Bay Zoltán Nonprofit Ltd. for Applied Research, Szeged, Hungary 4 Agricultural Biotechnology Centre, Godollo, Hungary 2 * E-mail: kereszt.attila@brc.mta.hu Abstract Rhizobium-legume symbiosis results in the formation of root nodules where intracellular bacteria reduce atmospheric nitrogen and supply ammonia for plant growth in exchange for energy and carbon sources. This interaction used to be considered mutually beneficial. This view has, however, changed drastically upon the discovery that certain plants exploit their bacterium partners by directing them into an irreversible, terminal differentiation with no chance to return to the free-living state. The mechanism of plant dominance has been elucidated in Medicago truncatula where ~500 antimicrobial-like peptides, related to the effectors of innate immunity, have adapted and evolved for symbiosis. These peptides are produced in the symbiotic cells and are targeted to the bacteria provoking genome amplification, extreme cell elongation, increased membrane permeability and loss of cell division capacity. The combined action of the peptides keeps the bacteria viable but uncultivable and necessitates the function of the bacterial BacA protein, which is also essential for the establishment of chronic intracellular infection by intracellular mammalian pathogens. To identify rhizobial pathways affected by NCR peptides we applied biochemical and gene expression approaches. The possible intracellular targets of NCR peptides were investigated by affinity chromatography using various tagged peptides. Protein complexes were analysed by SDS-PAGE separation followed by LC-MS/MS identification. To identify transcriptome changes, we performed sequencing of RNA from free-living and symbiotic rhizobia challenged with nodule-specific peptides. The identified pathways either individually or in combination could lead to the observed bacterial phenotypes, the decreased cell division and induced terminal differentiation. VI ICLGG - Abstracts S-SAD09 Multi-locus molecular phylogeny and allelic variation in a transcription factor gene suggest the multiple independent origins of kabuli chickpea 1 1 1 1 Penmetsa RV *, Carrasquilla-Garcia N , Bergmann EM , Vance LC , Castro 1 2 2 1,3 1 2 2 BM , Dubey A , Gujaria N , Sarma BK , Kassa MT , Kaashyap M , Gaur PM , 1,4 1 1,5 5 6 Datta S , Baek JM , Woodward JE , Farmer AD , Coyne CJ , Wettberg 7,8 2 1 EJB , Varshney RK , Cook DR * 1 University of California at Davis, Department of Plant Pathology, Davis, USA International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 3 Banaras Hindu University, Department of Mycology and Plant Pathology, Varanasi, India 4 Indian Institute of Pulses Research (IIPR), Kanpur, India 5 National Center for Genome Resources, Santa Fe, USA 6 USDA-ARS, Western Regional Plant Introduction Station, Pullman,USA 7 Florida International University, Department of Biological Sciences, Miami, USA 8 Center for Tropical Plant Conservation, Fairchild Tropical Botanic Garden, Coral Gables, USA 2 *Email:rvpenmetsa@ucdavis.edu & drcook@ucdavis.edu Abstract To examine molecular diversity in wild crop relatives and the cultivated gene pool of chickpea we genotyped a set of 98 wild annual and 224 cultivated accessions with a 768 feature assay that monitored SNPs in low-copy orthologous loci. Analyses of the resulting multi-locus genotyping data - which is deeper for both molecular markers and for wild relatives - recapitulated the taxonomic relationships between cultivated chickpea and its wild crop relatives. We identified gene flow both within the primary gene pool, and between the primary and secondary gene pools. Extant hybrids of the primary and secondary gene pools could serve as genetic bridges for introgression of desirable traits from the secondary pool into cultivated chickpea. Cultivated accessions typically had high levels of admixture but could be resolved into coherent geographic origin-based clades from low-admixed subsets. Contrary to the traditional two-way subdivision of chickpea into ‘desi’ and ‘kabuli’ types, our multi-locus analyses finds ‘kabuli’ to be polyphyletic. Moreover, from candidate pathways, syntenic relationships and genetic association we identified a candidate transcription factor for the B locus of chickpea that governs several ‘kabuli’ traits. Three of five of its variant alleles result in frameshift alterations, whose loss-of-function effect is reflected in the attenuated expression of transcriptional targets in petals and seed coats of ‘kabuli’ but not ‘desi’ types. The absence of nesting among variant alleles at the candidate gene, and the pattern of their occurrence within germplasm together suggest that the ‘kabuli’ type arose independently multiple times during chickpea’s crop evolutionary history. VI ICLGG - Abstracts The same or different? Flowering on time in the model legume Medicago 1 1 1 2 3 S-SAD10 1 Yeoh C , Zhang L , Jaudal M , Mysore K , Ratet P , Putterill J * 1 University of Auckland, Auckland, New Zealand Samuel Roberts Noble Foundation, Ardmore, USA 3 ISV, CNRS, Paris, France 2 *E-mail:j.putterill@auckland.ac.nz Abstract The transition from vegetative to floral development is a crucial step in sexual reproduction. Internal and external signals control the timing of flowering. While the relative importance of these different signals may differ between plants, they are integrated by conserved floral decision makers - such as the florigen FLOWERING LOCUS T (FT) which acts on downstream genes to promote floral development. Like Arabidopsis, the model legume Medicago truncatula (Medicago) is a eudicot long-day plant and pre-exposure of Medicago to extended cold (vernalisation) accelerates flowering. However, Medicago is missing a key flowering gene known from Arabidopsis and has 5 FT genes compared to just two in Arabidopsis. Thus we are using forward and reverse genetics to analyse Medicago flowering, with the ultimate goal to expand knowledge of flowering regulation and provide tools for modern plant breeding. Here, we will present the molecular identification and functional analysis of Medicago SPRING genes. These genes were identified by screening for mutants that flowered early and independently of vernalisation. The SPRING genes were cloned by a combination of positional cloning and Tnt1 tagging. We will also discuss our progress with investigation of the functions of candidate flowering time genes and their downstream targets and present a working model of Medicago flowering time control. VI ICLGG - Abstracts Theme: Evolution and Diversity DAVID JOHN BERTIOLI University of Brasilia Campus Universitario Darcy Ribeiro Brazil djbertioli@gmail.com Main research interests are peanut genetics and genomics, the evolution of legume genomes, the characterization of agronomically desirable alleles from peanut’s wild relatives and their introgression into cultivated peanut. Selected Publications: 1. Bertioli DJ, Seijo G, Freitas FO, Valls JFM, Leal-Bertioli SCM, Moretzsohn MC. (2011) An overview of peanut and its wild relatives. Plant Genetic Resources: Characterization and Utilization, 1-16. 2. Nielen S, Campos-Fonseca F, Leal-Bertioli S, Guimarães P, Seijo G, Town C, Arrial R, Bertioli D. (2010) FIDEL—a retrovirus-like retrotransposon and its distinct evolutionary histories in the A- and Bgenome components of cultivated peanut. Chromosome Research, DOI 10.1007/s10577-009-9109-z. 3. Leal-Bertioli SCM, José ACFV, Alves-Freitas DMT, Moretzsohn MC, Guimarães PM, Nielen S, Vidigal B, Pereira RW, Pike J, Fávero AP, Parniske M, Varshney R, Bertioli DJ. (2009) Identification of candidate genome regions controlling disease resistance in Arachis. BMC Plant Biology, 9:112. 4. Bertioli D, Moretzsohn M, Madsen LH, Sandal N, Leal-Bertioli S, Guimarães P, Hougaard BK, Fredslund J, Schauser L, Nielsen AM, Sato S, Tabata S, Cannon S, Stougaard J. (2009) An analysis of synteny of Arachis with Lotus and Medicago sheds new light on the structure, stability and evolution of legume genomes. BMC Genomics, 10:45. 5. Guimarães PM, Garsmeur O, Proite K, Leal-Bertioli SCM, Seijo G, Chaine C, Bertioli DJ, D`Hont A. (2008) BAC libraries construction from the ancestral diploid genomes of the allotetraploid cultivated peanut. BMC Plant Biology, 8:14. VI ICLGG - Abstracts The repetitive component of the A genome of peanut (Arachis hypogaea L.) and its role in the erosion of genome sequence similarity over time 1* 1,2 2,3 4 I-EAD01 5 Bertioli DJ , Vidigal B , Nielen S , Ratnaparkhe M , Bailey J , Soraya CM 2 4 2 5 4 Leal-Bertioli , Changsoo K , Guimarães PM , Schwarzacher T , Paterson A , 5 2 Pat H-H , Araujo ACG 1 University of Brasilia, Brasília, Brazil Embrapa Genetic Resources and Biotechnology, Brasilia, Brazil 3 International Atomic Energy Agency, Vienna, Austria 4 The University of Georgia, Athens, USA 5 University of Leicester, Leicester, UK 2 *E-mail: djbertioli@gmail.com Abstract Cultivated peanut (A. hypogaea L.) is a very important tropical crop, valued both for subsistence and as a cash crop. It has a large genome of about 2.8 GB with a high repetitive content. It is an allotetraploid of recent origin with an AABB-type genome. Its most probable diploid ancestors are A. duranensis (Krapov. & W.C. Greg.) and A. ipaënsis (Krapov. & W.C. Greg.). The evolutionary divergence of these two species is estimated at about 3.5 million years ago, more recent than the divergence of the two genome components of the allotetraploids cotton and soya, estimated at about 5-7 and 13 million years ago respectively. Sequence analysis of 12 genomic regions from A. duranensis totalling 1.26 Mb allowed the isolation of ten abundant long terminal repeat (LTR) retro transposons. These retro transposons accounted for more than one third of the analysed A genomic regions. The most abundant, named Feral, was nonautonomous and apparently parasitic on the transposon FIDEL. This pair of transposons accounted for one sixth of the analysed sequence. The next most abundant retrotransposon, Pipa was also non-autonomous. Pipa carries an enigmatic ORF that has no obvious similarities to any known protein, but is under purifying evolutionary selection. Also, to study the evolutionary dynamics of the genome sequences, two of the A genome regions were compared to their B genome homeologous regions. This showed conserved segments of high sequence identity punctuated by predominantly indel regions without significant A-B genome similarity. There was a marked tendency for the variable regions to be repetitive. We suggest that much of the sequence divergence of the A and B sub-genomes of cultivated peanut can be explained by the activity of a relatively few retrotransposons that have been highly active in the last 3.5 million years. VI ICLGG - Abstracts JEFF DOYLE Professor Plant Biology and Plant Breeding and Genetics College of Agriculture and Life Sciences (CALS) Cornell University USA jjd5@cornell.edu Jeff Doyle is a Professor in the departments of Plant Biology and Plant Breeding & Genetics at Cornell University. His research focuses on systematics and comparative genomics of legumes, particularly wild species in the genus Glycine, to which soybean belongs. A particular focus of the lab is polyploidy—whole genome duplication—an evolutionary phenomenon that has shaped the genomes of all flowering plants, and has been particularly important in the evolution of Glycine, which includes a number of recently formed allopolyploids. Additional interests include the evolution of nodulation. Selected Publications: 1. Ashfield T, Egan AN, Pfeil BE, Chen NWG, Podicheti R, Ratnaparkhe MB, Ameline-Torregrosa C, Denny R, Cannon S, Doyle JJ, Geffroy V, Roe BA, Saghai Maroof MA, Young ND, Innes RW. (2012). Evolution of a complex disease resistance gene cluster in diploid Phaseolus and tetraploid Glycine. Plant Physiology 159:336-354. 2. Cannon SB, Ilut DC, Farmer AD, Maki SL, May GD, Singer SR, Doyle JJ. (2010). Polyploidy did not predate the evolution of nodulation in all legumes. PLoS One 5(7):e11630. doi:10.1371/journal.pone.0011630. 3. Coate JE, Luciano AK, Seralathan V, Minchew KJ, Owens TG, Doyle JJ. (2012). Anatomical, biochemical and photosynthetic responses to recent allopolyploidy in Glycine dolichocarpa. American Journal of Botany 99:55-67. DOI: 10.3732/ajb.1100465. 4. Doyle JJ. 2011. Phylogenetic perspectives on the origins of nodulation. (2011). Molecular Plant Microbe Interactions 24:1289-1295. DOI: 10.1094/MPMI-05-11-0114. 5. Doyle JJ, Luckow MA. (2003). The rest of the iceberg: legume diversity and evolution in a phylogenetic context. Plant Physiology 131:900-910. VI ICLGG - Abstracts Diversity and evolution of the perennial Glycine allopolyploid complex I-EAD02 Doyle JJ Department of Plant Biology, Cornell University, Ithaca, USA E-mail: jjd5@cornell.edu Abstract Among the approximately 30 species of Glycine subgenus Glycine, the wild perennial relatives of the cultivated soybean, are a group of species with diploid chromosome numbers of 2n = 78 or 80. Crossing and phylogenetic data have shown that these species are allopolyploids, formed from extant diploid members of the subgenus. We have refined our understanding of several of these species using phylogenomic and coalescent approaches, confirming their relative recency and their genomic origins. We have studied the evolution of these fixed hybrid allopolyploids for a range of morphological, anatomical, biochemical, and physiological characters, and find general patterns that suggest consistent responses across polyploids of independent origins. Transcriptomic analyses have been conducted to measure the response of the overall transcriptome to polyploidy and to distinguish the contributions of homoeologous genomes to the expression of all genes and specifically of genes involved in processes such as photoprotection. We are developing the perennial soybean allopolyploids as a model system for recent polyploidy in plants. VI ICLGG - Abstracts S-EAD03 Using high-throughput sequencing data to speed the domestication of Apios americana - a potential new legume crop Cannon SB*, Belamkar V Iowa State University, Ames, USA *E-mail: steven.cannon@ars.usda.gov Abstract Apios americana, a species in the Phaseoleae tribe, is native to central and eastern North America. Apios produces edible tubers that were a staple of American Indian groups. The tubers are palatable and verstatile, with a potatolike starch and ~16% crude protein (about three times that of potato). Apios is unusual as a “warm-season legume” for its tolerance to winter cold and to flooding. Yields can be high, with per-plant tuber yields above 3 kg. Apios tubers also contain abundant amounts of the 7-O-glucosylglucoside of genistein, which is predominant in the putative cancer preventive activity of soybeans. Ongoing work is to characterize, at the at the morphological, molecular, and nutritional levels, diverse lines from wild sources and from a breeding effort in the 1980s, for use as a potential new crop. High-throughput sequencing, on a limited budget, has led to a high-quality reference transcriptome (of 62,614 contigs, with average length 818 nt), characterization of expression patterns by tissue, identification of several candidate genes for initiation of tuber formation, and detailed diversity characterization of a portion of the breeding collection. Analysis of sequence variation shows Apios to be highly diverse, with generally high levels of heterozygosity in this outcrossing species. Breeding objectives include large tuber size, high yields, short internodes in stolons, and dwarfing in the above-ground habit. Dense resequencing data may enable association analysis and selection for markers around domestication-syndrome genes in this promising potential crop. VI ICLGG - Abstracts Legume and symbiont phylogeography of South Florida 1,2, 1,2 3 S-EAD04 1,2 von Wettberg EJB *, Warschefsky E , Duyck F , Scharnagl K , 1,2 2 Sanchez V , Jestrow B 1 Florida International University, Miami, USA Fairchild Tropical Botanic Garden, Coral Gables, USA 3 Ecole Nacional Superieure Agronomique de Toulouse, France 2 *E-mail: evonwettberg@fairchildgarden.org Abstract When legumes disperse beyond their current range distributions, they are likely to undergo population bottlenecks, arrive without their rhizobial and mycorrhizal symbionts, and face differing soils and climates. In tropical southern Florida, where land has risen from ancient seabed in recent geological time, colonization by legumes has occurred both from temperate mainland North America and from the large, geologically old tropical islands of the Caribbean like Cuba and Hispaniola. Like southern Florida, the Bahamas have geologically young land that is former seabed, but colonization has only occurred over seawater because the Bahamas have never been connected to North America. Although colonization of southern Florida from mainland North America does not require crossing the sea as colonization from the Caribbean does, climate and soils differ more between southern Florida and the rest of North American than they do Cuba or Hispaniola. We expect a greater population bottleneck in Caribbean legume taxa than North American taxa in South Florida, while both should have large bottlenecks in the Bahamas. Rhizobia should also undergo population bottlenecks in colonizing recently formed land like Florida and the Bahamas. We examine these expectations in several legumes, with a focus on species of Chamaecrista from North America and the Caribbean. We find strong bottlenecks in the colonization of both Florida and the Bahamas, and stronger bottlenecks than expected in North American taxa. Ongoing work is examining patterns of potential bottlenecks in rhizobial species, and the extent to which host-symbiont networks shifted across different regions. VI ICLGG - Abstracts Theme: Harnessing Germplasm Resources CLL GOWDA Director Grain Legumes Research Program International Crops Research Institute for the Semi-Arid Tropics India c.gowda@cgiar.org C L Laxmipathi Gowda is Director of Grain Legumes Program at ICRISAT, Hyderabad, India. Dr Gowda received B.Sc (Ag) from University of Agricultural Sciences, Bengaluru; M.Sc (Ag) from GB Pant University of Agriculture and Technology, Pantnagar; and Ph.D. from Indian Agriculture Research Institute, New Delhi, India. Dr. Gowda has worked with chickpea breeders in 30 countries who have released 68 high-yielding varieties that have made significant impacts on farmers’ fields. As Cereals and Legumes Asia Network (CLAN) Coordinator, he has strengthened R&D capacity of 12 countries in Asia. Dr Gowda has been honored by Bangladesh, China, India, Nepal, Philippines, Thailand and Vietnam. Along with his colleagues, he initiated the Public-Private Partnership for Hybrid Parents Research at ICRISAT. Dr. Gowda has >360 publications, including 116 Journal Articles, 80 books/book chapters, 11 research bulletins, and 153 conference papers. He is an Editor of Food Security, Journal of Crop Science and Biotechnology and International Journal of Crop Production. Selected Publications: 1. Gowda CLL, Saxena KB, Srivastava RK, Upadhyaya HD, Silim SN. (2011). Pigeonpea: From an orphan to leader in food legumes. In Biodiversity in Agriculture: Domestication, Evolution & Sustainability (Gepts P, Bettinger RL, Brush SB, Famula TR, McGuire PE, Qualset CO and Damania AB, eds.). University of California, Davis, USA: Cambridge University Press. ISBN-13: 9780521170871. P361-373. 2. Gowda CLL, Upadhyaya HD, Dronavalli N, Singh S. (2011). Identification of large-seeded highyielding stable kabuli chickpea germplasm lines for use in crop improvement. Crop Sci. 51:198209. 3. Upadhyaya HD, Dronavalli N, Gowda CLL, Singh S. (2011). Identification and evaluation of chickpea germplasm for tolerance to heat stress. Crop Sci. 51: 2079-2094. 4. Upadhyaya HD, Salimath PM, Gowda, CLL, Singh S. (2007). New early-maturing germplasm for utilization in chickpea improvement. Euphytica 157:195-208. 5. Upadhyaya HD, Dwivedi SL, Gowda CLL, Singh S. (2006). Identification of diverse germplasm lines for agronomic traits in a chickpea (Cicer arietinum L.) core collection for use in crop improvement. Field Crops Research 100: 320-326. VI ICLGG - Abstracts Genetic resources and genomics for enhancing the efficiency of crop improvement in grain legumes I-HGR01 Gowda CLL*, Upadhyaya HD, Gaur PM, Saxena KB International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India. *E-mail: c.gowda@cgiar.org Abstract Grain legumes have great potential in alleviating protein hunger and malnutrition among poor people in developing countries, being the primary source of dietary protein. Besides their importance for human and animal nutrition, legumes fix nitrogen and are important components in the cropping systems for improving and sustaining soil fertility. Globally, ~1.1 million grain legume accessions are conserved in different genebanks, of which ICRISAT genebank holds ~50,000 accessions of cultivated and wild relatives of chickpea, pigeonpea, and groundnut from 133 countries. These genetic resources are reservoirs of many useful genes for the present and future crop improvement programs globally. ICRISAT’s strategic research on core (10% of entire collection) and mini core (10% of core or 1% of entire collection) collection approach has provided an entry point for crop scientists to utilize these resources in crop improvement. Evaluation of mini core collections of chickpea, pigeonpea and groundnut have resulted in identification of traitspecific germplasm for important agronomic traits (early maturity, large-seed, and high yield) as well as resistant/tolerance sources for various biotic (diseases and insect pests) and abiotic (drought, water-logging, heat, low temperature, and salinity) stresses and nutrition-related traits (oil, protein, O/L ratio) for use in breeding programs. In addition, vast genomic resources such as BAC libraries, ESTs, SSRs, DArT clones, and SNPs have been developed in chickpea, pigeonpea, and groundnut which will accelerate the molecular characterization of germplasm accessions and help in identification of genetically diverse trait-specific germplasm accessions to develop new cultivars with a broad genetic base. Based on these resources, dense genetic maps of these legume species have been developed and molecular markers linked with several traits of interest have been identified. Marker-assisted breeding approaches have already been initiated for some traits in chickpea and groundnut, and would lead to enhanced efficiency and efficacy of crop improvement. VI ICLGG - Abstracts BERT VANDENBERG Professor Plant Sciences University of Saskatchewan Canada bert.vandenberg@usask.ca Albert Vandenberg is a professor in the Department of Plant Sciences at the University of Saskatchewan. He currently holds the NSERC Industrial Research Chair in Lentil Genetic Improvement. His research group is actively engaged in many aspects of lentil genetics and genomics with a focus on developing a SNP marker based platform for genetic improvement of lentil. Major program goals are developing understanding of the genetic basis for improvement of yield, disease resistance, herbicide tolerance, lodging tolerance, and seed quality. The lentil breeding program at the UofS has produced a steady stream of improved varieties which now account for 35% of world lentil production and consumption. Dr. Vandenberg is actively involved in pulse industry development through participation in grower organizations, extension and international marketing and research projects with industrial partners. Selected Publications: 1. Vail SL, Vandenberg A. (2011). Genetic control of interspecific-derived and juvenile resistance in Lentil to Colletotrichum truncatum. Crop Science 51:1481-1490. 2. Tahir M, Baga M, Vandenberg A, Chibbar RN. (2012). An assessment of raffinose family oligosaccharides (RFO) and sucrose concentration in genus Lens. Crop Sci. 52:1713 – 1720. 3. Tullu A, Diederichsen A, Suvorova G. and Vandenberg A. (2011). Genetic and genomic resources of lentil: status, use and prospects. Plant Genetic Resources: Characterization and Utilization 9:19–29. 4. Vail S, Strelioff JV, Tullu A. and Vandenberg A. (2012). Field evaluation of resistance to Colletotrichum truncatum in Lens culinaris, Lens ervoides, and Lens ervoides × Lens culinaris derivatives. Field Crops Research 126: 145–151. 5. Yuan HY, Lulsdorf M, Tullu A, Gurusamy V, Vandenberg A. (2011). In vivo grafting of wild Lens species to Vicia faba rootstocks. Plant Genetic Res 9: 543-548 VI ICLGG - Abstracts Interspecific convergence of lentil genetics and genomics (ICLGG) 1 1 1 1 1 2 I-HGR02 2 Bett K , Fedoruk M , Tullu A , Saha S , Yuan HY , Sharpe A , Ramsay L , 1 1 1 1 1 Sari E , Banniza S , Podder R , Hajikamiloglu B, Kannan U , Tahir M , Chibbar 1 1 3 3 4 1 R , Vail S , Cook DR , Penmetsa V , Farmer A and Vandenberg A 1 University of Saskatchewan, Saskatoon, Canada National Research Council Canada, Saskatoon, Canada 3 Department of Plant Pathology, UC Davis, California, USA 4 NCGR, Santa Fe, NM, USA 2 *E-mail: bert.vandenberg@usask.ca Abstract The genus Lens has seven species, only one of which has been domesticated but several of which appear to be genetically accessible through hybridization and embryo rescue. Phenotypic traits of interest found in the wild germplasm include multiple disease resistances, seed appearance, nutritional quality and herbicide tolerance. We have developed extensive intraspecific and interspecific genetic resources which are being phenotyped and genotyped. This genetic and genomic information base will allow us to introgress genes of interest that affect the productivity of the lentil crop. Our intent is to expand the genetic base of the lentil crop which will lead to improvement of crop productivity and value for producers and consumers. VI ICLGG - Abstracts S-HGR03 Legumes germplasm subsets to select diverse trait-specific lines for use in improvement programs Upadhyaya HD*, Sharma S, Reddy KN, Singh S, Varshney RK, Gowda CLL 1 International Crops Research Institute for Semi-Arid Tropics (ICRISAT), Hyderabad, India *E-mail: h.upadhyaya@cgiar.org Abstract Plant genetic resources are essential for developing high yielding cultivars. About 1.1 million accessions of grain legumes have been conserved globally. ICRISAT’s genebank conserves about 50, 000 accessions of cultivated and wild relatives of chickpea, groundnut and pigeonpea from 133 countries. However, only a limited number (<1%) of accessions have been used in crop improvement, mainly due to lack of knowledge on traits of economic importance. The genebank databases do not have information that meets breeders’ needs for trait-specific, genetically diverse and agronomically desirable accessions that can be used as parents in breeding programs. To meet such needs, multilocational replicated evaluation is required which is not feasible with large germplasm collections. The mini core collections (10% of core or 1% of entire collection) serves as one of the efficient and convenient options for needed evaluation and identification of accessions with desirable traits, including resistance to abiotic (soil salinity/ acidity, drought, temperature, etc.) and biotic (pest and diseases) stresses and for nutritional quality traits. Using the mini core collection approach, the researchers at ICRISAT and in national programs have been able to identify diverse sources of resistance/tolerance for many biotic and abiotic stresses, and for agronomic and quality traits in chickpea, groundnut, and pigeonpea. This is expected to enhance the use of germplasm in crop improvement. Molecular characterization of the mini core collections have resulted in identification of genetically diverse trait-specific germplasm lines meeting the needs of breeders for use in developing high yielding cultivars with a broad genetic base. VI ICLGG - Abstracts Coupling genome resources to biological traits and breeding objectives in cowpea (Vigna unguiculata (L.) Walp) S-HGR04 Close TJ* University of California, Riverside, California, USA *E-mail: timothy.close@ucr.edu Abstract Cowpea (Vigna unguiculata (L.) Walp) is a primary source of protein in SubSaharan Africa where it is grown for fresh and dry grains, and for foliage. Cowpea is grown also in parts of Asia, Latin America, South America, Southeastern USA and California. Cowpea is a warm season legume with soybean, common bean and pigeon pea in the Phaseoleae tribe, sharing extensive genome similarities with these relatives. Core genomic resources of cowpea now include a draft genome assembly and 10X physical map of African breeding parent IT97K-499-35. These have been significantly coupled, mainly via an Illumina GoldenGate assay and minimal tiling path BAC sequencing, to a genetic linkage map composed of about 1100 SNP loci. The SNP map was developed as a consensus from more than 1000 individuals in 11 recombinant inbred populations. Sequences are available for BLAST via harvest-blast.org and can be retrieved via harvest-web.org. The linkage map with BAC information is available through the Windows software HarvEST:Cowpea (http://harvest.ucr.edu). A transition to marker-assisted breeding and mechanistic studies of cowpea trait determinants is underway as nearby markers of traits continue to emerge, and as candidate genes surface. Excellent SNP markers now are available for resistance to Striga, bacterial blight, Fusarium races 3 and 4, aphids, thrips, root-knot nematode, components of tolerance to water-limited environments including delayed senescence and recalcitrance to ashy stem blight, and various morphological and developmental traits. Through partnerships with NARS breeders these new resources are being integrated into African breeding programs to accelerate the development of improved varieties. VI ICLGG - Abstracts S-HGR05 Development of extra short duration genotypes in greengram (Vigna radiata L. Wilczek) 1, Pratap A *, Gupta DS, Rajan N 1 Indian Institute of Pulses Research (IIPR), Kanpur, India *E-mail: adityapratapgarg@gmail.com Abstract Greengram is economically one of the most important pulse crops of the Vigna group. It’s wider adaptability, low input requirements and shorter maturity duration (60-75 days) provide a scope of its cultivation in all seasons across different parts of India. Terminal heat stress during spring and summer and pre-harvest sprouting due to excess moisture during rainy season considerably reduce its productivity. If the life cycle of greengram is reduced by 10-15 days without significant yield penalty, it could be saved from above problems in the north and central India, the major mungbean growing areas. Keeping this in view, two extra short duration genotypes of greengram were developed at Indian Institute of Pulses Research, Kanpur. The genotype IPM205-7 was developed from the cross ‘IPM02-1 x EC398889’ following pedigree method of breeding while IPM409-4 was developed from ‘PDM288 x IPM03-1’, registered by NBPGR as INGR11043 and INGR11044, respectively. Resistant to mungbean yellow mosaic virus, both the genotypes have short and erect plants. The leaves are dark green, ovate and medium-sized with greenish purple veins while the flowers are light yellow in colour. The pods are borne above the canopy and are small and black on maturity while the seeds are shining green. Both the genotypes have been found to mature significantly early (46-48 days) during summer as well as rainy season as compared to the earliest maturing check varieties (PDM139 and Sona Yellow). These genotypes can be directly released as cultivars after their multilocation evaluation in formal chain besides using them as donors for transfer of earliness in agronomically superior backgrounds. VI ICLGG - Abstracts Present status of cultivation, constraints, level of on-farm improvement and breeding requirements of cowpea (Vigna unguiculata L. Walp) based on survey in Monaragala district of Sri Lanka 1,* 1 S-HGR06 1 Millawithanachchi MC , Bentota AP , Weerasinghe P , Saranasinghe 1 1 1 2 SNK , Prathapasinghe NT , Hettige DHNW , Kumari RAC 1 Grain Legumes and Oil Crops Research and Development Center Angunakolapellassa, Sri Lanka 2 Extension and Training Service, Uva Province, Sri Lanka *E-mail: withanamcm@yahoo.com Abstract On-farm survey was conducted to investigate present status of cowpea cultivation, major associated constraints, farmers’ adaptation for in-situ improvement, germplasm availability, seed flow network, and the future breeding requirements of crop with respect to three agriculture instructor regions of Monaragala district in Sri Lanka. Due to scarcity of water of surveyed area, only 22 % considered rice as main crop. Among the rest 62 % considered cowpea as the main crop. Adaptation of farmers to recommendations such as land preparation, fertilizer application, spacing and cultivation of recommended varieties were at minimal level. Most of farmers still practiced slash and burn system for land preparation. Due to high risk of this type of cropping system, return to the expense was uncertain. In-situ adaptation of farmers to select seeds from existing population based on desirable characters was 32%, and discarding based on undesirable characters was 46%. Farmers’ poor attention to keep the uniformity of the seed affected quality of the seeds. Due to the low selection pressure on the available germplasm and introductions from seed collecting centers and marketing personnel has created variability in existing populations of cowpea. Thirty two uniform lines were developed at the research centre from the collected samples of the region. High yielding, medium duration, drought and pest and diseases tolerance and glossy appearance were the major criteria demanded by the rainfed farmers from a cowpea improvement program in Sri Lanka. Further farmers give more priority to white seed color followed by red seed colour. VI ICLGG - Abstracts Theme: Abiotic Stress irector HENRY T. NGUYEN National Center for Soybean Biotechnology University of Missouri, Columbia, USA nguyenhenry@missouri.edu Henry Nguyen lab is pioneering molecular mapping and genetic dissection of drought tolerance in plants. Current efforts are focused on comparative genomics and genetics of drought tolerance in soybean. He has served on various NSF and USDA panels and served as Chairman for the Molecular Genetics and Cell Biology section of Crop Science journal and organizer/chairperson of the Abiotic Stress Workshop at the International Plant and Animal Genome Conference held annually in San Diego, CA. He has been invited to numerous international conferences and workshops to present results and discuss strategies for water and temperature stress tolerance research in relation to basic genomics and application to crop improvement. Serving on the steering and program committees for the International Congress on Drought Tolerance in Plants. Selected Publications: 1. Joshi T, Patil K, Fitzpatrick MR, Franklin LD, Yao Q, Cook JR, Wang Z, Libault M, Brechenmacher L, Valliyodan B, Wu X, Cheng J, Stacey G, Nguyen HT, Xu D. (2012). Soybean Knowledge Base (SoyKB): a web resource for soybean translational genomics. BMC Genomics. Suppl 1:S15. 2. Ha J, Abernathy B, Nelson W, Grant D, Wu X, Nguyen HT, Stacey G, Yu Y, Wing RA, Shoemaker RC, Jackson SA. (2012). Integration of the draft sequence and physical map as a framework for fenomic research in soybean (Glycine max (L.) Merr.) and wild soybean (Glycine soja Sieb. and Zucc.). G3 2:321-329. 3. Schmutz J, Cannon SB, Schlueter J, Ma J, Mitros T, Nelson W, Hyten DL, Song Q, Thelen JJ, Cheng J, Xu D, Hellsten U, May GD, Yu Y, Sakurai T, Umezawa T, Bhattacharyya MK, Sandhu D, Valliyodan B, Lindquist E, Peto M, Grant D, Shu S, Goodstein D, Barry K, Futrell-Griggs M, Abernathy B, Du J, Tian Z, Zhu L, Gill N, Joshi T, Libault M, Sethuraman A, Zhang XC, Shinozaki K, Nguyen HT, et al. (2010). Genome sequence of the palaeopolyploid soybean. Nature 463:178-183. 4. Wang Z, Libault M, Joshi T, Valliyodan B, Nguyen H, Xu D, Stacey G, Cheng J. (2010). SoyDB: a knowledge database of soybean transcription factors. BMC Plant Biol. 10:14. 5. Wu X, Ren C, Joshi T, Vuong T, Xu D, and Nguyen HT. (2010). SNP discovery by high-throughput sequencing in soybean. BMC Genomics. 11:469 VI ICLGG - Abstracts Exploitation of root system architecture and plasticity for improving drought resistance in grain legumes I-ABS01 Nguyen HT University of Missouri, Columbia, USA E-mail: nguyenhenry@missouri.edu Abstract Improved agricultural practices and development of cultivars for wide range of growing environments helped steady increase of global crop production and yield. Meeting the food demand of worlds growing population requires faster rate of yield increase for major crops. Drought is one of the major climatic conditions affecting yield and stability of crop production. The root system plays a vital role in plant adaptation and productivity under water-limited environments. Deeper and proliferate root system help extract enough water and nutrients under these environmental conditions. Our research program focuses on an integrated genetics and genomics approach to dissect molecular processes from transcript to phenome, in order to achieve increased water capture and use efficiency. Also, establishment of early seedling vigor and better root system architecture contribute towards enhanced crop productivity. We have screened and identified soybean lines which exhibit genetic diversity in root system developmental plasticity in response to water stress, in order to enable physiological and genetic analyses of the regulatory mechanisms involved. We have conducted extensive gene, protein and metabolite expression profiling to dissect molecular processes and genes associated with the root system responses under water deficit conditions. Functional characterization of key candidates, engineering of selected genes through translational genomics pipeline, and testing these transgenic plants to study associated physiological mechanism are in progress. This presentation will highlight the importance of beneficial root traits and recent discoveries towards better root system architecture in soybean for enhanced stress resistance and yield. VI ICLGG - Abstracts MARTIN CRESPI Institut des Sciences du Végétal (ISV)-CNRS, France crespi@isv.cnrs-gif.fr Our laboratory is interested in deciphering the mechanisms of action of different ribo regulators and their impact in plant growth and development by combining cell biology with genetic, genomic and molecular approaches in Arabidopsis and Medicago truncatula. The remarkable developmental plasticity of roots, their precisely defined gradient of differentiation, their capacity to form de novo meristems (lateral organs) from differentiated cells and their amenability for gene expression analysis, make them an attractive model to reveal mechanisms involving non-coding RNAs in adaptive developmental regulations to the environment. Hence, we focus our studies on the impact of post-transcriptional processes mediated by miRNAs and other non-coding RNAs in root responses to environmental stresses. Our results support the notion that a variety of non-coding RNAs (both long and small) through their interaction with specific regulatory genes (such as transcription factors or components of the epigenetic machinery) may modulate the developmental adaptation of the root system to environmental conditions. Selected Publications: 1. Zahaf O, Blanchet S, de Zelicourt A, Alunni B, Plet J, Laffont C, de Lorenzo L, Imbeaud S, Ichanté J-L, Diet A, Badri M, Zabalza A, González EM, Delacroix H, Gruber V, Frugier F, Crespi M. (2012) Comparative transcriptomic analysis of salt adaptation in roots of contrasting Medicago truncatula genotypes. Molec. Plant doi: 10.1093/mp/sss009. 2. Khan GA, Declerck M, Sorin C, Hartmann C, Crespi M, Lelandais-Brière C. (2011) Micro RNAs as regulators of root development and architecture. Plant Mol. Biol. 77: 47-58. 3. Ariel F, Diet A, Verdenaud M, Gruber V, Frugier F, Chan R, Crespi M. (2010) Environmental regulation of lateral root emergence in Medicago truncatula requires the HD-Zip I transcription factor Mt HB1. Plant Cell 22(7): 2171-83. 4. Jouannet V, Marin E, Herz A, Lokerse AS, Weijers D, Vaucheret H, Nussaume L, Crespi MD, Maizel A. (2010) miR390, TAS3 ta-siRNAs and their ARF targets define an auto-regulatory network quantitatively controlling lateral root growth. Plant Cell 22(4):1104-1. 5. de Lorenzo L, Merchan F, Laporte P, Thompson R, Clarke J, Sousa C, Crespi M. (2009) A novel plant LRR receptor kinase regulates the response of Medicago truncatula roots to salt stress. Plant Cell 21:668-680. VI ICLGG - Abstracts Transcriptional networks regulating legume root architecture adaptation to different environmental conditions 1 1 1,2 1 1 I-ABS02 1 De Zelicourt A , Zahaf O , Ariel F , Diet A , Lelandais C , Bustos P , Sallet 3 2 1 4 5 5 1 E , Chan R , Hartmann C , Badri M , Zabalza A , González EM , Gruber V , 3 1 1 Gouzy J , Frugier F , Crespi M 1 University of Paris VII, Paris, France Universidad Nacional del Litoral, Santa Fe, Argentina 3 LIPM (Laboratoire des Interactions Plantes Micro-organismes), CNRS-INRA, Castanet-Tolosan, France 4 Laboratory of Legumes, Centre of Biotechnology of Borj Cedria Hammam-Lif, Tunisia 5 Universidad Pública de Navarra, Pamplona, Spain 2 *E-mail: crespi@isv.cnrs-gif.fr Abstract Evolutionary diversity can be driven by the interaction of plants with different environments. Molecular bases involved in ecological adaptations to abiotic constraints can be explored using genomic tools. In legumes, root system architecture is determined by the formation of two different organs, lateral roots and symbiotic nitrogen-fixing nodules. Legumes are major crops worldwide and soil salinity is a main stress affecting yield in these plants. Comparative transcriptomic analyses using a genotype of the model legume Medicago truncatula sampled from salty soils, TN1.11, and the A17 reference line, identified several differentially salt-regulated TFs. Certain TFs are targeted by microRNAs, and we additionally identified, using deep sequencing of small RNA libraries, miRNAs differentially regulated in response to a salt stress and/or between root and nodules. We then selected several salt-regulated TFs for functional analyses. Among those, an HD-Zip TF (MtHB1) was shown to control lateral root emergence in response to salt through the direct modulation of an auxin-dependent LOB-domain TF. In addition, a NAC-type TF (MtNAC969), showing a contrasting salt regulation between roots and nodules, was linked both to root salt stress response and to symbiotic nodule senescence. Finally, over-expression in roots of the A17 genotype of the bHLH-type TF most differentially regulated between genotypes improved significantly root growth under salt stress. Despite the global complexity of the differential transcriptional responses, we propose that changes in the regulation TF expression may be linked to the adaptation of M. truncatula to saline soil environments. VI ICLGG - Abstracts GÜNTER KAHL Professor, Plant Molecular Biology Frankfurt University Germany kahl@em.uni-frankfurt.de Since 1973 Professor of Plant Molecular Biology, Molecular BioSciences, Biocenter of Frankfurt University (Germany) Visiting Professor in France, Poland, Japan, USA, Costa Rica (CATIE), Syria (ICARDA), Mexico, Venezuela, Austria (University Vienna). Organization of Molecular Biology Courses in Austria, Belgium, Brazil, Chile, Costa Rica, Malaysia, Mexico, Nigeria, Pakistan, Sri Lanka, Syria, Venezuela. Expert Missions for IAEA, FAO, World Bank, EU and UNESCO in Australia, Chile, Hongkong, India, Malaysia, Mexico, Pakistan, Sri Lanka, Syria, Tunisia, Europe. Organization of the workshop “Mutation Screening” at the Plant and Animal Genome (PAG) Conference in San Diego (USA) each year. Main Research Activities: Plant Genome Analysis, Fungal Genome Sequencing, Plant and Human Gene Expression Profiling. More than 250 scientific publications in peer-reviewed science journals. A total of 15 scientific books published, two more in 2012. Company responsibilities: Co-founder and Chief Scientific Officer, SME GenXPro, Frankfurt Innovation Center Biotechnology, specializing on (1) deep sequencing of human, animal, fungal and plant transcriptomes with patented deepSuperSAGE technology, discovery of novel genes and mutations and (2) technology development. VI ICLGG - Abstracts Salinity-induced expression landscape: Fast reply transcripts and transcript variants for oxygen burst and signal transduction in chickpea 1, 2 3 4 I-ABS03 5 Kahl G *, Molina C , Rotter B , Zaman-Allah M , Drevon JJ , Horres 1 , 5 3 1 3 R , Amenc L , Fatnassi N , Khan F , Winter P 1 University of Frankfurt, Frankfurt, Germany University of Kiel, Kiel, Germany 3 GenXPro GmbH, Frankfurt, Germany 4 International Crops Research Institute (ICRISAT), Hyderabad, India 5 INRA, Montpellier, France 2 *E-mail: kahl@em.uni-frankfurt.de Abstract Soil salinity limits crop production worldwide. Plants evolved several strategies to adapt to salinity stress to some extent, amongst them avoidance and tolerance. Both adaptations are relatively complex and therefore not well understood. But it goes without saying that breeding for durable tolerance requires a substantial improvement of our knowledge of the underlying molecular processes. We therefore began to characterize the salt stress transcriptome and the various sub-transcriptomes, especially transcriptomes encoding fast-reacting proteins, first-line defense proteins such as generators and detoxifiers of oxygen radicals, second-line ion exclusion and osmotic pressure adjustment proteins, and proteins catalyzing reactions in several signal transduction pathways. Chickpea plants were exposed to moderate salt stress of 25 mM NaCl in hydroaeroponics systems and the complexity of their root and nodule transcriptomes determined after an only 2 hours impact of salt. Many of the reactive genes (e.g. those encoding various aquaporins) are transcribed in anti-sense polarity, still others into multiple (splice) variants, that are time- and tissue-specifically regulated, and still others are down-regulated to various extents. The massive and complex data are composed to a gene expression panorama, whose complexity leads the researcher to his limits. VI ICLGG - Abstracts S-ABS04 An integrated framework combining crop physiology, genetics, and crop simulation modelling to improve drought resistance across legumes in the semi-arid tropics 1* 2 3 4 4 5 6 Vadez V , Halilou O , Belko N , Polania J , Rao IM , Sinclair TR , Soltani A , 1 2 Ratnakumar P , Hamidou F 1 International Crops Research Institute for Semi-Arid Tropics (ICRISAT), Hyderabad, India 2 International Crops Research Institute for Semi-Arid Tropics (ICRISAT), Niamey, Niger 3 Centre d’Etude Régional pour l’Amélioration de l’Adaptation à la Sécheresse, Thiès- scale, Sénégal 4 CIAT, Palmira, Colombia 5 North Carolina State University, Raleigh, USA 6 Gorgan University of Agricultural Sciences and Natural Resources, Gorgan ,Iran * E-mail: v.vadez@cgiar.org Abstract Drought stress is the main yield limiting factor across Asian and African semiarid tropics. Conventional breeding has contributed to major progress in developing varieties better adapted to drought, like early lines. However further progress is hampered by the usual large genotype-by-environment interactions in most field trials. A more comprehensive approach is required to revitalize the development of drought adapted materials. Many biotechnological solutions to drought have been proposed (transgenic, candidate gene, etc...), but have had limited concrete success. Their major drawback has been in not putting realistic water limitations at the centre of all considerations and designs of biotechnological work. While the genes are many, plant traits contributing to crops’ adaptation to relevant water limitation are few only, and revolve around the critical need to match water supply and demand. These relate in part to conservative water use, higher/better use of soil water, or success of reproduction/remobilization. However, traits themselves are highly dynamic during crop development: they interact among each other and with the environment. Further progress in breeding more drought resilient grain legume cultivars implies an understanding of plant traits affecting yield under drought, and of trait interactions. In addition, the phenotypic evaluation of germplasm/breeding material is limited by the number of locations and years of testing. Crop simulation modelling is a powerful tool to navigate the complexity of biological systems and predict the effect on yield and probability of success of specific traits across drought scenarios. We will present this integrated approach, using examples from on-going work. VI ICLGG - Abstracts Identification of drought-induced transcription factors in peanut (Arachis hypogaea L) 1 Dang PD , Chen CY 1, 2, *, Holbrook CC S-ABS05 3 1 National Peanut Research Laboratory, Dawson, Georgia, USA Auburn University, Auburn, USA 3 Crop Genetics and Breeding Research Unit, Georgia, USA 2 *E-mail: cyc0002@auburn.edu Abstract Transcription factors play key roles in the regulation of genes involved in normal development as well as tolerance to biotic and abiotic stresses. Specific transcription factors that are induced in peanut under drought conditions have not been identified. The objectives of this study were to compare gene-expression patterns of various transcription factors of a drought tolerant versus a susceptible peanut genotype under drought conditions and to identify transcripts that were regulated in a drought dependent manner. Twelve putative transcription factors were identified and real-time PCR analysis were performed which resulted in the identification of three unique transcripts in which ahERF1 was highly induced in recovery stage; and ahERF7 and ahERF8 were highly induced by drought and returned to nominal levels after recovery. These sequences contain DNA binding domains that are present in APETALA2/Ethelene Responsive Factors (AP2/ERF) family of transcription factors which have been shown to be induced by stress. Induction levels and patterns of gene-expression of ahERF1, ahERF7 and ahERF8 may be used to select plants that may have higher drought tolerance. VI ICLGG - Abstracts Theme: Pathogenesis and Disease Resistance KARAM SINGH Chief Research Scientist, CSIRO Plant Industry; Winthrop Professor, Institute of Agriculture, The University of Western Australia Australia Karam.Singh@csiro.au Karam Singh holds a joint appointment between CSIRO Plant Industry as a Chief Research Scientist, and the University of Western Australia as a Winthrop Professor in the areas of Molecular Plant Pathology and Crop Genomics. His major interests are the regulation of plant stress gene expression, plant defence responses to insect pests - particularly aphids, and fungal pathogens - particularly rizochtonia solani and legume genomics with a focus on lupins. Selected Publications: 1. Guo S-M, Kamphuis LG, Gao L-L, Klingler JP, Lichtenzveig J, Edwards OR, Singh KB (2012). Identification of distinct quantitative trait loci associated with defense against the closely related aphids Acyrthosiphon pisum and A. kondoi in Medicago truncatula. Journal of Experimental Botany, In Press. 2. Gleason CA, Huang S, Thatcher LF, Foley RC, Anderson CR, Carroll AJ, Millar AH, Singh KB (2011). Complex II has a key role in production of mitochondrial-derived reactive oxygen species influencing plant stress gene regulation and defence. Proc Natl Acad Sci USA www.pnas.org/cgi/doi/10.1073/pnas.1016060108 . 3. Gao LL, Hane J, Kamphuis LG, Foley RC, Shi B-J, Atkins CA. and Singh KB. (2011). Development of genomic resources for the narrow-leafed lupin (Lupinus angustifolius): construction of a bacterial artificial chromosome (BAC) library and BAC-end sequencing. BMC Genomics 2011, 12:521. 4. Anderson JP, Lichtenzveig J, Gleason C, Oliver RP, Singh KB. (2010). The B-3 ethylene response factor MtERF1-1 mediates resistance to a subset of root pathogens in Medicago truncatula without adversely affecting symbiosis with rhizobia. Plant Phys 154: 861-873. 5. Sappl PG, Carroll AJ, Clifton R, Lister R, Whelan J, Millar AH, Singh KB. (2009). The glutathione transferase gene family displays complex stress regulation and co-silencing multiple genes results in altered metabolic sensitivity to oxidative stress. Plant J 58: 53-68. VI ICLGG - Abstracts Resistance to sap-sucking insect pests and soil borne necrotrophic fungal pathogens in a model legume, Medicago truncatula 1,2,* 1,2 1 1 1,3 I-PAD01 1 Singh KB , Kamphuis L , Gao L , Hane J , Klingler JP , Guo S , 1,4 1 3 1,2 Lichtenzveig J , Zulak K , Edwards OR , Anderson J 1 CSIRO Plant Industry, Floreat, Western Australia, Australia University of Western Australia, Crawley, Australia 3 CSIRO Ecosystem Science, Floreat, Western Australia, Australia 4 Curtin University, Western Australia, Australia. 2 *Email: Karam.singh@csiro.au Abstract We are interested in two areas of biotic stress research in legumes. One is on aphids and related phloem-feeding insects, which cause severe plant damage, through feeding activities and as vectors of plant viruses. A focus has been on resistance to bluegreen aphid (BGA) where we have fine mapped a dominant gene conditioning resistance called AKR (Acyrthosiphon kondoi resistance). Targeted approaches to identify signaling pathways and genomic approaches involving transcriptomics and metabolomics are helping understand how this gene confers aphid resistance, through the identification of signaling pathways, transcription factors and key metabolites important for AKR mediated resistance. Progress is also being made in identifying other aphid resistance genes in M. truncatula including single dominant resistant genes against spotted alfalfa aphid (SAA) and pea aphid (PA). We have also characterised moderate forms of resistance to BGA, PA, SAA and cowpea aphid that are genetically more complex and mediated through multiple QTLs. Another area involves soil borne necrotrophic fungal pathogens which are causal agents of numerous important diseases. One pathogen is Rhizoctonia solani, with a major focus on R. solani AG8, the causal agent of root and crown rot resulting in bare patch of legumes, cereals and brassicas. This broad host range suggests that pathogenesis and host defence mechanisms may be different to those employed during more specific plant/pathogen interactions. Good progress has been made through studying the mechanisms involved in partial resistance in M. truncatula and in deciphering the genome, proteome and transcriptome of R. solani AG8. VI ICLGG - Abstracts VALÉRIE GEFFROY IBP, INRA Université de Paris Sud France valerie.geffroy@u-psud.fr Valérie Geffroy is a researcher from INRA (Institut National de la Recherche Agronomique) working at the Institute of Plant Biology (IBP) in Orsay (France). Her research focuses on the study of the evolution of disease resistance genes in legume, particularly common bean. Most of her efforts have focused on resistance to the fungus Colletotrichum lindemuthianum and to the bacteria Pseudomonas syringae pv phaseolicola. Selected Publications: 1. Ashfield T, Egan AN, Pfeil BE, Chen NWG, Podicheti R, Ratnaparkhe MB, Ameline-Torregrosa C, Denny R, Cannon S, Doyle JJ, Geffroy V, Roe BA, Maroof MAS, Young ND, Innes RW. (2012). Evolution of a complex disease resistance gene cluster in diploid Phaseolus and tetraploid Glycine. Plant Physiol 159: 336-354. 2. Chen NWG, Sevignac M, Thareau V, Magdelenat G, David P, Ashfield T, Innes RW, Geffroy V. (2010). Specific resistances against Pseudomonas syringae effectors AvrB and AvrRpm1 have evolved differently in common bean (Phaseolus vulgaris), soybean (Glycine max), and Arabidopsis thaliana. New Phytol 2010, 187:941-956. 3. David P, colas des Francs-Small C, Sévignac M, Thareau V, Macadre C, Langin T, Geffroy V. (2010). Three highly similar formate dehydrogenase genes located in a cluster of NBS-LRR are differentially expressed under biotic and abiotic stresses in Phaseolus vulgaris. Theoretical and Applied Genetics. 121: 87-103. 4. David P, Chen NWG, Pedrosa-Harand A, Thareau V, Sévignac M, Cannon SB, Debouck D, Langin T, Geffroy V. (2009). A nomadic subtelomeric disease resistance gene cluster in common bean. Plant physiology. 151: 1048-1065 5. V. Geffroy, C. Macadré, P. David, A. Pedrosa-Harand, M. Sévignac, C. Dauga, T. Langin. (2009). Molecular analysis of a large subtelomeric NBS-LRR family in two representative genotypes of the major gene pools of Phaseolus vulgaris. Genetics. 181: 405-419. VI ICLGG - Abstracts Evolution of subtelomeric disease resistance gene clusters in common bean (Phaseolus vulgaris) 1 1 1 1 1 I-PAD02 1 Chen NWG , Richard M , David P , Sévignac M , Thareau V , Pflieger S , 1 3 2 2 3 Alunni B , Ribeiro T , Ashfield T , Innes RW , Pedrosa-Harand A , 1,4* Geffroy V 1 Institut de Biologie des Plantes, Université Paris Sud, UMR-CNRS 8618, Orsay, France 2 Department of Biology, Indiana University, Bloomington, Indiana, USA 3 Laboratory of Plant Cytogenetics, Federal University of Pernambuco, Recife, Brazil 4 INRA, Unité Mixte de Recherche de Génétique Végétale, Gif sur Yvette, France *Email: Valerie.geffroy@u-psud.fr Abstract In the genome of common bean (Phaseolus vulgaris; Pv), large disease resistance gene (R) clusters are most often localized to the end of linkage groups (LG). This is true for the Co-x, Co-2 and B4 R gene clusters, localized at one end of LG-B1, LG-B11 and LG-B4, respectively. R genes and QTLs conferring resistance to a diverse selection of pathogens including fungi (Colletotrichum lindemuthianum, Uromyces appendiculatus) and bacteria (Pseudomonas syringae) have been mapped to these clusters. Sequencing of B4 and Co-2 clusters revealed that they each contain more than 40 CoiledCoil-Nucleotide-Binding-Site-Leucine-Rich-Repeat (CNL) genes. The origin of these CNL sequences was investigated through phylogenetic analysis which revealed that B4 and Co-2 CNL genes are closely related while belonging to nonhomologous chromosomes 4 and 11. Our results suggest that B4 CNL are derived from the Co-2 cluster via an ectopic recombination event. FISH analyses revealed that both R clusters are located in a similar genomic environment, adjacent to terminal knobs (heterochromatic blocks) and associated with a 528-bp subtelomeric satellite repeat, referred to as khipu. For both R gene clusters, khipu is interspersed between CNL sequences and is also present in the adjacent knobs. Khipu is specific to the Phaseolus genus and present on most Pv chromosomal termini, indicating the existence of frequent ectopic recombination events between Pv subtelomeric regions. To study the recent evolution of the Co-2 R cluster, we used a combination of sequence, genetic and cytogenetic analyses in genotypes of distinct geographical origins (Andean and Mesoamerican). Our results show that subtelomeres are rapidly evolving regions of the Pv genome and correspond to favorable niches for R gene proliferation. VI ICLGG - Abstracts S-PAD03 Molecular insight into the early defense signal modulation in chickpea upon Fusarium oxysporum f. sp. ciceris attack Gupta S, Das S* Bose Institute, Kolkata, India *E mail: sampa@bic.boseinst.ernet.in Abstract Chickpea, the most important pulse crop of India suffers from massive yield loss annually due to high susceptibility to Fusarium oxysporum f. sp.ciceris. The pathogen invades the root and colonizes vascular tissue at the root-stem transition zone preventing the translocation of essential solutes leading to wilting of the plant. Pathogenic variability leading to breakdown of naturally selected resistance is the main hindrance for breeders. Although, some chickpea genotypes show convincing resistance against the pathogen, the information on resistant gene/(s) are limiting. This prompted to generate transcriptomic information by triggering defense responses upon pathogen inoculation at molecular level. The present chickpea- Fusarium case study based on detailed investigation on pathogen progression using functional genomic approach contradicted the pre established classical theories of HR mediated programmed cell death and down stream defense signaling in plantpathogen interactions. The present study documented that hypersensitivity and reactive oxygen species generation failed to impart host defense in compatible interaction. On the contrary, the defense related gene(s) played a critical role in conferring natural resistance in resistant host. Pathogen invasion led to altered expression of early defense responsive transcripts relating to secondary metabolism, kinases, protease inhibitors, lipases, peptide hormones etc. Sugars were also found to act as signal generators and transmitters maintaining intercellular links. Functional characterization of above genes shall provide new insight to develop better understanding of chickpea- Fusarium inter-chemistry. VI ICLGG - Abstracts Gene expression revealed during the interaction of resistant Arachis stenosperma and Meloidogyne arenaria 1,* 2 3 1 S-PAD04 1 Guimarães PM , Morgante CV , Roberts PA , Silva -Jr OB , Arrais L , Leal1 1 1 4 1 Bertioli SC , Araujo ACG , Moretzsohn MC , Bertioli D , Brasileiro ACM 1 EMBRAPA Genetic Resources and Biotechnology Brasilia, Brazil EMBRAPA Semi –Arid, Petrolina, Brazil 3 University of California Riverside, CA 92521, California-USA 4 University of Brasilia, Brasilia, Brazil 2 *E-mail: patricia.guimaraes@embrapa.br Abstract The plant-parasitic root-knot nematodes have evolved sophisticated strategies for exploiting plants with high impacts in agriculture worldwide. The wild species Arachis stenosperma shows high levels of resistance to M. arenaria and other parasites. A genome-wide overview of differential gene expression during this plant–nematode interaction at different time points (3, 6 and 9 DAI) and healthy roots was conducted using Illumina Hi-Seq 2000. Eight cDNA libraries produced a total of 38´241´633 reads with an average sequence size of 200 bp and were assembled into 44,133 contigs. These unique sequences were grouped into eight clusters corresponding to their expression profile estimated by RPKM values. We found that a comparable number of genes were up- and down-regulated during the resistance response and genes belonging to the same family were often differentially regulated. Defencerelated genes (calmodulin, patatin, DC1, RGAs and PR), transcription factors and components of hormonal signaling pathways (ARP, CKX) were further characterized using qRT-PCR for validation of their expression profile. This study aimed to characterize genes regulated during early response of resistant Arachis to M. arenaria, which in association with gene-expression analysis at the cellular level, in planta gene function validation and biochemical data, will provide a set of candidate genes for plant transformation aiming nematode resistance in peanut. VI ICLGG - Abstracts Theme: Genomic Resources and Trait Mapping SACHIKO ISOBE Head, Applied Plant Genomics Kaduza DNA Research Institute (KDRI) Japan sisobe@kazusa.or.jp Sachiko Isobe had been a conventional red clover breeder for thirteen years. Currently she is a molecular biologist, lab head of Applied Plant Genomics of Kazusa DNA Research Institute. The research group has performed genome sequencing and molecular genetics for various model and crop plants including ten legume species, Lotus japonicus, groundnut, soybean, azuki bean, pea, faba bean and four clover species. Her research has focused on developing DNA markers, linkage map construction, QTL identification and molecular breeding. Selected Publications: 1. Nakaya A, Isobe S (2012). Will genomic selection be a practical method for plant breeding? Annals of Botany doi:10.1093/aob/mcs109. 2. Isobe SN, Hisano H, Sato S, Hirakawa H, Okumura K, Shirasawa K, Sasamoto S, Watanabe A, Wada T, Kishida Y, Tsuruoka H, Fujishiro T, Yamada M, Kohara M, Tabata S (2012). Comparative genetic mapping and discovery of linkage disequilibrium across linkage groups in white clover (Trifolium repens L.) G3 2:607-617. 3. The Tomato Genome Consortium (2012). The tomato genome sequence provides insights into fleshy fruit evolution, Nature 485, 635-641. 4. Shirasawa K, Koilkonda P, Aoki K, Hirakawa H, Tabata S, Watanabe M, Hasegawa M, Kiyoshima H, Suzuki S, Kuwata C, Naito Y, Kuboyama T, Nakaya A, Sasamoto S, Watanabe A, Kato M, Kawashima K, Kishida Y, Kohara M, Kurabayashi A, Takahashi C, Tsuruoka H, Wada T, Isobe S (2012). In silico polymorphism analysis for the development of simple sequence repeat and transposon markers and construction of linkage map in cultivated peanut. BMC Plant Biol 12:80. 5. Koilkonda P, Sato S, Tabata S, Shirasawa K, Hirakawa H, Sakai S, Sasamoto S, Watanabe A, Wada T, Kishida Y, Tsuruoka H, Fujishiro T, Yamada M, Kohara M, Suzuki S, Hasegawa M, Kiyoshima H, Isobe S (2012). Large-scale development of EST-derived SSR markers and diversity analysis in Arachis spp. Molecular Breeding 30:125-138. VI ICLGG - Abstracts Identification of optimum DNA markers sets for MAS in red clover with a novel approach based on ‘Adaboost’ algorithm 1 2 3 4 3 I-GRTM01 5 Nakaya A , Sharma TR , Boller B , Klimenko I , Kölliker R , Rana JC , 6 6 6 6 6 Shirasawa K , Hirakawa H , Sato S , Tabata S , and Isobe S * 1 Niigata University, Niigata, Japan. Himachal Pradesh Agricultural University, Palampur, India 3 Swiss Federal Research Station for Agroecology and Agriculture, Zurich, Switzerland 4 All-Russian Williams Fodder Crop Research Institute, Moscow Region, Russia 5 National Bureau of Plant Genetic Resources, Shimla, India 6 Kazusa DNA Research Institute, Chiba, Japan 2 *E-mail: sisobe@kazusa.or.jp Abstract Red clover (Trifolium pratense L.) is a diploid forage legume species, widely cultivated in cool and temperate regions of the world. Genomic selection (GS) has been expected to be an effective approach for marker assisted selection (MAS) in forage species, because it is based on the idea of recurrent selection, which has been traditionally employed in forage breeding programs. However, the operation cost of GS is still expensive due to necessity of genome wide genotyping data of both training and breeding populations. In this study, we demonstrated identification of optimum DNA marker sets for MAS of QTLs with a novel approach based on ‘Adaboost’ algorithm. An Advanced Intercross Line (AIL) population was generated from crosses between six individuals originating from Switzerland, Russia and Japan and used as training and validating populations. Approximately 200 plants each were established in 2010 and 2011 in the fields of India, Russia, Swiss and Japan. Seed yields, flowering characteristics and morphological traits were evaluated in each region. The each set of individuals investigated in different regions and years was regarded as a sub-population. Genotype data was obtained for all the sub-populations with 768 SNP and 75 SSR markers. The marker sets (20 ≤) relating to the investigated traits were selected in each sub-population based on the Adaboost algorithm. Cross validation of the selected marker sets within and between sub-populations indicated the availability of the novel approach. VI ICLGG - Abstracts ANA MARIA TORRES IFAPA Córdoba Spain anam.torress.romero@juntadcandatucia.es Ana M Torres is Senior Research Scientist at IFAPA (Institute for Agricultural and Fisheries Research and Training) in Córdoba, Spain, with more than 20 years of experience in legume molecular genetics. Her main research interests are: Classical and marker-assisted breeding, Genetic map development in different legumes (faba bean, pea and chickpea), Quantitative Trait Loci (QTL) analysis for disease resistance, yield and quality parameters with the final aim of using molecular markers to tag genes of interest, Comparative genetics of crop legume plants. Her research group has led the way in the development of the faba bean genetic maps available so far in the crop. Research has focused primarily on the identification of gene(s) and QTLs controlling resistance to broomrape (Orobanche crenata), ascochyta blight (Ascochyta fabae) and rust (Uromyces viciae-fabae), as well as on yield components and nutritional aspects such as tannins and vicine-convicine content. As a result, several molecular markers for marker assisted selection (MAS) are already available in this crop. Dr. Torres has participated in national and European projects on legumes, acting as task coordinator in st faba bean molecular breeding and comparative mapping. She was co-organizer of the 1 FABA BEAN International Workshop on faba bean Breeding and Agronomy, held in Córdoba in 2006. Recently, she was elected vice-president of the Spanish Association for Legumes (AEL). Selected Publications: 1. Cruz-Izquierdo S, Avila C, Satovic Z, Palomino C, Gutierrez N, Ellwood S, Phan H, Cubero J, Torres AM (2012). Comparative genomics to bridge Vicia faba with model and closely related legume species: stability of QTLs for flowering and yield-related traits. Theor Appl Genet (in press). 2. Gnanasambandam A, Paull J, Torres AM, Kaur S, Leonforte T, Li H, Zong X, Yang T. and Materne M (2012). Impact of molecular technologies on faba bean (Vicia faba L.) Breeding strategies. Special issue of Agronomy (ISSN 2073-4395). Impact of Genomics Technologies on Crop Breeding Strategies (in press). 3. Torres AM, Avila CM, Stoddard FL, Cubero JI, (2011). Faba bean. In: Kole, C. (series ed.); Pérez de la Vega., Torres AM., Cubero JI. and Kole C. (ed.). Genetics, Genomics and Breeding in Crop Plants: Cool Season Food Legumes. Science Pubs Inc., New Hampshire, Jersey, Plymouth. 4. Fondevilla S, Almeida NF, Satovic Z, Rubiales D, Vaz Patto MC, Cubero JI, Torres AM. (2011). Identification of common genomic regions controlling resistance to pinodes M., earliness and architectural traits in different pea genetic backgrounds. Euphytica 182:43-52. 5. Duc G, Link W, Marget P, Stoddard FL, Torres AM, Cubero JI. (2011). Genetic Adjustment to Changing Climates: Vicia faba. In: Crop Adaptation to Climate Change, First Edition (2011) by Shyam S. Yadav, Bob Redden, Jerry S. Hatfield, Hermann Lotze-Campen and Anthony Hall (eds). John Wiley & Sons, Ltd. Published 2011 by Blackwell Publishing Ltd. VI ICLGG - Abstracts Genomics research for faba bean (Vicia faba L.) breeding applications 1 2 3 1 1 4 I-GRTM02 1, Avila C , Satovic Z , Madrid E , Ocaña S , Ruiz MD , Cubero JI , Torres AM * 1 IFAPA-Alameda del Obispo Córdoba, Spain Faculty of Agriculture Svetosimunska, Zagreb, Croatia 3 University of Córdoba, Córdoba, Spain 4 CSIC-Instituto de Agricultura Sostenible Córdoba, Spain 2 *E-mail: anam.torres.romero@juntadeandalucia.es Abstract Domesticated in ancient times, faba bean (Vicia faba L.) is now a major food and feed legume, particularly in Eurasia and North Africa. Despite being one of the cytogenetically best characterized plant species, subsequent progress in faba bean genomics has lagged behind other legume crops. Comprehensive genomic and post genomic resources are being developed but the very large and repetitive faba bean genome has so far restricted its sequencing and hampered genetic research and breeding applications. There are over 10 mapping populations established to study the inheritance of agronomic traits in different locations. Different molecular marker sets have been used to construct saturated linkage maps and identify genes/QTLs controlling resistance to crenate broomrape and several fungal major diseases. Recently, markers linked to growth habit and nutritional value of faba bean seeds were also identified. Despite these significant efforts, complete translation of marker discovery to faba bean breeding is still to be achieved. Recent efforts offer great promise for the development of highly accurate selective breeding tools. Thus, transcriptomic analysis (SuperSAGE and cDNA libraries) are enriching the scarce faba bean ESTs libraries providing additional resources for refining the maps with functional markers. Moreover, a combination of candidate gene and colinearity studies is being pursued to identify genes underlying agronomically important traits including seed size, flowering time, disease resistances and plant architecture. Finally, a faba bean functional consensus map is being constructed integrating all the genes and QTLs previously published. These new tools may help researchers to find candidate genes to accelerated improvement of future faba bean cultivars. VI ICLGG - Abstracts S-GRTM03 Generation of ultradense genetic maps for peanut A and B genome RIL populations and a tetraploid mapping population 1 1 1 2 2 Froenicke L , Beitel C , Scaglione D , Pandey MK , Upadhyaya HD , 3 3 3 2 Moretzsohn MC , Guimaraes P , Leal-Bertioli SCM , Varshney RK , 4 1 Bertioli D , Michelmore RW * 1 University of California Davis, Davis, California, USA International Crops Research Institute for Semi-Arid Tropics (ICRISAT), Hyderabad, India 3 Embrapa Recursos Genéticos e Biotecnologia, Brasília, Brazil 4 Catholic University of Brasília, Brasília, Brazil 2 *E-mail: rwmichelmore@ucdavis.edu Abstract We are generating ultra-high density genetic maps through genotyping by low coverage, shotgun sequencing. These maps are generated through sequencing of diploid and tetraploid peanut mapping populations and of a peanut diversity panel. They will assist the reference genome assembly and provide the foundation for GWAS and QTL mapping studies. RIL mapping populations of the diploid wild progenitor species of both the peanut A and B genomes are analyzed as well as a tetraploid mapping population that consists of a cross of cultivated peanut with a synthetic amphidiploid of the two progenitor species and a cultivated peanut RIL panel phenotyped for agricultural traits. The diversity panel consists of 300 accessions selected on the basis of SSR genotyping data to represent the diversity of the peanut germplasm collection at ICRISAT. In contrast to previous mapping-bysequencing efforts, the maps are generated for species without an existing reference genome using a new genome mapping pipeline. This pipeline has been established by analyzing the Arachis B genome. The gene space of the parental lines of each RIL population (20x coverage) and of each RIL (1x coverage) and each accession of the diversity panel accessions are being sequenced using an Illumina HiSeq 2000. The SNPs identified in the diversity panel will be analyzed for linkage disequilibrium and the LD data will be used to refine the genetic bins generated from the RIL segregation data. VI ICLGG - Abstracts Deep sequencing of chickpea transcriptome for gene discovery, marker development and gene expression studies S-GRTM04 Garg R, Patel RK, Jhanwar S, Priya P, Bhattacharjee A, Yadav G, Bhatia S, Chattopadhyay D, Tyagi AK, Jain M* National Institute of Plant Genome Research (NIPGR), New Delhi – 110067, India. *E-mail: mjain@nipgr.res.in Abstract Chickpea ranks third among the food legume crop production in the world. However, the availability of limited genomic resources has been an important constraint in chickpea improvement. The generation of large-scale ESTs/transcripts is a very useful approach for gene discovery and marker development. We sequenced the transcriptome of chickpea (Cicer arietinum L.) using short-read Illumina and long-read Roche 454 platforms. We have assessed the effect of sequence quality, various assembly parameters and programs on the final assembly output. The optimization of de novo assembly via hybrid assembly of long-read and short-read primary assemblies generated a set of 34,760 transcripts with an average length of 1,020 bp representing about 4.8% (35.5 Mb) of the total chickpea genome. We identified 4,111 simple sequence repeats (SSRs) in the transcriptome, which could be developed as functional molecular markers in chickpea. Furthermore, we assigned putative function to more than 73% and one or more GOSlim terms to 71.9% chickpea transcripts. A total of 1,851 (5.33%) transcripts encoding members of different transcription factor families were identified. Based on various BLAST searches, we identified two sets of lineage-specific genes, including 741 genes that are legume specific and 3,632 genes as chickpea specific. We also investigated tissue-specific expression of chickpea transcripts based on RNA-seq data to identify the genes involved in various biological processes. A web resource, Chickpea Transcriptome Database (CTDB), has been developed for access of the transcriptome data. This study provides rich genomic resources for chickpea and will accelerate functional genomic studies and breeding programmes in chickpea. VI ICLGG - Abstracts S-GRTM05 Mungbean genome-wide SNP detection by Illumina HiSeq 1 1 1 2 3 Van K , Kang Y , Han K-S , Gwag J-G , Moon J-K , Lee S-H 1,* 1 Seoul National University, Seoul, Korea National Academy of Agricultural Science, Suwon, Korea 3 National Institute of Crop Science, Suwon, Korea 2 *E-mail: sukhalee@snu.ac.kr Abstract Mungbean (Vigna radiata (L.) Wilczek), a self-pollinated diploid plant with 2n = 22 chromosomes, is an important legume crop with high quality amino acid profiles. Sequence variation at the whole-genome level was examined by comparing two mungbean cultivars, Sunhwanokdu and Gyeonggijaerae 5, using Illumina HiSeq sequencing data. More than 40,000 million reads from both mungbean cultivars were generated to a sequencing depth of 72x. After de novo assembly of Sunhwanokdu contigs by ABySS1.3.2. (N50=9,958 bp), those longer than 10 kb were aligned with Gyeonggijaerae 5 reads using the Burrows-Wheeler Aligner. SAMTools was used for retrieving single nucleotide polymorphisms (SNPs) between Sunhwanokdu and Gyeonggijaerae 5, defining the lowest and highest depth as 5 and 100, respectively, and sequence quality as 100. Of the 305,504 single base changes identified, 40,503 SNPs were considered as unusual single substitutions (or hetero). Among the remaining 265,001 SNPs, 65.9% were transitions and 34.1% (90,422 cases) were transversions. Sunhwanokdu contigs longer than 10 kb and sharing at least 80% sequence identity with common bean expressed sequence tags (ESTs) as determined with est2genome were selected. Only SNPs positioned in the putative exon regions were used for validation of mungbean SNPs. Using seven mungbean cultivars from various origins in addition to Sunhwanokdu and Gyeonggijaerae 5, most of the SNPs identified by bioinformatics tools were confirmed by Sanger sequencing. These SNP markers could enrich the current EST-based molecular resource and might be of value for the construction of a transcript map of mungbean. VI ICLGG - Abstracts Establishment of a Lotus japonicus gene tagging population using the endogenous retrotransposon LORE1 1,2 1 1 2 2 S-GRTM06 2 Fukai E *, Soyano T , Umehara Y , Nakayama S , Hirakawa H , Tabata S , 2 1 Sato S , Hayashi M 1 National Institute of Agrobiological Sciences, Tsukuba, Japan Kazusa DNA Research Institute, Kisarazu, Chiba, Japan 2 *E-mail: efu@affrc.go.jp Abstract The model legume Lotus japonicus has facilitated dissection of the molecular mechanisms governing symbiotic nitrogen fixation with rhizobia. From this model legume, we had identified a transpositionally active LTR retrotransposon family designated as LORE1 (Lotus Retrotransposon 1). Using this endogenous LTR retrotransposon, we established a gene tagging population of L. japonicus. The population was composed of 2,450 plant lines from which a total 4,532 flanking LORE1 sequence tags were recovered by pyrosequencing. The two-dimensional arrangement of the plant population, together with use of multiple identifier sequences in the primers used to amplify the flanking regions, made it possible to trace insertions back to the original plant lines. The large-scale detection of new LORE1 insertion sites revealed a preference for genic regions, especially in exons of protein coding genes, an interesting feature to consider interaction between host genomes and chromoviruses, a class of retrotransposon widely distributed in plants to which LORE1 belongs. Forward screening of the symbiotic mutants from the population succeeded to identify five symbiotic mutants of known genes. These data suggested the robustness of LORE1 as a genetic tool. For the further accumulation of mutated loci, now we continue to work on increasing the population. Utilizing this mutant population together with other resources will accelerate molecular biological studies in legumes. The information of insertion sites is available from http://www.kazusa.or.jp/lotus, and the seeds are available from The National BioResource Project Lotus/Glycine (http://www.legumebase.brc.miyazaki-u.ac.jp/index.jsp) by request. VI ICLGG - Abstracts Theme: Genomics - Assisted Breeding JEAN-MARCEL RIBAUT Director Generation Challenge Programme (GCP) Mexico j.ribaut@cgiar.org Jean-Marcel Ribaut, PhD, Director of the Generation Challenge Program (GCP) in Mexico is responsible for leading and coordinating a large network of partners in modern crop breeding for food security. JeanMarcel’s scientific background is in plant physiology and genetics, with main research interest being understanding the genetic basis and underlying physiological and metabolic pathways that influence plant performance under abiotic stress– particularly drought – as well as innovations in molecular breeding. He holds a PhD in Plant Physiology from Lausanne University, Switzerland, and has cumulative experience in agriculture biotechnology and plant breeding, project and finance management and policy formulation, as well as leadership skills for dispersed global R&D teams. Jean-Marcel has a particular interest in promoting modern breeding methods to hasten crop improvement in the developing world. He believes in true partnership and solid capacity building to overcome some of the bottlenecks in R4D, with the developing-country partners as key actors and leaders in the research arena. Prior to becoming GCP Director in 2005, Jean-Marcel worked at CIMMYT, Mexico, where he held positions of increasing responsibility, rising to Deputy Director of the Genetic Resources Programme in 2003 and Biotechnology Group Leader in 2004. Selected Publications: 1. Delannay X, McLaren G, Ribaut J-M (2012). Fostering molecular breeding in developing countries. Molecular Breeding 29:857-873. DOI 10.1007/s11032-011-9611-9. 2. Ribaut J-M, de Vicente MC, Delannay X (2010). Molecular breeding in developing countries: challenges and perspectives. Current Opinion in Plant Biology 13:1–6. 3. Varshney RK, Glaszmann JC, Leung H, Ribaut J-M (2010) More genomic resources for less studied crops. Trends in Biotechnology 28:452-460. 4. Sawkins MC, Farmer AD, Hoisington D, Sullivan J, Tolopko A, Jiang Z, Ribaut J-M (2004). Comparative Map and Trait Viewer (CMTV): an integrated bioinformatic tool to construct consensus maps and compare QTL and functional genomics data across genomes and experiments. Plant Molecular Biology 56: 465-480. 5. Ribaut J-M, Hoisington DA (1998). Marker J-M, Hoisington DA (1998). Marker-assisted selection: new tools and strategies. Trends in Plant Science 3: 236-239. VI ICLGG - Abstracts Innovative approaches to increase tropical legume productivity in Africa and South Asia 1* 2 3 4 5 Ribaut J-M , Bertioli DJ , Raatz B , Roberts P , Vadez V , Varshney RK 1 Diop NN I-GAB01 1,5 , 1 GCP c/o CIMMYT, Km 45 Carretera México-Veracruz, El Batán, Texcoco, Estado de México, CP 56130, Mexico 2 Universidade de Brasília, Campus Darcy Ribeiro, L.2 Norte, CP 70910-100 Brasilia, Brazil 3 CIAT, Km 17 Recta Cali-Palmira, CP A.A. 6713, Cali, Colombi 4 UC- Riverside, 900 University Ave., Riverside, CA 92521-0124, USA 5 International Crops Research Institute for Semi-Arid Tropics (ICRISAT), Hyderabad, India *E-mail: j.ribaut@cgiar.org Abstract The global agricultural research agenda must take advantage of recent progress made in biotechnologies to enhance the agricultural knowledge base, and provide innovative solutions. To address this challenge, the Generation Challenge Programme in partnership with CGIAR Centers, Universities and National Programmes facilitates the application of modern breeding approaches using genetic resources and genomic tools to improve the productivity of groundnuts, cowpeas, common beans and chickpeas for SubSaharan Africa and South Asia. Access to new alleles from wild relatives to increase the level of disease resistance in cultivated groundnuts was made possible through the development of syntetics. New diversity has been created through recombination, and the development and characterisation of Multiparent Advanced Generation Inter-cross populations (MAGIC, with 8 parental lines) is ongoing for beans, cowpeas and chickpeas. These populations will provide researchers with a valuable long-term resource with the power to detect numerous QTLs for target agronomic traits and resolve them at or near the level of individual genes, while also providing in the short term a valuable resource to identify new genotypes for prebreeding. There has been considerable effort over the last years to develop suitable genomic resources for these legumes and building on these new resources, several QTLs have been identified through genetic studies for biotic and abiotic stresses that are now introgressed into elite germplasm through Marker-Assisted Backcrossing. Marker-Assisted Recurrent Selection (MARS) to improve drought tolerance is ongoing for both cowpeas and chickpeas. To support these different molecular breeding efforts aiming at increasing tropical legume productivity, an Integrated Breeding Platform to access data, germplasm, tools and services is emerging from a collective effort coordinated by GCP with the support of the Bill & Melinda Gates Foundation. VI ICLGG - Abstracts N NADARAJAN Director Indian Institute of Pulses Research (IIPR) India director@iipr.ernet.in th Prof. (Dr.) Nagasamy Nadarajan was born in Tamilnadu, India on 16 Dec 1951; holds a Ph.D degree with advance training on Marker Assisted Selection from Australia with FAO fellowship. As Director of IIPR, he coordinates the entire pulses research programmes of India such as MAS, transgenics, hybrids, prebreeding, varietal development, development of appropriate crop production and protection technologies at national level apart from International collaboration with CGIAR Institutes for frontier areas of research on chickpea, pigeonpea, lentil, lathyrus and mungbean. Under his leadership, many special programmes/ projects of National and International nature have been initiated; the results of some of the program are already focused. He has developed and released 20 crops varieties of rice, cotton, and pulses etc., which are popularly grown in India. His cited pioneering research work are tagged important 18 QTLs for drought tolerant traits in rice using SSR markers, development of four male sterile lines and 35 restorers in pigeonpea. He is the recipient of three International and six National Honours and Awards for his outstanding contribution in the field of Plant breeding and Genetics. A good biometrician with sound knowledge on molecular markers guided 41 students. He has visited several countries either as country expert on pulses or as a honorary fellow. He has in his credit 10 books, 33 book chapters and 187 research papers of National and International repute. Selected Publications: 1. Manickavelu A, Nadarajan N, (2006). Drought tolerance in rice: Growth Regul. 50:121-138. Ganesh SK, Gnanamalar RP, Chandra Babu R morphological and molecular genetic consideration. Plant 2. Manickavelu A, Nadarajan N, Ganesh SK, Ramalingam R, Raguraman S, Gnanamalar RP (2006). Organogenies induction in rice callus by cyanobacterial extracellular product. African J Biotech. 5 (5): 437-439. 3. Thiyagu K, Jayamani P, Nadarajan N. (2008). Pollen pistil interaction in inter-specific crosses of vigna sp. Cytologia 73 (3): 251-257. 4. Nadarajan N, Chaturvedi SK. (2010). Genetic options for enhancing productivity of major pulses. Journal of Food Legumes. 23(1): 1-8. 5. Kumar J, Pratap A, Solanki RK, Gupta DS, Goyal A, Chaturvedi SK, Nadarajan N, Kumar S. (2011) Genomic resources for improving food legume crops. The Journal of Agriculture Science. 150: 289-318. VI ICLGG - Abstracts Genomics enabled molecular breeding of pulse crops I-GAB02 Nadarajan N Indian Institute of Pulses Research (IIPR), Kanpur, India E-mail: director@iipr.ernet.in Abstract Crop improvement programs for pulse crops have now got the much-required fillip with the availability of wide range of genomic resources including the whole genome sequences of pigeonpea. With significant investments ingenomics research of these crops in recent years, few pulse crops of India viz. chickpea and pigeonpea have now several thousand polymorphic molecular markers, genetic maps and in few cases agronomically important QTLs have also been mapped in the chromosomes. This has paved the way to integrate the strong breeding infrastructure at ICAR Institutes and SAUs with the latest technological advancements for a paradigm shift in pulses breeding programs. Towards this effort, at IIPR-Kanpur, network projects on molecular breeding in chickpea, pigeonpea, lentil and mungbean has already been initiated in collaboration with several national and international labs. There is urgent need to increase the number of genome-wide polymorphic SSR markers in crops like lentil, blackgram and greengram. Recent advances in comparative genomics and gene orthology have opened the possibility of utilizing the genetic information across related genera/species. As a complement to comparative genetic mapping, we analyzed the transferability of microsatellite markers across different taxa viz. Lens, Cajanus, Lathyrus, Vigna and Phaseolus. We have also developed novel set of EST derived SSR markers from chickpea and pigeonpea to add to the marker repertoire. For estimating the genetic diversity in the germplasm of these crop species, several types of molecular markers have been employed and this information has been useful to breeders to select the appropriate parental combinations for crossing programs. MABC programs for incorporating wilt resistance and MARS for transferring draught tolerance in chickpea has started showing results. Similarly in case of pigeonpea, mapping of wilt resistance and fertility restoration QTLs are expected to help develop high yielding and disease resistant cultivars. VI ICLGG - Abstracts S-GAB03 Molecular marker platforms for chickpea, lentil, pea and common bean breeding 1* 2 1 1 3 3 Taran B , Bett K , Sindhu A , Marwan , Ramsay L , Sharpe A , Warkentin 1 1 T , Vandenberg A 1 University of Saskatchewan, Canada University of Saskatchewan, Canada 3 National Research Council, Saskatoon, Canada 2 *E-mail:bunyamin.taran@usask.ca Abstract The rapid expansion of pulse crop production in Canada combined with shorter rotations requires that the crops have a wider range of adaptation while maintaining high yield and acceptable quality. New tools are needed to facilitate rapid incorporation of multiple traits into new pulse crop cultivars. We recently developed a 1,536 SNP GoldenGate array for each of chickpea, lentil and pea and 768 SNP array for common bean. The SNPs were derived from sets ranging from 2,500 SNPs in chickpea to 8,000 SNPs in pea, which were originally identified using 454-sequencing technology. The SNPs were selected for lentil and pea to cover the entire genome based on their homologous locations on the Medicago truncatula chromosomes. The chickpea and bean SNPs were chosen based on distribution across their recently sequenced genomes. The SNPs were used to generate linkage maps of several mapping populations for each crop, and to characterize the diversity of elite breeding lines and germplasm accessions in each crop as a foundation for association analysis. Using a set of 94 RILs of the core mapping population for each crop, we were able to map 276 SNPs in common bean, 749 SNPs in chickpea, 477 SNPs in lentil, and 255 SNPs in pea. Population structure analyses of elite breeding lines and germplasm accessions identified three subpopulations each for chickpea and common bean, and eight subpopulations in pea. The utility of the core maps for QTL analyses and panel genotypes for association analyses of major agronomic traits, phenology, disease resistance and micronutrients concentration will be presented. VI ICLGG - Abstracts Marker-assisted breeding for drought tolerance traits in chickpea 1,* 1 1 1,2 1 S-GAB04 1 Gaur PM , Sivakumar C , Thudi M , Tripathi S , Jukanti AK , Srinivasan S , 1 3 4 5 6 Krishnamurthy L , Gangarao NVPR , Kimurto P , Fikre A , Jayalakshmi V , 7 1* Mannur DM , Varshney R 1 International Crops Research Institute for Semi-Arid Tropics (ICRISAT), Hyderabad, India 2 Indian Agricultural Research Institute, New Delhi, India 3 ICRISAT, Nairobi, Kenya 4 Egerton University, Egerton, Kenya 5 Debre Zeit Agricultural Research Centre, Debre Zeit, Ethiopia 6 Regional Agricultural Research Station, Nandyal, India 7 Agricultural Research Station, Gulbarga, India *E-mail: p.gaur@cgiar.org; r.k.varshney@cgiar.org Abstract Chickpea (Cicer arietinum L.) is predominantly grown as a rainfed crop on residual soil moisture after the rainy season and suffers heavy yield losses from terminal drought. Root traits, particularly rooting depth and root biomass, are known to play an important role in avoidance of terminal drought through more efficient extraction of available soil moisture. A genomic region that contained QTL for root traits and several other drought tolerance related traits, contributing up to 36% phenotypic variation, was mapped on Linkage Group IV based on extensive genotyping and phenotyping data generated on two RIL populations (ICC 4958 × ICC 1882; ICC 283 × ICC 8261). This genomic region flanking by SSR markers TAA 170 and ICCM 0249 was introgressed in three popular cultivars (JG 11, KAK 2 and Chefe) through MABC. Twenty-two BC3F4 progenies derived from the crosses that involved JG 11 as recipient parent and ICC 4958 as donor parent were evaluated at three locations in India (Patancheru, Nandyal, Gulbarga), one location in Ethiopia (Debre Zeit) and one location in Kenya (Koibatek) during 2011-12. Several promising progenies were identified at each location and in each growing condition (rainfed/irrigated). The progenies which performed better than JG 11 at all locations included two under rainfed conditions (5 to 24% higher yield than JG 11) and four under irrigated conditions (2 to 25% higher yield than JG 11). These results are very encouraging and demonstrate the effectiveness of marker-assisted breeding in improving terminal drought tolerance in chickpea. VI ICLGG - Abstracts S-GAB05 SNP discovery and validation for genomic-assisted breeding of faba bean (Vicia faba L.) 1 1 1 1,2 3 3 Cottage A , Webb A , Hobbs D , Khamassi K , Maalouf F , Ogbannaya F , 4 5 6 1 1 Stoddard F , Duc G , Link W , Thomas JE ,O’Sullivan DM * 1 National Institute of Agricultural Botany, Cambridge, UK Institut Nationale Agronomique de Tunisie, Tunis- Mahrajène, Tunisia 3 ICARDA, Aleppo, Syria 4 University of Helsinki, Finland 5 INRA-Dijon, France 6 University of Göttingen, Germany 2 *E-mail: donal.osullivan@niab.com Abstract The objective of this study was to build a medium-density genetic map of gene-based markers in the breeder-friendly KASPar format to facilitate trait dissection in faba bean pre-breeding programmes. To this end, we sequenced 10-day old seedling transcriptomes from two contrasting inbred lines, Albus (white-flowered, stem nematode susceptible) and BPL10 (coloured flower, stem nematode resistant), and were able to identify over 40,000 putative SNPs in overlapping alignments of 14,000 shared Albus:BPL10 contigs. KASPAR assays were designed for 887 loci, of which 85% (757) showed robust amplification and clear clustering patterns on a test panel of 33 inbred lines, many of which are parents of mapping populations. We present here the validation data for the 757 highest quality faba bean SNP assays, a first draft consensus genetic map locating a majority of these, and demonstration of use of this marker set to tag traits of interest with cheap and reliable SNP assays. VI ICLGG - Abstracts QTL mapping in AB-QTL and CSSL populations derived from the cross between a cultivated peanut (Arachis hypogaea L.) variety and a synthetic wild amphidiploid 1* 2 1 3 S-GAB06 4 Fonceka D , Tossim H-A , Rivallan R , Faye I , Leal-Bertioli SCM , Valls 4 3 5 1 1 1 JFM , Moretzsohn MC , Bertioli DJ , Glaszmann J-C , Courtois B , Rami J-F 1 CIRAD, UMR AGAP, Montpellier, France 2 ISRA/Ceraas, Route de Khombole, Thiès Escale, Senegal 3 ISRA, Centre National de Recherche Agronomique, Bambey, Sénégal 4 Embrapa Recursos Genéticos e Biotecnologia, Brasilia, Brazil 5 Universidade de Brasília, Campus Universitário, Brasília, Brazil * E-mail: daniel.fonceka@cirad.fr Abstract Cultivated peanut is an allotetraploid harbouring limited diversity at the DNA level. Peanut wild relatives represent an important source of novel alleles that could be used to broaden the genetic basis of the cultigen. For this perspective, in peanut, new population structures have to be developed in order to decipher the molecular basis of interesting traits coming from wild species. We present here our results on QTL mapping in AB-QTL and CSSL populations using a wild synthetic amphidiploid as donor. Using the AB-QTL population we conducted a QTL study for several traits involved in peanut productivity and adaptation. We mapped a total of 95 QTLs. About half of the QTL positive effects were associated with alleles of the wild parent. We then produced a set of 122 Chromosome Segment Substitution Lines (CSSL) that offers an extensive coverage of the cultivated peanut genome with generally a unique fragment per line and overlapping fragments between successive lines. As a first example of use, four morphological traits were evaluated on a subset of 80 CSSLs. The lines x traits significant associations were assigned to 42 QTLs. A detailed analysis of QTL involved in growth habit showed that this trait is complex and governed by several QTLs having different effects. The comparison of the number of QTLs mapped for growth habit between CSSLs and AB-QTL showed the higher efficiency of the CSSL population and its interest for QTLs validation, breeding and gene discovery. VI ICLGG - Abstracts Special Session: Young Scientists in Legume Genetics and Genomics YS-01 Genetic variation of parental environmental effects on seed transcriptome and transgenerational epigenetic inheritance of offspring response to salinity in M. truncatula 1* 1 2 1 Vu W , Chang P , Moriuchi K , Friesen ML 1 University of Southern California, Los Angeles, USA University of California, Davis, USA 2 *E-Mail: wvu@usc.edu Abstract Parental environmental effects, a form of transgenerational plasticity, are emerging as a viable evolutionary strategy for plants and other organisms with limited dispersal. Parental environmental effects are a cross-generational transmission of parental experience to optimize offspring response to the environment. Since parental environmental influence is large in seed character, one mechanism by which parental environmental experience is transmitted to the offspring is through the deposition of long-lived RNA transcripts in seeds. We find that parental environmental effects are genotype dependent and are prevalent in early development and fade later in the life cycle. To determine the extent of genetic variation of parental environmental effects on the level of gene expression in response to salt stress, we sequenced the mature seed transcriptome of four selfed M. truncatula inbred genotypes grown in saline and non-saline environments. Out of 9,321 genes detected in our seed transcriptome, we found 1,362 genotype-dependent parental environmentally responsive genes. In agreement with genotypic variation in offspring traits, we observe minimal overlap of parental environmental responsive genes among the four genotypes. Gene ontology enrichment analysis of the total genes expressed in our seed transcriptome revealed significant enrichment of terms related to stress response, epigenetics. Some of the most interesting genotype by parental environment transcripts code for factors involved in RNA-directed DNA methylation pathway. This pathway is known to be sensitive to environmental factors and has been well documented to mediate de novo methylation in plants and studies have suggested its involvement in transgenerational epigenetic inheritance. VI ICLGG - Abstracts Genome-wide sequence analysis and expression profiling using RNASeq implicates soybean (Glycine max) HD-Zip gene family members in dehydration and salt stress responses 1 2 3 3 2 YS-02 2 Belamkar V , Weeks N , Bharti AK , Farmer AD , Graham MA , Cannon SB * 1 Iowa State University, Iowa, USA Iowa State University, Iowa, USA 3 National Center for Genome Resources, New Mexico, USA 2 *E-mail: scannon@iastate.edu Abstract Homeodomain leucine zipper (HD-Zip) transcription factors are unique to plants, with diverse roles in growth, development, and response to environmental stress. HD-Zip genes have been well characterized in Arabidopsis thaliana, but are yet to be elucidated in soybean. In this study we identify all members of the HD-Zip gene family in soybean, and characterize their expression under dehydration and salt stress. Homology searches identify 101 HD-Zip genes in the soybean genome. Phylogeny reconstruction coupled with domain and gene structure analyses using soybean, Arabidopsis, rice, grape, and Medicago homologues enables placement of these sequences into four previously described subfamilies. HD-Zip genes are distributed over all twenty soybean chromosomes. Of the 101 HD-Zip genes, 96 exist as paralogous pairs, with the duplicates having being retained following paleopolyploidy events. An RNA-Seq experiment performed to study differential gene expression at 0, 1, 6 and 12 hr in the roots of soybean cv. ‘Williams 82’ under dehydration and salt stress identifies 22 differentially expressed genes (DEGs). Several of these DEGs are located in QTLs previously reported for yield-related traits under abiotic stress in soybean. Screening of HD-Zip gene promoters identifies numerous cis-elements that are overrepresented in the promoters of DEGs in soybean under both stresses, consistent with the role of HD-Zip in abiotic stress responses. A candidate gene hypothesized to be involved in leaf nastic responses under water stress has been selected for functional characterization. VI ICLGG - Abstracts YS-03 Genome structure, evolution and diversity in the genus Phaseolus 1, 2 1 2 3 4 5 Iwata A , Abernathy B , Radke B , Findley SD , Schmutz J , Vallejos CE , 1 6 1 Li Y , Geffroy V , Jackson SA * 1 University of Georgia, Athens, USA Purdue University, West Lafayette, USA 3 University of Missouri, Columbia,USA 4 HudsonAlpha Institute for Biotechnology, Huntsville, USA 5 University of Florida, Gainesville, USA 6 Institut de Biotechnologie des Plantes (IBP), Université Paris Sud, Batiment Orsay, France 2 *E-mail: sjackson@uga.edu Abstract The legume family (Fabaceae) is the third largest family of higher plants, including a number of agriculturally important crops. Recent progress in genome sequencing of multiple legume species has provided useful resources to study chromosome biology in combination with fluorescence in situ hybridization (FISH) technology. Here, we present examples of chromosome studies in legumes such as common bean (Phaseolus vulgaris) and its relatives to better understand genome organization and evolution at the chromosomal level. Using genome sequences and bacterial artificial chromosome (BAC) end sequences, centromeric satellite repeats were identified and characterized in common bean. FISH showed their unique chromosomal distributions and indicated local or chromosome-specific homogenization of the repeats. These repeats were also used to construct FISH-based karyotype map on mitotic metaphase chromosomes to identify all chromosomes simultaneously. Chromosome-specific BACs were mapped using FISH to integrate linkage maps with physical chromosomes. FISHbased karyotyping was performed in several common bean accessions and relatives; comparison of these karyotype maps showed the conservation and diversity of the genome at the chromosomal level. This data contributes to our understanding of genome evolution, chromosomal structural variation, chromosome behavior, and breeding strategy in common bean. VI ICLGG - Abstracts Functional genomics of M. truncatula seed development 1* 1 1 1 YS-04 1 2 Verdier J , Torres-Jerez I , Bandyopadhyay K , Wang M , He J , Buitink J , 1 1 3 1 4 5 Sumner L , Tang Y , Young N , Dixon RA , Harada JJ , Thompson RD , 1 Udvardi MK 1 The Samuel Roberts Noble Foundation, Ardmore, USA INRA PMS, Angers, France 3 University of Minnesota, St Paul, USA 4 University of California, Davis, USA 5 INRA UMRLEG, Dijon, France 2 *E-mail: javerdier@noble.org Abstract Legume seeds are a primary source of proteins, lipids, carbohydrates, secondary metabolites, and minerals for humans and other animals. Few regulatory genes have been discovered that control legume seed development and storage metabolism. Therefore, we have begun to identify and functionally characterize genes potentially involved in seed storage metabolism and seed yield in the model legume, Medicago truncatula. To identify and select candidate genes, we used different approaches: (i) an in silico gene regulatory network for seed development, which linked over four hundred seed-expressed transcription factor genes to their potential target genes including metabolic genes; (ii) a forward genetic screen of a Tnt1insertion mutant population, which revealed mutant lines displaying differences in seed size and yield; (iii) a genome-wide association study using resources developed by the Medicago HapMap project, which has revealed sequence polymorphisms associated with distinct seed composition phenotypes such as carbohydrate, protein, lipid, secondary metabolite and mineral contents; and finally (iv) a detailed analysis of three ecotypes with contrasting seed storage protein content, which may lead to new ways to improve the protein yield in legumes. The combination of these approaches has led to the discovery of genes controlling seed storage metabolism, seed yield and more broadly seed quality in legumes. So far, thirty genes were selected for reverse genetics or ectopic expression studies in M. truncatula and are now being characterized such as, for instance, a MYB gene, which acts as a positive regulator of tannin biosynthesis in seeds and has been used for biotechnological applications. VI ICLGG - Abstracts YS-05 Deciphering the role of stress responsive genes in chickpea (Cicer arietinum L.) 1 1 1 1 1 Himabindu K , Sharma HC , Krishnamurthy L , Adivi S , Vijay P , Farmer 2 1 1 1,3* AD , Shah T , Gaur PM , Varshney RK 1 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 2 National Centre for Genome Resources (NCGR), Santa Fe, USA 3 Generation Challenge Programme (GCP), c/o CIMMYT, Mexico DF, Mexico. *E-mail: r.k.varshney@cgiar.org Abstract Chickpea, an important legume crop is seriously challenged by several abiotic and biotic stresses. Understanding the mechanisms that regulate the expression of stress-related genes is a fundamental issue and is necessary for the genetic improvement of chickpea. In separate studies based on in silico expression analysis of Illumina GA and Sanger sequencing data generated from drought and Helicoverpa challenged tissues of contrasting parental genotypes provided differentially expressed stress-responsive genes that are involved in stress signaling pathways. This work presents deciphering the role of in silico identified stress-responsive genes for conferring resistance to drought stress and Helicoverpa infestation. A set of 111 and 55 candidate genes for insect resistance and drought tolerance, respectively, were targeted for experimental validation using quantitative real time-PCR (qRT-PCR). Total RNA of root (drought) and leaf (Helicoverpa) tissues from stressed and unstressed plants were analysed in the context of tolerant/sensitive genotypes. qRT-PCR analysis confirmed in silico predicted expression analysis for a total of 24 genes. This set of genes includes 15 known stress-responsive genes and 9 novel unreported genes. In terms of cross-talking between genes under these two stresses three genes encoding for mannitol dehydrogenase, NADPH quinine oxidoreductase and cytochrome P-450 showed differential expression. Emerging evidence suggests that stress signaling pathways regulated by ABA pathway, Oxidative phosphorylation pathway as well as ROS signaling pathways play key roles in the crosstalk between abiotic and biotic stress signaling. The identified genes could be used to improve resistance to important constraints affecting chickpea production. VI ICLGG - Abstracts Gene diversity and marker development in narrow-leafed lupin (Lupinus angustifolius L) 1,2,3,* 4 4 4 YS-06 4 Nelson MN , Hane JK , Berger JD , Williams AH , Kamphuis LG , 2,3 2,3 2,4 Clements JC , Erskine W , Singh KB 1 School of Plant Biology, Faculty of Natural and Agricultural Sciences, The University of Western Australia, Crawley, Australia 2 The UWA Institute of Agriculture, Faculty of Natural and Agricultural Sciences, The University of Western Australia, Crawley, Australia 3 Centre for Legumes in Mediterranean Agriculture, Faculty of Natural and Agricultural Sciences, The University of Western Australia, Crawley, Australia 4 CSIRO Plant Industry, Floreat, Australia *E-mail: matthew.nelson@uwa.edu.au Abstract Narrow-leafed lupin (Lupinus angustifolius L.) is the most important grain legume in Australia. It is unique in that it is the only crop species to be th domesticated in Australia in the 20 Century. A large national effort is currently underway to develop a comprehensive reference genome sequence for the lupin cultivar ‘Tanjil’. Alongside the genome sequencing effort, we are characterising gene variation in narrow-leafed lupin by sequencing the transcriptomes of a panel of 16 representative breeding lines, wild germplasm and parents of reference mapping populations. This gene variation resource will be mined to develop markers linked to – or underlying – traits that are important to the Australian lupin breeding programme. These markers will be used for marker-assisted selection of key agronomic traits and to assist the efficient introduction of new genetic diversity into the relatively shallow lupin breeding pool. VI ICLGG - Abstracts YS-07 Application of marker assisted selection to improve drought tolerance in adapted Kenyan chickpea varieties 1 3 1 1 Kosgei AJ*, Kimurto PK , Gaur PM , Mulwa RK , Towett BK , Gangarao 2 2 3 3 3 NVPR , Silim S , Sivakumar C , Thudi M , Varshney RK 1 Egerton University, Egerton, Kenya International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Nairobi, Kenya 3 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 2 *E-mail: alicesongio@yahoo.com Abstract Chickpea (Cicer arietinum L.) is an important pulse legume that provides cheap source of protein to both urban and rural families who cannot afford expensive animal protein sources. In Kenya chickpea production is still low and is produced mainly in dryland areas but is also being introduced in dry highlands as a relay crop. Its production is mainly under rain-fed conditions and drought is a major production constraint. There is need to improve the adapted Kenyan chickpea to withstand drought periods especially in dry areas that are usually underutilized. The objective of the study was to improve the adapted varieties to drought by introgressing drought tolerance quantitative trait loci (QTL) for roots using marker assisted backcrossing. Two donor parents ICC 8261 and ICC 4958 that have better RLD, root mass and length were crossed each to two adapted varieties, ICCV 92944 and ICCV 4958 in Egerton University, Kenya. The resulting 20 F1s were planted and genotyped using simple sequence repeat (SSR) markers to identify true crosses. The successful true crosses are being backcrossed to the recurrent parents to obtain BC1F1. This will be repeated to BC3F1 and selfed to BC3F3. Field evaluation of BC3F3 will be done in two locations, Koibatek Agricultural training centre and Kenya Agricultural Research Institute (KARI) – Marigat under well watered and water stress conditions, three replications each. The root traits will be evaluated in PVC at Egerton University under rainout shelter. It is anticipated that this study enhance introgressing drought tolerant QTLs for roots into Kenyan varieties for improved yield production in semiarid areas. VI ICLGG - Abstracts QTL analysis and heritability of seed size and shape in lentil (Lens culinaris Medic.) YS-08 Fedoruk M*, Vandenberg A, Bett K University of Saskatchewan, Saskatoon, Canada *E-mail: m.fedoruk@usask.ca Abstract The seed size of lentil is an important trait because it determines the market class, it can influence quality and yield when lentils are milled, and it can affect cooking time. For this study we evaluated a mapping population, segregating for seed size and shape, derived from a cross between the small red cultivar CDC Robin and the large green breeding line 964a-46. The objectives were to determine the heritability and identify the genomic regions controlling seed size and shape. The population was grown in two years at two different locations in Saskatchewan, Canada. Three different seed parameters were measured using sieves: seed diameter, thickness and plumpness. Days to flowering was also measured to determine if it had any effect on seed morphology. Each individual in the population was genotyped using a 1536 Illumina Golden Gate lentil SNP assay, along with 5 morphological, 12 SSR and 83 KASP SNP markers. The heritability estimates were high for seed diameter and seed plumpness (0.92 and 0.94, respectively) and moderate for seed thickness and days to flowering (0.70 and 0.47, respectively). A linkage map was constructed consisting of seven linkage groups with an average distance of 2 cM between markers. QTL analysis revealed major QTLs on four linkage groups, suggesting it is possible to select for seed morphology traits in lentil. VI ICLGG - Abstracts Theme: Nutrition and Quality RICHARD D. THOMPSON Research Director Institut National de la Recherche Agronomique (INRA)-Dijon France thompson@dijon.inra.fr Trained in biochemistry and molecular biology at the Universities of Birmingham and Edinburgh, UK, (Ph.D 1979). Isolated genes encoding wheat storage proteins at the BBSRC Plant Breeding Institute, Cambridge, (1978-1986). Studied regulation of gene expression in maize endosperm at the Max-Planck Institute for Plant Breeding Research, Cologne, Germany, (1986-2002). Habilitation in Genetics at Cologne University (1993). Unit Director, INRA Research Unit for Grain Legume Genetics and Ecophysiology (UMR LEG), Dijon, France, (2002-2010). In recent years we’ve been focussing on the genetic control of seed development and composition in legumes, using the model species Medicago truncatula, and more recently, Pea (Pisum sativum). By applying proteomics and transcriptomics approaches, clusters of coregulated genes corresponding to discrete steps in seed filling have been identified. We’ve also revealed extensive compartmentalisation of metabolism and the corresponding gene expression between different seed tissues. To narrow down candidates for mutant analysis, we have selected genes that co-locate with QTLs for traits such as 1000-seed weight and seed composition. The genes of particular interest to us are expressed in the endosperm, but affect the development and maturation of the entire grain. Selected Publications: 1. Zuber H, Davidian J-C, et al. (2010). The seed composition of Arabidopsis mutants for the group 3 sulfate transporters indicates a role in sulfate translocation within developing seeds. Plant Physiology 154(2): 913-926. 2. Zuber H, Davidian J-C, et al. (2010). Sultr4;1 mutant seeds of Arabidopsis have an enhanced sulphate content and modified proteome suggesting metabolic adaptations to altered sulphate compartmentalization. BMC Plant Biology 10(1): 78. 3. LeSignor C, Savois V, et al. (2009). Optimizing TILLING populations for reverse genetics in Medicago truncatula. Plant Biotechnology J. 7 : 430-441 4. Repetto O, Rogniaux H, et al. (2008). Exploring the nuclear proteome of Medicago truncatula at the switch towards seed filling. Plant J. 56 : 398-410. 5. Verdier J, Kakar K, et al. (2008). Gene expression profiling of M. truncatula transcription factors identifies putative regulators of grain legume seed filling. Plant Mol. Biol. 67: 567-80. VI ICLGG - Abstracts Dissecting the role of the endosperm in regulating Medicago truncatula seed filling 1 1 2 I-NAQ01 1 Noguero M , D’Erfurth I, Le Signor C , Vernoud V , Verdier J , Aubert G , 3 4 5 1 1 1,* Buitink J , Gouzy J , Prosperi J-M , Burstin J , Gallardo K , Thompson RD 1 1 INRA, UMR Génétique et Ecophysiologie des Légumineuses, Dijon, France The Samuel Roberts Noble Foundation Ardmore, France 3 INRA, UMR Physiologie Moléculaire des Semences, Angers, France 4 INRA Laboratoire des Interactions Plantes Micro-organismes, Toulouse, France 5 UMR Diversité et adaptation des plantes cultivées, Montpellier, France 2 * E-mail: thompson@dijon.inra.fr Abstract We are using genomics approaches with Medicago truncatula to identify key genes determining legume seed yield and composition. Analyses of the proteome and transcriptome of the component tissues of the developing seed revealed extensive compartmentalization of gene expression and metabolic activities within the seed (Gallardo et al, 2007). Using a TF (Transcription Factor) qRT-PCR platform (Verdier et al., 2008) and the Affymetrix Gene Chip (Benedito et al, 2008), TFs specific for each seed tissue were identified, along with putative target genes. These TFs have been located on the M. truncatula genetic map and correlations between map positions of TF loci and QTLs for protein quantities and other seed phenotypes were detected. Two genes, both specifically expressed in the developing endosperm, have received particular attention. One of the genes encodes a DOF class transcription factor. The mutant is severely affected in endosperm development and the embryo is arrested at the globular stage. The few seeds reaching maturity have a significantly reduced seed weight. The dof1147 seed phenotype can be partially reverted by auxin treatment. The second gene encodes an endosperm-specific subtilase (SBT1.1), whose chromosomal position coincides with a seed weight QTL. Association studies in ecotype collections support linkage between the SBT1.1 locus and seed weight variation. Sbt1.1 mutants of pea and M. truncatula also have decreased weight and surface area at maturity, due to reduced numbers of cotyledon cells. The possible mechanisms of action of these two genes in the endosperm will be discussed. VI ICLGG - Abstracts CLAIRE DOMONEY Project Leader, John Innes Centre Norwich United Kingdom claire.domoney@jic.ac.uk Dr Domoney’s research interests lie in defining and understanding the genetic control of legume seed composition that is relevant to food and feed end uses and to the development of improved crops that promote sustainable agriculture. She coordinates a number of UK legume consortium projects, including PCGIN (Pulse Crop Genetic Improvement Network) and QDiPS (Understanding Quality Determinants in Pea Seeds to improve market opportunities that promote sustainable agriculture), and manages a work programme within ProtYield (Protein content vs yield in legumes: releasing the constraint). In her research, the effects of natural and induced mutations in a number of genes that impact on protein and starch quality in pea seeds and on overall yield are being studied. Among these, there is a focus currently on senescence induced genes that are significant to nitrogen reassimilation in seeds and crop end use. Selected Publications: 1. Clemente et al. (2012). Brit J Nutr 108 (in press) 2. Clemente A, Sonnante G, Domoney C (2011). Bowman-Birk inhibitors from legumes and human gastrointestinal health: current status and perspectives. Current Protein and Peptide Science 12:358-373. 3. Chinoy C, Welham T, Turner L, Moreau C, Domoney C (2011). The genetic control of seed quality traits: effects of allelic variation at the Tri and Vc-2 genetic loci in Pisum sativum L. Euphytica 180: 107–122. 4. Clemente A, Moreno FJ, Marín-Manzano MC, Jimenez E, Domoney C (2010). The cytotoxic effect of Bowman-Birk isoinhibitors, IBB1 and IBBD2, from soybean (Glycine max) on HT29 human colorectal cancer cells is related to their intrinsic ability to inhibit serine proteases. Molecular Nutrition Food Research 54:396-405. 5. Vigeolas H, Chinoy C, Zuther E, Blessington B, Geigenberger P, Domoney C (2008). Combined metabolomic and genetic approaches reveal a link between the polyamine pathway and albumin 2 in developing pea seeds. Plant Physiology 146:174-182. VI ICLGG - Abstracts Exploiting novel mutations affecting seed composition and visual traits in Pisum sativum L. (pea) for improved food and feed uses 1 1 1 4 2 I-NAQ02 1 Moreau C , Chinoy C , Knox M , Christodoulou V , Warkentin T , Wang TL , 3 4 1 Ellis N , Isaac P , Domoney C * 1 John Innes Centre, Norwich, UK University of Saskatchewan, Saskatchewan, Canada 3 Aberystwyth University, Ceredigion, UK 4 Norwich Research Park, Norwich, UK 2 *E-mail: claire.domoney@jic.ac.uk. Abstract Pea (Pisum sativum L.) is an important cool season legume crop produced mainly in temperate regions. The crop is exploited for food and feed uses, as well as for its positive contribution to sustainable agriculture. The continuous improvement of seed traits by conventional marker-assisted breeding relies on the characterization of the genetic control of the relevant traits, as well as the exploitation of novel sources of mutations within the underlying candidate genes. Germplasm resources, together with a number of mutagenised populations, are providing a wealth of such variation, assisted by rapid advances in methods for systematically screening and characterizing variant candidate genes associated with quantitative trait loci (QTL). Natural and induced mutations in genes implicated in the control of seed protein quality, starch/sugar content and chlorophyll content have been identified. Included here, for example, are novel variants of genes encoding pea seed albumin 2, trypsin inhibitor, starch-branching enzyme, ADP-glucose pyrophosphorylase and a number of genes that underlie QTL for seed and yield traits, including chlorophyll content. In the last case, genes involved in the control of chlorophyll degradation are being tested using a transient leaf assay to enable rapid functional analysis and to understand the basis for quantitative variation in the expression of variant alleles. The combined data will assist greatly with the adoption and selection of superior alleles of relevant genes in breeding for improved legume food and feed crops. VI ICLGG - Abstracts S-NAQ03 Genetic analysis of seed carbohydrates in chickpea (Cicer arientinum L.) 1 1 2 1 1 2 Gangola MP , Khedikar YP , Thudi M , Wang R , Jaiswal S , Gaur PM , Baga 1 2 1,* M , Varshney RK , Chibbar RN 1 Department of Plant sciences, College of Agriculture & Bioresources, University of Saskatchewan, Saskatoon, SK, Canada 2 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad,India *E mail: ravi.chibbar@usask.ca Abstract Chickpea (Cicer arietinum L.) is an important pulse crop primarily as a source of protein, complex carbohydrates and dietary fiber in vegetarian diets. Half to two-thirds of the seed dry weight consists of carbohydrates, one of the most versatile storage components affecting both the extrinsic and intrinsic seed quality. The predominant seed storage carbohydrate is starch which in chickpea has higher amylose concentration compared to cereal starches. Increased amylose amylose concentration in chickpea seeds is a positive factor for human health especially in controlling type 2 diabetes and cardiovascular diseases. Compared to other grain crops, legume seeds differ by higher concentration of soluble carbohydrates belonging to the raffinose family oligosaccharides (RFO) including raffinose, stachyose and verbascose. Detailed analyses of a chickpea germplasm collection (156 genotypes) assembled at ICRISAT revealed that in greenhouse seeds accumulated significantly lower (1.58–4.67 mmoles/100 g) total RFO compared to field grown (2.08-5.83 mmoles/100 g) genotypes indicating the role of RFO in stress tolerance. Stachyose was the major RFO in chickpea seeds followed by raffinose and verbascose. ANOVA revealed a significant effect (P<0.001) of genotype (G), environment (E) and G x E on several chickpea seed constituents. Diversity Array Technology (DArT) analysis by 380 polymorphic markers showed that a germplasm collection as a highly structured population with eight different sub-populations. RFO concentration and type was associated with 15 DArT loci mapped on linkage groups LG1, LG2, LG4 and LG6 on a chickpea reference genetic map. The significance of these observations to develop reduced RFO chickpea genotypes will be discussed VI ICLGG - Abstracts Utilizing molecular mapping and transcriptome analysis to elucidate development of seed and nodule for enhancing yield and nutrition in chickpea S-NAQ04 Gaur R, Gupta S, Pradhan S, Shah N, Kant C, Kumar S, Bhatia S* National Institute of Plant Genome Research, New Delhi, India E-mail: sabhyata_bhatia@nipgr.res.in Abstract Chickpea (Cicer arietinum) is the third most important legume crop in India, especially valued for its nutritive seeds. It is imperative that the crop be studied in detail so as to develop strategies for improvement in yield. One such method for crop improvement is molecular breeding, which requires well-saturated linkage maps, especially those based on expressed sequence tag (EST)derived genic molecular markers (GMMs). Therefore, ESTs were generated in large number from chickpea seeds and utilized for the development of novel EST-derived functional markers which included EST-SSRs, intron targeted primers (ITPs), expressed sequence tag polymorphisms (ESTPs) and single nucleotide polymorphisms (SNPs). Moreover, large scale discovery of genome wide SNPs was done using the Roche 454 platform. Development and validation of high throughput genotyping assays was carried out to build a high resolution genetic linkage map of chickpea comprising of more than a thousand molecular markers. Although seeds are an obvious choice for study to enhance nutritive value, production of high quality protein rich food is extremely dependent upon availability of sufficient nitrogen. Root nodules are the most important tool of a legume for fixing atmospheric nitrogen in soil thereby making a larger reservoir of the element available to plants. Thus, seed and root nodule tissue, at various stages of their development, was used to generate transcriptomes of chickpea on the Roche 454 platform. Reads from respective tissue were filtered and assembled into contigs using Newbler 2.5.3 software. After annotation, a preliminary homology based comparison of the reads generated from seed tissue with total chickpea transcriptome showed a very small fraction of the transcripts to be specific to seed. A similar comparison was also done to determine genes and TFs specific to particular stages of seed development. These seed-specific TFs and genes may be putatively responsible for controlling seed yield or nutrition in chickpea. A similar analysis in root nodules gives an idea about the genes that may be specific to nodule development. VI ICLGG - Abstracts Theme: Translational Genomics DOUGLAS R. COOK Professor Plant Pathology College of Biological Sciences University of California, Davis USA drcook@ucdavis.edu Research in the Cook laboratory has three focuses: legume comparative genomics, functional analysis of the interaction between legumes and rhizobium, and population genomics of salt adaptation in Medicago truncatula. Our comparative genomics work has emphasized comparison of conserved orthologous genes across species, with the objective of inferring the genetic structure at ancestral nodes. Within this comparative structure, we are interested in the syntenic relationships among NBLRR disease resistance genes, their evolution, and functional characterization. Our work on symbiotic nitrogen fixation aims to identify the genetic, biochemical and cellular factors that govern nitrogen fixation. This work focuses at two levels: biochemical and cell biological analysis of Nod factor signaling in Medicago truncatula, and genetic and phenotypic analysis nitrogen-related traits in chickpea and wild relatives. In the later work we are particularly interested in the potential impact of domestication and breeding on nitrogen fixation. Our work on population genomics focuses on natural populations of Medicago truncatula in Tunisia and Portugal. By combining field ecology with greenhouse phenotyping and genome resequencing we have identified candidate pathways and genes for salt adaptation. Selected Publications: 1. Kassa MT, Penmetsa RV, Carrasquilla-Garcia N, Sarma BK, Datta, S, Upadhyaya HD, Varshney RK, von Wettberg EJB, Cook DR (2012). Genetic patterns of domestication in pigeonpea (Cajanus cajan (L.) Millsp.) and wild Cajanus relatives. PLoS ONE 7(6):e39563. 2. Young ND, et al. (2011). The genome sequence of Medicago truncatula and the evolution of nodulation. Nature 480:520-4. 3. Varshney RK, et al. (2011). Draft genome sequence of pigeonpea (Cajanus cajan), an orphan legume crop of resource-poor farmers. Nature Biotechnology 30(1):83-9. 4. Haney CH, Riely BK, Tricoli D, Cook DR, Ehrhardt DW, Long SR (2011). Symbiotic rhizobia bacteria trigger a change in localization and dynamics of the Medicago truncatula receptor kinase LYK3. Plant Cell 23:2774. 5. Riely BK, He H, Venkateshwaran M, Sarma B, Schraider J, Ane JM, Cook DR (2011). Identification of legume RopGEF gene families and characterization of a Medicago truncatula RopGEF mediating polar growth of root hairs. Plant Journal 65:230-243. VI ICLGG - Abstracts Harvesting legume genomes for agronomic traits 1 1 1 I-TLG01 1 2 Penmetsa RV , Rosen BD , Carrasquilla-Garcia N , Baek, JM , Kim DH , 1 1 2 1 3 Bergmann E , Vance L , Kashyap M , Greenspan A , Scharnagl K , 3 1 2 2 4 5 Sanchez V , Jinliang G , Upadhyaya H , Gaur P , Yasin M , Kaur J , Singh 5 3 2 1 1, S , von Wettberg E , Varshney RK , Bruening G , Cook DR * 1 University of California, Davis, California, USA International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 3 Florida International University, Florida, USA 4 RAK College of Agriculture, Sehore, India 5 Punjab Agricultural University, Ludhiana, India 2 * E-mail: drcook@ucdavis.edu Abstract We have used comparative genomics to link many of world’s most important crop legumes and we have initiated analysis of phenotypic and genetic diversity within cultivated species and their wild progenitors. Our goal is to combine these data sets to identify the molecular-genetic bases of agronomic and/or domestication-related traits in legumes. We have focused on genes regulating disease resistance in cowpea and pigeonpea and on the impact of domestication on nitrogen-related traits including symbiotic nitrogen fixation in chickpea. To a lesser extent we have also analyzed traits controlling anthocyanidin synthesis, shoot architecture and flowering time in chickpea, each of which have been significantly influenced by the processes of domestication and/or breeding. In several cases we have identified the molecular basis of these agronomic traits, establishing the function of candidate genes either by reverse genetics or by means of association genetic analyses. In other cases, we are developing data sets that implicate specific genes and/or genetic pathways as being causal to phenotypic differences, with the eventual goal of understanding their molecular bases and evolutionary histories. Our long-term interest is to use this strategy to leverage information across species and dissect the molecular-genetic basis of nitrogen-related traits in crop legumes, especially those traits that have been modified during the course of domestication. Initial evidence suggests domestication related impacts on nitrogen sensitivity, nitrogen-responsive root architecture, and potentially on mesorhizobial strain selectivity. Progress on developing this strategy and its application to the study of nitrogen fixation in chickpea will be presented VI ICLGG - Abstracts T J HIGGINS Honorary Research Fellow Commonwealth scientific and Industrial Research Organisation (CSIRO)- Plant Industry Australia tj.higgins@csiro.au TJ Higgins is an Honorary Fellow at CSIRO Plant Industry. He works on protecting food legumes from insect damage, researching the application of gene technology for plant improvement. His current research is focused on international agriculture with particular emphasis on Africa and India and he has a special interest in public awareness of science. He is a Fellow of The Australian Academy of Science. Selected Publications: 1. Chen H, Bodulovic G, Hall PJ, Moore A, Higgins TJV, Djordjevic MA, Rolfe BG (2009). Unintended changes in protein expression revealed by proteomic analysis of seeds from transgenic pea expressing a bean -amylase inhibitor gene. Proteomics 9, 4406-4415. 2. Washida H, Sugino A, Kaneko S, Crofts N, Sakulsingharoj C, Kim D, Choi S-B, Hamada S, Ogawa M, Wang C, Esen A, Higgins TJV, Okita TW (2009). Identification of cis-localization elements of the maize 10-kDa -zein and their use in targeting RNAs to specific cortical endoplasmic reticulum subdomains. The Plant Journal 60, 146-155. 3. Acharjee S, Sarmah BK, Kumar PA, Olsen K, Mahon R, William J, Moar WJ, Moore A, Higgins TJV (2010). Transgenic chickpeas (Cicer arietinum L.) expressing a sequencemodified cry2Aa gene. Plant Science 178, 333-339. 4. Watkins PR, Huesing JE, Margam V, Murdock LL, Higgins TJV (2011). Controlling plant responses to the environment:biotic stress-insects, nematodes and other pests. In “Plant biotechnology and agriculture: Prospects for the 21st century”: A. Altman and P.M. Haegawa eds., Elsevier Press (in press). 5. Campbell PM, Reiner D, Moore AE, Lee R-Y, Epstein MM, Higgins TJV (2011). Comparison of the α-amylase inhibitor-1 from common bean, Phaseolus vulgaris varieties and transgenic expression in other legumes - post-translational modifications and immunogenicity. J. Agric. Food Chem. 59, 6047-6054. VI ICLGG - Abstracts Protecting cowpeas from field and storage pests using gene technology I-TLG02 Higgins TJ CSIRO Plant Industry, Canberra, Australia E-mail:tj.higgins@csiro.au Abstract Cowpeas (Vigna unguiculata) are important in both crop rotations and as good sources of nutrients for humans and their livestock especially in subSaharan Africa. As a component of crop rotations cowpeas provide a disease break for subsequent crops and also provide biologically fixed nitrogen for the same crops. World production of cowpeas shows only a very slight positive trend over the last 30 years mostly due to productivity gains in the industrial world. Productivity in developing countries is half that in the developed countries largely due to abiotic and biotic constraints. For example, globally, there is a total of 6 million tonnes of cowpea produced each year. But yield could be doubled if the insect pests that cause losses in production or during storage could be controlled. Gene technology can lead to the introduction and deployment of new insect protection traits that will enhance production and storage. Several technical hurdles have been solved for the use of gene technology in cowpea such as plant transformation with genes encoding resistance to caterpillars and bruchids. Other hurdles such as biosafety testing, consumer awareness, regulatory approval, trait introgression and a seed dissemination system are already beginning to be tackled. VI ICLGG - Abstracts GENGYUN ZHANG Vice President Beijing Genomics Institute (BGI) Shenzhen China zhanggengyun@genomics.cn Gengyun Zhang obtained his Ph.D. degree at Rutgers University in 2003. Currently he is the VP of BGI, who is in charge of Life Science Division of BGI. His research interests are combining molecular and conventional plant breeding techniques together to accelerate the development of new cultivars, especially focusing on whole genome based molecular marker assisted selection systems for rice, foxtail millet and soybean, based on high throughput sequencing and data analysis capacities of BGISZ. Selected Publications: 1. Zhang G, Liu X, Quan Z, et al. (2012). Genome sequence of foxtail millet (Setaria italica) provides insights into grass evolution and biofuel potential. Nature Biotechnology. doi:10.1038/nbt.2195. 2. Chia Jer-Ming, Song C, Bradbury PJ……Zhang G, Xu Y, Ware D. (2012). Maize HapMap2 identifies extant variation from a genome in flu Nature Genetics. doi:10.1038/.2313. 3. Lam Hon-Ming, Xu X …. Jun Wang, Sun SS, Zhang G. (2010). Resequencing of 31 wild and cultivated soybean genomes identifies patterns of genetic diversity and selection. Nature Genetics 42, 1053–1059. VI ICLGG - Abstracts Whole genome molecular marker assisted crop breeding based on translational genomics approach in BGI I-TLG03 Gengyun Zhang BGI-Shenzen China E-mail: zhanggengyun@genomics.org.cn Abstract Recent quick advance of sequencing technology makes us easier to elucidate whole genome genotype of a crop strain, which is the largest obstacle for traditional breeders to accelerate cultivar improvement. A whole genome based molecular marker assisted selection system could be quickly established based on high throughput sequencing and bioinformatics platforms, even for a neglected crop. The practice of whole genome based MAS in BGI indicated that, with existing affordable low cost technologies, super cultivars with improved yield potential, better suited to the climate stresses, higher nutritional quality, greater ability to grow in problem soils, and improved tolerance to pests and diseases can be developed to meet unexpected needs through exploiting a huge range of potentially valuable genes in the species. Platforms in BGI could provide practical solutions for developing countries to develop and apply their own molecular breeding systems economically. VI ICLGG - Abstracts S-TLG04 Groundnut breeding at ICRISAT: from tools of genetics to genomics 1* 1, 2 Janila P , Varshney RK 1 Nigam SN 1 1 1 1 Pandey MK , Vadez V , Sudini HK , Patne N , 1 International Crops Research Institute for Semi-Arid Tropics (ICRISAT), Hyderabad, India 2 CGIAR Generation Challenge Programme (GCP), Mexico *E-mail: p.janila@cgiar.org Abstract The breeding program for genetic enhancement of groundnut has begun in 1976 at International Crops Research Institute for the Semi-Arid Tropics (ICRISAT). So far over 10,000 crosses were made using germplasm, breeding lines and interspecific derivatives. Since 1986, a total of 138 improved varieties of groundnut, bred at ICRISAT were released by national partners in 36 different countries of Asia and Africa. The adoption of improved varieties resulted in increased yields and production and thus improved the livelihoods of small families in semi-arid tropical regions, engaged in groundnut cultivation. At ICRISAT’s Asia program, traditional breeding procedures based on the principles of genetics along with the physiological tools and screening for pest and diseases were largely used in the development of improved varieties of groundnut. Significant progress has been made to breed varieties with high yield potential, short maturity duration, resistance to foliar fungal diseases, drought tolerance, enhanced oil content and traits for food uses. Since both, empirical and trait-based approaches were equally efficient in breeding for drought tolerance; a combination of these approaches was adopted. With the development of molecular tools in groundnut during the last decade, it is now possible for integrated marker assisted selection (MAS) in groundnut breeding for improved efficiency and optimal utilization of time and resources. The introgression of QTLs for rust and late leaf spot resistance and mutant FAD alleles (for enhanced oil quality) are under progress. Further, initiatives have been taken for the development of genomic tools for improvement/selection of traits such as, drought tolerance, high oil content, aflatoxin resistance and fresh seed dormancy. VI ICLGG - Abstracts A search for novel sources of resistance in Medicago truncatula to improve fungal resistance in crop legumes 1,* 1 1 1 S-TLG05 2 Uppalapati SR , Ishiga Y , Mittal S , Wensheng Li , Schultheiss H , Sumner 1 1 1 L , Dixon RA , Mysore KS 1 Plant Biology Division, Samuel Roberts Noble Foundation, Ardmore, USA BASF Plant Sci GmbH, Ctr Agr, Limburgerhof, Germany 2 *E mail: srinivasaraou@yahoo.com Abstract Improvement of fungal resistance through biotechnology in crop legumes is limited by the pools of available candidate genes, genotypes, and the labor involved in positional or map-based cloning of the identified gene(s). We have adapted Medicago truncatula as a model legume and have been applying both genetic and functional genomic approaches to identify novel genes involved in host and nonhost fungal resistance. To identify candidate genes to engineer cotton root resistance in alfalfa, using M. truncatula and a combination of global expression analyses we have identified that Phymatotrichopsis omnivora targets flavonoid and lignin pathways. Surprisingly, however, downregulation of hydroxycinnamoyl CoA: shikimate hydroxycinnamoyl transferase (HCT), caffeoyl CoA 3-O-methyltransferase (COMT) and caffeic acid methyltransferase (CCoAOMT ) resulted in increased tolerance with restricted growth of P. omnivore and Fusarium oxysporum f. sp. medicaginis. Metabolite and gene expression profiling revealed that the induced tolerance in these lignin modified plants may result from increased accumulation and/or spillover of flux towards the (iso) flavonoid pathway. In addition, we have setup large scale forward genetic screens to identify mutants which would lead to the identification of novel genes that confer resistance of M. truncatula to Phakopsora pachyrhizi. We identified an inhibitor of rust germ tube differentation1 (irg1) mutant. Cytological and chemical analyses revealed that the inhibition of rust preinfection structures in irg1 mutants is due to complete loss of the abaxial epicuticular wax crystals and reduced surface hydrophobicity. Furthermore, the potential of using M. truncatula as model host or nonhost will be discussed. VI ICLGG - Abstracts Posters Theme: Next Generation Genomics P-NGG05 ISMU: Integrated SNP mining and utilization pipeline for accelerated breeding 1, # Azam S 1,# 1 1 1 1,2,* , Rathore A , Shah T , Amindala BP , Telluri M , Varshney RK 1 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 2 Generation Challenge Program (GCP), Mexico DF, Mexico *E-mail: r.k.varshney@cgiar.org #Authors contributed equally Abstract Detection of variants from Next Generation Sequencing (NGS) data is often complicated and requires various methodologies and multiple steps. An effort has been undertaken to include all the steps in a user-friendly pipeline which we defined as Integrated SNP Mining and Utilization (ISMU) pipeline. After pre-processing the raw data, the pipeline provides users a range of options for alignment using tools such as Bowtie, BWA, SOAP, Maq and NovoAlign. The tools which provide alignments in SAM format, are then processed for SNP prediction with SAM tools, otherwise existing packages within the programs such as SOAPsnp and cns2snp (Maq) are used. The Coverage based Consensus Calling (CbCC) (Azam et al, 2012, AM J BOT, 99:186–192) approach is integrated with all alignment tools. The output after analyzing NGS data through this pipeline include list of SNPs between any combination of genotypes. Tablet and Flapjack have also been integrated into the pipeline to allow users to visualise the alignment and SNPs respectively. For further validation on array based SNP platforms, users can select SNPs based on confidence score or PIC values with flanking sequences in a standard format for developing genotyping (KasPar and Golden Gate) assays. The pipeline has been developed to run on 64-bit Linux and is available at “http://hpc.icrisat.cgiar.org/NGS/”. The server edition is written in CGI Perl while standalone version is developed in Java. In the next phase of development, we will be adding genomic selection (GS), genome wide association studies (GWAS) and marker assisted recurrent selection (MARS) modules to utilize SNPs data for breeding programs. VI ICLGG - Posters RNAseq transcriptome de novo assembly in pea: a new genomic resource for pea genetics and genomics 1 1 2 1 1 P-NGG06 1 Alves-Carvalho S , Aubert G , Cruaud C , Savois V , Klein A , Jacquin F , 3 1 1 2 3 1 Carrère S , Martin C , Boucherot K , da Silva C , Gamas P , Gallardo K , 3 1,* Gouzy J , Burstin J 1 UMR AgroEcology, Dijon, France GENOSCOPE, Evry, France 3 UMR LIPM, Auzeville, France 2 *E-mail: burstin@dijon.inra.fr Abstract Because of their high protein seeds and their unique ability to establish a symbiosis with soil nitrogen fixing bacteria, legumes should play a central role in future sustainable agriculture. Pea appears a strategic crop for temperate regions of the world. However, molecular tools are lacking to enhance markerassisted selection and/or gene cloning in pea. A first step towards this aims was to develop a Unigene set for pea. Using Next-Generation sequencing technologies, we sequenced pea cDNA libraries for 20 different plant organs, stages, and nutrition condition, and miRNA from two seed stages. One billion reads corresponding to ca 100 Gb were produced. These sequences were de novo assembled and a full-length Unigene set was produced. These data revealed the complexity of transcriptomes and allowed developing the first Gene Atlas of pea to identify interesting expressional candidates. VI ICLGG - Posters P-NGG07 Combining genomics, association genetics, and ecology to uncover genes and adaptation in Medicago 1,2 1 3 3 4 5 Cordeiro MA , Moriuchi KS , Vua W , Friesen ML , Mun J-H , Choi H-K , vonWettberg 3 1* Nuzhdin SV , Cook DR 1 University of California Davis, Davis, USA Instituto de Tecnologia Química e Biológica, Oeiras, Portugal 3 University Southern California, Los Angeles, USA 4 Rural Development Administration, Suwon, South Korea 5 Dong-A University, Busan, South Korea 6 Florida International University, Miami, USA 7 Fairchild Tropical Botanic Garden, Coral Gables, USA 2 *E-mail: drcook@ucdavis.edu Abstract Medicago truncatula (Mtr) is a model for molecular, ecological, and evolutionary biology genomics, association genetics, and field ecology to examine the molecular basis a salinity tolerance in natural populations from saline and non-saline environments in Tu Previous work on Tunisian Mtr identified candidate genes that assort with soil origin populations are adapted to salinity using avoidance mechanisms (i.e. earlier reprod exposure to toxic salinity levels). To test whether mechanisms of salinity adaptation bet Portuguese populations are the same, a field reciprocal transplant experiment was con in environments that naturally differ in salinity. The mechanisms of adaptation be diverge: Tunisian salt adapted populations evolved avoidance mechanisms, while Portu populations evolved tolerance mechanisms (i.e., ion balance). Full genome re-sequen lines is being used to identify candidate genes based on association with traits, so Genome comparisons of Tunisian and Portuguese saline adapted genotypes will serve and independently evolved adaptations to soil salinity. Additionally, natural hybrid popul and M. litorallis (Mli) were identified, genotyped and phenotyped in a field experiment genotyping data was used to identify hybrid individuals and quantify the extent of ad results indicate that hybrid zones are stable and potentially play an important role in transfer and salinity adaptation of Mtr. VI ICLGG - Posters Transcriptome assembly and SNP identification in Pongamia pinnata (L.) Pierre using RNA-seq 1 2 3,* Wegrzyn JL , Loopstra CA , Harry DE , Kinlaw CS P-NGG08 3 1 University of California Davis, Davis, USA Texas A&M University, Texas, USA 3 TerViva BioEnergy, Inc, Oakland, USA 2 *E-mail: david.harry@terviva.com Abstract Pongamia, or pongam, (Pongamia pinnata (L.) Pierre, is a familiar tree in many parts of India. Outside of India, it is native to Southeast Asia and also occurs naturally in Australia. Pongamia’s oil-rich seeds have sparked considerable attention as a source of high quality bio-oil. We seek to better understand traits and biochemical pathways contributing to oil production and quality as well as adaptive vigor in varying environments. Thus, we have begun to develop DNA markers for implementing marker accelerated breeding. We conducted Next Generation (NextGen) sequencing using RNA-seq to simultaneously characterize expressed genes and identify candidate single nucleotide polymorphisms (SNPs). RNAs were isolated from leaves of 72 seedlings and consolidated into eight pools of nine individuals. From eight sequencing lanes on an Illumina HiSeq 2000 (100 bp paired-end reads), we generated ~40 Gbp of sequence. After various high-stringency quality control steps, we processed 77.8M reads through the Trinity and Oases/Velvet assemblers, yielding ~57K and ~64K contigs of >300 bp. The final assembly encompassed 56,691 unique contigs (“unigenes”) with an average length of 705 bp. Sequence alignments with several reference genomes, including Glycine max, coupled with functional comparisons such as GO, revealed an expected assortment of expressed genes. In addition, we identified ~98K candidate SNPs, with a frequency of about 1 in 400 bases. As for other datapoor species, we conclude that RNA-Seq will be an effective tool for transcriptome assembly and SNP discovery to build infrastructure for marker accelerated breeding. VI ICLGG - Posters P-NGG09 Genomic resources for functional analysis in Medicago truncatula and related crop species 1,2* 5 5 5 1,2 1,2 Debellé F , Belser C , Aury JM , Labadie K , Roux B , Sallet E , Carrere 1,2 5,6,7 3 4 1,2 1,2 S , Wincker P , Schiex T , Julier B , Gamas P , Gouzy J 1 INRA, Laboratoire des Interactions Plantes-Microorganismes, Castanet Tolosan, France 2 CNRS, Laboratoire des Interactions Plantes-Microorganismes, Castanet Tolosan, France 3 INRA, Biométrie et Intelligence artificielle, Castanet Tolosan, France 4 INRA, Unité de Recherche Pluridisciplinaire Prairies et Plantes Fourragères, Lusignan, France 5 Génoscope, Institut de Génomique CEA, Evry, France 6 CNRS, Evry, France 7 Université d’Evry, Evry, France *E-mail: frederic.debelle@toulouse.inra.fr Abstract Medicago truncatula, a model legume for the analysis of root symbioses, is phylogenetically close to important legume crops such as alfalfa, clover, pea or faba bean. A complete reference genome sequence of M. truncatula is thus of great importance for functional genomic analysis in this species and related ones. In order to complement the current genome assembly produced by the International Medicago truncatula Genome Sequencing Initiative we developed a whole genome sequencing approach based on Illumina paired end and mate pair technology supplemented by other available resources (BAC sequences, BES). We assembled the M. truncatula A17 genome sequence in large scaffolds, anchored to chromosomes by genetic and optical mapping, and covering >80 % genome and > 95 %genes (< 70 % in the published Mt3.5 assembly). This sequence was annotated using an improved Eugene pipeline integrating in particular paired and oriented Illumina RNA seq data representing genome expression in various tissues and organs. The M. truncatula genome sequence was used as a reference to sequence the gene space of M. sativa (alfalfa), an autotetraploid and heterozygous species highly syntenic to M. truncatula. The M. sativa sequences were generated by Illumina paired end and mate pair technology and used to characterize gene structure and polymorphism in this important forage crop. The resources developed in this program will benefit functional approaches involving for example transcriptomics or non-coding RNA analysis as well as the identification and mapping of genes or QTL of interest in M. truncatula and related species. VI ICLGG - Posters Small transcriptome deep sequencing reveals novel microRNA differentially regulated by specific hormonal environments in Medicago truncatula P-NGG10 * Eyles RP , Williams PH, Ohms SJ, Weiller GF, Djordjevic MA, Imin N Australian National University, Canberra, Australia *E-mail: rod.eyles@anu.edu.au Abstract Development of root architecture is dependent on the interplay of key phytohormones such as auxin and cytokinin. Recently, microRNA (miRNA) have also emerged as major regulators of plant development and in Medicago Truncatula have been shown to contribute to the control of nodulation and root meristem formation and maintenance. How miRNA interact with the hormonal environment within the root is, however, largely unknown. Using deep sequencing we compared the small transcriptome of hormone treated explant tissue of root forming callus (auxin treated), non-root forming callus (auxin/cytokinin treated) and meristem containing root tip tissue. A strict set of criteria, based on expression pattern and bioinformatic prediction were used to predict miRNA involved in root development. We predicted a total of 78 novel miRNA. Of these, 21 are conserved in plants but have not previously been identified in M.truncatula while 45 have not been described in plants before. Many are predominantly expressed in the root tip, confirming a strong role for miRNA control of the root meristem. We found strong expression of miR156, miR159 and miR172. Null mutants of each of these are known to cause acute developmental defects in aerial tissue, but to date have no known function in root. Expression patterns and target prediction indicate it is probable that many miRNA identified are responsive to specific hormonal regimes and are therefore likely to be involved in root development. Preliminary functional analysis has identified novel miRNA showing evidence of involvement in nodulation and early root formation. VI ICLGG - Posters P-NGG011 Genomic resources for functional analysis in Medicago truncatula and Chickpea genomic resources: development and utilization for linkage map construction Gaur R, Gupta S, Khajuria Y, Bhatia S* National Institute of Plant Genome Research, New Delhi, India *E-mail: sabhyata_bhatia@nipgr.res.in Abstract Chickpea (Cicer arietinum L.), an important legume crop, valued for its nutritive seeds that serves as an important source of plant-based dietary protein (approx 20-30%) especially in developing countries. Despite its agricultural value and continuous demand, the production is seriously challenged by several biotic and abiotic stresses. Recently chickpea genomics research has significantly progressed as evidenced by the development of modern tools such as molecular markers, linkage maps, and genomic resources like ESTs and gene expression studies that facilitated breeding programs for stress tolerance and yield improvement. With the aim to enhancing genomic resources in chickpea, a set of 278 genomic STMS markers were developed using conventional and enriched methods. Moreover, from 3 different cDNA libraries, ESTs were generated and exploited to develop different types of genic molecular markers such as EST-SSRs, ITPs, ESTPs and SNPs. Linkage analysis was performed using both types of markers in an intra-specific (ICCV2 x JG-62) and inter-specific (ICC4958 x PI489777) mapping populations to construct linkage maps. Besides, from the next generation sequencing, genome wide SNP identification was carried out. These SNPs were validated using high-throughput GoldenGate genotyping platform and used to generate a dense genetic linkage map of chickpea. Production of this high-density SNP based map has facilitated anchoring and orienting the scaffolds generated by whole genome sequence assembly data. Moreover, this map was also used to study syntenic relationship between chickpea and other legume crop. In addition, the sequence information generated from the cDNA libraries, has allowed performing gene expression studies of target genes involved in seed developmental processes. VI ICLGG - Posters Sequencing, annotation and analysis of the narrow-leaf lupin genome Kamphuis LG 1,2* 1 1 3 , Hane JK , Gao L , Nelson M , Singh KB P-NGG12 1,2 1 CSIRO Plant Industry, Wembley, Australia The UWA Institute of Agriculture, the University of Western Australia, Crawley, Australia 3 School of Plant Biology, the University of Western Australia, Crawley, Australia 2 * E-mail: lars.kamphuis@csiro.au Abstract Narrow-leaf lupin (NLL) is the main grain legume grown in Australia and forms an important part of sustainable farming systems, reducing the need for nitrogenous fertilizer, providing valuable disease breaks and boosting cereal yields. We have generated a BAC library of the NLL cultivar Tanjil, comprising 111,360 clones with an average insert size of 99.7Kbp, with an estimated average 12X genome coverage. We generated ~ 14,000 BAC-end sequences covering ~ 1% of the NLL genome. We have performed tissue-specific transcriptome sequencing for three cultivars with sequenced tissues from roots, stems, leafs, flowers and seeds, which will aid the annotation and identification of the gene-rich regions. From this dataset we have identified several groups of genes that are tissue specific. This information is useful for accelerating the genetic mapping of traits that are strongly associated with certain tissues. We have adopted high-throughput molecular marker techniques for assaying gene-based SNPs and SSRs in NLL, which we will use to generate dense genetic maps from the polymorphisms identified from the transcriptome datasets. Finally, in partnership with the Grains Research and Development Cooperation and the Beijing Genome Institute we embarked on the sequencing of the NLL genome. We have developed a preliminary draft genome of NLL cultivar Tanjil, based on 30X coverage with a total assembly size of 521Mb. Together these resources will significantly improve and accelerate NLL breeding programmes, especially since NLL has only been ‘domesticated’ for little more than 50 years. VI ICLGG - Posters P-NGG013 Next-Gen sequencing of coding and non coding transcriptome of chickpea root nodule for discovery of genes and novel miRNA Kant C, Shah N, Kumar S, Bhatia S * National Institute of Plant Genome Research, New Delhi, India *E-mail: sabhyata_bhatia@nipgr.res.in Abstract A hallmark trait of legumes is their ability to develop root nodules and to fix N2 in symbiosis with compatible Rhizobia, leading to a terrestrial input of 40-50 million tons of nitrogen per year. Chickpea (Cicer arietinum L.) is widely grown across the Indian subcontinent and an important source of protein and plays an important role in the farming system by fixing atmospheric nitrogen (140 kg N / hectare). This study reports the deep sequencing of the coding and non-coding transcriptome of chickpea root nodule. For this, chickpea root tissues inoculated with Mesorhizobia ciceri and collected at different stages, were utilized. cDNA libraries were prepared and sequenced on Roche 454 Titanium platform. Quality filtering of the raw data was done using online available NGS QCToolkit_v2.1 and assembly was done using Newbler 2.5.3 software using various standardized parameters for minimum overlap identity and Kmer. These unigenes were searched for sequence similarity using BlastX with uniprot database. Nodule specific and stage specific genes were identified. Transcription factor genes from the data were also identified. Since small RNAs have emerged as an important regulatory component in a number of developmental processes including legume-Rhizobia symbiosis. We sequenced a smallRNA library from root nodules of chickpea using the Illumina GA II X platform. Conserved miRNA were identified based on similarity to miRNA of other plant species present in miRBase, and putative novel miRNAs were identified based on potential hairpin loop forming precursors from available plant databases. Validation of the putative novel miRNA by northern blotting and stem loop RT-PCR is underway. The conserved miRNA and putative novel miRNA identified in this study will help in identifying role of miRNA in chickpea-Mesorhizobium symbiois. VI ICLGG - Posters Decoding transcriptional reprogramming in blackgram during MYMIVinfestation P-NGG14 Kundu A, Patel A, Pal A* Bose Institute, Kolkata, India *E-mail: amita@bic.boseinst.ernet.in Abstract Plant responses to pathogenic infection involve large-scale transcriptional reprogramming, however, fine-tuning of such reprogramming in response to different pathogens varies. Information on transcriptional modulation during the occurrence of yellow mosaic disease (YMD) is meager, despite its serious consequences of yield penalty in grain legumes. YMD is caused by Mungbean yellow mosaic India virus (MYMIV). In order to gain insight into the early transcriptional events of MYMIV-reaction, and to identify the key players in imparting MYMIV-resistance, EST analysis by suppression subtractive hybridization was performed employing a MYMIV-susceptible cultivar (T9) and a resistant inbred line (VMR84) of blackgram. Distinct transcript profiles were observed in the two genotypes following MYMIV-stress. Transcripts that were modulated during susceptible reaction in T9 were associated mainly with primary metabolism and photosynthesis, while in VMR84, categories for primary metabolism, stress/defense and signal transduction were enriched. Identification of NBS-LRR genes indicates efficient pathogen recognition and activation of the defense related signaling pathways. Differential regulation of calcium and protein kinases in the signaling cascade was observed. MYMIVinduced accumulation of transcripts for ROS homoeostasis, HSPs and WRKY transcription factors provide evidences for their important role in host defence arsenal. Interestingly, MYMIV-resistance is associated with specific modulation of transcripts encoding PR proteins and genes involved in salicylic acid biosynthesis resulting in the activation of the systemic acquired resistance. Differential temporal expressions of selected transcripts were validated through qPCR. We conclude that blackgram resistance against MYMIV comprised of a highly adaptive regulatory system that regulates at different levels even at early stages of MYMIV-infection. VI ICLGG - Posters P-NGG015 Resequencing in a mapping population 1,* 2 Lightfoot DA , Town C , Chan A 2 1 Southern Illinois University, Carbondale, USA J. Craig Venter Institute, Rockville, USA 2 *Email: ga4082@siu.edu Abstract Investments in soybean cv. Forrest systems biology have provided many resources: (i) a 50K SNP genetic map; (ii) three RIL populations (96>n>975); (iii) ~200 NILs; (iv) 115,220 BACs and BIBACs; (v) a physical map; (vi) 4 different MTP sets; (vii) 25,123 MTP BAC end sequences (BES) (viii) a map of duplicated regions; (ix) a map of homoeologous regions; (x) transcript abundance measurements (xi) transformation; (xii) RNAi; (xiii) a TILLING resource; (xiv) conserved synteny maps; (xv) proteome maps; (xvi) and metabolomics. Genes isolated include <i>Rhg4</i> and <i>rhg1</i>, <i>Rps5</i>, <i>Rps1</i> and <i>Rfs2, Rfs3, Rfs1</i>, <i>GmNark, GmNod</i>. The most diverse 67 RILs and 6 NILs were barcoded and sequenced by Illumina. Approximately 25.5 million 100 bp reads were generated. Alignments produced a mean of 12,628 contigs of length 104 bp with 19x coverage. A total of 7,755 SNP positions were identified. To date 309 SNP positions were identified in the RILs and 153 in the NILs. These SNPs have read coverage from the majority of individuals (>50% individuals) and segregate close to 1:1 in the population. In addition a pair of F5:7:16 NILs were analyzed with 52,000 SNPs. Heterozygosity was detected at a 10 fold lower frequency than heterogeneity. Genomic features associated with increased heterozygosity included members of the resistance gene families. Recombination rates varied across a 20 fold range by chromosome and by regions of chromosomes. The SoyGD portal at http:/soybeangenome.siu.edu integrates the chromosome map with the genome sequences that identifies complete genes, a partial genome annotation and many thousands of SNPs. VI ICLGG - Posters Application of NGS tools in assembly and prediction of resistance analogues for screening pigeonpea germplasm against fusarium wilt 1,* 1 1 1 1 1 P-NGG16 2 Marla SS , Gahoi S , Grover M , Singh M , Singh N , Kumar R , Rai A , 1 Bansal KC 1 National Bureau of Plant Genetic Resources, New Delhi, India National Agricultural Bioinformatics Grid, IASRI, New Delhi, India 2 * E-mail: soma.marla@nbpgr.ernet.in Abstract Pigeanpea (Cajanus cajan) is a major grain legume providing nutritional security in diet of population from Indian subcontinent, South East Asia and East Africa. Fusarium wilt is a major disease hampering efforts of legume breeders in improvement of Pigeanpea productivity. We analyzed, nearly 13 mln raw sequence reads (454 GS-FLX; Narendra K. Singh et al., 2012) of popular variety ‘Asha’. So far analysis of long reads (< 400 bp) employing various Next Generation Sequencing Tools enabled assembly of 56 contigs and in silico prediction of nearly 100 disease resistance analogues. The predicted gene annotation revealed presence of known disease resistance domains such as ARC-NBS-LRR, Transmembrane, Kinases. Comparative Genome analysis of Glycine max, Medicago trancatulum, field bean and Pigeonpea revealed some syntenic relations and presence of a few disease resistance analogues. Primer sets were designed for the obtained RGA sequences for their PCR amplification from genomic DNA extracted from 31 land races and varieties of pigeonpea conserved at National Bureau of Plant Genetic Resources, New Delhi. Analysis is being carried out employing PCR screening to identify probable Fusarium wilt resistant pigeonpea genotypes. VI ICLGG - Posters P-NGG017 spliceR: Detecting and quantifying allele-specific-expression from RNAseq data 4, 5* 1 2 3 6 Mukherjee S , Johnson GR , Scott PS , Farmer AD , May GD , Lamkey 4, 4,5 KR Beavis WD 1 Department of Crop Sciences University of Illinois, Urbana, USA Corn Insects and Crop Genetics Research Unit, USDA-ARS, Ames, USA 3 National Center for Genome Resources, Santa Fe, USA 4 Department of Agronomy Iowa State University, Ames, USA 5 Bioinformatics and Computational Biology Program Iowa State University, Ames, USA 6 Pioneer Hi-Bred International, Inc., Johnston, USA 2 *E-mail: shreymuk@iastate.edu Abstract Allelic specific expression (ASE) is a vital factor in phenotypic variability and for the development of complex traits. Some genes display allelic disparity in gene expression that is transmitted by Mendelian or non-Mendelian inheritance and this discrepancy may be associated to effects like heterosis, variation in yield, uniformity in plants and complex traits and diseases in animals. It is of great interest to study how genetic and epigenetic modifications lead to transcriptional variation and how transcriptional variation affects the phenotype. Differential allele expression may be controlled by changes to the nucleotide sequence and regulatory elements, such as single nucleotide polymorphisms (SNPs), insertions and deletions, and studies indicate that these variations are rampant across the plant genomes and tissues. Such variants in the coding regions of genes may alter the structure and function of the gene product. Recent studies have shown that preferential expression of alleles is widespread in mammals. Non-imprinted autosomal genes exhibit allelic imbalance at the transcript level in mouse hybrids (Cowles et al., 2002) and humans (Yan et al., 2002), and such expression produces proteins associated with diseases. Hence a solid understanding of classification and functional annotation of allele-specifically expressed genes is vital to recognize the extent of functionally important regulatory variation. This will help us identify candidate haplotypes and the correlation between their genetic sequences and heterotic traits. The physiological vigor and variations in general health of an organism is strongly associated with the extent of variation of parental gametes. In our study we combine sequence variants and transcript counts from next-generation sequence data to develop a novel “personal haplotyping” method (and a software pipeline spliceR) in order to study allele-specific expression and identify alleles that are preferentially expressed across genetic backgrounds and levels of inbreeding. VI ICLGG - Posters Chickpea interactome for signaling components: A case study of MAPK cascade P-NGG18 Purayannur S, Kumar K, Verma PK* National Institute of Plant Genome Research, New Delhi, India *E-mail: praveen_verma@nipgr.res.in Abstract Chickpea (Cicer arietinum) is an important legume crop but genome wide interaction analysis of the signaling components involved in stress responses has not been performed. Mitogen activated protein kinases (MAPKs) form a crucial eukaryotic signaling module that play significant roles in various cellular processes. We identified 16 MAPKs and 7 mitogen activated protein kinase kinase (MEK) of chickpea using EST sequences from public database and cDNA pyrosequencing. These MAPKs and MEKs were separately classified into four groups based on phylogeny and established Arabidopsis classification. The MAPKs had 2, 5, 1 and 8 members in groups A, B, C and D respectively. All the MAPK members had the signature motif of 'T-x-YxxTRWYRAPEL' in activation loop and 12 protein kinase sub-domains. In MEKs group A, B, C and D had 3, 1, 1 and 2 members respectively that shows signature motif of 'YMSPER'. We generated entry clones for these 23 members and subsequently cloned them in GAL4 based yeast two-hybrid (Y2H) destination vectors. The Y2H interaction analysis in all possible combinations was carried out. Many novel interactions were elucidated by this approach. Future work is focused on elucidating the possible roles of this cascade in chickpea response to Ascochyta rabiei infection. VI ICLGG - Posters P-NGG19 Transcriptome sequencing in winged bean: assembly, annotation, and marker discovery 1 1 2 3 Sharma S , Shetty P , Doyle JJ , Sathyanarayana N , Egan AN 4,* 1 Sir M Visvesvaraya Institute of Technology, Bangalore, India Cornell University, Ithaca, USA 3 Sikkim University, Gangtok, India 4 East Carolina University, Greenville, USA 2 *E-mail: egana@ecu.edu Abstract Winged bean (Psophocarpus tetragonolobus (L.) DC) is similar to soybean in use and high protein content, with the added advantage of producing better yield than soybean under drought conditions. Lack of genomic resources has hindered application of modern breeding tools in this little-known legume crop. In this study, we have analyzed transcriptome sequences of two P. tetragonolobus genotypes for their utility in marker discovery and gene function annotation. About 8M reads obtained through 454 pyrosequencing were assembled using several publicly available assemblers, and a de novo assembly of 47,057 contigs was generated. The average contig length was 731 bp. A total of 96% contigs showed high similarity and probable homology with sequences in the non-redundant database suggesting robustness of the assembly. In silico analysis identified 7,046 SSRs among 5,874 contigs, and 142 SNPs between the two genotypes. Functional characterization by Gene Ontology analysis revealed close similarities with a recent study done on the Chickpea transcriptome. About 85% of the contigs were assigned at least one GO term. Additionally, functional annotation by KAAS generated 20 complete pathway maps. Nearly 4% of the contigs showed similarity to known plant transcription factor families in the plant transcription factor database. The first reference transcriptome assembly along with molecular markers and gene annotations reported in this study for winged bean will pave the way for further investigation and utility of modern computational techniques in breeding and improvement of this potential legume crop. VI ICLGG - Posters Nucleotide sequence variations on conserved regulatory sequence motifs between Glycine max and G. soja 1 1 1 1 1 1 1 P-NGG20 1,2 Shim S , Yoon MY , Kim SK , Lee T , Kang YJ , Kim MY , Van K , Lee S-H * 1 Department of Plant Science and Research Institute for Agriculture and Life Science, Seoul National University, Seoul, Korea 2 Plant Genomics and Breeding Institute, Seoul National University, Seoul, Korea *E-mail: sukhalee@snu.ac.kr Abstract Recently, comparative genomics has been vigorously studied because wholegenome draft sequences of plant species including several legumes, such as Glycine max, G. soja, Cajanus cajan and Medicago truncatula, are available. The majority of the comparative genomics studied synteny of coding region sequences because variations on those regions could contribute to speciation and evolution. However, the non-coding regions were also the important phenotypic regulator and it suggested that comparative analysis on these regions is also worthwhile. In particular, short sequence motifs in the promoter regions tend to be highly conserved, though these sequences are not encoded. In this study, we predict the conserved short sequence motifs by BLASTN algorithm using dicot promoter database on Softberry (http://www.softberry.com). A total of 37,396 conserved short sequence motifs on to 2 kb upstream sequences were identified in 46,367 high confident genes of G. max (cv. Williams 82). We also detected single nucleotide polymorphisms (SNPs) in regulatory and genic regions of 7 G. max landraces and 7 G. soja cultivars. SNPs were retrieved by SamTools software after mapping short reads were produced by Illumina HiSeq 2000. Among 46,367 high confident G. max genes, 15 candidate genes for comparing transcriptional level between G. max and G. soja and 2 candidate genes for comparing transcriptional level between intra-species were selected by the number of SNP variations in regulatory region containing conserved short sequence motif. No SNP variation was detected in all of these 17 putative genes of genic region and 17 genes were verified by RT-PCR methods. VI ICLGG - Posters P-NGG21 Comparison of protein sequence variation in ZIP transporters isolated from natural populations of Medicago truncatula 1,* 2 Stephens BW , Grusak MA , Mills WR 1 1 University of Houston Clear Lake, Houston, USA USDA-ARS Children’s Nutrition Research Center at Baylor College of Medicine, Houston, USA 2 *E-mail: stephensb@uhcl.edu Abstract Iron (Fe) and zinc (Zn) are required by plants and must be obtained from the soil and redistributed throughout the plant in order to maintain optimal growth. The ability of plants to maintain Fe and Zn homeostasis is important due to its participation in many diverse and essential processes. These processes are inhibited if an organism is unable to maintain adequate concentrations of Fe and Zn; however, they can be detrimental to organisms when present in excess. Therefore, the concentration of both Fe and Zn must be controlled by transporters that are responsible for uptake, efflux and compartmentalization within the plant. There are several families of metal transporters responsible for maintaining homeostasis within the plant. The ZIP transporters are one family of divalent metal transporters that includes members in plant, animal, and microbial species, with all having similarity to the ZRT (zinc-regulated transporter) and IRT (iron-regulated transporter) genes from Saccharomyces cerevisiae. In Medicago truncatula, two predicted ZIP proteins have been identified that transport Zn and Fe (MtZIP5 and MtZIP6), one that transports Fe (MtZIP3) and two that transport Zn (MtZIP1 and MtZIP2). We have identified haplotypes of MtZIP1and MtZIP5 from a population of approximately 300 single plant lines of M. truncatula arising from accessions obtained through the USDA National Plant Germplasm System (NPGS). Additionally, 51 single seed descent accession lines from the Core Subset of M. truncatula are currently being screened for sequence variation in MtZIP genes. VI ICLGG - Posters Identification of conserved and novel miRNAs in chickpea (Cicer arietinum L.) by high-throughput sequencing of small RNA libraries 1 2 3 3 Jagadeeswaran G , Zheng Y , Kudapa H , Varshney RK , Sunkar R P-NGG22 1 1 Department of Biochemistry and Molecular Biology, Oklahoma State University, Stillwater, USA 2 Institute of Developmental Biology and Molecular Medicine and School of Life Sciences, Fudan University, Shanghai, China 3 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India Abstract Recent discovery of microRNAs (miRNAs) with regulatory roles has uncovered an active role for RNA in regulating gene expression besides passive intermediary role between genes and proteins. Approximately 21-nt long processed miRNAs are loaded into RNA-induced silencing complex (RISC), and the RISC causes largely sequence-specific transcript cleavage, thus contributing to post-transcriptional gene regulation in plants. Thus, the identification of entire set of miRNAs and their targets is important to unravel the complex miRNA-mediated regulatory networks controlling development and stress responses. Chickpea (Cicer arietinum L.) is one of the very important legume crops but the miRNA component is unknown in this plant species. To identify miRNAs in chickpea, small RNA libraries were constructed from leaves and flowers and sequenced using Illumina analyzer. Bioinformatic analysis of the small RNA reads led to the identification of 189 miRNA loci represented by 123 known (highly conserved as well as conserved only in few related plant species) miRNA homologs belonging to 44 miRNA families in chickpea. In addition, we have identified 29 novel miRNAs that are supported by the cloning of miRNA star sequences and 205 novel candidate miRNAs. Approximately 250 genes have been predicted as targets for miRNAs in chickpea. This information not only offers resources for better understanding of the posttranscriptional gene regulation mediated by the miRNAs but also contributes to the annotation of genome sequencing project that is currently underway. VI ICLGG - Posters P-NGG23 Isolation of differentially expressed genes (DEGs) of potential agronomic importance from groundnut 1 1 1 1 2 1 Wang CT , Tang YY , Wang XZ , Wu Q , Yu ST , Ding YF , Fang CQ 1 1 Shandong Peanut Research Institute, China Sandy Land Amelioration and Utilization Research Institute of Liaoning, Fuxin, China 2 *E-mail: chinapeanut @126.com Abstract Differential expression analysis was used to identify genes potentially related to resistance to low temperature at seeding stage and bacterial wilt (BW), and high oil or protein content in groundnut. Three suppression subtractive hybridization cDNA libraries were constructed for a chilling tolerant cultivar A4 with mRNA isolated from seeds imbibed at 2℃ or 15℃ for 1h, 6h or 24h. Sequencing 500 randomly picked clones from each library resulted in 7 contigs and 1 singlet, 14 contigs and 4 singlets, and 73 contigs and 120 singlets, respectively. Fourteen transcripts were selected based on BLAST2FO annotation for further analyzing their expression by qRT-PCR, and found that relative expression ranged from 2.85 to 12.85. Complete cds of LEA, Oleosin, NAC and MYB were obtained using RACE. Seeds of Rihua 1 (a variety 9 -1 resistant to BW) inoculated with (10 cfu ml ) BW suspension or simply imbibed in water, were studied for differential gene expression using TM Genefishing . A total of 25 DEGs were isolated. Expression of genes encoding cyclophilin and ADP-ribosylation factor were studied and full length cDNAs obtained. Similarly, 27 and 40 DEGs were obtained for high oil and protein content respectively, using developing seeds of chemical mutants and their wild types. Expression of 3 genes for oil/protein content was studied. VI ICLGG - Posters Whole - transcriptome analysis between UV-B -resistant and –susceptible soybean (Glycine max) 1 2 1 1 Yoon MY , Kim KD , Shim SR , Jang HJ , Lee SH P-NGG24 1,3, * 1 Department of Plant Science, Seoul National University, Seoul, Korea Center for Applied Genetic Technologies, Univ. of Georgia, Athens, USA 3 Research Institute for Agriculture and Life Sciences, Seoul National University, Seoul, Korea 2 *E-mail: sukhalee@snu.ac.kr Abstract With planet-wide environments and climate changes, such as stratospheric ozone depletion, biotic and abiotic effects are increasing on plants. Lots of researches are being carried out to understand these external effects lead in terms of plant metabolism and gene expression. Ultraviolet-B (UV-B; 280320nm) is one of most important factors in environmental and climate changes. As stratospheric ozone depletion increases, more UV-B penetrates to the earth’s surface. Also, the importance of sustainable and adaptable crop breeding for climate changes is growing bigger and bigger. Soybean (Glycine max) is the most important crop in the world as food, energy and animal feeds. However, soybean gene and expression analysis on UV-B irradiation is insufficient. It is necessary to search and develop stable traits and genes in soybean plants. The main purposes of this study are identification of UV-B resistant gene and development of sustainable and adaptable soybean crop. This research is focused on gene expression pattern analysis of UV-B – resistant and – susceptible soybeans after these soybeans were identified by ultraviolet-B irradiation. Next-generation sequencing technology and bioinformatics tools were used for understanding UV-B effects on genomic DNA and gene expression by low coverage of whole-genome sequencing and transcriptome analysis. VI ICLGG - Posters P-NGG25 Transcriptome analysis of chickpea seed for discovery of novel genes Pradhan S, Gaur R, Shah N, Bhatia S* National Institute of Plant Genome Research, New Delhi, India *E-mail: sabhyata_bhatia@nipgr.res.in Abstract Chickpea (Cicer arietinum) is the third most important legume crop in India, especially valued for its nutritive seeds. In order to explore the possibilities of improvement in the nutritional quality of the seeds, it is necessary to have a clear understanding of the regulatory mechanisms involved in the process of seed development. This has now become feasible with the advent of NGS technology. Here, we have used the Roche/454 platform to generate a transcriptome of chickpea using seed tissue at various stages of development. We generated a total of 3.8 GB of data. The reads were filtered using NGS QC Toolkit v 2.1 and assembled into contigs using Newbler 2.5.3 software with default parameters. Transcripts were annotated and categorised according to their functions i.e. biological, molecular and cellular processes. A preliminary homology based comparison with total chickpea transcriptome showed a very small fraction of the transcripts to be specific to seed development. A similar comparison was also done to determine genes and TFs specific to particular stages of seed development. These seed-specific TFs and genes should facilitate the identification of regulatory networks that are important for the progression of seed development. Also, the Illumina GA II X platform was used to mine for miRNA families involved in seed development. After screening for rRNA, tRNA, snRNA, snoRNA and miRNAs already reported in the miRBase, some putative novel miRNA sequences were discovered which are being characterized to determine their role in seed development. VI ICLGG - Posters Theme: Symbiosis and Development P-SAD11 Mapping the gene for no-nodulation mutation (PM233) in chickpea (Cicer arietinum L.) 1 2 1 1 Ali L *, Madrid E , Millan T , Gil J , Rubio J 3 1 Dept of Genetics, University of Cordoba, Cordoba, Spain Institute for Sustainable Agriculture, Cordoba, Spain 3 Area de Mejora y Biotecnología, IFAPA, Córdoba, Spain 2 *E-mail: alatifa23bhr@yahoo.com Abstract The legume/rhizobial symbiosis plays a crucial role in the introduction of fixed nitrogen into both agricultural and natural systems. Development and study of nodulation (Nod) mutants is essential to dissect the transduction pathway of Nod-factors. Several Nod mutants have been obtained in chickpea, one of them was a stable no-nodulation mutation in PM233 mutant. Selection of heterozygous plants after seven successive self-pollination generations, derived from the cross between the Spanish kabuli landrace CA2139 (Nod+) and the mutant PM233 (Nod-), permitted to select two pairs of Near Isogenic Lines (NILs) for this trait. The screening of microsatellite markers distributed across different linkage groups (LG) in the chickpea genetic map showed that the NILs vary in markers located in LG 5. Phenotypic evaluation for presence/ absence of nodules and genotyping for STMS markers in a F 2 derived from the cross between a pair of NILs confirmed the location in LG 5 of the nodulation gene. VI ICLGG - Posters Ethylene response factor (ERF) transcription factors of the B-3 subgroup include master regulators of ethylene signalling and mediate resistance to root pathogens without adversely affecting rhizobial symbiosis 1,2 1,3 Anderson JP , Lichtenzveig J , Oñate-Sánchez L 1,4 , Singh KB P-SAD12 1,2,* 1 CSIRO Plant Industry, Floreat, Australia The University of Western Australia, Crawley, Australia 3 Curtin University of Technology, Bentley, Australia 4 Current address: Universidad Politécnica de Madrid, Madrid, Spain 2 *E-mail: karam.singh@csiro.au Abstract Fungal diseases continue to cause major problems for legume production worldwide. Translation of findings from model species such as Medicago truncatula and Arabidopsis holds promise for addressing some of these industry issues. Our previous studies on ERF transcription factor, AtERF14, indicate it is a master regulator required for ethylene- and pathogen– responsive expression of other ERFs and defence genes, such as PDF1.2. AtERF14 Knock-out lines displayed enhanced susceptibility to the rootinfecting Fusarium oxysporum. Homologs of AtERF14 were studied in the model legume Medicago truncatula during interactions with the root infecting fungus, Rhizoctonia solani, which causes significant diseases to numerous legumes including soybean, lupin and alfalfa. Despite extensive germplasm screens in many crops, no strong genetic resistance has been identified, suggesting alternative strategies for resistance are required. A specific induction of B-3 subgroup ERFs was associated with moderate resistance to R. solani in Medicago. Over-expression of B-3 ERFs in hairy roots increased resistance to R. solani and Phytophthora medicaginis, but not to root knot nematode indicating that targeting specific regulators of ethylene defence may enhance resistance to an important subset of root pathogens. Moreover, overexpression of B-3 ERFs enhanced disease resistance without apparent impact on symbiotic interactions with rhizobium in Medicago genotype A17, while over-expression in skl reduced the hypernodulation phenotype. This suggests that under normal regulation of nodulation, enhanced resistance to root diseases can be uncoupled from symbiotic plant-microbe interactions in the same tissue and ethylene/ERF regulation of nodule number is distinct from the defenses regulated by B-3 ERFs. VI ICLGG - Posters P-SAD13 Phenotypic characterization of mungbean (Vigna radiata) rhizobial isolates for abiotic stress performance Bansal M*, Kukreja K Chaudhary Charan Singh Haryana Agricultural University, Hisar, India *E-mail: monabansal30@gmail.com Abstract Nighty one rhizobial isolates were obtained from the nodules of mungbean (Vigna radiata) crop grown in different parts of Haryana. Authenticity of rhizobia was done by plant infection test under sterilized conditions. All the rhizobial isolates were characterized for their growth performance against different abiotic stresses including salinity (1-4% NaCl), drought (10, 15, 20% 0 PEG6000) and high temperature (35, 40, 45 C). About 21% of rhizobial isolates were found to be halotolerant, capable of growing at 4% NaCl concentration. Only 10% of rhizobial isolates were drought tolerant which were growing at 20% PEG6000 concentration. All the rhizobial isolates were 0 capable of growing at temperature of 35 C whereas only 2% of rhizobial 0 isolates were found to grow at high temperature i.e. 45 C. The salt and drought tolerant rhizobial isolates are excellent models to study the resistance mechanism(s), and to elucidate the role of genetics of NaCl and drought tolerance. The salt tolerant, drought tolerant and high temperature tolerance patterns found among the mungbean rhizobial isolates are reflecting the environmental stresses pressure predominant in their locations and are very good examples of the importance of using efficient rhizobial isolate for plant successful inoculation. VI ICLGG - Posters The molecular basis of floral transition in shoot apical meristem of soybean P-SAD14 Bhalla PL*, Wong CE, Jung C-H, Singh M B Plant Molecular Biology and Biotechnology Laboratory, The University of Melbourne, Victoria, Australia *E-mail: premlb@unimelb.edu.au Abstract Flowering and seed set underpin most of the agriculture production. The development of the shoot apical meristem into floral meristem is a major event in a plant’s life that has to be precisely timed to ensure reproductive success. Many plants use the change in day length ie photoperiod as signals for flowering. Though understanding of floral initiation and timing is critical for breeding climate adapted highly productive crops, there have only been limited studies on floral initiation in legume species. This is particularly true for soybean, a major oilseed legume crop used for human and animal feed. The transition to flowering is characterized by a shift of the shoot apical meristem from leaf production to the initiation of a floral meristem. Our aim is to characterize the molecular events leading to floral transition in soybean shoot apical meristem. We used genomics approaches to examine flowering regulatory genes in the soybean and also made comparison with the most studied model plant, Arabidopsis. This has led to identification of number of transcripts showing differential expression profiles during the floral transition. Further analysis implicates various molecular processes, especially hormonal pathways, critical to the floral transition during soybean development. Our study provides an essential genomic resource for functional analyses of the soybean flowering pathway, facilitating future biotechnology research and efforts into breeding robust high-yielding crop varieties. VI ICLGG - Posters P-SAD15 Oxygen binding by leghemoglobin, a late nodulin is modulated by phosphorylation and interaction with another late nodulin Bhar K, Ghosh A, Bhowmik D, Chakrabarti R, Chakraborty S, Gupta R, Siddhanta A* University of Calcutta, Kolkata, India *E-mail: asiddhanto@yahoo.com Abstract The symbiotic interaction between legumes and rhizobia consists of two sets of genes namely early and late nodulin genes that are expressed in root hair cells of the legumes. There are a number of late nodulin gene products found so far in legumes. Among them, Leghemoglobin (LegH) maintains anaerobic environment by sequestering O2 inside the nodule and thus supports nitrogen fixation. Very little is known about the regulation of LegH during nodulation. Furthermore, so far there is no known network between these late gene products. To address this, in order to find interacting partners we have generated (His)6-tagged LegH and some other late gene products from a cDNA library derived from 21 day post-infection nodules of Lotus japonicus(lj). Using affinity pull down assay, we have tested pair wise interaction between these (His)6-tagged late gene products. Our result shows that LegH specifically interacts with a late gene product, PITP-Nlj16. PITP-Nlj16 is a novel gene product that shares a two-domain structure consisting of an Nterminal Phosphatidylinositol Transfer Protein (PITP)-like domain joined to a Cterminal domain composed of amino acid sequences identical to, or highly related to, late nodulin LjNOD16 (Nlj16) whose function is yet unknown. Our data clearly reveal that the C-terminal Nlj16 domain but not the N-terminal ‘PITP’ domain is necessary to bind LegH. Most significantly, our result also 2+ shows that a Ca /Cam kinase from L.japonicus mediated serine phosphorylation of Nlj16 domain reduced this interaction. The oxygen binding by leghemoglobin is also modulated by the interaction with Nlj16. VI ICLGG - Posters Characterisation of candidate Glycine max symbiosome membrane proteins 1,* 1 1 1 1 1 P-SAD16 2 Brear E , Clarke V , Loughlin P , Qu Y , Chen L , Overall R , Day D , Smith 1 P 1 University of Sydney, Sydney, Australia Flinders University, Adelaide, Australia 2 *Email: ella.brear@sydney.edu.au Abstract The legume-rhizobium symbiosis has great importance to society as a source of high protein food and for enriching soil nitrogen. Legumes house their symbiotic bacteria, termed bacteroids, in a new plant organ called the nodule. Within infected nodule cells the bacteroids are encapsulated by the plant derived symbiosome membrane, which forms an interface for nutrient exchange. The primary exchange involves the supply of fixed nitrogen produced by the bacteroids, in return for energy and metabolites derived from the plant. Protein transporters embedded within the symbiosome membrane facilitate this exchange. My research focuses on characterising proteins, identified during proteomic analysis of isolated symbiosome membrane. Two of the candidates that I am studying are possible metal transporters. Nodulin 21 is a potential iron transporter and PLAC8 candidates contain a domain common to known cadmium, zinc and calcium transporters. The third candidate, DUF588, may be involved in vesicle trafficking to the symbiosome membrane. Both Nodulin 21 and DUF588 are highly and specifically expressed in mature, nitrogen fixing nodules. Preliminary data from silencing DUF588 expression using artificial microRNA suggests that DUF588 plays an important role in nodule development. Currently I am confirming candidate localisation to the symbiosome membrane using reporter gene fusions and characterising candidate function using yeast complementation. VI ICLGG - Posters P-SAD17 Medicago truncatula CYP716A12 is a multifunctional oxidase involved in the biosynthesis of hemolytic saponins 1 1 2 1 1 2 Carelli M , Biazzi E , Panara F , Tava A , Scaramelli L , Porceddu A , Graham 3 1 1 2 3 1 2, N , Odoardi M , Piano E , Arcioni S , May S , Scotti C , Calderini O * 1 CRA-Fodder and Dairy Productions Research Centre, Lodi, Italy Institute of Plant Genetics, Perugia, Italy 3 NASC, School of Biosciences, University of Nottingham, UK 2 * E-mail: ornella.calderini@igv.cnr.it Abstract Saponins are a group of glycosidic compounds common in legumes including the model species Medicago truncatula. They have been characterised for several health benefits for humans and for their biological activity against plant pathogenic microorganisms. In Medicago saponins belong to two main subclasses depending on their hemolytic activity. Understanding the genetics and biochemistry of saponins represents an important goal for the exploitation of their properties in both food and feed. In the last years a combination of transcriptomic, bionformatic and genetic analysis in M. truncatula discovered several P450 monoxygenases and glycosyltransferases (GTs) as candidate genes for the two main steps in the saponin synthesis: oxidation of the βamyrin ring and glycosylation of the sapogenins. We report the first genetic analysis of a P450 gene (CYP716A12) involved in saponin synthesis in M. truncatula. Genetic and biochemical approaches proved that CYP716A12 is responsible for an early step in the saponin biosynthetic pathway. In fact mutant plants for CYP716A12 were unable to produce hemolytic saponins and were thus named lacking hemolytic activity (lha). In vitro enzymatic activity indicated that CYP716A12 catalyzes the oxidation of β-amyrin and erythrodiol at the C-28 position, yielding oleanolic acid. Transcriptome changes in the lha mutant showed a modulation in the main steps of triterpenic saponin biosynthesis, highlighting further candidate genes in the saponin pathway. VI ICLGG - Posters Toward understanding bud outgrowth in red clover (Trifolium pratense): a morphological, physiological and molecular approach 1,2 1 3 3 P-SAD18 1,2 Van MA , Roldán-Ruiz I , Ruyter-Spira C , Bouwmeester HJ , Van BE , 1,* Cnops G 1 Institute for Agricultural and Fisheries Research, Melle, Belgium Ghent University, Gent, Belgium 3 Wageningen University, Wageningen, Netherlands 2 *E-mail: gerda.cnops@ilvo.vlaanderen.be Abstract Mixed grass-clover grasslands are an essential element of sustainable agriculture. The presence of clover in the mixture reduces nitrogen fertilizer application needs and results in improved nutritional value (high levels of protein and polyunsaturated fatty acid levels). Plant architecture may have a strong influence on agronomic important traits such as forage yield, re-growth capacity, seed yield and persistence in red clover. We have done an in-depth study of the architecture of a limited number of red clover genotypes with contrasting branching phenotypes based on the number of branches, the outgrowth of the main axis and the growth habit (erect/prostrate). Detailed morphological analysis revealed differences in the number of buds, the number of branches and the position and timing of bud outgrowth in the different genotypes. The bud outgrowth percentage (defined as percentage of buds that grow out into branches) was also significantly different among genotypes. We then determined the role of branching hormones on bud outgrowth using isolated single node fragments. These results were linked to expression levels of genes involved in branching according to literature. In addition, we determined concentrations of endogenous strigolactone in intact plants of the various genotypes. The results demonstrate that both bud initiation and bud outgrowth are important to explain branching differences in red clover and that the strigolactone biosynthesis-perception pathway is a relevant candidate for further analysis and molecular breeding for high yielding and more persistent red clover varieties. VI ICLGG - Posters P-SAD19 Role of cyclops in an aeschynomeneae legume Arachis hypogaea 1 1,2 Das DR , Sinharoy S , DasGupta M 1,* 1 University of Calcutta, Kolkata, India. Present address: Samuel Roberts Noble Foundation, Ardmore, USA 2 * E-mail: maitrayee_d@hotmail.com Abstract Cyclops or IPD3 is a member of Common Sym pathway that is required for both nodulation and mycorrhization. It encodes a protein with a C-terminal coiled-coil motif and 2 nuclear localization signals that direct the protein to the nucleus, colocalizing with CCaMK, a Ca2+/calmodulin-dependent protein kinase essential for decoding calcium signatures in Sym Pathway. Functionally Cyclops is associated with intracellular colonisation or infection of the symbionts. In this report, we describe the role of Cyclops in an aeschynomeneae legume Arachis hypogaea, where rhizobial invasion bypass the infection-thread. Instead, the invasion occurs through epidermal cracks and the rhizobia are endocytosed directly by a cortical cell and thereby triggers for nodule development. AhCyclops is demonstrated to be a substrate of AhCCaMK suggesting the interaction between these two factors to be conserved in both infection thread as well as non infection thread legumes. RNA interference of Ahcyclops considerably decreased the number of nodules which is consistent with the coupling of bacterial internalization and nodule development in aeschynomeneae legumes. RNAi nodules were properly developed with proper dissemination of symbionts. However these nodules showed improper development of symbiosome as compared to the nodules developed in the vector transformed plants indicating the role of this factor in the process of symbiosome development. VI ICLGG - Posters A typical calcium dependent protein kinase (CDPK) in aeschynomeae legume Arachis hypogaea Ghosh B, Kundu A , Dasgupta M P-SAD20 1,* University of Calcutta, Kolkata, India *E-mail: maitrayee_d@hotmail.com Abstract In legumes, both a rapid calcium influx and a subsequent calcium spiking encode informational signals that are associated with the establishment of root nodule symbiosis. Forward genetics identified a Calcium/calmodulindependent protein kinase (CCaMK) that is involved in decoding these calcium signals. Several report indicates Calcium-dependent protein kinase (CDPKs) to also have a role in establishment of symbiosis. We have come across some interesting observations on a CDPK from Arachis hypogaea that indirectly indicates a specific role of this kinase in the legumes belonging to the aeschynomeneae clade. This CDPK contains a replacement of an invariant Lys86 for Glu86 in αC helix of the N-terminal lobe in kinase domain suggesting it to be an inactive kinase. Our observations however shows this kinase to be more active towards Manganese dependent autophosphorylation than its mutated counterpart where the Glu86 is reverted back to Lys86. This is an unique replacement that has not been reported for any other CDPKs. In fact no other protein kinase has been shown to have a replacement in this position. Genomic DNA sequence encoding this CDPK also had the replacement of Lys86 for Glu86 eliminating the possibility of RNA editing for this kinase. The gene turned out to be intron-less and this appear to be the first report of an intron less CDPK in plants. We sequenced this CDPK from another aeschynomene legume Aeschynomene aspera where also the gene was intronless with the unique substitution of the invariant Lys for Glu. This clade specific variant of CDPK could have a functional role in the clade specific processes of crack invasion and/or aeschynomenoid nodule development. VI ICLGG - Posters P-SAD21 Distinct Bradyrhizobial populations from Oryza sativa and Arachis hypogaea collected from an intercropped field 1 Guha S, Saha S, Sarkar M , Dasgupta M 1,* University of Calcutta, Kolkata, India *E-mail: maitrayee_d@hotmail.com Abstract Arachis hypogaea belongs to the Aeschynomeneae tribe that are basal Papilionoids from the nodulating clade. In this legume, rhizobial invasion bypass the intracellular infection-thread mediated processes that are governed by specific molecular dialogues and are adapted by most legumes. Instead, the invasion occurs through epidermal cracks and the rhizobia are endocytosed directly by a cortical cell. This mode of invasion leads to promiscuity and opportunistic colonizers are reported. Here we report the genetic diversity of rhizobial strains collected from the nodules of Arachis hypogaea cultivated in a field at 23°24”N,88°30”E. These were compared and contrasted with the collection of rhizobial strains from the roots of Oryza sativa that were collected from the same field where it was intercropped with Arachis. The objective was to check if there was an overlap and/or segregation between the strains that are symbiosis competent and those that are endophytic colonisers. In total, 48 culturable rhizobial strains were obtained from the Arachis nodules, almost all belonging to the genus Bradyrhizobium. Few non rhizobial strains like Pseudomonas, Rhodococcus were also detected as opportunistic nodule colonisers. From rice, 20 rhizobial strains were isolated which were also of Bradyrhizobium genus. Phylotyping the rice and Arachis isolates with marker genes of symbiosis, nitrogen fixation, and housekeeping genes would be discussed. VI ICLGG - Posters Flower colour in pea (Pisum sativum L.) 1, 2 2 P-SAD22 3 Hofer J *, Moreau C , Ambrose M , Kite G , Ellis THN 1 1 Institute of Biological, Environmental & Rural Sciences, Aberystwyth University, Aberystwyth, UK 2 Department of Crop Genetics, John Innes Centre, Norwich, UK 3 Jodrell Laboratory, Richmond, UK *E-mail: jmh18@aber.ac.uk Abstract Flavonoids are polyphenolic secondary metabolites that are involved in pigmentation, defence, fertility and signalling in plants. Understanding the genetic regulation of flavonoid production in legumes will help to identify breeding targets. Crop legumes for human consumption often have white flowers and are low in tannins, while those grown as forage for animals are usually coloured and contain tannins which protect against bloat. Recently progress has been made in identifying the genes underlying flower colour variation in pea. The B gene encodes flavonoid 3’ 5’ hydroxylase, a structural enzyme in the flavonoid biosynthetic pathway. The A and A2 genes were found to encode, respectively, a basic helix-loop-helix (bHLH) transcription factor and a WD40 repeat protein, both of which are predicted to be components of the conserved Myb-bHLH-WD40 transcription factor complex that regulates flavonoid biosynthesis in plants. The gene encoding the Myb component of this complex, as well as genes at other loci involved in pigment production, such as Ce and Cr, remain to be identified. We report here on the preliminary characterisation of pigmentation mutants found in a screen of pea lines generated by fast neutron bombardment. These include a white flowered mutant that is allelic to neither a nor a2 and new alleles at the cr locus. VI ICLGG - Posters P-SAD23 Revisiting Medicago truncatula nodule development by RNA-seq and laser micro-dissection analyses 1 1 1 1 1 1 1 Jardinaud F , Roux B , Sallet E , Rodde N , Sauviac L , Capela D , Moreau S , 1 1 2 1 1 Catrice O , Timmers T , Faraut T , de Carvalho-Niebel F , Debellé F , Schiex 3 1 1 1 1,* T , Bruand C , Gouzy J , Gamas P 1 Laboratoire des Interactions Plantes Microorganismes (LIPM) CNRS/INRA – Auzeville, France 2 Laboratoire de Génétique Cellulaire, INRA - Auzeville, France 3 Unité de Biométrie et Intelligence Artificielle, INRA - Auzeville, France *E-mail: pascal.gamas@toulouse.inra.fr Abstract To have a comprehensive and integrated view of the genes involved in symbiotic nodule development, we set up methods that allow simultaneous gene expression profiling of both plant and bacterial partners. We analyzed, using High Depth RNA-Seq approaches the plant and bacterial transcriptomes of whole and laser-dissected root nodules (six regions) as well as roots or bacterial cultures, using Sinorhizobium meliloti 2011-Medicago truncatula as a model system. We opted for strand-specific RNA-seq techniques that allowed to identify, in addition to protein coding mRNAs, short and long non-coding RNAs (ncRNAs) not previously predicted by automatic genome annotation. A reliable framework for quantitative transcriptome analyses was provided by sequencing of S. meliloti strain 2011 genome, M. truncatula genome sequence data completion, and development of bioinformatic tools. Lowly expressed transcripts were identified thanks to the power of High Depth RNA-Seq coupled with laser capture micro-dissection. We also gained a spatial and genome-wide view of gene expression. Results were validated by analyzing a series of symbiotic genes previously characterized by other methods. Interestingly, we discovered genes likely to be involved in the regulation of key stages of nodule development, like meristem formation or plant cell differentiation. In addition we identified antisense ncRNAs that are differentially regulated in roots and nodules and may be involved in the regulation of certain symbiotic genes via novel mechanisms. Using those novel approaches, we gained a global view of genome expression in model plant and bacteria, a quantitative estimate of the relative abundance of various plant and bacterial transcripts in different symbiotic tissues, and discover new regulators of gene expression associated with defined stages of nodule development. VI ICLGG - Posters Legume nodule development and nutritional signalling influenced by the membrane localised soybean basic helix-loop-helix transcription factor, SAT1 1 2 1 P-SAD24 1 Chiasson DM , Loughlin PC , Mazurkiewicz D , Mohammadidehcheshmeh M , 3 4 5 1 3 1 Fedorova E , Okamoto M , Glass ADM , Smith S , Bisseling T , Tyerman SD , 6 1 Day DA , Kaiser BN * 1 School of Agriculture Food and Wine, University of Adelaide Waite Campus, Urrbrae, Australia 2 School of Biological Sciences, University of Sydney, Sydney, Australia 3 Laboratory of Molecular Biology, Department of Plant Sciences, Wageningen University,Wageningen, Netherlands 4 The Australian Centre for Plant Functional Genomics, University of Adelaide Waite Campus, Urrbrae, Australia 5 Department of Botany, University of British Columbia, Vancouver, Canada 6 Flinders University, Adelaide, Australia *E-mail: brent.kaiser@adelaide.edu.au Abstract SAT1 is a membrane bound basic helix-loop-helix transcription factor first identified in soybean (Glycine max) through its ability to rescue growth of an ammonium transport yeast mutant (26972c). SAT1 expression is enhanced in nitrogen-fixing legume nodules, displays phosphorus responsiveness and is diurnally regulated. Loss of SAT1 activity disrupts nodule growth and nitrogen fixation and when colonised by arbuscular mycorrhizal fungi (AM), phosphorus homeostasis is disrupted. SAT1 is localised at the symbiosome, plasma membrane and endoplasmic reticulum and upon posttranslational modification, released to the nucleus. Transcriptional targets of SAT1 include nutritional related genes including an uncharacterised major facilitator protein capable of ammonium transport, homologs of which reside next to SAT1 chromosomal loci across many plant species including soybean, Medicago, Arabidopsis and maize. VI ICLGG - Posters P-SAD25 Relatedness of drought with nitrogen fixation in chickpea (Cicer arietinum L.) 1 1 1 1 1 Kaashyap M , Krishnamurthy L , Rathore A , Purushothaman R , Anil V , 1 1 1 2 2 Parupalli S , Gaur PM , Upadhyaya HD , Penmetsa RV , Cook DR , Varshney 1 RK * 1 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 2 University of California Davis, Davis, USA *E-mail: r.k.varshney@cgiar.org Abstract Biological nitrogen fixation (BNF) is complex physiological feature having multiple environmental interaction effects such as drought which severely hampers crop yield. Domestication and extensive breeding have led to adaptation of chickpea to their needs for nitrogen fixation according to different environmental conditions like drought. To elucidate this factor, 3 Cicer species (arietinum, reticulatum, echinospermum) were subjected to interactions amongst drought and N-fixation conditions in split-spilt plot design. Main-plot treatment were three water regimes (well watered, drought at early stage, drought at late stage), sub-plot treatments being control, added nitrogen, added rhizobium while sub-sub plots were 12 chickpea genotypes. Based on root-shoot allocations as fitness parameters, drought tolerant and susceptible genotypes were recorded for their BNF efficiencies in stress. Significantly, genotypes could interact within main plot and sub-plots resulting in depleted shoot biomass but high root biomass in stressed conditions. Unlike susceptible, drought tolerant genotypes (ICC 8261, ICC 4958) could make high shoot biomass but less root biomass with added nitrogen. Phenological trends in term of shoot allocations suggest that none of the genotypes could efficiently establish symbiosis to avail biological nitrogen under stressed conditions with added rhizobium. As added nitrogen dosage will always be triggering sensitivity to nodulation gene responses, it will be indicative to identify the parental genotypes which may efficiently fix nitrogen under drought stress. In accomplishment, conserved N-fixation gene orthologs from Medicago, Lotus are amplified in drought tolerant and susceptible parents to identify useful SNPs in this locus for mapping N-fixing hotspots underlying vicinities of drought QTLs. VI ICLGG - Posters Examination of relationship of rhizobium sequences and chickpea traits for efficient nitrogen fixation 1 1 2 2 P-SAD26 3 Kim DH , Kaashyap M , Greenspan A , Penmetsa RV , von Wettberg E , 1 1 1 1 4 4 5 Gaur PM , Rathore A , Upadhyaya HD , Das RR , Singh S , Kaur J , Yasin M , 2 1,6, Cook DR , Varshney RK * 1 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 2 University of California-Davis, Davis, USA 3 Florida International University, Miami, USA 4 Punjab Agricultural University, Ludhiana, India 5 Rajmata Vijayaraje Scindia Krishi Vishwa Vidyalaya (RVSKVV), Sehore, India 6 Generation Challenge Programme, c/o CIMMYT, Mexico DF, Mexico *E-mail: r.k.varshney@cgiar.org Abstract Legumes have symbiotic interaction with specific nitrogen-fixing bacteria, called rhizobium. To characterize phenotypic plasticity for nitrogen fixation across evolutionary gradient in Cicer species, 18 cultivated chickpeas (C. arietinum) and 12 wild accessions (C. reticulatum, C. bijugum, C. echinospermum, C. judaicum, C. pinnatifidum and C. yamashitae) were phenotyped for fitness parameters in three treatments i.e. control, rhizobium and nitrogen at Patancheru, Ludhiana and Sehore in India. Cultivated chickpeas showed less nodule number / weight but more shoot weight in rhizobium treatment. Cultivated chickpea provided with improved inoculums performed equivalent to nitrogen treated plants but wild chickpea showed similar shoot weight in all three treatments. Statistical analyses suggest less symbiotic efficiency in cultivated chickpea with indigenous rhizobium which means that wild chickpea can make efficient symbiosis with indigenous rhizobium but cultivated chickpea do not. Interestingly, wild chickpeas did not utilize added nitrogen so there was no difference in nitrogen treatment. Rhizobial sequences from C. reticulatum showed the highest sequence diversity and OTU (Operational Taxonomic Units). These results show chickpeas that have more diverse rhizobium might have high possibility to have bigger nodules and shoot. To validate this results, a genetic gradient panel of 12 genotypes comprising of 6 C. arietinum (3 primitives, 3 elites), 3 accessions each of C. reticulatum and C. echinospermum was selected for undertaking field experiments to examine correlations between genotypes and nitrogen fixation-related traits and sequence analysis is in progress. VI ICLGG - Posters P-SAD27 Identification of a MAPK gene involved in MYMIV-resistance in Vigna mungo: In silico and biochemical characterization Patel A*, Kundu A, Chaudhuri S, Pal A Bose Institute, Kolkata, India *Email: anjupatl@bic.boseinst.ernet.in Abstract Mitogen-activated protein kinases (MAPK) are essential components of stress signaling in plants. It transduces pathogenic signals to downstream molecules leading to cellular responses involved in plant immunity. We have identified a MAPK homologue in the defence signaling pathway during incompatible interaction between Vigna mungo and the pathogen, Mungbean Yellow Mosaic India Virus (MYMIV). In this study, we have isolated and partially characterized a stress-responsive VmMAPK gene, the first to be reported in V. mungo. Full length cDNA was isolated using RNA ligase mediated rapid amplification of cDNA ends (RLM-RACE) technology which showed the clone to be 1,557 bp in length with an open reading frame (ORF) of 1,119 bp. The ORF encodes a 372-amino-acid polypeptide with a calculated molecular weight of 42 kDa. In silico sequence analysis indicated the isolated gene is a member of serine threonine protein kinase family with a signature motif of TEY residue, the putative phosphorylation site. Time course expression analysis through qPCR under MYMIV-stress indicated a marked induction of this transcript with its highest accumulation at 12 hours post infection. Phylogenetic analysis has revealed that VmMAPK is closely related to MAPK homologue of cowpea and soybean. The cDNA clone was subcloned in the expression vector pET32a and the protein was purified from the soluble fraction of total protein. Purified recombinant VmMAPK showed autophosphorylation activity in presence of 2+ Mg ions in a concentration dependent manner. Further biochemical analyses are underway. VI ICLGG - Posters Transcript profiling of the common bean – Rhizobium symbiosis under oxidative stress 1, 2 1 1 P-SAD28 1 Ramirez M *, Guillén G , Fuentes SI , Iñiguez LP , Aparicio-Fabre R , Rivera 1 1 1 3 3 P , Salazar E , Encarnación-Guevara S , Panzeri D , Castiglioni B , 3 3 3 1 3 1 Cremonesi P , Strozzi F , Stella A , Girard L , Sparvoli F , Hernández G 1 Centro de Ciencias Cenómicas - Universidad Nacional Autónoma de México, Cuernavaca, México 2 Instituto de Biotecnología - Universidad Nacional Autónoma de México, Cuernavaca, México 3 Istituto di Biologia e Biotecnologia Agraria, Milano, Italy *E-mail: mario@ccg.unam.mx Abstract A wide range of environmental stresses lead to an elevated production of reactive oxygen species (ROS) in plant cells thus resulting in oxidative stress. The biological nitrogen fixation in the legume - Rhizobium symbiosis is at high risk of damage from oxidative stress. Common bean (Phaseolus vulgaris) active nodules exposed to the herbicide Paraquat (1,1 '-Dimethyl-4, 4'bipyridinium dichloride hydrate) that generates ROS accumulation, showed a reduced nitrogenase activity and ureide content. We analyzed the global gene response of stressed nodules using the Bean CombiMatrix Custom Array 90K, that includes probes from some 30,000 expressed sequence tags (EST). A total of 4,280 ESTs were differentially expressed in oxidative stressed bean nodules; of these 2,218 were repressed. These genes were grouped in 44 different biological processes as defined by Gene Onthology. Analysis with the PathExpress bioinformatic tool, adapted for bean, identified five significantly repressed metabolic pathways related to carbon/nitrogen metabolism that is crucial for nodule function. The expression platform for transcription factor (TF) genes based in qRT-PCR revealed that 67 TF genes were differentially expressed in oxidative stressed nodules. The expression analysis of fifteen oxidative stress responsive genes from the bacteroids in stressed nodules revealed that only few genes showed significant changes in their expression level. VI ICLGG - Posters P-SAD29 Selective recruitment of mRNAs and miRNAs to polyribosomes in response to rhizobia infection in Medicago truncatula 1,* 1 2 3 1 Reynoso MA , Blanco FA , Bailey-Serres J , Crespi M , Zanetti ME 1 Instituto de Biotecnología y Biología Molecular, FCE-UNLP CCT-CONICET, Argentina 2 Department of Botany and Plant Sciences, Center for Plant Cell Biology, University of California, Riverside, USA 3 Institut des Sciences du Végétal, CNRS, Gif sur Yvette, France *E-mail : mauricio_a_reynoso@yahoo.com.ar Abstract Translation of mRNAs is a key regulatory step that contributes to the coordination and modulation of eukaryotic gene expression during development or adaptation to the environment. mRNA stability or translatability can be regulated by the action of small regulatory RNAs (sRNAs), which control diverse biological processes. Under low nitrogen conditions, leguminous plants associate with soil bacteria and develop a new organ specialized in nitrogen fixation, the nodule. To gain insight into the translational regulation of mRNAs during nodule formation, the association of mRNAs and sRNAs to polysomes was characterized in roots of the model legume Medicago truncatula during the symbiotic interaction with Sinorhizobium meliloti. Quantitative comparison of steady-state and polysomal mRNAs for fifteen genes involved in nodulation identified a group of transcripts with slight or no change in total cellular abundance that were significantly up-regulated at the level of association with polysomes in response to rhizobia. This group included mRNAs encoding three receptors like kinases required either for nodule organogenesis, bacterial infection or both and transcripts encoding GRAS and NF-Y transcription factors. Quantitative analysis of sRNAs in total and polsyomal RNA samples revealed that mature microRNAs (miRNAs) were associated with the translational machinery, notably, miR169d/l, which targets the NF-YA/HAP2 mRNA. Upon inoculation, levels of miR169d/l pronouncedly decreased in polysomal complexes, concomitant with increased accumulation of NF-YA/HAP2 protein. These results indicate that both mRNAs and miRNAs are subjected to a differential recruitment to polysomes and expose the importance of selective mRNA translation during root nodule symbiosis. VI ICLGG - Posters Early events during rhizobial invasion and colonisation of a crackinvasion supported legume Arachis hypogaea P-SAD30 Saha S, DasGupta M* University of Calcutta, Kolkata, India * E-mail: maitrayee_d@hotmail.com Abstract A common rhizobial invasion strategy is through epidermis by root hair curling and infection thread (IT) formation. The alternate mode of rhizobial invasion is known as “crack invasion”. This is a characteristic feature of approximately 25% of all legume genera, where rhizobia directly access the cortical cells through epidermal “cracks”. While detail features associated with infection thread mediated invasion has been worked out, very little is known about inception of nodulation following crack invasion. Arachis hypogaea is a representative member of the aeschynomeneae tribe that are supported by crack invasion. Here we report the initial events during rhizobial invasion of Arachis hypogaea that led to development of their characteristic aeschynomenoid nodules. Tufted rosettes of long root hairs occur in axils of young lateral roots of Arachis hypogaea. We demonstrate that crack invasion in Arachis is mediated by intracellular transport of symbionts through rosette type root hair followed by their accumulation near lateral root bases. There was however no sign of infection threads in the rossette hairs. The early events during rhizobial invasion and colonization in Arachis hypogaea would be discussed. VI ICLGG - Posters P-SAD31 Regulation of symbiosis receptor kinase 1 1,4 1 1 1 Samaddar S , Sinharoy S , Dutta A , Saha S , Paul A , Bhattacharya 1 3 3 3 3 3 A , Chien K , Goshe MB , Banerjee A , Das A , Chakrabarti S , DasGupta 1,* M 1 University of Calcutta, Kolkata, India. North Carolina State University, Raleigh, USA 3 Indian Institute of Chemical Biology, Kolkata, India 4 The Samuel Roberts Nobel Foundation, Ardmore, USA 2 *E-mail: maitrayee_d@hotmail.com Abstract SYMRK is a leucine-rich-repeat (LRR)-receptor kinase that mediates intracellular symbioses of legumes with rhizobia and arbuscular mycorrhizal fungi. Present analyses indicate that this gene is not involved in recognising rhizobial specificity. It rather participates in signalling events associated with developmental programs that are central for intracellular accommodation of symbionts. It is paramount to understand the exact biochemical functions and regulation mechanisms of SymRK because it is a receptor kinase that appears to govern the entire symbiotic process through various interacting proteins/substrates. Here we report characterisation of SymRK from Arachis hypogea (AhSymRK), a basal papilinoid from the nodulating clade. Although annotated as a Ser/Thr kinase, sequence homology suggests SymRK to be a member of Tyrosine Kinase-like (TKL) family. Recent reports on other plant RLKs like AtBRI1, AtBAK1, MtLyk3 etc., identified sites of tyrosine phosphorylation which are emerging to be ‘more abundant than expected’ in plants. We hereby demonstrate, for the first time, tyrosine phosphorylation on AhSymRK which is also the first report on the regulatory role of tyrosine phosphorylation amongst the plant RLKs. This phosphorylation functions as a lock to maintain a catalytic ‘off’ state of this kinase. Our observations suggest dephosphorylation to be a central trigger for turning on the downstream symbiotic responses mediated through SymRK. VI ICLGG - Posters Comparative genomic analysis of soybean flowering genes P-SAD32 Singh MB*, Chol-Hee Jung, Wong ACE, Bhalla PL Plant Molecular Biology and Biotechnology Laboratory, University of Melbourne, Parkville, Australia *E-mail: mohan@unimelb.edu.au Abstract Flowering is one important agronomic trait that determines crop yield. Soybean is a major oilseed legume crop used for human and animal feed. Legumes have unique vegetative and floral complexities. Our understanding of the molecular basis of flower initiation and development in legumes is limited. Here, we address this by undertaking using a computational approach to examine flowering regulatory genes in the soybean genome in comparison to the most studied model plant, Arabidopsis. For this comparison, a genomewide analysis of orthologue groups was performed, followed by an in silico gene expression analysis of the identified soybean flowering genes. Phylogenetic analyses of the gene families highlighted the evolutionary relationships among these candidates. Our study identified key flowering genes in soybean and indicates that the vernalisation and the ambienttemperature pathways seem to be the most variant in soybean. A comparison of the orthologue groups containing flowering genes indicated that, on average, each Arabidopsis flowering gene has 2-3 orthologous copies in soybean. Our analysis highlighted that the CDF3, VRN1, SVP, AP3 and PIF3 genes are paralogue-rich genes in soybean. Furthermore, the genome mapping of the soybean flowering genes showed that these genes are scattered randomly across the genome. A paralogue comparison indicated that the soybean genes comprising the largest orthologue group are clustered in a 1.4 Mb region on chromosome 16 of soybean. Furthermore, a comparison with the undomesticated soybean (G. soja) revealed that there are hundreds of SNPs that are associated with putative soybean flowering genes and that there are structural variants that may affect the genes of the light-signalling and ambient-temperature pathways in soybean. Our study provides a framework for the soybean flowering pathway and insights into the relationship and evolution of flowering genes between a short-day soybean and the long-day plant, Arabidopsis. VI ICLGG - Posters P-SAD33 Identification & functional characterization of Medicago truncatula mutants with impaired symbiotic nitrogen fixation 1 2 1 1 1 2 Sinharoy S , Liu C , Torres-Jerez I , Pislariu C , Wang M , Breakspear A , 2 1 3 2 1 Guan D , Jiangqi W , Benedito V , Murray J , Udvardi M * 1 The Samuel Roberts Noble Foundation, Plant Biology Division, Ardmore, USA John Innes Centre, Cell and Developmental Biology, Norwich, UK 3 Division of Plant & Soil Sciences, West Virginia University, Morgantown, USA 2 *E-mail: mudvardi@noble.org Abstarct Transcriptomic studies have revealed that thousands of plant genes are involved in symbiotic nitrogen fixation (SNF). To understand SNF more deeply we undertook both forward and reverse genetic approaches to determine gene function, using the Medicago truncatula Tnt1-insertion mutant population. We constructed hypothetical gene regulatory networks consisting of TFs and their putative targets, based on correlation analysis of spatio-temporal gene expression data. Thirty four mutant lines with insertions in sixteen different genes were identified. Further detailed investigation has been undertaken with a C2H2-TF (MtNERP), bHLH-TF, WRKY and some of the non TF targets. Analysis of flanking sequence tags in one fix- mutant revealed an insertion in a putative membrane localized Cystathionine Beta Synthase domain-containing gene (MtCBS1). The insertion was found to co-segregate with the nodulation phenotype, and transgenic expression of MtCBS1 in the mutant restored the wild-type phenotype. Detailed phenotyping of the cbs1 mutant revealed a primary defect in infection of root hairs, and when infection threads (ITs) did penetrate into the nodule, very few cells were infected. The mutant exhibited excessive root hair curling in response to S. meliloti with an unusual shepherd’s crook structure, and most infections aborted after formation of infection foci. The few ITs generated from these foci were usually defective. MtCBS1 expression was induced in root hair cells during the early stages of infection by rhizobia and after Nod factor treatment. MtCBS1 promoter-GUS studies showed that MtCBS1 expression is restricted to the distal part of the invasion zone and to infected root hair cells. VI ICLGG - Posters Origin and evolution of polymeric gynoecium in legumes: evidences from Pisum floral mutants P-SAD34 Sinjushin AA* M.V. Lomonosov Moscow State University, Moscow, Russia *E-mail: asinjushin@mail.ru Abstract Although flowers of legumes (Fabaceae) are known as having single carpel, some representatives are characterized with polymeric gynoecium. Most of them belong to tribes Swartzieae (Papilionoideae subfamily) and Ingeae (Mimosoideae subfamily). Garden pea (Pisum sativum L., Papilionoideae) normally has unicarpellate flower but some of its mutants produce supernumerary carpels. Studies on these mutants may shed light on possible developmental and evolutionary mechanisms of gynoecia formation in given family. Generally, four different mechanism can provide increase of carpel number, viz. flower fasciation (increase of floral meristem size), fusion of few independent floral meristems, homeotic transformation of any organs into carpels, and increased intensity of flower axis proliferation (superposition of flower and inflorescence developmental programs). No pea (or other legumes) mutants are known with flower fasciation; despite Arabidopsis (Brassicaceae) and other model plant species, stem fasciation is not associated with flower abnormalities in legumes. However, in fasciated pea mutants flower fusion may occur. In this case multicarpellate flowers may arise with carpels positioned valve-to-valve (as in normal multicarpellate flowers of Swartzia p.p., Swartzieae tribe). Homeotic transformation occurs in stamina pistilloida (stp) mutants with additional carpels oriented with their ventral sutures towards the floral centre (as in normal flowers of Archidendron, Ingeae tribe, or Acacia celastrifolia, Acacieae). The cochleata mutants seem to produce additional carpels due to ectopic flower proliferation. All revealed cases find natural similarity within family and may reconstitute independent evolutionary origin of multicarpelly in legumes. VI ICLGG - Posters P-SAD35 Identification of transport proteins on the symbiosome membrane in soybean 1 1 1 1 1 3 3 Clarke V , Loughlin P , Qu Y , Chen C , Brear E , Castelli J , Ludwig M , 1 2 1,* Overall RL , Day DA , Smith PMC 1 University of Sydney, Sydney, Australia Flinders University, SA, Australia 3 University of Western Australia, Crawley, Australia 2 *E-mail: penny.smith@sydney.edu.au Abstract The symbiosome membrane (SM) is the interface between the plant and bacteria in the legume-rhizobium symbiosis. This plant-derived membrane separates the symbiotic form of the rhizobia, the bacteroid, from the plant cytoplasm and controls both the type and amount of solutes exchanged. In exchange for nitrogen fixed by the bacteroid, the plant provides reduced carbon, probably in the form of malate, to the bacteroids, as well as other solutes including iron, sulphate and zinc. We are investigating transport proteins on the SM that regulate this exchange in soybean nodules using a proteomic approach. Candidates that potentially encode malate or metal transporters are a major focus. Proteomic analysis of the SM involved trypsin digestion of total SM proteins, separation of peptide fragments by liquid chromatography (LC), and analysis with tandem mass spectrometry (MS/MS). Of the 71 proteins identified in all three biological replicates, 33 are predicted to be integral membrane proteins. A number of proteins known to be associated with the + SM, such as nodulin 26, H -ATPase, symbiotic sulphur transporter (sst1), Rab7, nodulins 24 and 53, were identified. Many of the identified SM proteins are highly expressed in nodules compared to other tissues. We are currently assessing the importance of some integral membrane proteins that are potential malate or metal transporters or of unknown function through an RNAi approach, and confirming the location of proteins on the SM using reporter gene fusions. Candidate transporters confirmed to be localised to the SM will be functionally characterised in heterologous expression systems. VI ICLGG - Posters Drought and elevated temperature negatively impact translocation of photosynthate into developing seeds and alter the expression of seed coat specific transporters in soybean (Glycine max) 1 1,2 Leisner CP , Ainsworth EA , Yendrek CR P-SAD36 1,* 1 University of Illinois, Urbana, USA USDA ARS, Urbana, USA 2 *E-mail: cyendrek@illinois.edu Abstract Within the next 30 years, the world population is projected to increase by over two billion people, requiring substantial improvements in crop productivity to meet global food demands. In addition to this challenge, agricultural output must also improve within the context of a changing growth environment. Two important climate change parameters that are expected to increase in frequency and magnitude, drought and elevated temperature, are well know to negatively impact reproductive development and ultimately crop yield. In this study we characterized the physiological response of soybean to drought and high temperature stress during seed filling in plants grown at the Free Air Concentration Enrichment field site in Champaign, Illinois. Net assimilation of CO2 and stomatal conductance were significantly decreased by both drought and elevated temperature. In addition, total nonstructural carbohydrate content in leaves was decreased by 23.9% in drought and by 28.2% in elevated temperature, indicating altered photosynthate supply to the developing seeds. Seed number was significantly decreased by both drought and temperature stress, while individual seed weight was not. In order to identify stressresponsive targets involved with regulating the movement of solutes into the developing seeds, we isolated total RNA from seed coat tissue and used RNAseq analysis to identify transcripts that were differentially expressed in drought and elevated temperature. The gene encoding cell wall invertase, which can increase sink strength by generating a high hexose-to-sucrose gradient in the apoplast between the seed coat and developing cotyledons, was highly up-regulated by both drought and temperature stress. In addition, the transcript abundance of several sucrose transporters was also upregulated. Thus, despite lower carbohydrate content from the source leaves in plants exposed to drought and high temperature, sucrose transport into the seeds was maintained, perhaps as a mechanism to ensure healthy individual seed size. VI ICLGG - Posters P-SAD37 Symbiotic performance of chickpea genotypes and their comparative flavonoids profiling Swarnalakshmi K*, Saha S, Pooniya V, Saxena AK Indian Agricultural Research Institute (IARI), New Delhi, India *E-mail: swarna_bga@yahoo.com Abstract Chickpea (Cicer arietinum L.) is an economically important pulse crop and is capable of assimilating atmospheric dinitrogen due to the presence and activities of Mesorhizobia in the roots. However, variable rresponse to Mesorhizobial inoculation ranging from 9-33% over uninoculated control was observed under field conditions. This differential response is attributed by plant genotype and soil type. Symbiotic process of chickpea genotype Mesorhizobium strain is depending on chemical signals present in the root exudates. These signal molecules, particularly flavonoids mediate nodulation process of Mesorhizobium in chickpea plant. Therefore attempts were made to identify the optimum combination of chickpea genotype-Mesorhizobium to obtain the maximum benefit of symbiotic nitrogen fixation and better growth of the plant. Chickpea genotypes representing different agro climatic regions were evaluated for nodulation potential in terms of Chlorophyll content, specific nodule weight and N uptake under IARI field during rabi 2010-12. Two genotypes showing high and low nodulation potential (BG 256 and ICC 4948) under field conditions were selected for their comparative flavonoids analysis in root exudates. For the collection of root exudates, chickpea seeds were surface sterilized using 0.1 % mercuric chloride and then rinsed several times with sterile distilled water and grew on perforated plate in a glass cylinder containing hydroponic system for two weeks. Ethyl acetate fraction of root exudates were analysed for flavonoids content. HPLC analysis revealed difference in flavonoid profile in high nodulation and low nodulating genotypes. Genistein (905 ppm) and formononetin (3134 ppm) were present in high nodulating genotype (BG 256) which is below detectable limit in the root exudates of low nodulating genotype (ICC 4948) which might be contributing difference in nodulation and N fixation. However LC-MS analysis is required to indentify other unknown flavonoids compounds present in the high nodulating genotype. Chemotactic interaction of Mesorhizobium strains with these flavonoids also requires further study. . VI ICLGG - Posters Negative regulation of CCaMK is essential for symbiotic infection in L. japonicus 1,# 2,# 1,# 3 1 P-SAD38 4 Liao J , Singh S , Hossain MdS , Andersen SU , Ross L , Sato S , Tabata 4 3 2 1,5,* S , Stougaard J , Parniske M , Szczyglowski K 1 Agriculture and Agri-Food Canada, Ontario, Canada Biocenter University of Munich (LMU), Munich, Germany 3 University of Aarhus, Denmark 4 Kazusa DNA Research Institute, Chiba, Japan 5 University of Western Ontario, Ontario, Canada # Authors contributed equally 2 * E-mail: krzysztof.szczyglowski@agr.gc.ca Abstract In contrast to pathogenic interactions, where the rapidly stimulated evolution of 2+ intracellular Ca partakes in signalling for plant defence, the peri-nuclear calcium spiking is part of a signal transduction process required for the intracellular colonization of plant roots by arbuscular mycorrhiza (AM) fungi 2+ and rhizobial symbionts. During both symbioses, Ca spiking is thought to be 2+ decoded by the same Ca and calmodulin (CaM)-dependent protein kinase (CCaMK). Deleterious mutations in CCaMK prevent root infections by both microsymbionts and also abolish nodule primodia formation in the root cortex during root nodule symbiosis (RNS) with rhizobia. These observations indicate that CCaMK has an essential signalling role in all of these processes. To gain insight into how CCaMK diversifies the action of calciummediated signals during symbiosis, we have analysed the L. japonicus ccamk14 mutant. Normal root infection by AM fungus and M. loti was impaired in ccamk-14, while no effect on nodule primodia formation was detected. Importantly, ccamk-14 specifically restricted the cortical but not the epidermal 337 infection during RNS. We show that the underlying mutation replaced serine 337 (S ), which we determined to be an in vitro autophosphorylation site within 337 the CaM binding region of the L. japonicus CCaMK. The S residue is conserved across angiosperm CCaMK and by testing discrete substitutions at this site we show that it participates in a negative regulation of CCaMK-activity, which is required for the cell-type-specific integration of symbiotic signalling a during infection . VI ICLGG - Posters P-SAD39 WUS-box mediated repression is central to STF regulated leaf blade development in Medicago truncatula 1 1 2 Lin H , Niu L , Mysore KS , Tadege M 1,* 1 Institute for Agricultural Biosciences, Oklahoma State University, Ardmore, USA 2 The Samuel Roberts Noble Foundation, Ardmore, USA *E-mail: million.tadege@okstate.edu Abstract The WOX class homeobox gene, STENOFOLIA (STF), is critically required for lateral outgrowth of the leaf blade in Medicago truncatula evidenced by severely affected narrow leaf stf mutant phenotypes. The equivalent mutation in Nicotiana sylvestris causes the lam1 phenotype where leaf blades are reduced to vestigial strips. STF/LAM1 promotes blade outgrowth by activating cell proliferation. The mechanism of this activation of cell proliferation is unknown. Using site-directed mutagenesis of the STF protein and transgenic complementation of the lam1 mutant, we identified that the WUS-box near the C-terminus of the STF protein is required for STF function in blade outgrowth. We found that two aminio acid substitutions in the WUS-box of STF (STFm1) abolish to a great extent its ability to complement the lam1 mutant. Fusing a repressor domain, SRDX, to STFm1 fully restores complementation while fusing an activator domain, VP16, to STFm1, if any, enhances the lam1 phenotype, suggesting that STF acts mainly as a repressor in blade outgrowth using its WUS-box as repressor domain. The WUS-box is conserved in the modern (WUS) clade of Arabidopsis and other species WOX genes except WOX7, but not in the intermediate and ancient clades. Consistent with this, all the modern clade members except WOX7 can complement the lam1 mutant when expressed under the STF promoter, but the intermediate and ancient clades cannot, suggesting evolutionary modification of function by acquisition of the WUS-box. We will present data suggestive of repressor complex formation at the adaxial/abaxial junction required for leaf blade outgrowth and female fertility. VI ICLGG - Posters Dissecting the metabolomic plasticity during seed development to characterize biochemical regulatory networks in soybean 1* 1 2 2 3 1 Valliyodan B , Shi H , Farmer A , May GD , Xu D , Stacey G , Nguyen HT P-SAD40 1 1 National Center for Soybean Biotechnology and Division of Plant Sciences, University of Missouri, Columbia, USA 2 National Center for Genome Resources, Santa Fe, USA 3 Computer Science Department, University of Missouri, Columbia, USA *E-mail: valliyodanb@missouri.edu Abstract Integrated ‘omics’ studies will contribute towards a systems approach to key processes such as seed development, which will lead to the genetic improvement of yield and seed composition. Thus, by studying gene products, a direct correlation between developmental related or stress related response at the level of specific peptides, and/or metabolites can be made. This will lead to crop improvement either through breeding or transgenic efforts. We have conducted metabolomics and transcriptomic analysis during seed developmental stages and in matured seeds to catalogue major metabolite and correlated transcript changes during seed development and maturation in soybean; both under controlled and water deficit conditions. Soybean variety Williams 82 was grown under controlled conditions and the drought stress was imposed by withholding water. The water relations parameters were monitored and seeds were harvested at the right developmental stages and processed for the analysis. We have used Gas Chromatography - Mass spectrometry (GCMS) and Liquid chromatography – Mass spectrometry (LC-MS/MS) for the chromatographic separation and identification of metabolites. Next generation sequencing based transcriptome analysis was conducted using the same tissue. We have identified developmental related and drought response related metabolite changes across the developmental stages. Integration of metabolite data with the transcripts helped identify key regulation of C-N partitioning and carbon flux during seed development. This approach helps dissect biochemical regulatory pathways associated with seed development and stress in soybean and major findings will be discussed. Several TFs regulating selected key pathways were tested in-planta to characterize and confirm functions. This will be applied further to develop soybean with better yield and seed composition. All information are populated in the soy metabolite data base (SoyMetDB) and the comprehensive soybean database, SoyKB. Various components of these databases also will be presented. VI ICLGG - Posters P-SAD41 A conserved basis for photoperiod adaptation in several temperate legumes 1 1 1 1 1 1 Weller JL ,Liew LC ,Hecht V , Rajandran V , Ridge S , Schoor JKV , Laurie 2 3 4 3 4 2 R , Wenden B , Dalmais M , Rameau C , Bendahmane A , Macknight RC , 5 Isabelle L-J 1 University of Tasmania, Australia University of Otago, New Zealand 3 INRA Versailles, France 4 CNRS Evry, France 5 INRA Mons, France 2 *E-mail: jim.weller@utas.edu.au Abstract Cool-season legumes are among the world’s oldest crops but the molecular basis for domestication and phenological adaptation in these species is not well understood. We surveyed a diverse collection of pea (Pisum sativum) germplasm and identified wide variation in flowering time and photoperiod responsiveness, supporting the conclusion that wild ancestors of cultivated pea had an obligate long-day flowering response. Genetic analyses have defined three major loci controlling photoperiodic flowering in global pea germplasm collections. The HR gene is an ortholog of Arabidopsis ELF3, with a single, widespread functional variant conferring altered circadian rhythms and the reduced photoperiod response associated with the early-flowering spring habit. A second major locus, QTL6, acts with HR to confer the ancestral, obligate long-day-requiring winter habit and is also likely to have contributed to domestication-related changes in flowering phenology and growth habit. Mutations at a third locus, SN, also affect circadian rhythms and completely eliminate photoperiod responsiveness, and have a restricted distribution within cultivated germplasm. We are also examining whether these loci may have a conserved role in other legumes. In lentil (Lens culinaris), photoperiod-insensitive early flowering is important for adaptation to low latitudes in southern Asia, while in the ornamental species sweet pea (Lathyrus odoratus), distinct winter and spring variants significantly extend the flowering season relative to conventional summer-flowering types. In both species we have identified HR/ELF3 mutations contributing to early flowering and reduced photoperiod sensitivity. These results suggest that mechanisms for flowering time adaptation may be conserved across temperate long-day legumes, and provide impetus for comparative studies of flowering in legumes generally. VI ICLGG - Posters Aspartate aminotransferase is located in the Lupinus angustifolius generich region highly syntenic to Glycine max and Medicago truncatula 1,* 1 2 2 P-SAD42 3 Wyrwa K , Ksiazkiewicz M , Zielezinski A , Karlowski WM , Podkowinski J , 1 1 Naganowska B ,Wolko B 1 Institute of Plant Genetics, Poznan, Poland Laboratory of Computational Genomics, Adam Mickiewicz University, Poznan, Poland 3 Institute of Bioorganic Chemistry, Polish Academy of Sciences, Poznan, Poland 2 *E-mail: kwyr@igr.poznan.pl Abstract Lupinus angustifolius is an important grain crop cultivated in sustainable farming. As a nitrogen fixing plant, the narrow-leafed lupin contributes to organic matter to soil, resulting in increased yields of successive crops. The key enzyme in primary nitrogen assimilation is aspartate aminotransferase (AAT). The molecular probe was designed on the basis of the L. angustifolius AAT sequence (GenBank: L29258). Primers were anchored in the external th th parts of 10 and 11 exons. The hybridization of probe with the nuclear genome L. angustifolius BAC library tagged 5 clones. These BACs were subjected to restriction fingerprinting and grouped into one contig covering approximately 200 kb. Fluorescent in situ hybridization with the BAC clones as molecular probes (BAC-FISH) on metaphase chromosomes confirmed the integrity of this contig and proved its single locus localization in the narrowleafed lupin genome. The representative BAC clone was selected for sequencing to survey genomic neighborhood surrounding the aspartate aminotransferase gene. The annotation of the BAC sequence demonstrated that AAT is located in gene-rich region of Lupinus angustifolius, almost completely devoid of repetitive DNA. Besides AAT, several genes were -80 identified within given e-value cut-off 1e , e.g.: ein3-binding f-box protein, chromatin remodelling complex, BRI1-KD interacting protein, vacuolar transporter chaperone and aberrant root formation protein. This region was mapped by AAT sequence-specific molecular marker in the linkage group NLL03 of the narrow-leafed lupin genetic map. The comparative genomics alignment approach revealed a high level of synteny of Lupinus AAT gene-rich region to homologous soybean and Medicago truncatula genome segments. VI ICLGG - Posters Theme: Evolution and Diversity P-EAD05 Diversity and linkage disequilibrium features in an IITA soybean panel: consequences for association genetics 1,* 2 3 2 Agrama H , Belzile F , Boahen S , Dion P , Sonah H 2 1 International Institute of Tropical Agriculture (IITA), Lusaka, Zambia Département de phytologie, Université Laval, Québec, Canada 3 International Institute of Tropical Agriculture (IITA), Nampula, Mozambique 2 *E-mail: h.agrama@cgiar.org Abstract Understanding the basis of complex quantitative traits of economic importance is a major tactic behind the whole progress in plant breeding. During the past decade, association mapping, commonly applied in human and animal genetics, has shown greater promise and power of mapping complex quantitative traits in plants as compared to traditional linkage mapping. Germplasm diversity is the mainstay for crop improvement and genetic dissection of complex traits. A collection of 200 IITA soybean lines (Glycine max L) lines were analyzed by genotyping by sequencing to identify SNP markers associated with nodulation traits. Understanding genetic diversity, population structure, and the level and distribution of linkage disequilibrium in target populations is of great importance and a prerequisite for association mapping. In this study, genome-wide association mapping using models controlling both population structure (Q) and relative kinship (K) were performed to identify the marker loci/QTLs underlying the naturally occurring variations of nodulation in an IITA soybean panel. Efforts to identify novel variability in these genes may present opportunities to improve soybean natural nodulation quality. VI ICLGG - Posters SSR markers for determining genetic identities and diversity among promising lines of desi and kabuli chickpea (Cicer arietinum L.) P-EAD06 Shukla N, Babbar A*, Prakash V Jawaharlal Nehru Krishi Vishwa Vidhyalaya, Jabalpur, India *E-mail: anitababbarjnkvv@gmail.com Abstract Use of molecular markers has emerged as a powerful and efficient approach to complement traditional plant breeding for improving crops. Mapping of wilt resistance genes in chickpea is difficult because of minimal polymorphism; however, it has been facilitated to great extent by the development of simple sequence repeats (SSR) markers that have revealed significant interspecific and intraspecific polymorphism. In the present investigation, twenty SSR markers were associated with Fusarium wilt resistant loci were analyzed for the polymorphism and genetic diversity among thirty chickpea promising lines. Out of these 20 markers, 10 belong to the linkage group 4 and 10 from the linkage group 2. In both LG 4 and LG 2 markers, eighteen markers found polymorphic and two markers were found monomorphic. Polymorphism was studied based on polymorphic information content (PIC) and genetic similarity ranged from 0.51 to 0.87 and from 0.69 to 0.97, respectively, among chickpea promising lines. Out of all markers, TA 146 showed highest polymorphism (PIC value 0.891) as well as genetic diversity (0.898). To assess the genetic diversity among the thirty chickpea genotypes for twelve morphological traits were grouped into two major clusters in cluster analysis. JG 19 showed highest similarities (96.77%) with JG 21 and similarly MP-JG 99-115 showed with both JG 17 and JG14-11 (96.77%). Whereas, ICCV 07110 and ICC 16216 showed lowest similarity (69.35%) with MP-JG 03-115 and JG 1-14, respectively. This study may serve as a basis for marker assisted selection and other agronomically important genes in future. VI ICLGG - Posters P-EAD07 Harnessing genetic diversity for black gram (Vigna mungo L.) improvement based on morphological and molecular approach 1,* 1,2 1 1 3 Shridevi BM , Salimath PM , Ontagodi TP , Shridevi O , Biradar S , 4 5 3 Vijaykumar AG , Basavanagoud K , Naik VR 1 Department of Genetics and Plant Breeding, University of Agricultural Sciences (UAS), Dharwad, India 2 UAS, Shimoga, India 3 AICRP on wheat, UAS, Dharwad, India 4 AICRP on MULLaRP, UAS, Dharwad, India 5 Department of Agril. Entomology, UAS, Dharwad, India *E-mail: bmshree.4795@gmail.com Abstract Blackgram (Vigna mungo (L.)) is an important short-duration pulse crop cultivated in tropical to sub-tropical countries where it is mainly grown in summer season. Objective of this study was to characterize ninety five diverse blackgram genotypes including three standard checks viz; TAU-1, BDU-4 and Manikya. Based on the diversity analysis using ten productivity triats, 95 genotypes were grouped into 14 clusters. Maximum number of genotypes (39) were found in cluster-II. Characters like number of pods per plant and plant height contributed greatly towards divergence. Intra cluster distance was 2 highest in cluster XI (D =15.9) and inter cluster distance ranged from 7.71 to 40.43 which indicated wide genetic variability. Considering genetic distance of individual genotype from others, KU-5-573 and KU-98-40-2; KU-5-573 and G-1 were found to be highly divergent and also complementing each other for different traits. Accordingly, the crosses KU-5-573 X KU-98-40-2 and KU-5-573 X G-1 have been suggested to combine different productivity traits. The 95 genotypes were also subjected to molecular diversity using RAPD markers. The level of polymorphism generated among the genotypes was around 40.42 percent. On an average 7.05 bands per primer were produced. The dendrogram constructed from the pooled data revealed three distinct clusters. The primer OPA-11 gave the highest (57.14) polymorphism whereas, lowest (12.5 %) polymorphism was given by the primer OPE-07. The diversity coefficient ranged from 87 to 99 indicating a close genetic similarity in all the 2 genotypes analyzed. Here the grouping of genotypes based on D analysis and DNA fingerprinting was not concurrent. These differences may be due to the limited number of primers used for the analyses, besides inherent low genetic diversity reported for legumes in general and blackgram in particular. VI ICLGG - Posters Current scenario of diversity of races of Fusarium oxysporum f.sp. ciceri and their sources of resistance in India P-EAD08 Chaudhary RG*, Dhar V, Gurha SN, Trivedi S, Singh A Indian Institute of Pulses Research (IIPR), Kanpur, India *E-mail: rgc_1234@yahoo.com Abstract Wilt caused by Fusarium oxysporum f.sp. ciceri (Foc) is one of the serious diseases in India inflicting over 10% yield loss annually. Disease is prevalent in all the chickpea growing areas of India. Development of stable wilt resistant varieties is possible only when the current knowledge about the races existing in chickpea growing regions and their resistant donors are known. Studies in this direction have been made at the Indian Institute of Pulses Research, Kanpur for past one decade. A total of 326 isolates of Foc collected from 12 major chickpea growing states were categorized into 27cultural and morphological groups. Differential reaction of 59 representative isolates from these 27 groups revealed existence of seven Foc races in the country besides 18 ungrouped isolates. Data revealed presence of all the 5 races each in Uttar Pradesh, 7 in Rajasthan and Madhya Pradesh, 3 in Haryana and Gujarat, 2 each in Andhra Pradesh, Maharashtra and Punjab and one each in Delhi, Chhattisgarh and Karnataka. Massive screening of chickpea genotypes in sick-plot at IIPR, Kanpur could identify more than 100 chickpea genotypes resistant to wilt. In recent years, screening against individual races under controlled conditions showed 25 chickpea genotypes as resistant to race 1, 29 against race 2,19 against race 3, 24 against race 4, 30 against race 5 and 26 against race 6. Genotypes IPC nos.2004-3,2004-8,2004-34,2004-52,200515,2005-19,2005-24 and KGD 1255 showed resistance to all six races. Based on the information generated so far, a race distribution map has been attempted. VI ICLGG - Posters P-EAD09 Assessment of genetic diversity among garden pea cultivars grown in India using molecular marker (RAPD) Arul S, Dhar S*, Choudhary H Indian Agricultural Research Institute, New Delhi, India *E-mail: shridhar@iari.res.in Abstract Garden pea (Pisum sativum L. subsp. hortense (Neilr.) Asch. & Graebn.) is an important leguminous vegetable cultivated for green pods throughout the world. Crop improvement mainly depends on the extent of heritable diversity existing in the available germplasm. Genetic diversity was assessed among 28 garden pea genotypes with sixteen RAPD primers which generated 79 (73.15%) polymorphic fragments. The number of alleles ranged from 2 to 12 with an average of 6.75 per primer. Maximum similarity was observed between AP 3 vs. Arkel and Arkel vs. Glossy (89%) and the lowest between VP 266 vs. VP-233 and GP 6 vs. GP 207(43%). Arkel and AP-3 are of early maturity and Glossy is a mutant of Arkel. GP-6 is an afila genotype and resistant to powdery mildew, while GP207 having normal leaf and susceptible. All the accessions were grouped into 6 clusters which size varied from 16 (Cluster I) to 1 (Cluster III, VI). The cluster I included mostly early genotypes (PSM 3, AP 3, PSM 4, Arkel, VL 7, Glossy, E 6 , GP 17, GP 18, GP 19 and GP 901). However, some medium maturing genotypes (Bonneville, GP 473, GP 471, VL 3 and VP 233 ) were also grouped in cluster I. Hara Bona (tall and pulse type) was grouped separately in cluster III. The polymorphism information content (PIC) was highest for the RAPD primers OPN14 (0.74) and HU11 (0.64) which reflects that OPN 14 was found to be highly informative for diversity analysis and it may be used for fingerprinting of genotypes also. . VI ICLGG - Posters Preliminary results of E2 gene distribution among soybean maturity groups P-EAD10 Djordjevic V*, Tomicic M, Mikic A Institute of Field and Vegetable Crops, Novi Sad, Serbia *E-mail: vuk.djordjevic@ifvcns.ns.ac.rs Abstract The genotypes of soybean, a short-day plant, are divided in several maturity groups according to photoperiod and temperature requirements. Several maturity loci, designated as E loci, have been characterized by classical and molecular methods. E2 gene, ortholog of GIGANTEA, is responsible for time to flowering and time to maturity. Dominant allele delays soybean flowering and its effect, under different environments, is stable. It is a well characterized gene, but its distribution among germplasm and different maturity groups is not known. A preliminary set of 78 genotypes, originating from North America and Europe, was tested on the presence of dominant (E2) or recessive allele (e2). Over all, from 000 to III maturity group a frequency of dominant allele is low (p=0.22). In 000 and 00 maturity groups, all tested genotypes (n=15) have recessive allele. Low frequencies of dominant allele were observed in 0 (0.13) and I (0.12) maturity group. In later maturity groups, (II and III) there were observed significantly higher frequencies of dominant allele (0.36 and 0.46). A clinal variation of gene frequencies among maturity groups was observed. Significant differentiation between maturity groups in allele frequencies was found, based on gST values (0.439). Also, a fixation index (F ST=0.76) indicates strong differentiation among soybean maturity groups, based only on E2 gene. VI ICLGG - Posters P-EAD11 Chloroplast DNA phylogeny of the Asian Vigna species (subgenus Ceratotropis, Fabaceae) 1* 2 Javadi F , Ye TT , Yamaguchi H 3 1 Kyushu University, Fukuoka, Japan Ministry of Agriculture and Irrigation, Yezin, Myanmar 3 Tokyo University of Agriculture, Kanagwa, Japan 2 * E-mail: fjavascb@kyushu-u.org Abstract The Subgenus Ceratotropis is originally circumscribed as a group of the Asian species of the genus Vigna. It consists of 21 species, and eight of these are used for human and animal food. The 21 known wild species distributed widely in South Asia, the Himalayan highlands, Southeast Asia, and East Asia. East Asian and Southeast Asian Vigna species occur naturally in temperate and subtropical regions, while Indian subcontinental species are mainly confined to tropical regions. However, the interspecific and geographical relationships among them are poorly understood and mainly focus on the limited geographical regions. Based on the improved species sampling in comparison to that of previous studies, the present work aims to clarify those relationships. Four chloroplast intergenic spacers regions (psbD-trnT, trnT-trnL, trnT-trnE, and petA-psbJ) have been sequenced for 42 accessions representing the 18 species as well as four taxa used as outgroups. Both parsimony and Bayesian analyses have been performed on the combined data set. Phylogenetic analysis of four DNA regions confirmed that Asian Vigna is composed of three clades that correspond to species distributions. The temperate clade consists of V. minima, V. nepalensis, and V. angularis. The subtropical clade comprises the V. nakashimae-V. riukiuensis-V. minima subclade and the V. hirtella- V. exilis- V. umbellata subclade. The tropical clade contains two subclades: the V. trinervia-V. reflexo-pilosa-V. trilobata subclade and the V. mungo-V. grandiflora subclade. The evidence presented here clearly demonstrates the geography and ecological habitat as important factors in shaping phylogenetic relationships of the Asian Vigna. VI ICLGG - Posters Small scale duplication events in crop genome 1 1 1 Kang YJ , Kim MY , Van K , Lee S-H P-EAD12 1,2 1 Department of Plant Science and Research Institute for Agriculture and Life Sciences,Seoul National University, Seoul, Korea 2 Plant Genomics and Breeding institute, Seoul National University, Seoul, Korea *E-mail: sukhalee@snu.ac.kr Abstract Several studies regarding duplication events on plant genome have reported that they took major parts in evolutions including divergence, speciation, and also domestication. Previous researches surveyed the gene contents on the duplicated regions and identified that highly connected genes were wellconserved after large-scale duplication events and those genes are easily decayed after small scale duplication events; however, the genes involved in secondary metabolism and response to biotic stimulus were not decayed after large and small scale duplication. From those observations, we assumed that the small scale duplication is on-going event, which might contribute real-time adaptation process against various environmental stresses and affect to agricultural traits including abiotic and biotic stresses. In the present study, we gathered tandemly duplicated genes from whole genome sequenced crops including soybean, rice, maize, common bean, and Medicago and model plant Arabidopsis. Gene ontology analysis was conducted to understand the functions of tandemly duplicated genes. The majority of those genes belong to the GO categories of cellular process, metabolic process and response to stimulus. Interestingly, approximately 50% tandemly duplicated gene of legume genome including soybean, common bean, and Medicago showed the involvement of responses to stimulus. Ks values among the tandemly duplicated gene clusters were also analyzed to figure out the activity of the tandem duplication process. It was observed that the legume genome have more recent activity of tandem duplication than Arabidopsis genome. VI ICLGG - Posters P-EAD13 Assessment of genetic diversity in kabuli chickpea genotypes in relation to seed size using SSR markers 1 1,* 1 2 1 1 Choudhary S , Kaur J , Chhuneja P , Sandhu JS , Singh I , Singh S , Sirari 1 A 1 Punjab Agricultural University, Ludhiana, India Indian Council of Agricultural Research, New Delhi, India 2 *E-mail: jagskaur@gmail.com Abstract Chickpea (Cicer arietinum L.) is an important pulse crop which ranks third in world legume production. Seed size in kabuli chickpea is an important trait for trade, productivity and adaptation. Hence, breeding for seed size for kabuli types is of major importance. The assessment of genetic diversity has significance not only for crop improvement but also for efficient management and conservation of germplasm resources. Chickpea has comparatively smaller genome size (750 Mbp) and shows only low levels of genetic polymorphism. A large number of polymorphic markers are required for studying diversity or linkage analysis in this crop. Amongst the various markers, SSR markers are highly polymorphic, more reproducible and distributed throughout the genome and can provide practical information for selection of parental material to formulate breeding strategies. In the present study, out of 42 SSR markers used, 30 revealed 71 alleles ranged from 2 to 4 with average of 2.37 alleles per locus across the 15 chickpea genotypes varied in seed size. The Polymorphic Information Content (PIC) values ranged from 0.125 (CaSTMS5) to 0.721 (TA146) with an average of 0.415. The genetic dissimilarity index between genotypes ranged from 0.091 to 0.514. Cluster analysis using Unweighed Pair-group Method of Arithmetic average method classified genotypes into two major groups comprised seven genotypes each while one large-seeded genotype GLK26171 showed divergence from clustered genotypes. As expected the minimum dissimilarity (the highest similarity) was observed between GLK24096 and GLK24092 (both smallseeded) having a common female parent in their parentage. The maximum dissimilarity was observed between GLK24092 (small-seeded) and GLK26173 (medium-seeded). VI ICLGG - Posters Tracing back the history of soybean by estimating the divergence time between Glycine max and G. soja based on whole-genome low coverage sequencing data 1 1 1 1 1 Kim SK , Kang YJ , Yoon MY , Kim MY , Van K , Lee S-H P-EAD14 1,2* 1 Department of Plant Science and Research Institute for Agriculture and Life Sciences,Seoul National University, Seoul, Korea 2 Plant Genomics and Breeding Institute, Seoul National University, Seoul, Korea *E-mail: sukhalee@snu.ac.kr Abstract Because of domestication process of soybean, there have been major phenotypic chances in seed size, pod length, and plant architecture. As soybean (Glycine max) is known for its high nutritional value of oil and protein, people have been domesticating and cultivating by selecting one specific character trait based on the needs of people. Importantly, tracing back in time where G. max and G. soja have diverged plays an important role in studying of genetic diversity and in investigating the common ancestor of soybean. We sequenced 6 G. max and 6 G. soja using Illumina’s Hiseq 2000 with a low coverage sequencing technology to estimate the divergence times among genotypes and populations. A total of the 12 genotypes were sequenced at the average depth of 6.5 and resulted 892.5 Mb and 903.3 MB consensus sequences with the coverage of 91.54% and 92.65% for G. max and G. soja, respectively. Whole-genome SNP analysis showed that G. max had lower frequency of polymorphism (~0.1%) than G. soja (~0.25%). A high number of SNPs in introns were identified among 6 G. soja genotypes (133,329 SNPs) twice more than among 6 G. max (53,134 SNPs). Almost an equal number of SNPs between G. max and G. soja were discovered in 5’UTR and exon regions, however, different SNP numbers in coding regions and 3′UTR. The nonsynonymous rate of G. max showed lower Ks value as 0.0015 compared to G. soja (0.0018), suggesting nonsynonymous mutations may have caused by selection and changed phenotypic characteristics in wild soybean. VI ICLGG - Posters P-EAD15 Prediction of wild soybean (Glycine soja Sieb. and Zucc.)-specific genes by homology using deep sequenced soybean accessions 1 1 1 1,2 Lee T , Bae A , Rastogi K , Lee S-H * 1 Department of Plant Science and Research Institute for Agriculture and Life Sciences,Seoul National University, Seoul, Korea 2 Plant Genomics and Breeding Institute, Seoul National University, Seoul, Korea *E-mail: sukhalee@snu.ac.kr Abstract Wild soybean (Glycine soja Sieb. and Zucc.) is considered to be the closest plant of soybean (G. max). But, they have very different morphology in seed size, shattering and so on. These phenotypic differences might be regarded as domestication syndrome occurred by G. soja-specific genes. These genes might be useful in soybean breeding and can distribute domestication related research. So, we predicted G. soja-specific genes using four G. soja deepsequenced genome (KS-04, KS-05, CS-12, CS-14). Because whole-genome sequences of G. soja were obtained by re-sequencing method, it is difficult to predict G. soja-specific genes. For resolving this matter, we retrieved both unmapped reads from the mapped files, which contained aligned G. soja reads to G. max reference genome sequence using Samtools (http://samtools.sourceforge.net/). And, both unmapped reads were assembled into contigs using ABySS genome assembler ver.1.3.3 (http://www.bcgsc.ca/platform/bioinfo/software/abyss/). We analyzed these contigs using homology based method using BLAST tools (http://blast.ncbi.nlm.nih.gov/). Firstly, we compared between contigs and nonredundant protein database using BLASTX, and then we chose the contigs matched with domestication related and agronomically useful proteins such as disease resistance proteins and stress tolerance proteins. Finally, we selected contigs, which have not homologous with G. max using BLASTN as G. sojaspecific gene candidates. As a result, we were able to obtain eight G. sojaspecific gene candidates and further study is in progress for amplifying putative G. soja-specific gene using designed primers to confirm prediction. VI ICLGG - Posters Genome-wide characterization of non-reference transposons reveals evolutionary propensities of transposons during soybean diversification and domestication 1,2 1 1 1 3 4 4 5 P-EAD16 5 Tian Z , Mao S , Zhao M , Du J , Cannon SB , Liu X , Xu X , Qi X , Li M-W , 5 1, Lam H-M , Ma J * 1 Department of Agronomy, Purdue University, West Lafayette, USA State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Beijing, China 3 United States Department of Agriculture-Agricultural Research Service, Corn Insects and Crop Genetics Research Unit, Ames, USA 4 BGI-Shenzhen and the Key Laboratory of Genomics of the Minister of Agriculture, Shenzhen, China 5 State Key Laboratory of Agro biotechnology and School of Life Sciences, The Chinese University of Hong Kong, China 2 *E-mail: maj@purdue.edu Abstract Preferential accumulation of transposable elements (TEs), particularly LTRretrotransposons (LTR-RTs), mostly in nested clusters in recombinationsuppressed pericentromeric regions, seems to be a general pattern of TE distribution in many flowering plants. However, whether such a pattern was formed primarily by preferential insertions into pericentromeric regions or by selection against insertions of TEs into euchromatin remains obscure. We have recently investigated TE insertions in 31 resequenced wild and cultivated soybean genomes and detected 34,152 unique TE insertions that are not present in the soybean reference genome. The majority of these non-reference TE insertions were found in single accessions, suggesting their recent occurrence. Profiling of these insertions against the soybean reference genome revealed a consistent chromosomal distribution pattern of the nonreference LTR-RT insertions and those present in the reference genome, whereas the distribution patterns of the non-reference DNA TE insertions and the accumulated DNA TEs were significantly different. The densities of the non-reference LTR-RT insertions were found to negatively correlate with the rates of local genetic recombination, but no significant correlation between the densities of the non-reference DNA TE insertions and the rates of local genetic recombination was detected. Different from the accumulated LTR-RTs, >60% of which were found within TE sequences, the non-reference LTR-RTs show non-nested patterns of integration. Together, these observations suggest that, although purifying selection plays an important role in purging TE DNA from the host genome, distinct insertional preferences are primary factors that lay foundations for different distribution patterns of LTR-RTs and DNA TEs in the soybean genomes. VI ICLGG - Posters P-EAD17 Assessment of genetic variability for economic traits among Velvet bean (Mucuna pruriens L.) accessions Misra HO*, Lal RK, Dhawan SS, Mishra LN, Kumar B, Dhawan OP Central Institute of Medicinal & Aromatic Plants, Lucknow, India *E-mail: ho.misra@cimap.res.in Abstract Kewanch or Velvet bean (Mucuna pruriens L.) is an annual twining tropical legume of family Fabaceae (2n=22), whose seeds contain the catecholic amino acid L-Dopa (L-3, 4-dihydroxyphenylalanine), which is a neurotransmitter precursor, used for treatment of Parkinson’s disease, mental disorders; also as aphrodisiac. The efficiency of a plant breeding programme is determined by the amount of genetic variability for specific traits in the breeding population and the efficiency of selection. There was apparent lack of improved varieties of Kewanch. To enhance genetically seed yield with better medicinal value, an effort was made to clear understand the nature and amount of genetic variability present in accessions and the extent to which desirable morphometrical traits are heritable. The experimental material comprised of more than twenty accessions collected from different places of India and induced variability, was raised in replicated field trial at CIMAP, Lucknow in different years. Morphometric observations i.e. pods/pl., pod length, seeds /pod, seed yield /pl., L-Dopa content etc. were recorded. The PCR based DNA finger printing method (RAPD), using deca-nucleotide primers was also applied. There was a good extent of variation among accessions for seed colour, pod size, pods/pl., seed yield/pl. Seed colours recorded were black, dull black, dull black seeds with brown flacks or stripes, bright black, ash and white, slaty colour. The outer surface of pods was green, yellow and black tinged green; itching and non itching trichomes. Recorded data was statistically analysed to estimate allied genetic parameters. Concepts about non itching trichome type improved variety developed and genetic aspects will be discussed. . VI ICLGG - Posters Induced mutagenesis generated repeat expansions/contractions and transposition of MITE in the genome of groundnut. P-EAD18 Mondal S*, Badigannavar AM, D’Souza SF Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre, Mumbai, India *E-mail: suvenduhere@yahoo.co.in Abstract Groundnut (Arachis hypogaea L.) is one of the major oilseed crop grown in India. To generate more genetic variability in this crop a rust resistant breeding line (TG 66) was treated with gamma rays and sodium azide in single or in combination. A total of 74 true breeding mutants were isolated for different qualitative and quantitative traits from 20,619 M2 plants. It was found that combination doses of gamma rays and sodium azide generated more mutations than the single dose of each mutagen. Of the 74, 35 induced mutants were analyzed for genetic diversity and to reveal nature of mutations. Screening of 412 simple sequence repeat primer pairs and 100 AhMITE specific primer pairs revealed that a very low level of polymorphism (0.24 to 5.34%) between a mutant and the parent. Most of the SSR mutations were either repeat contraction or repeat expansion. The induced mutagenesis also revealed null mutations and point mutations in some of the polymorphic SSR loci. Of the 100 AhMITE primer pairs, 12 detected transposition event. Extent of expansion and/or contraction of repeat length suggested that the induced mutagenesis followed both the stepwise mutation model and infinite allele model of mutations. Further induced mutations in SSR loci did not show any linear relationship between repeat length and mutation frequency. The results showed the occurrence of more mutations at SSR loci in some of the rust susceptible mutants which grouped them distantly from the rest of the mutants and parent in a neighbor joining tree. VI ICLGG - Posters P-EAD19 Characterization of a new begomovirus and associated betasatellite causing leaf curl disease in French bean * Naimuddin , Akram Md, Pratap A, Yadav P Indian Institute of Pulses Research (IIPR), Kanpur, India * E-mail: naimk@rediffmail.com Abstract A leaf curl disease with symptoms typical of infection by begomoviruses was observed in French bean at Main Research Farm of the Indian Institute of Pulses Research, Kanpur, India during 2011 and 2012. The disease causes severe distortion of leaves and makes the plants unproductive. The disease was transmitted from infected to healthy plants of French bean through whitefly (Bemisia tabaci). The total DNA extracted from the leaves of five symptomatic plants (Fb1-Fb5) was subjected to rolling circle amplification (RCA) using Phi 29 DNA polymerase. The RCA products restrict digested with EcoRI yielded DNA fragments of ~2.7kb. One of the five isolates (Fb1) was cloned and sequenced. The complete sequence comprised 2741 nucleotides with genome organization typical of begomoviruses having two ORFs in virion-sense and five ORFs in complementary-sense separated by an intergenic region that had a conserved nonanucleotide sequence, TAATATTAC. Complete DNA-A sequence homology revealed that the Fb1 was most closely related to CLCuBV-[IN:Ban:05] with 81% nucleotide sequence identity. Based on the species demarcation criteria, Fb1is considered as a distinct begomovirus, has been named French bean leaf curl virus and designated as FbLCV[IN:KNP:12] (NCBI accession number JQ866297). Presence of the associated satellite DNA-β molecule was confirmed using primers Beta01/Beta02. The complete sequence of associated satellite DNA-β comprised 1379 nucleotides with single ORF (C1) and has 78% sequence identity with Papaya leaf curl betasatellite (HM143906). This appears to be a new begomovirus associated satellite DNA-β molecule and is named as FbLCB-[IN:KNP:12] (NCBI accession number JQ866298). VI ICLGG - Posters Phylogenetic analysis using Panzee retrotransposon based SSAP markers in pigeonpea P-EAD20 Patil P*, Datta S, Singh IP, Das A, Soren KR, Kaashyap M, Choudhary AK, Chaturvedi SK, Nadarajan N Indian Institute of Pulses Research (IIPR), Kanpur, India *E-mail: patilbt@gmail.com Abstract In the present study, specific primer designed to 5'LTR region of the pigeonpea retrotransposon viz. Panzee was successfully developed into SSAP markers and used for phylogenetic analysis. Total four primer combinations were screened on 21 pigeonpea genotypes (representing three gene pools), which produced 183 bands of which 166 (90.71%) bands were polymorphic with an average of 41.5 polymorphic bands per primer combination. The marker features like polymorphism information content (PIC), Marker index (MI) and resolving power (Rp) values were calculated for four primer combinations. PIC values ranged from 0.15 [Panzee-LTR/ EcoRI (+AAC)] to 0.26 [Panzee-LTR/ EcoRI (+AAG)], MI values ranged from 5.25 [Panzee-LTR/ EcoRI (+AAC)] to 14.25 [Panzee-LTR/ MseI (+CAC)] and RP values ranged from 10.14 [Panzee-LTR/ EcoRI (+AAC)] to 24.92 [Panzee-LTR/ MseI (+CAC)]. The average RP and MI values for four primer combinations 2 exhibiting strong positive correlation (r =0.99 p<0.05). A primer combination Panzee-LTR/ MseI (+CAC) was found most informative for phylogenetic analysis. NJ tree based on SSAP markers exhibited concordant relationships with high bootstrap values. One accession of Cajanus cajanifolius (ICP 15632) was placed closer to primary gene pool, confirming closest progenitor of cultivated pigeonpea. SSAP markers produced high rate of polymorphism in pigeonpea and considered to be best marker of choice for phylogenetic studies. VI ICLGG - Posters P-EAD21 Genetic divergence in germplasm lines of pigeonpea (Cajanus cajan L.) Dharmaraj PS*, Muniswamy S, Yamanura, Girish G, Sheela D Agricultural Research Station, Gulbarga, India *E-mail: psdharmaraj@yahoo.com Abstract Pigeonpea (Cajanus cajan L. Mill (sp)) is an important legume crop of rainfed agriculture including Indian subcontinent. In India, it is one of the very important grain legumes and occupies second position in area and production next to chickpea. It is mainly grown in the states of Maharashtra, Karnataka, Uttar pradesh, Madhya pradesh and Gujarat. Eighty Pigeonpea genotypes received from NBPGR are studied for genetic divergence using Mahalanobis 2 D statistic indicated wider genetic diversity. Genetic diversity was independent of their pedigree. Considerable variability was observed for days to 50% flowering, days to maturity, plant height, number of pods per plant, Pod bearing length, 100 seed weight and seed yield. The variability was low for number of primary branches per plant and test weight. Among the 7 characters studied, seed yield contributed the most (44.30%) towards the divergence of genotypes followed by number of pods per plant (22.37%), Pod bearing length and plant height. The genotypes were grouped into 7 clusters, maximum inter cluster distance being observed between the clusters III and VII (30.29). Hence, crossing of parents from these clusters would be desirable for combining earliness, short plant height, and more primary branches per plant with more pod number coupled with high seed yield. Sufficient amount of variability was observed in these genotypes, which indicated scope for selecting more diverse parents to produce high heterotic effects and desirable segregants. VI ICLGG - Posters Cross species amplification of Adzuki bean derived microsatellite markers in Asian Vigna species P-EAD22 Vinoth R, Srimathy M, Sathya M, Sowmini K, Muthiah AR, Jayamani P* Centre for Plant Breeding and Genetics, Tamil Nadu Agricultural University, Coimbatore, India *E-mail: jayamani1108@gmail.com Abstract Vigna is one of the important genus of grain legumes which forms the source of dietary protein. In recent years, molecular marker technology has greatly accelerated the breeding programs for the improvement of various crops. Among the different DNA markers, microsatellite or simple sequence repeats (SSRs) are the markers of choice for various genetic studies due to their codominant nature, loci specificity and high reproducibility. To date, only few reports are available on isolation and development of microsatellite markers in some of the Vigna species. Hence, the available SSR markers from other Vigna species should be validated for their transferability and utility in those species in which they are unavailable. In the present study, a set of 35 microsatellite primer pairs derived from adzukibean (Vigna angularis) were used to assess the transferability and tested for their ability to amplify microsatellite loci in 68 genotypes consisted of eight Asian Vigna species. The percentage of amplification varied for each genotype/ species. In urdbean and its progenitor species 100 per cent amplification was observed whereas, in mungbean and its related wild species, 94.3 per cent was recorded. The percentage of polymorphic markers obtained was more than 80 per cent. Total number of alleles observed was 87 and 98, respectively in mungbean and urdbean. The allele size varied from 90-350 bp. Further, a set of 169 SSRs (http://www.gene.affrc.go.jp/databases-marker_information_en.php) were used to amplify microsatellite loci in four different parents of two mapping populations of urdbean. The percentage of markers amplified was ranged from 87.6 to 88.2. Among the amplified markers, the polymorphic markers obtained were low and it ranged from 25.7 to 31.5 per cent. Total number of alleles observed was varied between two mapping populations and it was found to be 185 and 171, respectively in population I and II. Variation in allele size was also noticed between two populations and it ranged from 100-290 bp. These findings suggest that microsatellite markers derived from adzukibean could be used in molecular genetic diversity and genomic studies in mungbean, urdbean and also in other Vigna species and thus aid in their improvement. VI ICLGG - Posters P-EAD23 SNP genotyping using Illumina BeadXpress for germplasm diversity studies in chickpea and pigeonpea 1 1 1 1 1 Roorkiwal M , Sawargaonkar SL , Thudi M , Saxena RK , Upadhyaya HD , 1,2* Varshney RK 1 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 2 CGIAR Generation Challenge Programme (GCP), CIMMYT, Mexico DF, Mexico *E-mail: r.k.varshney@cgiar.org Abstract Single nucleotide polymorphisms (SNPs) have become an ideal marker system due to their greater abundance in the plant genomes. In cases like marker-assisted selection (MAS) where only few markers are required for genotyping a set of potential lines, a cost-effective SNP genotyping platform is required. In this scenario GoldenGate assays based on VeraCode technology using Illumina BeadXpress platform seems to be a cost-effective genotyping method. In the case of chickpea and pigeonpea, >10,000 SNPs have been identified using next-generation sequencing (NGS) technologies in earlier studies. Based on high polymorphism information content and assay designing tool (ADT) scores, two sets of SNPs one each for chickpea (96 SNPs) and pigeonpea (48 SNPs) were designed for custom Oligo Pool Assays (OPAs) for Illumina BeadXpress genotyping platform. These OPAs were used to genotype reference sets (ca. 300 genotypes each) of both the legumes. All SNPs were successfully scored in the reference set, with less than 1% missing data in case of chickpea and around 1-2% missing data in pigeonpea. The mean polymorphism information content (PIC) of SNP markers in case of chickpea was 0.31 and 0.32 in pigeonpea. No unique pattern of grouping in chickpea reference set was observed based on neighbour joining tree constructed. However, in the case of pigeonpea two major groups are observed with SNPs data. Our results indicate that Illumina BeadXpress based system provides a cost effective SNP genotyping method for selected set of potential SNP markers and can easily be used for diversity studies. VI ICLGG - Posters Characterizing rhizobial diversity in legumes from pine rockland soils in south Florida 1,2* Sanchez V 1,2 1 , Scharnalg K , Jayachandran K , von Wettberg E P-EAD24 1,2 1 Florida International University, Miami, USA Fairchild Tropical Botanic Garden, Coral Gables, USA 2 *E-mail: vsanc004@fiu.edu Abstract Pine rocklands are globally endangered ecosystems unique to South Florida, the Bahamas and Cuba. Due to karstic calcium carbonate-rich soil, these systems are limited in phosphorus and nitrogen, making symbiotic soil mutualisms critical to plant success. Pine rocklands contain the highest plant diversity of any other habitat in Florida, with over 400 native plants, a large number of which are endangered. These habitats are highly threatened by development and sea level rise. Legume-rhizobium interactions studies have not been previously conducted in pine rocklands, and the diversity of rhizobia across the remaining forest patches is unknown. Four leguminous species representing a range of crops, native species, and exotic species (Cajanus cajan-crop, Tephrosia purpurea- endangered, Chamaecrista fasciculata-native, and Abrus precatorius-invasive) will be use to trap rhizobia from four different pine rockland soils that will be either treated with saline water simulating sea level rise or distilled water. Planting initiated in late May, and harvesting of roots for nodules began in early July. DNA isolation and sequencing will follow immediately in order to identify rhizobial strains. It is expected to find differences in rhizobial communities based on their host plant, with crops and exotics utilizing a greater range of rhizobial strains, as well as an increase in the variation of rhizobia within each remaining forested area. Finally, it is expected that the non-saline treatment in one of the pine rocklands that was badly inundated by sea-water during Hurricane Wilma in 2005 to approximate saline treatments in the other pine rocklands. VI ICLGG - Posters P-EAD25 Single-nucleotide polymorphisms (SNPs) for diversity analysis in Cajanus germplasm 1 2 1 1,3 2 Saxena RK , von Wettberg E , Upadhyaya HD , Songok S , Sanchez V , 3 4 1,* Paul K , Cook DR , Varshney RK 1 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 2 Florida International University, Florida, USA 3 Egerton University, Egerton, Kenya 4 University of California, Davis, USA * *E-mail: r.k.varshney@cgiar.org Abstract This study was undertaken to estimate the levels of genetic variability both within and between diverse set of Cajanus lines including 56 breeding lines, 21 land races and 107 accessions from 18 wild species by using 1,616 single nucleotide polymorphism (SNPs) on KASPar assays. The results revealed 75.8% SNPs polymorphic, and more than 95% of these assays could be transferred to related Cajanus wild species. Genetic polymorphism was highest in the wild relatives (73.76%) followed by landrace (13%) and breeding lines (8.29%). STRUCTURE and Analysis of Molecular Variance (AMOVA) were used to partition variance among hierarchical sets of landraces and wild species at either continental scale or within India. STRUCTURE separated most of the domesticated germplasm from wilds, and separates Australian and Asian wild species. At the continental scale, 69% of the variation was found between landraces and wild species, and 31% within continents, with no variation among continents. Among Indian regions and states within regions, 36% of the variation was found between regions, and 64% within landraces or wilds within states and highest diversity was found in Andhra Pradesh (AP). These results indicate AP being the center of domestication, and potentially a center of diversification of pigeonpea, though sample sizes analyzed in this study were insufficient to have complete confidence in this conclusion. In addition to enhancing understanding about domestication and diversification of pigeonpea, this study also provides diverse lines and highly informative markers for their use in breeding programmes. VI ICLGG - Posters Pathogenic variability in Fusarium oxysporum f.sp. ciceri in India P-EAD26 Chaudhary RG, Dhar V, Singh A*, Trivedi S Indian Institute of Pulses Research (IIPR), Kanpur, India *E-mail: ainmishasingh1@gmail.com Abstract Fusarium oxysporum f.sp. ciceri, (Foc) is an important pathogen reducing chickpea productivity in many countries. Resistant varieties are the best option for the management of chickpea wilt. However, many-a-times, the pathogenic variability occurring in this pathogen results in the failure of resistant varieties. Seven races of Foc have so far been reported in the world, out of which race 1, 2, 3 and 4 are reported from India. The objective of the present study was to work out the pathogenic variability of 58 isolates of Foc representing 12 states of India. The study was conducted on 10 wilt differential chickpea genotypes earlier used by Haware and Nene at ICRISAT. Out of 58 isolates, 41 could be grouped into 7 races based on disease reaction on differential genotypes. The reaction of 7 isolates each matched with race 0 and 1, whereas 4 isolates each matched with race 2 and 3. The reaction of 6 isolates matched with race 4, of 9 isolates with race 5 while 4 isolates resembled with race 6. Thus, this study revealed that race 5 is present in 9 states followed by race 0 and 4 in 5 states each. Race 1, 2 and 3 was prevalent in 4 whereas race 6 in 3 states. These findings indicated the presence of 7 races of Fusarium oxysporum f.sp. ciceri in India. The reaction of remaining 17 isolates did not match with either of the reported races indicating the presence of some more variability in this country. VI ICLGG - Posters P-EAD27 Genetic diversity analysis in Cajanus species using SSR markers 1 1 1,* 2 1 Singh AK , Rai VP , Singh MN , Chand R , Ramdhari , Singh RM 1 1 Department of Genetics and Plant Breeding, Banaras Hindu University, Varanasi, India 2 Department of Mycology and Plant Pathology, Banaras Hindu University, Varanasi, India * E-mail: mnsbhu@yahoo.co.in Abstract Pigeonpea (Cajanus cajan L. Millspaugh) is one of the major grain legume crops of the tropical and subtropical regions. The genetic diversity among forty pigeonpea genotypes including four wild species viz., C. scarabaeoides, C. cajanifolius, C. volubilis and Rhynchosia rothii was analyzed using 12 polymorphic SSR markers. A total of 24 polymorphic bands with an average of two bands per primer were obtained. The polymorphic information content (PIC) ranged from 0.434 - 0.594 with an average of 0.523. The variation in genetic diversity among these cultivars ranged from 0.0 to 0.85. Cluster analysis based on Jaccard’s similarity coefficient using unweighted pair group method with arithmetic mean (UPGMA) revealed three distinct clusters comprising of 17, 20 and 03 genotypes, respectively. Cluster III consisted of all the four but C. scarabaeoides wild pigeonpea species. Principal components analysis (PCA) also revealed the similar results with that of UPGMA. The first, second and third PCs contributed 22.57, 20.47 and 15.43% of the variation, respectively; with cumulative variation of the first three PCs was 58.47%. It is interesting to note that most of the Fusarium wilt resistant genotypes were grouped together and six (MA-3, MA-6, MAL-23, MAL-13, MAL-31 and MAL34) out of eight Malaviya series pigeonpea genotypes were placed in cluster I, except for MAL-18 and MAL-23. VI ICLGG - Posters In-silico identification, sequential and structural classification of TIRNBS-LRR resistance gene family from Pigeonpea (Cajanus cajan) genome 1, 2 3 4 P-EAD28 3, Singh VK *, Singh S , Kayastha AM , Singh NK , Singh BD * 1 Centre for Bioinformatics, Banaras Hindu University, Varanasi, India Center of Bioinformatics, University of Allahabad, Allahabad, India 3 School of Biotechnology, Banaras Hindu University, Varanasi, India 4 National Research Centre on Plant Biotechnology, New Delhi, India 2 *E-mail: vinaysingh@bhu.ac.in, brahmadsingh@gmail.com Abstract Toll interleukin 1 receptor (TIR) domain is mainly involved in plant responses against various pathogens. This domain has been recognized at the Nterminus of the disease resistance proteins carrying the nucleotide binding (NB)-ARC domain (also called NBS) and leucine-rich repeats (LRR), these proteins confer resistance to various microbial pathogens, viruses, and invertebrates. Arabidopsis genome contains at least 200 proteins containing TIR domains. There are nearly 100 TIR-NBS-LRR class of resistance (R) genes in the Arabidopsis genome. The availability of Cajanus cajan (L.) Millsp. draft genome sequence gave us opportunity for the identification and classification of candidate’s gene families that are functionally important. In this study we focused on only TIR-NBS-LRR domain specific genes to investigate their numbers in the available draft genome sequence of C. cajan. Using bioinformatics approach we have successfully identified 373 TIR domain specific proteins from 510 best hits, out of which 135 TIR-NBS-LRR like genes were predicted within 186 best hits with available datasets. A total of 21 full length TIR-NBS-LRR genes were identified and verified using Interproscan with IPR000157,IPR002182 and IPR001611. Domain specific three dimensional predictions for TIR, NB-ARC and LRR were successfully done using Discovery studio 3.1 and the structures were deposited in Protein Model Database (PMDB) with PMDBID: PM0078097, PM0078232 and PM0078100. Structural classification of predicted models using SCOP and CATH servers revealed the classification lineage (3.80.10.10; 3.40.50.1820; 3.20.20.80; 3.40.50.300) having α-β class with α-β horseshoe, 3-Layer(aba) sandwich and Alpha-Beta barrel architecture. Topology of these models suggests that NBARC and TIR have TIM Barrel and Rossmann fold type topology, while LRR domain contains right-handed beta-alpha super helix. Gene ontology biological process shows their involvement in defense response, innate immune response and signal transduction. Cellular component analysis revealed that the identified genes are expected to be located in the membranes. Function elucidation of identified proteins using structural classification showed their role in ATP binding and transmembrane signaling receptor activity. VI ICLGG - Posters P-EAD29 Genetic diversity study among greengram [Vigna radiata (L.) Wilczek] genotypes using SRAP and RAPD Markers 1 Aneja B, Taunk J*, Yadav NR, Yadav RC , Kumar R CCS Haryana Agricultural University, Hisar, India *E-mail: jyotibiotech86@gmail.com Abstract Mungbean [Vigna radiata (L.) Wilczek] is an important protein source of low flatulence which is indispensable in Indian vegetarian diet. Iron and zinc; the two important micronutrients for human health are limiting in commercial mungbean cultivars. In the present study, a total of 62 primers including 29 Sequence related amplified polymorphism (SRAP) and 33 Random Amplified polymorphic markers (RAPD) were used to study DNA polymorphism among thirteen mungbean genotypes which are being used in crop improvement programme at CCSHAU. The study revealed 269 amplified bands with an average of 5.17 bands per primer out of which 263 were polymorphic and 6 were monomorphic. The similarity coefficients among different genotypes ranged from 0.56-0.85. At an arbitrary cut- off of 0.68 per cent similarity level, mungbean genotypes were categorized into two major clusters. Iron content (mg/kg) varied from 44.52 in GP-181 to 106.15 in Ganga-8 while zinc content (mg/kg) varied from 23.31 in Satya to 40.46 in ML776. High iron and high zinc genotypes did not cluster together; however SMH-99-1A and ML-776 (high iron and zinc content) made a separate group in cluster analysis. ML-776 and Satya, the two contrasting genetically diverse cultivars for micronutrient content have been used to develop mapping population and tagging genes/QTLs for micronutrient content. OPB7 primer and SRAP primer (me2em7) produced 900 bp and 700 bp amplified product respectively in ML776 that was found absent in Satya. OPB7 marker can be converted to Sequence characterized amplified region (SCAR) marker for differentiating the genotypes for micronutrient content. VI ICLGG - Posters Studies on genetic divergence and character association in elite lines of soybean (Glycine max (L.) Merrill) P-EAD30 Yadav A*, Tomar K, Singh K G.B.Pant University of Agriculture and Technology, Pantnagar, India *Email: yadav.aneeta@gmail.com Abstract The present investigation was carried out with 68 genotypes (including 62 elite lines + 6 check varieties) of soybean for eighteen quantitative and nine morphological characters with the objective to study genetic divergence and variability for yield, its components and their direct and indirect effect on seed yield. The mean squares due to genotypes were highly significant for all the 2 characters studied. Based on D statistic, the 68 genotypes were grouped into 13 clusters. The maximum numbers of genotypes (12) were grouped in cluster I and lowest (1) in cluster XI, XII and XIII. The maximum divergence was 2 observed between cluster X and XII. Based on D analysis, the potential parental combinations were found to be ABL-11 X ABL-62, ABL-15 X ABL-62, ABL-16 X ABL-62, ABL-36 X ABL-62, ABL-49 X ABL-62, ABL-66 X ABL-62, ABL-62 X ABL-43. Dry matter weight per plant contributed maximum towards genetic divergence followed by number of pods per plant and nodules mass weight per plant. Morphological characters revealed that leaf colour, pod colour, seed coat colour and hilum colour contributed more towards genetic divergence than pubescence and flower colour and nodule type. Highest coefficient of variation was observed for nodules mass weight per plant followed by nodules dry weight per plant and yield per plot. Based on divergence, variability and path coefficient analysis, number of nodules per plant, nodules mass and dry weight per plant, plant height, pods per plant and oil content were found to be important yield contributing characters for selection procedure in soybean. VI ICLGG - Posters P-EAD31 Genetic variation of Australian faba bean (Vicia faba) varieties revealed by EST-SSR markers 1, 2 3 Yang SY *, Bagheri A , Ahmad M , Paull JG 1 1 School of Agriculture Food and Wine, the University of Adelaide, Urrbrae, Australia 2 Department of Biotechnology and Plant Breeding, Ferdowsi University of Masshad, Masshad, Iran 3 Bayer CropScience, Horsham, Victoria, Australia. *E-mail: shi.yang@adelaide.edu.au Abstract Faba bean (Vicia faba) is one of most important grain legume crops in Australia. The industry has developed initially based on var. Fiord and then var. Fiesta VF, while in recent years a range of varieties has been available. Development and application of EST-SSR markers is popular in many species and they enable genetic variation analysis. The aim of this study was to develop EST-SSR markers to apply to Australian faba bean varieties, Fiord v Ascot and Fiesta v Farah, and reveal the genetic variation within and between pairs of related varieties. ESTs were searched from NCBI dbEST and 30 PCR primers were designed based on di- and tri-nucleotide SSR motifs. These primers were pre-tested on 10 genotypes of each of Cicer arietium, Lens culinaris, Vicia faba and Pisum sativum, and subsequently used to test 48 plants of each of these faba bean varieties. Seven of 30 primers tested in the four varieties revealed polymorphic bands. A total of 149 bands from these seven primers were found. The PIC value was between 0.33 and 0.83. A binomial test of the comparison of Fiord and Ascot showed that for primer 46 there was one polymorphic band and Fiord was more variable; while for primer 131 there was also one significant band but Ascot was more variable. For the comparison of Fiesta VF and Farah, there were two polymorphic bands for primer 46 and Fiesta was more variable; and there were three polymorphic bands for primer 95 within Fiesta and Farah. . VI ICLGG - Posters Can cross-species transferable molecular markers be useful on genetic diversity studies? 1* 1 2 3 P-EAD32 1 Leitão ST , Almeida NF , Šatović Z , Rubiales D , Fevereiro P , Vaz Patto 1 MC 1 Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal 2 Faculty of Agriculture, University of Zagreb, Zagreb, Croatia 3 Institute for Sustainable Agriculture, CSIC, Córdoba, Spain *E-mail: sleitao@itqb.unl.pt Abstract Nowadays underused in Europe, Lathyrus species were in the past widespread in Mediterranean regions. The superior atmospheric nitrogen fixation rate and potential in crop rotation, allied to the ability to tolerate salinity, drought and flooding, make some of these Lathyrus species highly potential for sustainable agriculture. Being member of the legume family Fabaceae, Lathyrus spp. could share genes, physiological processes and defense mechanisms with economically important related crops such as pea and lupines. Likewise, the molecular tools developed on these model species and cross-amplifiable on Lathyrus spp. could be used on them for different genetic studies, such as diversity analysis. Accordingly, the genetic diversity of an Iberian collection of 45 Lathyrus sativus (grass pea) and 45 L. cicera (chickling vetch) accessions was evaluated using 17 cross-species amplifiable molecular markers (ITAP, EST-SSR and gSSR), developed for Pisum sativum, Lupinus albus and Medicago truncatula. Homology of amplified fragments was confirmed by sequencing. In L. sativus, 2 loci did not amplify, 7 were monomorphic and 8 polymorphic, with an allele average of 3.75 (range 2-8) and a PIC average of 0.356 (range 0.024-0.792). For L. cicera, 7 loci were polymorphic and 10 monomorphic, with an allele average of 2.14 (range 2-3) and a PIC average of 0.287 (range 0.099-0.435). Four loci were polymorphic in both Lathyrus species. These results confirm the feasibility of interchange molecular markers for genetic studies, revealing a higher heterozygosity and a potential higher outcrossing rate of L. sativus in relation to L. cicera. VI ICLGG - Posters Theme Harnessing Germplasm Resources P-HGR07 Developement and evaluation of high yielding mungbean mutants 1,* 1 2 1 3 Vijaykumar AG , Soregaon CD , Giridhar G , Math GI , Ontagodi TP , 4 Shakuntala NM 1 AICRP on MULLaRP, MARS, Dharwad, India NAIP on Millets, Dharwad, India 3 Department of Genetics & Plant Breeding, Dharwad, India 4 Department of SST, Raichur, India 2 *E-mail: vijay_gpbd@yahoo.co.in Abstract Mungbean (Vigna radiata (L.) Wilczek) is the major pulse crop of South Asia. In Karnataka, it ranks second after chickpea and occupies an area of 4.01 lakh hectares with a production of 1.12 lakh tonnes. However, the productivity is much below (305 kg/ha) the national average (428 kg/ha), mainly due to nonavailability of high yielding, photo insensitive genotypes. Development of photo-insensitive genotypes that suit their cultivation across seasons i.e., kharif (June- September), spring (February – April) and summer (March-April) is very important to bridge the gap between potential and realized yield. In this context mutation breeding can serve as an efficient method for generating variability in the local varieties, and selecting desirable early and synchronously maturing lines suitable for mechanical harvesting. Keeping this in view, an attempt was made to enhance the variability and improve the yield potential of mungbean through the use of physical and chemical mutagens. An indeterminate local variety “Black greengram” was treated with gamma rays (150 & 200 gray), 0.25% EMS and combination of both gamma rays and EMS (150 gray+0.25 % EMS & 200 gray + 0.25% EMS). Mutants superior to parents/checks with respect to number of pods per plant, seed weight, numbers of seeds per pod, yield and seed protein content were identified based on their evaluation across two seasons and four locations. The mutants M-70-15 (1614 kg/ha), M-71-09 (1584 kg/ha), M-129-02 (1548 kg/ha) and M29-10 (1394 kg/ha) yielded higher than the checks LGG 460 (1323 kg/ha), Selection 04 (1274 kg/ha) and Black green gram (1107 kg/ha), while mutant M-70-03 gave significantly higher protein content (28.13%) with an yield of 1477 kg/ha. These mutants are determinate, non shattering, photo-insensitive and suitable for mechanised harvesting. VI ICLGG - Posters Vetches and grass pea as summer cover crop after barley in semi-arid regions 1, Alizadeh K *, Sabouri N P-HGR08 2 1 Mahabad Branch, Islamic Azad University, Mahabad, Iran Agriculture and Natural Resources Service, Malakan, Iran 2 *E-mail: khoshnod2000@yahoo.com Abstract There are limited forage resources and plant coverage during late summer in the semi-arid regions which can be alleviated by cultivation of suitable cover crops. Two vetches and one grass pea species were studied as summer cover crop after barley harvesting under irrigated condition in Mid July using a randomized complete block design with 6 replications. Amount of dry matter (DM), crude protein (CP), and percent of nitrogen (N), phosphorus (P) and potassium (K) in DM were determined when vetch reached flowering stage which coincided with the milky/initial creamy stages of barley. The combined analysis showed that main effect of cover crops on DM, CP and percent of N and P were significant (P < 0.01). Due to its running type as well as higher potential N and CP performance, smooth vetch cv. Maragheh was a recommended summer fallow crop for semi-arid areas. Present limitations and opportunities to develop summer cover crops are discussed in details. VI ICLGG - Posters P-HGR09 Landraces in chickpea (Cicer arietinum L) - use them or lose them Aski M*, Chaturvedi SK Indian Institute of Pulses Research (IIPR), Kanpur, India *E-mail: murali2416@gmail.com Abstract Landrace is a variety with a high capacity to tolerate biotic and abiotic stress, resulting in high yield stability and an intermediate yield level under a low input agricultural system. These landraces are the rich source of valuable traits which may be use full in meeting future challenges of food security. The Chickpea (Cicer arietinum L) is a major pulse crop covering area of 9.06 mha and producing 8.25 million tonnes of the grains in India. In the era of modern agriculture, chickpea is very much prone to genetic erosion. Efforts were made by Indian Institute of Pulses Research Kanpur (IIPR) India in collaboration with ICARDA (Syria) to prevent such genetic erosion by harnessing the locally adopted landraces in breeding programme. 100 chickpea land races collected from 56 countries were obtained from ICARDA and we evaluated for 13 different yield and yield related traits at IIPR Kanpur. This material will help adding more divergent parents in crossing programme and leads to generation of variability in chickpea breeding material. Coefficient of variation was found very high for all traits in studies. It was maximum for number of pods per plant (43.29%) fallowed by seed yield per plant (42.29%), seed yield per plot (40.11%) and number of primary branches per plant (29.62%). Remaining traits exhibited medium to low coefficient of variation. Cluster analysis revealed that 100 land races were grouped into 9 clusters. Cluster 8 was the largest cluster having 16 landraces most of them belonged to temperate regions and cluster 7 was the smallest cluster having only one landrace (ILC 9793) which belonged to country Turkey. Cluster analysis also revealed that the recorded diversity was actually due to the different gene pools, which might be due to varying degree of out breeding. Geographic relationships based on cluster analysis provided clues of introduction of the same germplasm in different areas and transgression of genes into different landraces. VI ICLGG - Posters Screening of fababean (Vicia faba L.) against diseases: harnessing resistant source for crop improvement P-HGR10 Bag MK*, Roy A, Prasad TV, Gangopadhyay KK, Phogot BS, Dutta M National Bureau of Plant genetic Resources, New Delhi, India *E-mail: manas.bag@gmail.com Abstract Fababean (Vicia faba L.), grown in many countries but an underutilized grain legume in India, is now increased importance for its high yield potential, richness in protein, highest total essential amino acids and high mineral contents particularly calcium and iron. But this crop is prone to infection by many diseases that limit popularization of its cultivation in India. This crop is very important in developing countries, where low input agriculture is practiced using simple technologies. Correct diagnosis of disease is fundamental to control and this has been facilitated by advances in pathogen identification which actually help in screening of the crop against those diseases for identifying resistant sources. 62 germplasm lines were grown during Rabi, 2011 - 2012 at NBPGR, New Delhi following standard agronomic practices for evaluation. Diversified diseases including seedling Blight (Rhizoctonia solani) root rot (Fusarium sp., Rhizoctonia solani), Sclerotinia stem rot (Sclerotinia sclerotiorum), Ascochyta leaf and pod spot. (Ascochyta fabae f.sp. fabae), powdery mildew (Microsphaera penicillate var. ludens) was recorded during the crop season. A mosaic disease suspected to cause by viruses is also noticed in a few germplasm. Chocolate spot, Ascochyta leaf and pod spot, two commonly occurring diseases in temperate region were observed only in a few accessions while occurrence of root rot and stem rot were predominant among the majority of the germplasm with high per cent disease severity. Screening of those germplasm following standard evaluation scoring scale identified 12 germplasm which were found free from diseases. This study will help in harnessing resistant sources against major diseases like root rot and Sclerotinia stem rot. VI ICLGG - Posters P-HGR11 Screening of lentil (Lens culinaris L.) and chickpea (Cicer arietinum L.) breeding lines in Alberta 1 1 2 2 2 Bandara MS , Kruger A , Vandenberg A , Warkentin T , Taran B 1 Alberta Agriculture and Rural Development, Alberta, Canada University of Saskatchewan, Saskatoon, Canada 2 *E-mail: manjula.bandara@gov.ab.ca Abstract In Canada, chickpea (Cicer arietinum L.) and lentil (Lens culinaris L.) producing areas are mainly confined to the Brown and Dark Brown soil zones in Saskatchewan (SK) and Alberta (AB) provinces. The total chickpea production area in Canada had been gradually declined from 480,000 ha in 2001 to 46,600 ha in 2004 due to adverse climatic conditions caused by erratic rainfall, epidemics of Ascochyta blight; and low global market price. The growing conditions in Southern AB, however, are warmer and drier than Southern SK. For over fifteen years, lentil production area in AB has been fluctuated between 5,000 and 8,000 ha due to lack of well-adapted cultivars. Improved cultivars with desirable and promising traits, including earlier maturity, Ascochyta blight resistance and drought tolerance, are needed for sustaining of both chickpea and lentil industries in AB. In 2001, a collaborative research project between CDC and Alberta Agriculture on cultivar development for these two crops was initiated. Consequently, lentil varieties such as CDC Redbow, CDC Rosebud and CDC Redcliff and chickpea varieties such as CDC Orion were developed and released. Further studies are being focused on the development of red lentil cultivars with herbicide resistance and desired starch profile. . VI ICLGG - Posters Productivity and diseases resistance improvement through transgressive breeding in black gram (Vigna mungo (L.) HEPPER) 1,* 2 3 4 P-HGR12 1 Basamma K , Salimath PM , Benagi VI , Suma Biradar , Patil Malagouda , 1 1 1 Timmanna O , Gangashetty PI , Sridevi BM 1 Dept. of Genetics and Plant Breeding, UAS, Dharwad, India Special Officer, UAS, Shimoga, India 3 Dept. of Plant Pathology, UAS, Dharwad, India 4 Wheat Improvement Scheme, UAS, Dharwad, India 2 *E-mail: basammapatil09@gmail.com Abstract Urdbean is an important grain legume with easily digestible protein and low flatulence contents. The present productivity level of this crop in India is very low. Susceptibility to a number of biotic and abiotic stresses is an important constraint responsible for low and unstable productivity. Yellow Mosaic Virus and Powdery Mildew are two important diseases causing considerable loss to the tune of 300 million dollars annually. Considering the importance of the crop, it is necessary that research efforts are directed for genetic enhancement of the crop for improving the productivity and resistance to diseases. In this context, identification of transgressive segregants for economically important traits such as yield and disease resistance is an important aspect of any practical breeding programme. The F2 populations of two diverse blackgram crosses were evaluated as well as the number of transgressive segregants during 2008 to 2010. TAU-1, a high yielding but powdery mildew and MYMV susceptible genotype was crossed to LBG-17 and BDU-4 which are resistant powdery mildew and MYMV respectively. The F 1, F2 and F3 populations were evaluated along with the parents for resistance to both diseases separately under artificial condition. The evaluation of F2 population revealed high variability for various productivity traits and generated transgressive segregation. Similarly, evaluation of F3 families further based on selections made in F2 confirmed the superiority of few families in both the populations. Twelve families showing resistance to powdery mildew and high yield and nine F3 families showing resistance to MYMV and high yield were identified. It is suggested to carry forward these promising selections further to develop high yielding and disease resistant lines. b VI ICLGG - Posters P-HGR13 Utility of peanut mapping populations developed at UAS Dharwad, India 1, 2 2 2 2 1 Bhat RS *, Sujay V , Mukri G , Khedikar YP , Sarvamangala C , Yeri SB , 2 2 2 2 2 Jadhav SS , Pujer SB , Narasimhulu R , Varshakumari , Azharudheen TPM , 3 2, 2 Varshney RK , Nadaf HL Gowda MVC 1 Department of Biotechnology, University of Agricultural Sciences, Dharwad, India 2 Department of Genetics and Plant Breeding, University of Agricultural Sciences, Dharwad, India 3 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India *E-mail: bhatramesh12@gmail.com Abstract Groundnut (Arachis hypogaea L.) is one of the principal oilseed legume crops of the world. In India, its low productivity is mainly attributed to the susceptibility of available cultivars to the biotic stresses. Breeding for resistance to these stresses through the development and application of genomic resources and markers has marked advantages over conventional breeding. In this context, development of mapping populations from parents differing for various traits of agronomical importance and resistance to biotic stresses is considered to be the key step, which can eventually be used for constructing the linkage map and QTL mapping to enable marker assisted breeding. A constant effort at UAS Dharwad, Karnataka, India has led to the development of seven recombinant inbred line mapping populations from the parents mainly differing for resistance to late leaf spot, rust, Asperigillus and Sclerotium rot. Phenotypic evaluation of these mapping populations over the years has revealed considerable phenotypic and genotypic coefficient of variation for the traits. Analysis of TAG 24 X GPBD 4 and TG 26 X GPBD 4 mapping populations at ICRISAT, Hyderabad could detect major QTLs for rust and late leaf spot resistance. These QTLs are being used for marker assisted breeding. VI ICLGG - Posters Agronomic response of promising mungbean (Vigna radiata L.) mutant lines to drought stress under arid areas P-HGR14 Rahayu S, Dewi AK, Zulhedi* Indonesian National Nuclear Energy Agency, Indonesia *E-mail: upikyuliasti@yahoo.com Abstract The objectives of this study were to develop superior genotypes to improve mungbean production andquality for food. Mungbean production is aimed at optimal use of unproductive or marginal land such as that of drought prone areas. Mungbean breeding through induced mutations was conducted at Center for the Application of Isotope and Radiation Technology, National Nucleare Indonesia. Through selection processes and direct screening for drought tolerance in Mataram direct 2mutant line and two check had been select in dry line.The mutant PSj S31 produced grain yields of 1.64ton/ha respectively.These yields were signifi cantly higher than the original parent Gelatik 6.02ton/ha control check. The mutant lines PSj S31 were drought tolerant genotypes having relatively higher amounts of total soluble sugars than control plants (gelatik and perkutut), these mutant lines were promising for further Mungbean breeding and development in accordance with attempts of increasing land productivity of drought prone areas. VI ICLGG - Posters P-HGR15 Wild Cicer species: A repository of Botrytis grey mould resistance 1,* 1 2 1 1 1 3 Kaur L , Sirari A , Singh M , Singh S , Kumar D , Singh I , Sandhu JS , 1 4 4 4 4 Kapoor K , Gaur PM , Pande S , Sharma M , Gowda CLL 1 Punjab Agricultural University (PAU), Ludhiana, Punjab, India National Bureau of Plant Genetic Resources (NBPGR), New Delhi, India 3 Indian Council of Agricultural Research, New Delhi, India 4 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 2 *E-mail: livinderk@rediffmail.com Abstract Botrytis grey mould (BGM) a devastating disease of chickpea (Cicer arietinum L.) in northern India causes significant yield losses in epidemic years. Resistance to BGM is very low in cultivated chickpea, however, this resistance was explored in Wild Cicer sp. BGM resistance was identified through host pathogen interaction in accessions of different wild Cicer species using cut twig technique. Out of 85 accessions, 16 accessions of C. pinnatifidum, 10 of C. judaicum possessed high degree of resistance (rating < 3.0), however, most of the C. reticulatum accessions were moderately susceptible (rating 6-7). In C. bijugum, three accessions each were resistant (rating < 3.0) and moderately resistant (rating 4-5). Inter and intraspecific genetic diversity in these accessions was demonstrated through molecular markers. A BGM resistant accession of C. pinnatifidum acc. 188, was crossed to highly susceptible variety ICCV 96030 and BGM resistant derivative lines in F9 were developed by selection and pedigree method. Four derivative lines, which were identified with C. pinnatifidum acc. 188 specific segments through molecular markers, were further used to develop mapping populations with susceptible line BG 256. Parental polymorphism was analysed using 120 SSR markers, of which 60 were polymorphic. These and additional markers can be used for construction of linkage maps using F2 population of above crosses. The BGM resistant derivative lines from interspecific cross (C. pinnatifidum acc. 188 and ICCV 96030) are being used as donors in breeding programme. VI ICLGG - Posters Inheritance of stipule spot pigmentation in faba bean (Vicia faba L.) P-HGR16 Khazaei H*, Stoddard FL University of Helsinki, Helsinki, Finland *E-mail: hamid.khazaei@helsinki.fi Abstract The stipule at the base of each leaf of faba bean (Vicia faba L.) carries an extrafloral nectary marked in most accessions by dark pigmentation (stipule spot pigmentation, SSP). The absence of pigment has been used elsewhere as a pre-flowering marker for the zero-tannin trait, but there is conflicting information concerning the inheritance of SSP in this species. The experiments were based on 4 inbred lines, of which Disco/2 was zero-tannin (zt2) with colourless stipule spots, ILB 938/2 was normal-tannin with colourless stipule spots, and both Aurora/2 and Mélodie/2 were normal-tannin with coloured stipule spots. Crosses Mélodie/2 × ILB 938/2, Mélodie/2 × Disco/2, ILB 938/2 × Aurora/2 and ILB 938/2 × Disco/2 (A, B, C and D, respectively) were prepared and advanced through single-seed descent in bee-proof glasshouse conditions. All F1 hybrid plants had coloured stipule spots, and in the F 2 generation, the ratio fit 3 coloured: 1 colourless in crosses A, B and C and 9:7 in cross D. In the F3 generation, the ratio fit 5:3 in cross A and 25:39 in cross D, and in the F4 generation, 9:7 in cross A. SSP was linked to the zero-tannin characteristics (white flower and colourless hilum) only in cross B. The results show that coloured stipule spot is dominant to colourless, and that colouration is determined by two unlinked genes. We propose the symbols ssp1 for the gene associated with zero tannin in Disco/2 and ssp2 for the gene not associated with tannin content in ILB 938/2. VI ICLGG - Posters P-HGR17 Herbal vaccination - a tool for tolerating different stresses and improving productivity of mungbean and blackgram 1, Kundagrami S *, Roy Choudhury D, Das M Institute of Agricultural Science, University of Calcutta, Kolkata, India *E-mail: skundagrami@gmail.com Abstract The productivity of mungbean and blackgram remain stagnant over a few years due to different biotic and abiotic stresses. So an attempt has been made to cope up different stresses and to improve seed yield through herbal vaccination of seeds as well as foliar application. Twenty herbal treatments in dust form were given to the seeds of two popular varieties, B1 (mungbean) and WBU108 (Blackgram)) @ 2gm/kg of seeds and stored for two months. During germination another pre-treatment @ 2gm of each treatment/litre of water was given. Laboratory screening for salinity tolerance were done by -1 nutrient culture method of germinated treated seeds at 5 dS m saline solution for seven days and laboratory screening for submergence tolerance were done by inserting the germinated treated seeds in submerged condition for seventy two hours followed by desubmergence for seven days in nutrient culture and their vigour were compared with the control. Screening for bruchid attacks o were done by infesting bruchid populations to the treated seeds in 35 C temperature and 80% relative humidity for six weeks in B.O.D. and infestation were compared over the control. In field trial the treated germinated seeds were sown in three replications and foliar applications of herbal treatments were done at 30 and 50 DAS (Days after sowing) and attack of pod borer, seed yield/plot and harvest index were recorded. Turmeric, chilli, ginger, hing, neem, tobacco and kalmegh powder were shown significance appreciable results in all of the cases over the control indicating immense scope for herbal vaccination as they are eco friendly and important component of sustainable agriculture. VI ICLGG - Posters Identifying superior salt tolerant cowpea (Vigna unguiculata L.Walp) genotype P-HGR18 Arulvadivookarasi K, Mini ML, Jayachandran MA Bharathidasan University, Tiruchirappalli, India *E-mail: msanushyas2005@gmail.com Abstract Cowpea (Vigna unguiculata L.Walp) is one of the ancient food legume crop cultivated in arid & semi-arid region. It is a drought tolerant crop. Soil salinity is an ever increasing threat for crop production where, it restricts the use of agricultural land for further cultivation. In year 2010, Central Soil Salinity Research Institute, Karnal, India has estimated that salt affected soils occupy an area of 953 Mha. The goal of the present study was to identify the superior salt tolerant genotype of cowpea cultivated under salt stress. Germination studies were conducted in eight different genotypes of cowpea by roll towel method. Seeds were allowed to germinate in five different concentrations of NaCl (0, 50, 75, 100 and125mM) solution. Leaf and root samples were th harvested on the 7 day after germination and the morphological and biochemical parameters were observed. The study showed significant differences in shoot length, root length, total soluble sugars, protein, proline, + + Na and K between genotypes. Salinity induced significant increase in total + soluble sugars, proline levels, and Na concentrations, while reduced the + accumulation of K ion concentration. Of the various genotypes (COCP 7, VBN 01, ACM 008, GC 0817, ACM 011, CP 16, ACM 05-07, DC 15) used for the study, the genotype ACM05-07 showed higher leaf protein and proline + contents while lower Na compared to other genotypes. The current study showed that ACM05-07 is the most salt tolerant genotype at germination stage. The selected genotypes have been subjected to MALDI-TOF to study the protein expression pattern between the tolerant and sensitive genotypes to further confirm the superiority of the genotype. VI ICLGG - Posters P-HGR19 Interspecific hybridization in breaking yield barriers in chickpea (Cicer arietinum L.) Mishra N*, Chaturvedi SK, Soren KR, Nadarajan N Indian Institute of Pulses Research (IIPR), Kanpur, India *E-mail: neelumishra111@gmail.com Abstract Interspecific hybridization has been recognized as important method to create variability and broaden genetic base. Four Interspecific crosses in chickpea involving Cicer reticulatum (IPCK2002-29 x ILWC21: 5 sps, GNG469 x ILWC21: 16 sps, IPC2008-57 x ILWC21: 13 sps) and Cicer echinospermum (IPCK2002-29 x ILWC245: 5 sps) were made during 2010-11 at IIPR and true F1 plants were identified using morphological and SSR markers (TR11 and TA76). Large amount of variations and heterobeltiosis was observed for yield attributes. Days to first pod ranged from 66 (IPCK2002-29 x ILWC21) to 116 days (IPCK2002-29 x ILWC245), 50% podding from 98 (IPCK2002-29 x ILWC21) to 126 days (IPCK2002-29 x ILWC245) whereas days to maturity exhibited narrow range (142-148 days). Three interspecific crosses produced typically prostrate plants except (GNG469 x ILWC21). Canopy width, which is the direct indicator of total biomass production in semi-spreading/prostrate types ranged from 31 (IPC2008-57 x ILWC21) to 94 cm. (IPCK2002-29 x ILWC 245) and primary branches per plant from 48 (IPCK2002-29 x ILWC245) to 7 branches (GNG469 x ILWC21). Similarly, the most important yield attribute, pods per plant ranged from 48 (GNG469 x ILWC21) to 179 pods (IPCK2002-29 x ILWC245) indicating vast variability with respect to important yield attributes. Systematic evaluation and intermating among desirable F 2 plants may likely to release more variability and will help in breaking undesirable linkages. . VI ICLGG - Posters K 1535, A high yielding Spanish bunch groundnut variety with multiple resistance for biotic and abiotic stresses developed at Agricultural Research Station, Kadiri, AP. India P-HGR20 Naik KSS, Rajesh AP, Vemana K, Venkateswarlu NC, Sampath Kumar D Chandrayudu E, Reddy DL, Narayana TS , Goud TES Agricultural Research Station, Acharya N.G. Ranga Agricultural University, Kadiri, India *E-mail: drkssnaik@yahoo.com Abstract Groundnut (Arachis hypogaea L.) is the most important edible oil seed crop of AP, India, it is grown mostly under rainfed conditions, where the crop exposured to devastating pests (Thrips and Jassids) diseases (leaf spots, PBND and PSND) and drought which are major threats for higher yield K 1535 a high yielding spanish bunch groundnut variety was developed at Agricultural Research Station, Kadiri by crossing very popular commercial groundnut variety Kadiri 6 with wild germplasm line NCAc 2242 (ICG5044) during kharif 2001. The F1 was advanced during kharif 2002 to F2 and selection (03X154001) was made in F2 generation during kharif 2003 and K1535 was evaluated in In the Station level kharif trials at Kadiri over 4 years (2004-2007). The mean pod and kernel yield were 1705 and 1077 kg/ha, with an increase of 28 and 18% over local check Vemana during kharif rainfed conditions. The mean -1 haulm yield was 2937 kgha which is a most desirable trait in the semi arid tracts for the survival of the livestock during drought years. Under favorable season the yield is comparable with the check Vemana but under severe drought conditions the mean pod and kernel yield of K1535 were 46.5 and 37.3% higher than Vemana respectively, showing its ability to resist drought. The Incidence of Peanut Stem Necrosis Disease ranged from 2.5 % to 6.0 % against 14.6 % to 25.0 % in check over three years of screening. Intensity of early leaf spot and late leaf spot ranged from 3.0 to 5.0 scale (Intermediate reaction) and 2.0 to 3.0 scale (resistant reaction) respectively. It has given good reaction against foliar diseases without defoliation even under high disease pressure. The thrips damage ranged from 2.0 to 3.0 and jassid 1.0 to 3.0 scale recorded against 5.0 and 4.0 scale respectively in check during the test period. While the leaf miner damage was ranged from 1.8 to 6.4% and defoliators 5.6 to 12.6% against check 9.2% and 14.8% respectively. b VI ICLGG - Posters P-HGR21 Breeding better mungbean (Vigna radiata (L.) R. Wilczek var. radiata) 1,* 2 ,2 2 2 Nair RM , Yang R-Y , Schafleitner R , Kenyon L , Srinivasan R , Easdown 1 2 2 W , Ebert AW , Hanson P 1 AVRDC, ICRISAT Campus, Hyderabad, India AVRDC, Shanhua, Tainan, Taiwan 2 *E-mail: ramakrishnan.nair@worldveg.org Abstract Mungbean (Vigna radiata (L.) R. Wilczek var. radiata) is cultivated on more than 6 million hectares in the warmer regions of the world and is one of the most important pulse crops in South and Southeast Asia. Mungbean protein and carbohydrates are easily digested and create less flatulence than other legume crops. Mungbean has a lower phytic acid content than chickpea, pigeonpea, soybean and other legumes; this compound, commonly found in cereal and legume crops, has a negative effect on mineral bioavailability in plant-based diets. The productivity of mungbean is relatively low (average about 400 kg/ha), similar to other pulse crops. Some strategies for developing better mungbean include broadening the genetic base of varieties, development of varieties with resistance to major diseases such as mungbean yellow mosaic disease and pests such as bruchids, enhancing the protein quality by improving the methionine content, and improving the iron and zinc content. Emphasis on thermo- and photo-insensitivity and tolerance to waterlogging and salinity will be vital to improve the crop’s ability to adapt to climate change. Active promotion of the nutritional value of mungbean along with its proven agronomic advantages will strengthen the case for further expansion in Asia and other parts of the world including sub-Saharan Africa. Improving the nutrient content and productivity of mungbean will also help to reduce malnutrition, especially among women and children in developing countries. VI ICLGG - Posters Induced male sterility and peroxidase isozyme in winged bean, Psophocarpus tetragnolobus (L) DC 1, P-HGR22 2 Ghanawat NA *, Dixit GB 1 Yashvantrao Chavan Institute of Science, Satara, India Shivaji University, Kolhapur, India 2 *E-mail: namdeoghanawat249@gmail.com Abstract Winged Bean, Psophocarpus tetragonolobus (L.) DC. Belonging to subfamily fabaceae of family leguminosae is underexploited tropical crop with promising economic value, a high protein crop and a possible soybean for the tropics. Male sterility resulted from heritable system is an interesting phenomenon useful in the plant breeding. In present investigations, seed samples of winged bean of Philippine accession procured from Dambala Institute of, Pallekele, KundasleSri Lanka were irradiated with 5, 10, 12.5, 15, 17.5, 20, 25, 30, 35, 60 and 40 Krad doses from Co source at BARC, Bombay (India) with dose rate at 2.8 Krad per minute and sown in the field. Forty eight M 4 families were screened for male sterility. One of these families showed segregation for male fertile and male sterile individuals. Male sterile plants were successfully isolated and studied for 15 different morphological, cytological and molecular level parameters. All parent plant, fertile male and sterile segregents were found similar with respect to Habit, plant height, and stem colour, presence of tubers, calyx colour, corolla colour, and stamen adhesion. However, as compared to parent, the male sterile segregants were found different viz. slow in plant senescence; Leaves-ovate, larger size, dark green, slightly leathery and thick, slow in senescence; reduction in number and staining intensity of peroxidases bands; anthers with late dehiscence, no pollen shedding, sticky pollen grains and high pollen sterility (95.9881 %). The results have been discussed in context to relevant earlier reports on male sterility. This male sterility may found helpful in breeding programmes in winged bean. VI ICLGG - Posters P-HGR23 Adaptation of blackgram (Vigna mungo L.) genotypes to paddy fallow areas 1,* Ontagodi TP , Salimath PM 5 4 Hanamaratti NG , Naik VR 1, 2 1 3 4 , Shridevi BM , Vijaykumar AG , Biradar S , 1 Department of Genetics and Plant Breeding, UAS, Dharwad, India UAS, Shimoga, India 3 AICRP on MULLaRP, MARS, UAS, Dharwad, India 4 AICRP on wheat, MARS, UAS, Dharwad, India 5 ARS, Mugad, UAS, Dharwad, India 2 *E-mail: timmu07uas@gmail.com Abstract Blackgram (Vigna mungo (L.)) is an important food legume grown as a sole crop under residual moisture conditions after the harvest of rice under dryland conditions. Aim of this study was to evaluate forty one blackgram genotypes in paddy fallow situations at Dharwad, Mundgod and Mugad of Karnataka state for five different characters to asses there performance and adaptability. Data was subjected to stability analysis following Eberhart and Russell (1966) model. Significance of main environmental and GxE interaction effects indicated the differential response of the genotypes to changing environments. Pooled analysis of variance revealed significant differences among genotypes and environments for all the characters (except 100 seed weight) indicating that genotypes and tested environments are diverse in nature, whereas genotype x environment interaction was also significant for all the characters suggesting that differential genotypic response to environments. Based on stability parameters genotypes viz., 488-15-6, Manikya and LBG-685 were found to be stable for majority of characters with higher mean performance. The genotypes 488-15-6 and KU-5-554 were high yielders across environments compared to check varieties TAU-1 and Manikya. The genotypes namely 2KU-64, KU-5-554, KU-5-542, 488-15-6, IC-436609, IC321892, IC-436535 and Manikya showed high mean seed yield per plant and were found to be stable across test locations. This information generated from the study provides a strong base to initiate breeding programme to develop improved stable varieties for paddy fallow situations which in turn will help in improving urdbean production in time and space.. VI ICLGG - Posters Biomass accumulation and seed yield of late planting chickpea (Cicer arietinum L.) in Delhi conditions 1 1 1 1 P-HGR24 1 Shah D , Bahuguna, RN , Sadhna, Sweta, Anand A , Khetrapal S , Kumar P , 2 2 1 1, Bhardwaj C , Kumar J , Deshmukh, PS , Madan Pal * 1 Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi, India 2 Division of Genetics, Indian Agricultural Research Institute, New Delhi, India *E -mail: madanpal@yahoo.com Abstract Rising atmospheric temperature under climate change scenario may affect the seed yield of late planting chickpea under Delhi conditions. A filed study was conducted during 2009-10 and 2010-11 with three staggered sowings viz. S1 (early November), S2 (late November) and S3 (mid December). S2 and S3 0 sown crops experienced low temperature during the vegetative stage (11.5 C average daily mean temperature). On the other hand, average daily mean temperature at mid-flowering and podding stages was observed 14. 6 to 21.2 0 0 C and 20.4 to 25.1 C, respectively for all three sown crops. Observations were recorded for membrane stability index and biomass assimilation during mid-flowering and podding stages. No significant changes were observed for membrane stability index in the plants of all three sowings, which indicated less sensitivity of chickpea genotypes to high temperature stress during flowering and podding under late sowing conditions. However, there were marked reductions in total biomass accumulation and partitioning in different plant parts in S2 and S3 sown crop plants. The reduction in plant biomass occurred due to sensitivity of S2 and S3 sown crops to low temperature during vegetative stage. Poor vegetative growth in late sown crops resulted in production of lower plant dry weight and seed yield per plant compared with normal sown crop. The study concludes that chickpea crop may be less 0 affected by 3-4 C increase in atmospheric temperature projected under future climate change scenario and rather may be benefitted from chilling effects in Delhi conditions. b VI ICLGG - Posters P-HGR25 Identification of apomixes in chickpea 1, 2 Panchangam S *, Suravajhala P , Mallikarjuna N 1 1 International Crops Research Institute for Semi - Arid Tropics (ICRISAT), Hyderabad, India 2 BioClues Organization, ICICI Knowledge Park, Secunderabad, India *E-mail:sameera.panchangam@gmail.com Abstract Apomixis or asexual reproduction by seeds is wide spread in Poaceae and Asteraceae families and in apomeiotic mutants of medicago which is a legume. Early Apomictic events were observed in chickpea as a result of wide cross utilizing incompatible Cicer species. Molecular information is now becoming available for chickpea and EST sequences are now available. It is not necessary that ESTs reveal the full proportion of the functional repertoire. Interlog based protein interaction analyses was employed to identify appomixis related genes in chickpea. Functional annotation of all proteins when searched against Arabidopsis thaliana protein sequences showed few plausible proteins, viz. MADS, MYB, C2C2-YABBY, SHP2, AP2, SPL which are involved in floral development and embryogenesis. Together with protein interaction network and orthology based similarity searches putative candidates for apomixes in chickpea have been identified. These when combined with wet lab experiments provide ample evidence that EST analysis plays an important role where sequences are not known and annotated. Further analysis of the transcript data might help to identify the factors which trigger or regulate apomictic gene expression. VI ICLGG - Posters Molecular characterization of minicore collection set of pigeonpea (Cajanus cajan (L.) Millsp) for sterility mosaic disease and fusarium wilt 1 1, 1 1 1 P-HGR26 1 Jaggal LG , Patil BR *, Talikoti MM , Naik PM , Narasimhan A ,Nadaf HL , 1 2 2 2 Patil K , Saxena RK , Upadhyaya HD ,Varshney RK 1 Department of Genetics and Plant Breeding, University of Agricultural Sciences, Dharwad, India 2 International Crops Research Institute for Semi-Arid Tropics (ICRISAT), Hyderabad, India *E-mail: patilbhuvaneshwara@gmail.com Abstract Pigeonpea (Cajanus cajan (L.) Millsp) is traditionally cultivated grain legume mainly for protein. Sterility mosaic disease (SMD) and fusarium wilt (FW) are the most destructive diseases with respect to yield in Asian and other developing countries. Simple sequence repeats (SSR) were employed to characterize minicore collections set of 135 pigeonpea accessions for host plant resistance against SMD and FW in Dharwad and Gulbarga using sick plot techniques. Based on per cent disease incidence 13 accessions comprising 6 resistant and 7 susceptible for SMD; 16 accessions comprising 8 resistant and 8 susceptible for FW were selected for molecular characterization using 100 crop specific SSR markers. Among these SSR markers, totally 59 markers generated 99 amplification products out of which 77 alleles were found polymorphic with an average of 1.68 alleles per marker for SMD. While, 68 markers generated 91 amplification products out of which 46 alleles were found polymorphic with an average of 1.34 alleles per marker for FW. The average polymorphic information content (PIC) value noticed was 0.18 for SMD and three markers viz., CcM0246, CcM0381 and CcM0399 generated three alleles each. The average PIC value noticed for FW was 0.08. The dendrogram constructed using DARwin 5.0 software distinguished the selected accessions for both SMD and FW into three clusters each. The most abundant motif’s in the present study were AT based (AT, AAT, TTAT) followed by TC class of repeats. The developed molecular marker resources will be useful for genetic diversity, mapping studies and molecular breeding applications. VI ICLGG - Posters P-HGR27 Genetic variation for morpho-physiological characters influencing seed yield in greengram (Vigna radiata (L.) Wilczek) Ashok KSM, Lavanya GR, Paul PJ*, Reddy PA, Suresh BG Department of Genetics and Plant Breeding, Sam Higginbottom Institute of Agriculture, Allahabad, India *E-mail: pronobpaul@gmail.com Abstract In a field experiment 83 genotypes of green gram were evaluated during kharif, 2010 to study the genetic parameters for different morpho-physiological characters and their correlation with the seed yield. Out of total 28 characters, harvest index, seed yield per plant, LA at 25 DAS, LAI at 25 DAS and RGR at 45 DAS showed high GCV and PCV whereas primary branches per plant, pod length, TDM at Harvest, TDM at 45 DAS, BMD at 45 DAS and CGR at 25 DAS showed high values of heritability. Harvest index, primary branches per plant, NAR, CGR at 45 DAS, LAI AT 25 DAS and LA at 25 DAS had high genetic advance. Seed yield per plant exhibited significant positive association with pod yield per plant, days to maturity, harvest index, pods per plant, seeds per pod, clusters per plant and RGR at 45 DAS-Harvest at genotypic level and with seeds per pod, 100- seed weight, pods per plant, pod yield per plant and clusters per plant at phenotypic level, should be given priority during selection for greengram for yield improvement. VI ICLGG - Posters Genetic resources of grain legumes in the Indian Himalayan region P-HGR28 Rana JC National Bureau of Plant Genetic Resources Regional Station, Shimla, India E-mail: ranajc2003@yahoo.com Abstract Grain legumes such as Phaseolus, Pisum, Lens, Vicia, and Vigna possess potential genes for resistance to biotic and abiotic stresses, adaptation and nutritional properties. Kidney bean has genetic variability for seed and pod colour, size, shape and its red colored small seeded strains are popular for their taste and cooking quality. Horse gram is adapted to rainfed & less fertile lands and carries lot of medicinal significance. Rice bean has high seed and green pod yield; possess resistance to major diseases of other Vigna species and a good substitute for V. mungo in the sub-Himalayan areas where incidences of foliar diseases are more. Among wild relatives, Vigna sublobata is highly resistant to bruchid and YVMV; V. vexillata has high protein and resistance to cowpea pod sucking bug and buchids; Cicer microphyllum occur in cold deserts may have genes for cold and drought, pod length and number of seeds/pod; Pisum sativum var. arvense have useful variability for powdery mildew resistance and cold tolerance; Cajanus platycarpa is closely related to C. cajan and is photo-insensitive, highly resistant to Phytophthora bight and reniform nematode, pod borer and Macrotyloma sar-garhwalensis has high protein content. Others species like Cicer macranthum, C. soongaricum, Lathyrus aphaca, Moghania vestita, M. bracteata, Mucuna capitata, Vicia sativa are also occurring here and needs more study. The outstanding properties of these genetic resources may lead to discovery of new genes for efficient harnessing of genetic resources of grain legumes in the Indian Himalayan region. b VI ICLGG - Posters P-HGR29 Morphological and molecular characterization of pigeonpea [Cajanus cajan (L.) millsp.] genotypes 1,* 1 2 Rani KJ , Oinam R , Anuradha Ch , Keshavulu K 1 1 Department of Seed Science and Technology, ANGRAU, Hyderabad, India. Institute of Biotechnology, ANGRAU, Hyderabad, India. 2 *E-mail: jhansisagar@yahoo.com Abstract Pigeon pea [Cajanus cajan (L.) Millsp] is an important protein rich pulse crop of the semi-arid tropics. Varietal characterization is essential to carryout scientific seed production and certification to enforce proper quality control. The objective of the present investigation was to characterize 40 genotypes of pigeon pea using morphological characters and molecular markers and to carry out the cluster analysis and grouping of genotypes by using 21 morphological traits and 54 SSR markers. Plant height, days to 50% flowering and maturity and 100 seed weight were more useful for characterization of genotypes. The genetic diversity analysis resulted in the grouping of the genotypes into seven clusters. Out of 54 SSR markers, 22 were polymorphic. The polymorphic information content for these primers ranged from 0.609 to 0.929.The cluster analysis based on Jacquard’s similarity coefficient using UPGMA grouped the genotypes into seven major clusters. The similarity coefficient among the genotypes ranged from 0.20 to 0.82. The cluster analysis based on morphological and SSR data did not show perfect congruence in clustering patterns. However, a very low correlation existed (r = 0.0117) and the SSR markers provided more definitive separation of clusters indicating their better discriminating power than morphological traits. Based on the cluster analysis diverse genotypes may be suggested for crossing programme and genotype specific characters or markers are useful for varietal identification. VI ICLGG - Posters Pyramiding wilt and drought tolerance for developing productive and stress tolerant chickpea genotypes following conventional approaches 1,3 2 3 P-HGR30 3 Salimath PM , Parameshwarappa SG , Shivakumar MS , Ontagodi TP , 4 4 3 3 5 Gaur PM , Upadhaya HD , Basamma K , Shridevi BM , Biradar SS 1 University of Agricultural Sciences (UAS), Shivamoga, India AICRP on Sesame and Niger, MARS, UAS, Dharwad, India 3 Department of Genetics and Plant Breeding, UAS, Dharwad, India 4 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 5 AICRP on Wheat, MARS, UAS, Dharwad, India 2 *E-mail: salimathpm@gmail.com Abstract Chickpea is an important cool season food legume contributing immensely in fighting malnutrition. Drought and wilt are the two major constraints limiting productivity. The objective of this study was to initiate a sound and scientifically based breeding programme to develop productive and stress tolerant chickpea genotypes. Critical evaluation of minicore collections of chickpea for the target traits helped in identifying ICC 13124 as a promising drought tolerant source. By virtue of its relatively higher root mass (21.4% more than ICC 4958 - a well known drought tolerant source), it showed relatively less yield reduction under drought (18.9%) as compared to that of ICC 4958 (26.9%). Based on desirable combining ability of ICC 13124 and its diversity with WR-315 a well known wilt resistant source, the cross was effected between than to develop a breeding population. A large F 2 population was evaluated for productivity and its component traits. A wide variation was -1 observed for yield plant (4.3 to 71.8g vs 31.2g of best parent), 100 seed -1 weight (13.8 – 34.2g vs 32.4g of best parent) and pods plant (20.0 – 102.7 vs 44.7 of best parent). Productive segregants were used to generate F 2-3 progenies. They were evaluated simultaneously for productivity, tolerance to drought and resistance to wilt, in separate experiments using appropriate environments. From a large number of progenies, two productive progenies with high level of drought tolerance and wilt resistance were identified. The study exemplifies the possibility of developing productive and stress tolerant lines by following systematic breeding approaches and selection schemes. VI ICLGG - Posters P-HGR31 Pre-breeding for grain legume improvement Sharma S*, Upadhyaya HD, Reddy KN, Singh S, Gowda CLL International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India *E-mail: Shivali.Sharma@cgiar.org Abstract Low use of germplasm (<1%), especially landraces and wild relatives in crop improvement is one of the most important constraints resulting in narrow genetic base of crop cultivars. However, the major limitation in utilizing these accessions in crop improvement is associated with their poor adaptability and other undesirable associations/linkage drag. This necessitates pre-breeding for the introgression of desirable genes from such un-adapted cultivated and wild germplasm into genetic backgrounds readily used by the breeders with minimum linkage drag. Trait-specific germplasm accessions for important biotic/abiotic stresses and for agronomic and nutrition-related traits in chickpea, pigeonpea, and groundnut have been identified using mini core (10% of core or 1% of entire collection) collection approach. Besides, wild Cicer species possessing high levels of resistance to Ascochyta blight, Botrytis grey mold, Fusarium wilt, and Helicoverpa pod borer; wild Cajanus species having sterility mosaic disease, Phytophthora blight, Fusarium wilt, pod borer, and pod fly resistance; and wild Arachis species having late leaf spot, and rust resistance have been identified. Pre-breeding activities using both cultivated and wild relatives and popular varieties have been initiated at ICRISAT to develop new genepools with high frequency of useful genes, wider adaptability, and a broad genetic base. Further, the availability of molecular markers will greatly assist in reducing linkage drag and increasing the efficiency of introgressions in pre-breeding programs. VI ICLGG - Posters Double synthetics of groundnut Shilpa K 1, 2 , Mallikarjuna N P-HGR32 1 1 International Crop Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 2 Jawaharlal Nehru Technological University, Hyderabad, India *E-mail: krishnashilpa.11@gmail.com Abstract Groundnut (Arachis hypogaea L.), an amphidiplopid, has two genomes A and B. There is some evidence now that one or a few doubling events of a diploid hybrid between A. duranensis (A genome) and A. ipaensis (B genome) gave rise to the tetraploid groundnut. Hence groundnut is susceptible to a range of disease and pests, owing to its narrow genetic base. A major initiative at ICRISAT to recreate new sources of tetraploid groundnut has resulted in the creation of a range of autotetraploids and amphidiploids by combining A with A or B with B genome species or between A and B genome species. More recently crosses were made between two tetraploids to create double synthetics which would have a minimum of three and a maximum of four Arachis species. Meiotic analysis of double synthetics showed minor variation in chromosome configuration in metaphase, anaphase, tetrad formation in some synthetics but in all of them pollen fertility was high, thus showing that creation of double synthetics is a good approach to create new sources of tetraploid groundnut. b VI ICLGG - Posters P-HGR33 Vascular tissue-specific down-regulation of lignin biosynthetic pathway genes improves the wood quality of Leucaena leucocephala for paper and biofuel production 1,2,* Singam P 1 , Narasu ML , Kavi PBK 2 1 Jawaharlal Nehru Technological University, Hyderabad, India Osmania University, Hyderabad, India 2 *E-mail: prashantsingam@gmail.com Abstract Lignin hinders the optimum utilization of cellulosic biomass in paper and biofuel industry. Reduction of lignin could be achieved through down-regulation of monolignol biosynthetic pathway genes. cDNA clones of cinnamoyl CoA reductase (LlCCR) and cinnamyl alcohol dehydrogenase (LlCAD) were isolated from Leucaena leucocephala, a leguminous pulpwood tree exclusively used for paper production in India. Sense and antisense constructs of LlCCR and LlCAD under CaMV 35S constitutive promoter were introduced in tobacco to up and down-regulate CCR and CAD genes. All down-regulated lines displayed stunted growth and development with significantly reduced lignin content. Contrarily, up-regulated lines exhibited robust growth, development and significant increase in lignin content. These observations support a role for CCR and CAD down-regulation in improving wood properties of L. leucocephala. Such attempts under constitutive promoters are deleterious to the plant due to pleotrophic effect. A 795 bp CCR promoter and 1882 bp CAD promoter were isolated from L. leucocephala by genome walking approach and sequence analysis revealed the presence of cis-elements like AC boxes, XYLAT, WRKY and MBS. Promoters were fused to β-glucuronidase (GUS) reporter gene in pBI101 and introduced in tobacco. Histochemical observations indicated expression of GUS in the vascular tissues of leaf, stem and root of transgenic tobacco. Down-regulating CCR and CAD genes under such vascular tissue specific-promoter could be sufficient for reducing the lignin content only in vascular tissues of L. leucocephala, overcoming the risks associated with reduction of lignin content across the whole plant.. VI ICLGG - Posters Pattern of variation in common bean (Phaseolus vulgaris L) genetic resources of Jammu and Kashmir 1,* 1 2 2 2 2 P-HGR34 2 Sofi PA , Zargar MY , Arifi U , Birjees J , Rozy R , Mudasir A , Suhail A , 2 2 Kumar P , Hafizullah 1 Regional Research Station (SKUAST-K), Sopore, India Faculty of Agriculture (SKUAST-K), Sopore, India 2 *E-mail: parvazesofi@gmail.com Abstract Two hundred eighty seven genotypes of Rajmash collected from different potential areas of variability and three checks viz., Shalimar Rajmash-1, Canadian Red and Selection-3 were evaluated in an Augmented Block Design during 2011. Morphological characterization revealed that the accessions could be grouped into two distinct classes for use category, pigmentation on the neck of flower and pod tip. Maximum numbers of classes were observed in case of pod colour (9) followed by Seed coat colour (8), primary color of wing petal (6), seed coat pattern (5) and primary color of standard petal (5). Highest frequency was recorded in case of stripes on neck of flower (91.38%) followed by pigmentation on neck of flowers (88.62%), pod colour at maturity (78.96%), pod colour (76.89%) and colour of stem (73.79%). Mean performances of accessions was less than the checks for days to flowering, days to maturity, seeds per pod, 100-seed weight and seed yield per plant whereas reverse was recorded for plant height, pods per plant and pod length. Analysis of variance revealed that there were significant differences between checks and accessions, between accessions and between checks for all the traits. It indicated presence of substantial amount of variation among the test entries. Among various diseases screened for, the most prominently recorded disease was bean yellow mosaic virus, followed by common rust, common blight, bean common mosaic virus, curly top virus and halo blight. The least recorded diseases were alternaria leaf spot, anthracnose and angular leaf spot. VI ICLGG - Posters P-HGR35 A new successful wide cross between C. cajan (L.) Millsp. and C. lanceolatus (WV Fitgz) van der Maesen 1, 2 Sandhya S 2 , Rao MV , Mallikarjuna N 1 1 International Crop Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 2 Bharathidasan University, Tiruchirapalli, India *E-mail: sandhya.thudumu@gmail.com Abstract The cultivated pigeon pea has a narrow genetic base. Wild relatives play an important role in the efforts to broaden its genetic base. The utilization of wild species from the secondary gene pool are important as they are presumably closely related leading to normal chromosome recombination. This helps in the transfer of useful genes to cultivated gene pool (Mallikarjuna et al., 2011). In this report Cajanus lanceolatus, a wild relative from secondary gene pool, native to Australia, has desirable traits like frost and drought resistance (Sateesh kumar, 1985), was successfully crossed and a range of F 1 progeny obtained. The resultant F1 hybrids set mature pods/seeds. Hybrids had intermediate morphology sharing traits of both the parents. All the hybrids flowered profusely. Some of the hybrids were completely male sterile and some were partially fertile with the pollen fertility ranging from 35% to 50%. Meiotic analysis of the sterile plants revealed that the break down in the microsporogenesis was at the post meiotic stage after the formation of tetrads. Fertile plants formed regular bivalents with normal disjunction except for occasional asynchrony at meiosis II division. Meiotic analysis of the plants will be discussed in detail. VI ICLGG - Posters Linking of genetic improvement programmes with seed systems for enhanced production of pulses in Madhya Pradesh 1,* 2 2 Rao SK , Gaur PM , Pandey S , Nigam SN P-HGR36 2 1 Jawaharlal Nehru Agriculture University, Jabalpur, India International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Patancheru, India 2 *E-mail: skrao_jnau@yahoo.co.in Abstract Increased production and productivity of pulses can be brought by the increased availability and adoption of improved varieties, seed and production technologies. 725 improved varieties of pulses have released but about 324 varieties are in seed production chain in the country. Reorganization and strengthening of genetic improvement programmes resulted in the development of pigeonpea (TJT 501 and JKM 189), mungbean (TJM-3), uradbean (JU-86) and chickpea (JG-130, JG-11, JGK-1, JG-63, JG-412, JKG2, JAKI 9218, JKG-9, JG-226, JG-14, JG-6), having resistance to major biotic stresses and better adaptability. Genetic improvement programmes linked with seed systems had increased the availability of quality seeds. Maintenance breeding programmes of 45 varieties strengthened the Breeder Seed Production Model seed system in chickpea in Vidisha and Sagar districts, Rainfed rice fallow chickpea seed system in Jabalpur, Satna, Rewa and Damoh in chickpea (86,620 qtls) seed village systems 553,815 qtl/year, in majority of the districts in pulses, farmers participatory seed production seed cooperatives District Poverty Initiative and watershed development had paid the devidents to increase the seed replacement rates (20 %)as well as productivity of pulses. Capacity building of 52,219 pulse seed growers made the farmers self reliance in quality seed. Impact of these studies has been discussed in the light of enhanced pulse production. b VI ICLGG - Posters P-HGR37 Heterosis, combining ability and gene action analysis for seed yield and its components in soybean (Glycine max (L.) Merrill) Tiwari G*, Puspendra, Singh K G. B. Pant University of Agriculture and Technology, Pantnagar, India *Email: gnjntiwari1@gmail.com Abstract Soybean is a dominant oilseed crop in the world trade accounting for about 25% of the world’s total oil and fats production. Exploitation of hybrid vigour in autogamous crop species like soybean is useful in improving yield potential of the existing cultivars by identifying potential cross combination on the basis of heterosis. Thirty soybean hybrids derived from crossing ten lines and three testers in a line x tester mating design along with their thirteen parents were evaluated to estimate heterosis, gene action and combining ability analysis for thirteen yield and yield component characters. The magnitude and direction of heterosis, heterobeltiosis and useful heterosis varied substantially from cross to cross and character to character. The cross AGS-129 x PS-1347 was found to have maximum useful heterosis, heterobeltiosis and average heterosis for seed yield per plant, dry matter weight per plant and number of pods per plant, hence selected as best cross from this study. Estimates of specific combining ability variances were higher than general combining ability (GCA) variances 2 2 2 for all the measured traits. Values of the ratio 2σ g/2σ g+2σ s (<0.30) indicated a preponderance of non-additive gene action invariably for all traits. The mean values of parents reflected their combining ability effects in general, and genotypes AGS-129, DS-74 and PS-1225 were good general combiners for yield and yield contributing traits. Based on the sca effects results, cross DS-74 x PS-1225 followed by AGS-129 x PS-1347 were found to be best cross- combinations for yield and its component characters. VI ICLGG - Posters Induced mutagenesis in cowpea (Vigna unguiculata L. Walp.) 1 2,* 1 3 P-HGR38 2 Kumawat KC , Raje RS , Ramakrishna K , Singh SJ , Geddam SB , Parihar 4 2 A ,Tyagi A 1 Department of Plant Breeding and Genetics, SKN College of Agriculture, Jobner, India 2 Division of Genetics, IARI, New Delhi, India 3 Associate Professor, ARS, Durgapura, India 4 Indian Institute of pulses Research, Kanpur, India *E-mail: rajers@rediffmail.com Abstract Induced mutagenesis was carried out in cowpea during kharif-2002 to kharif2004 to generate variability and to assess effectiveness of mutagens. Sodium Azide (SA) (0.3, 0.4, 0.5, 0.6, 0.7mM), EMS (0.15, 0.20, 0.25, 0.30 and 0.35%) and Hydroxylamine (0.096, 0.108, 0.12, 0.13 and 0.14%) were used to treat presoaked seeds of the 2 cowpea varieties (RC-19 and RC-101). M1 generation was evaluated in RBD design with 3 replications and M 2 families were evaluated as per the procedure outlined by Snedecor & Cochran and the outstanding superior M2 families were further evaluated in M3 generation in Augmented Randomized Complete Block Design with six blocks for seed yields and its components. All the 3 mutagens yielded a wide spectrum of morphological mutants in both the genotypes. Mutagen SA induced more number of superior families than the control in both the genotypes (30.6% and 33.6%). The mutagen EMS induced more number of superior families than the control in the RC-19 (26.1%) while Hydroxylamine induced 23.8% (in RC-19) and 21.3% (in RC-101) superior families than the control for seed yield per plant. The high estimate of heritability was recorded for biological yield/plant (with high GA as % of mean), number of clusters/plant and 100 seed weight (with moderate GA as % of mean). The most effective treatments were 0.096% Hydroxylamine (in RC-19) & 0.108% Hydroxylamine (in RC-101) which may be used to develop TILLING population. A mild relative mutagenic specificity among the three mutagens was observed in both the genotypes in relation to morphological mutations. VI ICLGG - Posters P-HGR39 Assessment of performance of RILs of chickpea (Cicerarietinum L.) derived from ICC 283 x ICC 8261 under rainfed and irrigated conditions 1, 2 1 1 1 Yasin M *, Gaur PM , Joshi P , Solanki S , Solanki MS , Saxena AK 1 1 R. A. K. College of Agriculture, R.V.S.K.V.V., Sehore, India International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 2 *E-mail: myasin23@gmail.com Abstract Chickpea is grown mainly in rainfed condition and terminal drought at maturity period resulted loss in seed yield. Two hundred eighty one recombinant inbred lines derived from ICC 283 x ICC 8261 were evaluated under rainfed and irrigated conditions to analyse diversity, variability and association. New transgressive variations was recorded for qualitative traits in RILs such as small leaflet size (32%), dark pink flower (2%), presence of stem pigmentation (30%), semi erect (14%) and prostrate (4%) plant growth habit, pea shape seed (37%), seed colour (28%) and few new recombination in kabuli with desi traits and vice versa also observed in many RIL which provides good selection possibility for drought tolerance traits. Significant variation was observed among RILs in rainfed and irrigated condition for seven yield attributes. In RILs wider range of variation was recorded as compared to their parents for days to flowering (43 to 83 day), days to maturity (106 to 128 days), plant canopy (27 to 53 cm), biological yield (7.83 to 38.5 g), harvest index (10.6 to 66.1%) and seed weight (6.1 to 34.8 g). In irrigated condition variation range was higher for most of the traits but in rainfed condition range for harvest index was higher that indicates few RILs were performing better under stress condition. PCV, GCV, h2, GA as % of mean magnitude and pattern for all the traits was similar in both rainfed and irrigated conditions. The best RILs was identified on the basis of selection indices under rainfed condition viz; ICCRIL-0064, ICCRIL0005, ICCRIL-0004, ICCRIL-0080, ICCRIL-0133, ICCRIL-0183, ICCRIL-0129, ICCRIL-0215, ICCRIL-0223 and for irrigated condition viz; ICCRIL-0004, ICCRIL-0185, ICCRIL-0025, ICCRIL-0129, ICCRIL-0062, ICCRIL-0197, ICCRIL-0233, ICCRIL-0238, ICCRIL-0109. VI ICLGG - Posters Response of soybean mutant lines to drought under in vitro culture and dry land P-HGR40 Yusati Z Indonesian National Nuclear Energy Agency, Indonesia E-mail: upikyuliasti@yahoo.com Abstract Response of soybean mutant lines to drought under in vitro culture and dry land, Improvement of growth duration of mutant lines soybean through induced mutation. Mutation technique combined with in-vitro culture through evaluation and somaclonal variation selection could increase genetic variation of Soybean (Glycine max). The objectives of this research are to evaluated responses of of soybean mutant lines to drogh under in vitro and dry land. Results revealed that G4 mutant line also produced more dry weight of plant than that of Pandermen national control of variety, when they were grown on medium containing PEG 15% concentration under in vitro condition. Based on the evaluation of physiology analysis of prolin and glucose results were identified G1 and G2 mutant lines of soybean had higher than control of parent variety (Muria). Selection of yield trials 6 mutant lines on dry land Nusa tenggara Barat (central Indonesia), the result showed G2 had higher yield than parent Muria and control national variety Panderment. b VI ICLGG - Posters P-HGR41 Development of pigeonpea (Cajanus cajana L) genotypes for changed climatic condition Muniswamy S*, Dharmaraj PS, Yamanura, Girish G, Sheela D Agricultural Research Station, Gulbarga, India *E-mail: munis01@rediffmail.com Abstract In the previous years there is delayed onset of monsoon and less rainfall with intermittent drought periods. Hence due to moisture stress long and medium duration varieties like ICPL-87119 (Asha), BSMR-736, ICP-8863(Maruti) performed very poorly. Farmers of northern Karnataka demanding pigeonpea variety for changed climatic condition. More than 80% of tur area is under rainfed condition. Therefore breeding of Pigeonpea genotypes that fits to changed climatic condition was initiated at ARS, Gulbarga by making cross between GC-11-39 x WRP-1. Pedigree method of breeding was followed after hybridization. In F2 generation, more than 800 individual plants were raised along with parents. Those, which are showed early maturity of 150 days duration, fusarium wilt resistance, high yield, seed size and were advanced to next generation. The selection for above traits continued up to F6 generation. Finally, 50 homozygous F6 families, which had early maturity, fusarium wilt resistance, high yield and seed size were selected and evaluated in station trails. The resultant genotype has been named as GRG-333 and same was sent for evaluation in AICRP trials for yield and yield attributes. The said variety performed well in multi locations as well. The main features of the variety GRG-333 is, it matures in 150-160 days, 100 Seed Weight 11.0 gms. Yield potentiality 14-16 Qtl/Ha, resistance to Fusarium wilt disease and it has very good dhal quality characteristics. VI ICLGG - Posters A case of hyper-variable spontaneous genetic variation for earliness, seed characters and other yield contributing traits in lentil (Lens culinaris Med.) P-HGR42 * Punia SS , Ram B, Dheer M, Verma P, Koli NR, Raghuvanshi A, Jain NK Agricultural Research Station (MPUA&T), Kota, India *E-mail: sumerspunia@hotmail.com Abstract The extent of genetic variation obtained naturally is very useful in self pollinated crops like lentil. Though, the valuable variation is very low in frequency. During regular seed production programme in lentil, a single plant with early maturity was isolated from a commercial population of DPL-62 in the year 2007-08 assuming that it’s a mutant of DPL-62 with earliness. Interestingly, the progenies of the isolated plant showed segregation in the next generation for maturity and seed traits. Presently the material has reached in fifth generation (2011-12). Huge variation has been observed for days to flowering (38-66 days), days to maturity (95-135), reproductive phase (55-80 days), plant height (35-65 cm), pods per plant (30-185), 100-seed weight (1.25-9.15 g), seed coat colour and other traits. Though, the population size was small, but the extent of variation observed for seed traits and other agro-morphological traits was very high representing the maximum diversity found in cultivated lentils. This kind of spontaneous variation happens due to recombination or mutation in large. Lentil being a self pollinated cleistogamous crop the possibility of natural recombination is negligible. Mutation at multiple loci may be the possible reason. In every generation segregation has been seen for major traits, still the material is segregating shows a kind of dynamic mutation. Presence of mobile genetic elements in the functional genome may be a reason. This genetic stock can play a crucial role in understanding the genetics of the target trait and improvement of small seed Indian lentils. VI ICLGG - Posters Theme Abiotic Stress P-ABS06 Transcriptome analysis in response to gradual water deficit in Arachis wild relatives 1, 2 1 1 1 Brasileiro ACM *, Morgante C , Leal-Bertioli SC , Araujo ACG , Pappas G , 1 1,3 1 3 1 Silva Jr OB , Silva AK , Martins ACQ , Bertioli D , Guimarães PM 1 EMBRAPA Recursos Genéticos e Biotecnologia. Brasília, Brazil EMBRAPA Semiárido, Petrolina, Brazil 3 Universidade de Brasília, Brasília, Brazil 2 *E-mail: ana.brasileiro@embrapa.br Abstract Peanut (cultivated tetraploid Arachis hypogaea) is an important food legume widely cultivated mainly in Asia, Africa and the Americas although its productivity is limited in drought-prone areas; therefore the development of drought-resistant varieties is a priority. Peanut has a narrow genetic diversity and is reproductively isolated from its wild diploid relatives due to ploidy differences. In contrast to peanut, wild relatives have higher genetic diversity and show adaptation to a range of environments thus constituting a rich source of allele diversity for resistance to biotic and tolerance to abiotic stresses. In this study, the transcriptomes of two wild diploids, A. duranensis and A. magna, representatives of the AA and BB peanut component genomes respectively, were analyzed under gradual water deficit. Two SSH libraries produced from roots of A. magna generated 759 Sanger ESTs with 249 Unigenes, whilst from A. duranensis roots, 380,601 Roche/454 reads resulted in12,792 Unigenes. Forty-six Unigenes were selected as drought-related candidates for validation by RT-qPCR by in silico analysis of all libraries. A total of 18 candidate genes showed significantly different levels of expression in stressed plants, confirming their involvement in drought response. To date, this is the first report on global gene expression profile of wild Arachis species submitted to an abiotic stress. Data produced in this study is a valuable resource for gene identification, characterization of new alleles and development of molecular markers in Arachis. VI ICLGG - Posters Drought physiological and morphological response patterns in common beans for QTL mapping Briñez B 1,2,* 2 2 2 3 , Rosa J , Esteves J , Bassi D , Blair MW , Benchimol LL P-ABS07 2 1 State University of Campinas (UNICAMP), Campinas, Brazil The Agronomic Institute (IAC), Campinas, Brazil 3 Cornell University, Ithaca, USA 2 *E-mail: borisbrinez@hotmail.com Abstract Drought is one of the major factors limiting common bean (Phaseolus vulgaris L.) production in the tropics. The objective of this study was to evaluate the morphological and physiological responses of segregant population derived from the Mesoamerican drought-tolerant SEA 5 crossed to Andean droughtsusceptible AND 277 for drought QTL mapping. The experiment was conducted under two greenhouse conditions applying contrasting water supply treatments in a complete randomized experimental design with four replicates. The first greenhouse was well-watered at 80% of the field capacity and the second greenhouse was under terminal stress conditions in the vegetative phase. A total of thirty watermark sensors (granular matrix sensor) were randomly installed at soil depths of 20 cm to monitor and measured soil water in the control and drought stress pots. After 20 days of water stress, the phenotypic analyses were performed collecting canopy temperature, chlorophyll level present in the leaf, stem biomass fresh weight, leaf biomass fresh weight, stem biomass dry weight, leaf biomass dry weight and leaf area for each genotype and replication. Performances of the RILs and heritability of the traits were evaluated. Analysis of variance showed significant differences for most of the traits evaluated in drought-stress conditions. In the nonstress environment the differences were principally presented among the recombinant inbred lines and not in the parents. The results showed that the population used and the physiological traits measured for drought tolerance should be accounted for an effective QTL mapping procedure. b VI ICLGG - Posters P-ABS08 Adaptability of Vigna mungo to the semi arid agroclimate, increasing drought tolerance through salicylic acid and Rhizobium - symbiosis Pandey S, Prakash T, Mathur S, Jindal R, Chakraborty D* Banasthali Vidyapith, Banasthali, India *E-mail: dciitkgp@gmail.com Abstract Intensive agriculture techniques prevalent currently are largely dependent on the application of chemical fertilisers, herbicides and pesticides. Viable environment-friendly alternatives to these substances have been in the forefront of research of which symbiotic relationships developed in legumes and the associated PGPRs (Plant growth promoting rhizobacteria) are thought to be the most promising. Vigna mungo, a tropical leguminous plant is widely cultivated in the Indian subcontinent is popular for its high protein content, antioxidants, vitamins and minerals. In this study the role of salicylic acid and Rhizobium in conferring tolerance to drought stress and increasing adaptability to the semi-arid agroclimate was elucidated. Plants were treated with salicylic acid prior to imposition of drought stress. The treated plants exhibited better physio-biochemical characteristics, enhanced activity of the antoxidant enzymes and phenylalanine ammonia lyase and were more tolerant towards drought stress over short and long duration. Plants without Rhizobium lacked proper growth and were vulnerable to mild drought stress. The study illustrates the importance of phenolic compounds to confer drought tolerance in V. mungo and the relevance of optimum growth conditions for soil microbes for better adaptability of the plant to the semi-arid region. VI ICLGG - Posters Genotypic variations for herbicide tolerance in chickpea 1,* 2 3 4 P-ABS09 1 1 Chaturvedi SK , Gaur PM , Tripathi S , Singh S , Mishra N , Aski M , 1 Nadarajan N 1 Indian Institute of Pulses Research (IIPR), Kanpur, India International Crops Research Institute for the Semi Arid Tropics (ICRISAT), Hyderabad, India 3 Indian Agricultural Research Institute (IARI), New Delhi, India 4 Punjab Agricultural University (PAU), Ludhiana, India 2 *E-mail: sushilk.chaturvedi@gmail.com Abstract Application of post-emergence herbicides is known to reduce weeds population and ensure better crop growth. Till date, no post-emergence herbicide could be recommended to control seasonal weeds in chickpea. Efforts have been made to search gene(s) conferring tolerance to postemergence herbicides. 509 accessions were screened during 2011/12 against Imazethapyr and 40 lines rescreened for confirmation of reaction at IIPR. Genotypes viz., ICC 1164, IPC 2010-81, ICC 1161, ICC 1205 and IPC 2008-59 were found most tolerant ones as these were almost not affected and only showed inhibited growth initially. ICC 5484 were found most sensitive as it showed leaf burning within 7 days, 100% plant mortality within 25 days of herbicide application, respectively without any regrowth. Similarly, 300 accessions were also screened against Metribuzin at IARI and PAU in 201112. Genotype ICC 1710 was found tolerant at IIPR and ICRISAT against Imazethapyr, and genotype ICC 1205 was tolerant against Metribuzin at IARI and PAU locations. Four genotypes viz., ICC 1205, ICC 1161, ICC 1164 and ICC 1398 exhibited tolerance against both herbicides (Imazethapyr and Metribuzin). The crosses among most tolerant and sensitive ones have been made to map and tag the herbicide tolerant gene(s). VI ICLGG - Posters P-ABS10 Screening and development of salt tolerant lines and management for improved productivity in chickpea (Cicer arietinum L.) * Das M , Kundagrami S Institute of Agricultural Science, University of Calcutta, Kolkata, India *E-mail: mou.19may@gmail.com Abstract Saline soils are an important ecological entity worldwide.Rabi cropping with pulse crops in such soil would be beneficial for farmers. Chickpea is valued for its agronomic importance and nutritive seeds but the crop is relatively salt sensitive among the legumes. In view of the above a three way approach was under taken. Firstly, seventy one chickpea cultivars were screened in laboratory water culture method as well as in field condition, two location each -1 of normal(Baruipur & Udaynarayanpur,0 dSm salinity) and saline -1 situation(Hingalgunge and Canning,4- 6 dSm salinity).Some high yielding salt tolerant cultivars like RSG 888,WBG 29,Anuradha 39/2,IC 268873,IC 268863,G 24,B108 were selected by lab and field screening. Secondly, three relatively salt sensitive cultivars (BGM 408, B 115, JG 315) were treated with gamma ray @ the dose of 0,10,20 and 30 Kr. Some mutant high yielding lines were selected from M3 generation and their salinity tolerance was tested in laboratory and as well as in field condition as stated earlier. Some promising high yielding salt tolerant mutant (M4) was selected. Proper management practices are essential for successful crop establishment in saline soil. So, lastly, seeds of chickpea variety Mahamaya 2 were treated with nineteen different herbal materials and their salinity tolerance was tested in laboratory and field condition. The treatments like turmeric, chilli, coriander, bittle vine, lemon, neem exerted beneficial effects on chickpea seeds when they were exposed to a given salinity stress. Salinity tolerances of the treated seeds were further examined by applying Hoagland’s nutrient solution. Growths of the plants were improved in salt stressed condition when supplemented with nutrient solution. VI ICLGG - Posters Study on drought tolerance in groundnut (Arachis hypogaea L.) P-ABS11 Pavithra DS, Manivannan N*, Vindhiya VP, Ganesamurthy K Tamil Nadu Agricultural University, Coimbatore, India *E-mail: nmvannan@gmail.com Abstract Fifty genotypes of groundnut including released varieties and advanced breeding lines were used to assess the drought tolerance. All these genotypes belong to Spanish bunch habit group. These genotypes were evaluated both in managed drought stress and non stress situations during post rainy season (Jan- April), 2011 at Oilseeds Farm, Tamil Nadu Agricultural University, Coimbatore, Tamil Nadu, India. The drought stress was imposed by withholding irrigation after 60 days after sowing for the drought stress trial. Observations were recorded for pod yield per plant and kernel yield per plant on randomly chosen five plants per entry per replication. The data were subjected to ANOVA and drought tolerance parameter viz., drought susceptibility index (DSI) for pod yield per plant and kernel yield per plant. The results indicated that genotypes differed significantly for pod and kernel yield per plant. Among the genotypes, 11 and 9 genotypes were observed as highly drought tolerant based on pod and kernel yield per plant respectively. A total of 6 and 10 genotypes recorded as highly susceptible to drought for pod and kernel yield per plant respectively. Among these genotypes, each three drought tolerant (ICGV 91114, K1375 and ICGV 02125) and drought susceptible (ICGV 01279, ICGV 98170 and ICGV 98175) genotypes were selected for hybridization programme to develop mapping populations. VI ICLGG - Posters P-ABS12 Discovery of conserved domains for development of functional markers and Identification of microRNAs from drought responsive expressed sequence tags (ESTs) in Chickpea (Cicer arietinum L.) 1,* 2 3 3 Mandape M , Kompelli SK , Dudhe MY , Kumar J , Meena HP 4 4 J , Afroz A 3,# , Chauhan 1 Post Graduate Institute, ANGRAU, Hyderabad, India Indian Agriculture Research Institute, New Delhi, India 3 Directorate of Oilseeds Research, Hyderabad, India 4 Banasthali University, Banasthali, India #Present Address: Directorate of Oilseeds Research, Hyderabad, India 2 * E- mail: mrinalimandape@gmail.com Abstract Domains can be thought of as distinct functional and structural units of protein. We have analyzed drought responsive expressed sequence tags (ESTs) for the identification of conserved domains in chickpea (Cicer arietinum L.). The occurrence of conserved domains, super families and multi domains having putative functions like insect pest resistance, improved quality parameters, resistance to abiotic stresses viz., temperature, drought, salinity or other evolutionary functions and physiogenic nature have been identified. Identified conserved domains or gene families from the ESTs could be utilized for the identification of gene-targeted markers and for development of functional markers to accelerate marker assisted breeding in chickpea. Also we conducted a mining of miRNA genes in chickpea using computational approaches. Based on homology and secondary structure analysis potential miRNA targets have been found for newly identified chickpea conserved miRNAs. These target genes included number of transcription factors that control plant growth, development and metabolic enzymes, as well as other genes involved in stress response. miRNAs and their target genes can initiate further study on chickpea miRNA regulation mechanisms and it can help in knowing more about the important role of genes. These findings suggest that computational analysis is a good alternative strategy for identifying new conserved domains, miRNA candidates and their target genes in plants. VI ICLGG - Posters The ecological and genomic basis of salinity adaptation in Tunisian Medicago truncatula 1,*,# 2,3 4 P-ABS13 4 Friesen ML , Eric JB, von Wettberg , Badri M , Moriuchi KSS , Barhoumi 4 5 1 5,6 1 4 F , Cuellar-Ortiz S , Chang PL , Cordeiro MA , Wendy TV , Arraouadi S , 4 4 4 7,8 4 5 Djébali N , Zribi K , Badri Y , Porter SS , Aouani ME , Cook DR Strauss 7 1 SY , Nuzhdin SV 1 Section of Molecular and Computational Biology, Department of Biological Sciences, University of Southern California, Los Angeles, USA 2 Department of Biological Sciences, Florida International University, Miami, USA 3 Center for Tropical Plant Conservation, Fairchild Tropical Botanical Garden, Coral Gables,Florida, USA 4 Centre of Biotechnology of Borj Cedria, Hammam-Lif 2050, Tunisia 5 Department of Plant Pathology, University of California Davis, Davis, USA 6 Instituto de Tecnologia Química e Biológica, Oeiras, Portugal 7 Department of Evolution and Ecology, University of California Davis, Davis, USA 8 Center for Population Biology, University of California Davis, Davis, USA #present address: Department of Plant Biology, Michigan State University, East Lansing, USA *E-mail: maren.l.friesen@gmail.com Abstract As our world becomes warmer, agriculture is increasingly impacted by rising soil salinity and we need to understand trade-offs that limit plant adaptation to salt stress to enable effective crop breeding. Salt tolerance is a complex plant phenotype and we know little about the pathways utilized by naturally tolerant plants. Legumes’ capacity for symbiotic nitrogen fixation makes them cornerstone species in agricultural and natural ecosystems but they are particularly vulnerable to salt since symbiosis is salt sensitive. Our studies of the model legume Medicago truncatula in natural field and controlled greenhouse settings demonstrate that Tunisian populations are locally adapted to saline soils. Whole genome re-sequencing of 39 wild accessions reveals that despite complex population structure and reduced sequence polymorphism within saline populations, a small number of candidate genomic regions assort non-randomly with saline source population. Saline alleles deviate more often from the range-wide allele frequency observed in the Medicago HapMap lines and they are more often derived relative to the sister species Medicago littoralis, consistent with the action of natural selection. These candidate regions for adaptation contain genes that regulate physiological acclimation to salt stress, such as abscisic acid and jasmonic acid signaling, including a novel salt-tolerance candidate orthologous to the uncharacterized gene AtCIPK21. Unexpectedly, these regions also contain biotic stress genes, including a NB-LRR gene, and flowering time pathway genes, including CONSTANS. We show that flowering time is differentiated between saline and non-saline populations and may allow salt stress escape. Results utilizing an independent set of lines in a single population and the role of rhizobium strains in salinity adaptation will also be presented. VI ICLGG - Posters P-ABS14 Role of arbuscular mycorrhizal fungi in the development of salt tolerance in Cajanus cajan (L.) Millsp. under salinity stress * Garg N , Pandey R Panjab University, Chandigarh, India * E-mail: gargneera@gmail.com Abstract Pigeonpea (Cajanus cajan (L.) Millsp.) is an important legume crop, grown in the tropics and sub-tropics. A pot experiment was conducted to examine the effects of three arbuscular mycorrhizal fungi, indigenous mycorrhizal mix (Mi), Glomus mosseae (Gm), Glomus intraradices (Gi), and a mix of G. mosseae and G. intraradices (Gm+Gi), on growth and nutrient acquisition of pigeonpea plants under salt stress. Two genotypes (Paras, Pusa-2002) were subjected to four levels of salinity (0, 40, 60 and 80 mM NaCl) with and without mycorrhizal inoculations. While salinity reduced plant biomass in both genotypes, mycorrhization caused an increase in root and shoot dry weights at all salinity levels. The mycorrhizal dependency of pigeonpea plants increased with an increase in salinity, with the sensitive genotype showing a higher dependency than the tolerant one. Mycorrhizal colonization decreased with increasing salt stress; Gi and Gm+Gi showed high colonization even at increased levels of + 2+ salinity. Mycorrhization alleviated salt-induced reduction of N, P, K and Ca uptake. Presence of AM in roots prevented excess uptake and translocation of + + Na with increasing levels of salinity, thereby maintaining lower Na shoot: root + ratios and a balanced mineral nutrition. The reduction in Na uptake with a + 2+ concomitant increase in N, P, K and Ca absorption may be important saltalleviating mechanisms in mycorrhizal pigeonpea plants. Different AMF species differed in their ability to minimize stress effects and promote growth. G. intraradices (Gi) alone or in combination with G. mosseae (Gm+Gi) showed highest efficiency in terms of growth and nutrient acquisition. VI ICLGG - Posters Responses of the jasmonate pathway upon stress: genes and metabolites 1,* 2 1 Grunz F , Rotter B , Kaiser C , Horres R 4 1 2 A , Kahl G , Winter P 1,2 1 P-ABS15 3 , Fatnassi N , Imtiaz M , Santino 1 Molecular BioSciences, Johann Wolfgang Goethe University, Frankfurt am Main, Germany 2 GenXPro GmbH, Frankfurt Innovation Center FIZ Biotechnology, Frankfurt am Main, Germany 3 International Center for Agricultural Research in the Dry Areas (ICARDA), Aleppo, Syria 4 Institute of Sciences of Food Production C.N.R. Unit of Lecce, Italy *E-mail: fgrunz@stud.uni-frankfurt.de Abstract Plants growing in less than ideal conditions are considered to be under stress. Suboptimal growth conditions can be caused by environmental challenges such as heat, cold, drought, lack of nutrients, high salinity, radiation, as well as exposure to toxic chemicals, and in crop plants these adversities account for a significant fraction of the discrepancy between potential and effective yield. The food legume chickpea (Cicer arietinum) is one of the most important agricultural crops worldwide, and since it is mostly grown in arid and semi-arid regions, potential and real harvest yields differ significantly, especially because of drought stress. As a consequence, improving dehydration stress tolerance in chickpea is important to optimize production. Due to its crucial role in water and mineral uptake, dehydration is first perceived by the root tips, where abiotic stresses elicit early signals affecting root growth and root-shoot communication. To gain new insights into the molecular mechanisms of drought tolerance we investigated dehydration stress-induced gene expression along with metabolite levels in root tips from drought-tolerant as well as -sensitive chickpea varieties, and found distinct expression patterns in both. Stress-induced expression of specific lipoxygenase Mt-lox1, hydroperoxide lyases Mt-hpl1 and Mt-hpl2, allene oxide synthase Mt-aos, and oxophytodienoate reductase Mt-opr was more persistent and also initiated at an earlier point of time in the drought-tolerant varieties, resulting in increased synthesis of jasmonic acid, a well-known stress signaling molecule. The levels of major oxylipin metabolites from the AOS branch of the oxylipin pathway were positively correlated with the respective gene expression patterns. b VI ICLGG - Posters P-ABS16 Screening of groundnut (Arachis hypogaea L.) genotypes for salinity tolerance under natural saline soils 1,* 1 2 1 1 Kuchanur PH , Bellakki MA , Katkar GD , Kattimani SH , Patil N , 3 1 4 5 1 Nadagouda BT , Balagavi SB , Badigannavar AM , Gowda MVC , Patil SG 1 University of Agricultural Sciences, Raichur, India University of Mysore, Mysore, India 3 University of Agricultural Sciences, Dharwad, India 4 Bhabha Atomic Research Center, Mumbai, India 5 Indian Council of Agricultural Research, Bangalore, India 2 * E-mail: prakashkuchanur@yahoo.co.in Abstract Peanut (Arachis hypogaea L.) is a major oilseed and food crop, grown across the world. Salinity in agricultural land is alarmingly increasing with irrational use of water resources. It affects plant growth at all stages and results in low yield. Therefore, our goal was to identify salinity tolerant groundnut genotypes. Three hundreds genotypes (minicore germplasm, Trombay Groundnut Lines H and commercial varieties) were evaluated under natural saline soils (p 8.4 -1 and ECe 4 dSm ) of Tunga Bhadra Project command area during summer 2009 and 2010. Most of the genotypes recorded very poor yield with low shelling percentage (SP). Among 300, the selected 36 genotypes were H -1 screened at Upper Krishna Project command area (p 8.6 and ECe 2 dSm ) during summer 2011. Among them, the pod yield ranged between 280 to 1482 -1 kg ha . The mean shelling per cent was 51 (70% is desirable) indicating that the kernel development is highly sensitive even at lower salinity. The plant mortality at 90 days after sowing (DAS) ranged from 10% to 49%. Germination percentage and radical growth (cm) of the genotypes were assessed using 125 and 150 mM NaCl solutions along with control (distilled water). The germination per cent and radical growth reduction at 150 mM over 125 mM ranged from -41.67 to 5.00 and –1.28 to 0.00, respectively. Based on pod yield, plant mortality, K/Na ratio and chlorophyll content at 55 and 70 DAS, GPBD5, ICG3673, TGS110 and ICG12189 were identified as relatively saline tolerant. Further, the identified lines could be used to understand the inheritance of salinity tolerance and identify QTLs for marker assisted selection. VI ICLGG - Posters Variability for root traits and their relation with drought tolerance in lentil 1,* 2 1 1 1 P-ABS17 3 Kumar J , Basu PS , Srivastava E , Chaturvedi SK , Nadarajan N , Kumar S 1 Division of Crop Improvement, Indian Institute of Pulses Research, Kanpur, India 2 Indian Institute of Pulses Research (IIPR), Kanpur, India 3 International Center for Agricultural Research in the Dry Areas (ICARDA), Aleppo, Syria * E-mail: jitendra73@gmail.com Abstract Lentil is one of most important pulse crops in South Asia, and invariably encounters terminal moisture stress, thus, leading to forced maturity and lower yield. Long and prolific root system is known to enhance capacity of the plant to extract water from the lower soil strata and thus help avoid the water stress. Therefore, in the present investigation, we assessed genetic variation for 14 traits among 43 lentil genotypes comprising improved varieties and promising breeding lines. The average root length at the 65-day plant stage ranged from 41.99 to 82.83 cm. Two genotypes (EC 208362, and VKS 16/11) for shorter root length and poor dry root weight (DRW) and three genotypes (DPL 53, JL 1 and IPLP 98/193) for longer root length and high DRW were identified with stable performance over the years. SPAD value showed significantly positive correlation with DRW (r=0.45**) and root length (r=0.44**) and thus, can be used as selection criterion for phenotyping root traits which are otherwise difficult to measure in fields. Under drought-prone environments, early flowering, pod initiation, maturity, and seedling vigour and high SPAD value, biological yield, and harvest index were identified as key traits for higher seed yield in lentil. Our results revealed significant genetic variability for these traits in lentil germplasm. It has been shown that Indian genotypes adapted to rainfed conditions have longer roots and higher DRW. Three donors (DPL 53, JL 1 and IPLP 98/193) identified for the root traits were either originated from or have in their ancestries at least one parent adapted to rainfed conditions. These donors can be utilized for the development of mapping population to identify QTLs associated with these traits for marker assisted breeding of drought tolerant high yielding varieties in lentil. VI ICLGG - Posters P-ABS18 Identification and annotation of abiotic stress responsive candidate genes in peanut ESTs Kumari A, Kumar A, Wany A, Prajapati GK, Pandey DM* Birla Institute of Technology, Ranchi, India *E-mail: dmpandey@bitmesra.ac.in Abstract Peanut (Arachis hypogaea L.) ranks fifth among the world oil crops and is widely grown in India, China, Nigeria and United States. Due to its large and unknown genome size, studies on genomics and genetic modification of peanut are still scanty compared to other model crops like arabidopsis, rice, cotton and soybean. Because of its favourable cultivation in semi-arid regions, study on abiotic stress responsive genes and its regulation in peanut is very much important. Therefore, we aim to identify and annotate the abiotic stress responsive candidate genes in peanut ESTs. Expression data of drought stress responsive corresponding genes and EST sequences were screened from dot blot experiments shown as heat maps and supplementary tables, respectively as reported by Govind et al. (2009). Some of the screened genes having no information about their ESTs in above mentioned supplementary tables were retrieved from NCBI. A phylogenetic analysis was performed to find a group of utmost similar ESTs for each selected gene. Individual EST of the said group were further searched in peanut ESTs (1,78,490 whole EST sequences) using stand alone BLAST. For the prediction as well as annotation of abiotic stress responsive selected genes, various tools (like Vec-Screen, Repeat Masker, EST-Trimmer, CAP3 and WISE2) were used and obtained result will be presented. . VI ICLGG - Posters Genotypic responses for biochemical, physiological and biological nitrogen fixation traits under water-deficit stress in advanced breeding lines of groundnut (Arachis hypogaea L.) P-ABS19 Chuni Lal*, Rupapara KV, Chikani BM, Gor HK Directorate of Groundnut Research, Junagadh, India *E-mail: chunilal@nrcg.res.in Abstract Drought stress remains an ever-growing problem that severely limits crop production worldwide and causes important agricultural losses particularly in arid and semiarid regions. Thirteen advanced breeding lines of groundnut were tested in summer 2012 at Directorate of Groundnut Research, Junagadh along with three checks under irrigated and simulated water-deficit conditions. Highly significant differences due to genotypes were observed for most of the traits. Withholding of irrigation had brought the plants under water-deficit stress as was revealed by the significant reduction in relative water content, RWC of leaves of stressed plants. Highly significant differences in variances due to irrigation treatments were found for specific leaf area, SLA; leaf growth rate, LGR; relative leaf growth rate, RLGR; net assimilation rate, NAR; RWC, number of active nodules, and contents of proline, wax and chlorophyll a, Chl a. Significant increase was observed under water-deficit stress for traits namely, NAR, proline, wax and Chl a. The remaining traits registered significant decrease in response to water-deficit conditions. Significant interactions due to genotypes x irrigation treatments were found for most of the traits. Though there was significant decrease in the magnitudes of traits related to biological nitrogen fixation under water-deficit stress, however, interactions due to genotypes x irrigation treatments observed for these traits suggest that some of the groundnut genotypes will have stable or even higher values under water-deficit stress conditions. Drought susceptibility index worked out on the basis of pod yields revealed that the advanced breeding line PBS 15031 was the most tolerant one followed by PBS 16038 and PBS 30055. VI ICLGG - Posters P-ABS20 Screening of algerian Medicago truncatula lines under salt stress 1,* 2 2 Laouar M , Lahreche NH , Abdelguerfi A 1 INRA/ENSA Algérie, El-Harrach Alger ENSA/INRA Algérie, Belfort El-Harrach 2 *E-mail: Laouar_m@yahoo.fr Abstract M. truncatula is a model plant of legume genomics. The study of the diversity of this species is important in this sense we study the behavior of this species under salt stress.34 lines, native of Algeria and parental lines of different genetic map, were grown on tubes (culture medium) during 37 days at different salt concentration (0, 50, 100, 150, 200 mM of NaCl).We estimate growth of all seedlings by measuring length and total biomass production of fresh and dry matter of the shoot and root. The relative water content (RWC) and some metabolites within ten lines and four increasing dose of salt are studied. The results obtained show a variability of response depending on the genotype and NaCl concentration applied. Indeed, the growth and development of all lines are affected by the concentrations 150 and 200 mM. 10 Algerian genotypes with TN1.11 and DZA315.16, tolerate the presence of 50 mM sodium chloride. 100 mM treatment has a negative effect on the growth of most lines, except for a few lines especially ALG405.340 and ALG331.252. The results showed that salt stress, leads in most lines studied to the accumulation of proline, a slight reduction of RWC, and a significant decrease in chlorophyll a, b, total and Carotenoïds. Medicago truncatula lines ALG331.252, TN1.11, ALG440.439, relatively salt tolerant have adapted different strategies of resistance according with difference rate of biochemical compounds synthesis (proline and chlorophyll pigments). VI ICLGG - Posters Role of a serine-threonine protein kinase from chickpea and its homologue in Arabidopsis in root development P-ABS21 Meena MK, Ghawana S, Chattopadhyay D* National Institute of Plant Genome Research, New Delhi, India *E-mail: debasis@nipgr.ac.in Abstract Calcium has emerged as a ubiquitous second messenger to a multitude of plant response to external stimuli and regulates a wide range of physiological processes. Apart from calcium dependent protein kinases (CDPK), a family of ser/thr protein kinases that also harbour calcium-binding domains; calmodulin (CaM) and CaM-like proteins, recently a group of calcium-sensing proteins have been identified in plants and named as calceneurin B-like proteins (CBL). Genetic and reverse genetic methods showed that different CBLs are responsible for different signaling processes. CIPKs (CBL-interacting protein kinases), a family of serine-threonine kinases represents targets of calcium signals sensed and transduced by CBL proteins. Mostly CIPKs are known to have role in abiotic stress (salt and dehydration) tolerance. Moreover, in depth study of CIPKs is restricted to Arabidopsis only. Two ESTs that code for two putative CBL-interacting protein kinases were identified in a population of ESTs, which showed high expression in the drought-tolerant chickpea cultivar in response to water deficit. These two CIPKs were named as CaPKS1 and CaPKS2. CaPKS1 was reported previously. CaPKS1 over-expressing transgenic tobacco showed longer root length phenotype compare to wild type. In this report we are discussing the possible role and function of CaPKS2. VI ICLGG - Posters P-ABS22 Root traits as a rapid and reliable screening technique for drought tolerance in chickpea (Cicer arietinum L.) 1,* 1 1 2 Bharathi S , Patil JR , Mannur DM , Basu PS , Singh NP 1 2 2 Agricultural Research Station, (UAS Raichur), Gulbarga, India Indian Institute of Pulse Research (IIPR), Kanpur, India *E- mail:bharathi_bup@rediffmail.com Abstract Drought is the most common abiotic stress limiting Chickpea production in the world. Ninety per cent of the world’s chickpea is produced in areas with receding soil moisture. Therefore, productivity is dependent on efficient utilization of available soil moisture through better root system. Thirty released chickpea varieties were grown in PVC cylinders (90 cm height) filled with vertisol with initial soil water content, 100% field capacity during 2011 postrainy season. The same chickpea varieties were also grown in an experimental field under rainfed condition. Genetic parameters were estimated for root and yield traits. The high range for seed yield (480 - 1400 kg/ha), root shoot ratio (0.4 - 2.03), root length (33 – 116.84 cm) and TDM (22.89 – 82.32 g/plant) indicated the variability among the entries. The observed narrow difference between the genotypic coefficient of variation (GCV) and phenotypic coefficient of variation (PCV) is due to the genetic makeup of the varieties showing lower 2 GxE interaction. Further, high heritability (h ) values for root weight/plant (84.25%), root shoot ratio (84.74%), 100 seed weight (82.72%) and shoot weight (81.80%) indicated the larger part of phenotypic variation is due to genotypic variance with less GxE interaction. The genetic advance values were higher for root shoot ratio (69.08) root weight/plant (58.68) shoot weight (49.03), 100 seed weight (39.07) and root length (35.48). Higher genetic advance values indicate better response to selection based on root traits. Thus, PVC cylinder system can be used as rapid phenotyping tool for root traits in large scale screening of germplasm. VI ICLGG - Posters Evaluation of chickpea genotypes for soil salinity tolerance in irrigated command area 1,* 1 Patil BS , Hegde VS , Kuchanur PH P-ABS23 2 1 IARI’s Centre for Pulses Improvement, Dharwad, India College of Agriculture, Raichur, India 2 *E-mail: bs_patil2000@yahoo.com Abstract Salinity is one of the major abiotic constraints in agriculture. About 20% of cultivated land is affected by salinity across all the continents (FAO 2004). With the implementation of major irrigation projects and other sources of irrigation it is predicted that, by 2050, 50% of all arable land will be salinized (Wang et al., 2003 Chickpea is one of the important grain legumes which is highly sensitive to soil salinity. Therefore, it is becoming increasingly important to produce cultivars tolerant to salinity stress for sustainable chickpea production. To identify suitable chickpea genotype for salt affected soils, field evaluation of one hundred chickpea cultivars was carried out under normal and two levels of soil salinity in irrigated command area. Effect of salinity on yield and yield related traits was studied. Large variation for grain yield was recorded in all the soil conditions. In normal soil, the grain yield ranged from 2.15 g/p (PUSA 212) -1 -1 to 19.97 g/p (PUSA 390). In moderate (1.80 dSm ) and high (3.0 dSm ) salinity condition, grain yield ranged from 0 to 9.22 g/p (ICC 92944) and 0 to 6.1 g/p (LBeG 7) respectively. About 40% of genotypes in moderate and 20% of genotypes high level of soil salinity recorded higher grain yield than previously identified salt tolerant genotype CSG 8962. Overall percent reduction of grain yield was 67% in moderate and 84.26% high salinity condition. Among the yield components, high reduction of pod and seed number was observed in both the levels of soil salinity. The results are in confirmation with Vandez et al., (2006). We found that the salinity tolerance was more related to the ability to maintain a large seed number than differences in seed size. The variation in the germplasm is sufficiently large to breed for salinity tolerance in chickpea. Further research is needed to understand the genetics of salinity tolerance and identify QTLs for marker assisted selection. VI ICLGG - Posters P-ABS24 Screening for drought tolerance through drought susceptibility Index of elite chickpea (Cicer arietinum L.) entries under natural conditions 1, 1 1 1 2 Patil JR *, Mannur DM , Bharathi S , Mahaboob S , Basu PS , Singh NP 2 1 Agricultural Research Station, Gulbarga, India Indian Institute of Pulse Research (IIPR), Kanpur, India 2 *E-mail: jrpatil_uas@yahoo.com Abstract Chickpea (Cicer arietinum L.) an important food legume grown in the semi-arid tropical suffers substantial yield loss due to drought at critical stage of the growing season. The aim of the investigation was to evaluate the genetic variability of notified varieties for drought tolerance. Thirty one released cultivars of chickpea were screened for drought tolerance in three year field experiments. The applied drought susceptibility index (DSI) considered the relations between seed yield under drought conditions (YD) and under adequate soil moisture (YIR) also their dependence on the drought severity index (DS). The application of DSI divided the cultivars into drought tolerant and drought susceptible group. The group of drought tolerant cultivars characterized by mean values of DSI < 1 comprised B 256 (0.71), Katila (0.72), JG 11(0.74), Saki 9516 (0.77) and Virat (0.77). The drought sensitive cultivars with DSI > 1 comprised of K850 (1.35), RSG 888 (1.33), RSG 143-1 (1.24), GCP 101 (1.20), ICCV 14800 (1.17), C214 (1.14) followed by Annigeri 1 (1.13). In general, the tolerant cultivars when compared with the susceptible ones were characterized by early maturity and better root system, which however was more stable both under conditions of drought and non stress conditions. Relative water content (RWC) varied from 16.13 to 50.0 under stress compared 4.55 to 33.33 under non stress indicating the variation in water status of plants under stress and non stress. The results clearly indicate that the DSI can be used as a tool for screening chickpea varieties/ lines for drought tolerance. VI ICLGG - Posters Cooler canopy temperatures at the mid-reproductive phase confers greater grain yields in chickpea (Cicer arietinum L.) under terminal drought P-ABS25 Purushothaman R, Krishnamurthy L* International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India *E-mail: l.krishnamurthy@cgiar.org Abstract Drought is the major constraint causing a yield loss of 25% worldwide. Contributory traits to drought tolerance may vary across locations and it is important to identify those traits with major contribution and high heritability for further breeding. Selection for grain yield can be deceptive as it is prone to environmental variations. Field studies were conducted during 2010-11 postrainy season to understand the contribution of canopy temperature (CT) to grain yield. Twelve genotypes, selected from the minicore chickpea germplasm for contrasts on drought tolerance, CT and rooting extent were studied. The 2 study was planted on 20 Nov 2010 on a Vertisol in 24 m plots both under drought stressed and optimally irrigated environments in three replicates. Both the ground cover and the CT were measured using an infrared camera (IRFLEXCAM, Infrared Solutions, Inc, USA) on 60, 70, 72 and 80 days after sowing (DAS) and the grain yield, shoot biomass and other yield components were measured at final maturity. Under non irrigated condition, ground cover range of the genotypes was narrow; i.e.0.88 to 0.98 on 80 DAS. Ground cover at 60 DAS alone influenced the grain yield and shoot biomass. CT was closely and negatively associated with the seed yield and shoot biomass production. Under nonirrigated condition, CT at 72 DAS explained 66% of the yield variation and under optimally irrigated conditions; it explained only 40 to 51% of the yield variation. These results confirm the importance of continued transpiration and roots ability to supply water under terminal drought. VI ICLGG - Posters P-ABS26 High seed number is associated with tolerance to water deficit at reproductive phase in chickpea (Cicer arietinum L.) Pushpavalli R 1, 2 1 1 2 1, , Zaman-Allah M , Rekha B , Rao MV , Vadez V * 1 International Crops Research Institute for the Semi-arid Tropics (ICRISAT), Hyderabad, India 2 Bharathidasan University, Tiruchirappalli, India * E-mail: v.vadez@cgiar.org Abstract Chickpea is often exposed to water deficit conditions especially during reproductive stage. Ten chickpea genotypes with comparable phenology and contrasting for seed yield under drought in the field were tested to identify traits associated with tolerance to drought during the reproductive stage in the glasshouse. The drought stress was initiated in two phases at the onset of flowering based on Normalized transpiration ratio (NTR) values; phase I (NTR = 1-0.5) and phase II (NTR = 0.49-0.11). The water stress (WS) clearly reduced total shoot biomass (12-54%), flower number (33-59%), pod number (31-73%), pod weight (15-63%), seed number (20-65%), seed weight (1970%), and 100-seed weight (0-12%). Reduction in seed number or seed abortion played a major role in yield losses along with flower and pod abortion, although the 100-seed weight suffered little. During phase I, the seed weight discriminated between tolerant and sensitive genotypes under WS conditions. Besides, the seed number produced during phase II significantly contributed to 2 final yield (r =0.63**) in water-stressed plants in contrast with well-watered 2 plants (r =0.05). These results suggested that high seed number rather than seed weight is critical for drought stress tolerance in chickpea during the reproductive stage. VI ICLGG - Posters Breeding for drought resistance and water-use efficiency in groundnut 1,* 1 1 1 P-ABS27 1 Vasanthi RP , Sudhakar P , Venkateswarlu O , John K , Latha P , Mohana 1 1 1, 2 KV , Thejaswi Y , Sireesha K , Reddy PV Nigam SN 1 Regional Agricultural Research Station, Tirupati, India International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 2 *E-mail: vasanthi.rrs@gmail.com Abstract Groundnut (Arachis hypogaea L.) is the major oilseed crop in Andhra Pradesh, India. In the state, it is being grown in an area of 1.9 million hectares with a -1 production of 1.7million tons and with a productivity of 924 kgha . 80% of the groundnut area is under rainfed situation. The average productivity of groundnut during kharif season under rainfed conditions varied in different -1 -1 districts from 301 kg ha (2006) to 1357 kg ha (2007) in different years. The fluctuations in productivity from year to year are mainly due to variation in rainfall pattern. Drought (moisture stress) is the major factor influencing the yield of rainfed groundnut. Depending on the stage and severity of the dry spell, yield reduction varies depending on the stage and severity of the dry spell. Hence, drought resistance is a complex trait governed by many genes and their interaction with the environment. To tackle drought, breeding programme was initiated in 2002 at RARS, Tirupati utilising the advanced breeding lines developed in ACIAR funded project. About 62 crosses were made in line x tester fashion and half-diallel fashion. About 160 breeding lines with different combinations of drought and water-use efficiency and desirable pod and seed features were selected in advanced generations and evaluated for yield and water use efficiency traits. Out of this, 16 lines with a good combination of yield, drought and water-use efficiency were isolated. From these, TCGS 1043, a breeding line with superior performance with respect to yield, drought tolerance, tolerance to peanut bud necrosis, stem necrosis, stem rot and dry root rot, high shelling out-turn of 75% in rainfed situation and 80% under irrigated situation was released as Dharani in July, 2012 by state varietal release committee after on-station and on-farm evaluation of more than five years. One more line, TCGS 1073 with water-use efficiency and high yield potential under irrigated situation with medium bold to bold pods is in minikit testing. Two more lines with high yield potential and water use efficiency with compact plant type are being seed increased for further evaluation for their suitability to high rainfall areas as well as for high input situations. VI ICLGG - Posters P-ABS28 In vivo mechanism of salicylic acid induced salt tolerance in chickpea (Cicer arietinum L.) seeds during germination * Raju D , Hoskote PV, Gurusamy S, Sarangapany N, Siddegowda R Directorate of Seed Research, Maunath Bhanjan, India * E-mail: dandyman2k3@yahoo.co.in Abstract Salinity is the major abiotic stress affected mainly germination and early seedling establishment of the crop plants. Generally, pulses that include chickpea were highly sensitive to salt stress. An experiment was conducted to demonstrate the differential performance of the saline sensitive (PG186) and saline tolerant (Karnal chenna) chickpea cultivar under salt stress and improve the germination and vigour of the sensitive cultivar using bioactive chemical Salicylic Acid (SA). Results found that, salt stress adversely affected the germination up to 90% in PG 186 cultivar, SA proved to enhance the germination up to 40% under 150 mM NaCl salt stress. The in-vivo mechanism of SA mediated stress tolerance were studied from its signal reception to transduction via estimating various antioxidative enzymes (AOE) that balances the H2O2 burst in cotyledon during germination. The unprimed PG 186 seeds accumulated more AOE in cotyledons than the SA primed seeds. Like SA primed seeds, unprimed seeds of Karnal chenna cultivar also were found to contain less AOE in the cotyledon and intern might have balanced to accumulate more H2O2 signal for enhanced germination. In order to find the effect of SA induced reduction in AOE on reserve protein mobilisation under salt stress, soluble protein content was estimated from both unprimed and SA primed seeds of PG 186 cultivar during pre and post sowing condition. The results found that, 8 days after sowing cotyledons of both unprimed and SA primed seeds contain same left over protein after mobilisation. However, prior to sowing, the initial protein content of SA primed seeds was more than the unprimed seeds revealing that, SA may not be mobilising more protein which were already present in the seeds during germination. Conversely, it signals the synthesis of stress tolerant protein during initial period of germination. These results showed that, SA negatively influenced the AOE and intern might have increased the H2O2 signal. This enhanced H2O2 signal under high salt stress might have induced the stress tolerant protein synthesis during first phase of germination and mobilised them during the final phase of the germination. VI ICLGG - Posters Computational analysis of Heat Shock Proteins (HSPs) to understand the molecular mechanism of heat stress tolerance in legumes and other plant species 1 1 1 2 2 P-ABS29 1, Singh N , Sahu TK , Singh UP , Gaikwad K , Singh NK , Rao AR * 1 Indian Agricultural Statistics Research Institute, New Delhi, India National Research Center for Plant Biotechnology, New Delhi, India 2 *E-mail: arrao@iasri.res.in Abstract Heat stress is one of the major abiotic stresses, mainly responsible for unstable and low seed yields in legume crops especially during flowering o stages. Therefore, high temperatures above 30 C often slow down the growth and adaptation of legumes worldwide. The HSPs are involved in folding and unfolding of other proteins and get expressed in the conditions of elevated temperature thereby inducing heat tolerance in the organisms. Members of the HSP protein family are strongly up-regulated by heat stress. So, to unravel the underlying molecular mechanism of heat stress in legumes, the primary and tertiary protein structure analysis of the HSP has been done here. The HSP sequences of the legume crops like, Cajanus cajan, Medicago truncatula, were collected form NCBI and NRCPB and the protein structures were predicted by in silico methods. Both sequence and structure alignment was performed to identify the important amino acid residues playing a key role in heat stress tolerance in legumes. Also, the HSP sequences and structure profiles of the legumes were aligned with that of other plant species like, Arabidopsis, Rice, Wheat and Maize to understand the evolutionary nature of the amino acid sequences. From this study, conserved residues responsible for heat stress tolerance have been identified. Besides, the phylogenetic relationship between the plants has also been identified based on HSPs. As pigeon pea is a heat tolerant crop, characterization of the nature and mechanism of thermotolerance in this crop will aid in better understanding of the abiotic response in other legumes. VI ICLGG - Posters P-ABS30 Genotypic differences in some physiological and biochemical parameters symptomatic for drought tolerance in groundnut (Arachis hypogaea L.) 1 1 2 1 Varakumar P , Naik JB , Reddy KKC , Sravani K , Krishna KG 1 1 1, Chandrasekhar A , Krishna VT , Reddy COP * 1 1 School of Life Sciences, Yogi Vemana University, Kadapa, India Agricultural Research Station, Kadapa, India 2 *E-mail: coreddy@yogivemanauniversity.ac.in Abstract Drought stress is the major abiotic stress that severely limits the productivity of the semi arid legume crop groundnut. Alteration of water relationships and formation of reactive oxygen species (ROS) are the two sources for the drought induced damage in groundnut. The extent to which genotype can circumvent or defense these physiological processes determines the degree of resistance to drought stress. In this study, eleven groundnut cultivars (Abhaya, K-1271, TCGS-888, JL-24, K6, Narayani, Prasuna, Rohini, TG47, TPT1 and TPT4) were subjected to drought stress by withholding water for 5 and 10days followed by rehydration. Our results indicate that the cultivar K-1271 is more tolerant to this stress, registering greater biomass and high yield associated with high antioxidant activity. Also, we found that a positive correlation of a positive correlation of relative growth rate (RGR) total and foliar with LRWC, and a negative one with the parameters malondialdehyde (MDA), antioxidants, and proline indicating the importance of lipid peroxidation as that determinant physiological process in selecting groundnut plants tolerant to drought stress. VI ICLGG - Posters Massive Analysis of cDNA Ends (MACE): low cost simultaneous genomewide gene expression profiling and SNP detection reveals allele-specific drought tolerance patterns in chickpea roots 1 2 3 2 2 P-ABS31 2,* Gómez N , Bokszczanin K , Kudupa K , Jüngling R , Krezdorn N , Rotter B , 3 1 2 Varshney RK , Kahl G , Winter P 1 Molecular BioSciences, Johann Wolfgang Goethe University, Frankfurt am Main,Germany 2 Frankfurt Innovation Center FIZ Biotechnology, Frankfurt am Main, Germany 3 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India *E-mail: rotter@genxpro.de Abstract Reducing the complexity of plant genomes prior to second generation sequencing is increasingly used for detection of SNPs and for genotyping. Here, we present a method and bioinformatics pipeline called “Massive Analysis of cDNA Ends” (MACE), which reduces the complexity of transcriptomes, by focusing the sequencing effort to the usually most polymorphic 3’ ends of transcripts. The method allows for simultaneous analysis of quantitative expression values of each transcript AND provides sufficient coverage of the 3’ ends to identify SNPs at 10-fold lower sequencing costs compared to RNAseq. Applying MACE to roots from parental and recombinant inbred lines from chickpea with different drought tolerances identified numerous high-coverage SNPs in relevant, differentially expressed genes, which may serve as markers in molecular breeding for drought tolerance, in the future. VI ICLGG - Posters P-ABS32 Screening and development of heat tolerant lines and management for improved productivity in lentil (Lens culinaris Medik.) * Roy Choudhury D , Kundagrami S Institute of Agricultural Science, University of Calcutta, Kolkata, India *E-mail: drc.gpb@gmail.com Abstract Due to climate change and global warming, the cultivation of cool season food legumes like lentil has now turned to be a big gamble for farmers. For sustainable food and nutritional security, it is necessary to develop high yielding, heat tolerant, early maturing lentil germplasms for changing climatic situation. Studies were conducted to screen 48 lentil germplasms in the 0 laboratory at 34 C temperature in plant growth chamber. Data of survival percentage and seedling vigour were recorded after one month of germination. This laboratory result was confirmed by cell membrane thermostability test by which some heat tolerant germplasms were screened. All the germplasms were further screened by field trial in optimum (mid of November) and late sown (end of December) condition. Yield, survival and maturity duration data were recorded. Some early maturing germplasms like Howrah local, Purulia local, Midnapore local 1, Sagardeep local and UP local were identified. Some high yielding, heat tolerant germplasms like Ranjan, Moitree, IC 201710, IC 208329 were finally isolated which supports our previous laboratory tests. In another experiment some advanced, early maturing, high yielding, heat tolerant mutant lines (M4) were generated by treating three popular lentil germplasms Subrata, WBL 81 and WBL 185 by gamma radiation of 0, 10, 20 and 30 Kr. Furthermore some management practices were developed to cope up with heat stress by treating the lentil seed with some herbal treatments like kalmegh, tea, coffee, garammasala, jeera and chili powder which depicted appreciable performance against heat stress both in laboratory and field condition. VI ICLGG - Posters A novel aldose reductase from resurrection plant: potential role in methylglyoxal detoxification and osmoprotection in blackgram Singh P, Sarin NB P-ABS33 * Jawaharlal Nehru University, New Delhi, India *E-mail: neerasarin@rediffmail.com Abstract Blackgram (Vigna mungo) is an important grain legume grown in the tropical and subtropical regions of the Indian subcontinent. The crop is subjected to a variety of abiotic and biotic stresses which are responsible for its poor productivity and among the abiotic stresses; the crop is notably susceptible to salinity and drought. We utilized the aldose reductase (ALDRXV4) gene isolated from Xerophyta viscose for engineering abiotic stress tolerance in an important legume;Vigna mungo (blackgram).Aldose reductases belong to the well-conserved aldo–keto reductase super family of enzymes and play numerous roles in the metabolism of steroids, sugars, and other carbonyls in plants and animals. Molecular analyses of independent transgenic lines performed by PCR, southern, RT-PCR and western blot revealed the stable integration and expression of the transgene in the progeny. These Southern positive transgenic plants showed 6-7 fold higher aldose reductase enzyme activity as compared to the untransformed control plants. The leaf discs from transgenic plants were able to tolerate upto 800mM of mannitol, 20% of polyethylene glycol (PEG) and 10mM of methylglyoxal stress as revealed by retention of more chlorophyll and delayed senescence as compared to the untransformed control plants. The transgenic plants were tolerant to higher and toxic levels of methylglyoxal (MG). These could also resist a long period of water deficiency and salt stress and exhibited improved recovery after rehydration.The health of the transgenic plants was assessed by reduction in lipid peroxidation-derived reactive aldehydes and better photosynthetic efficiency under drought and salt stress. Together, these results indicate that ALDRXV4 plays a key role in drought and salinity tolerance via ROS detoxification. VI ICLGG - Posters P-ABS34 Foliar application of calcium chloride and salicylic acid induce cross tolerance to high temperature in chickpea (Cicer arietinum L.) Shah D, Bahuguna RN, Khetarpal S, Anand A, Pal M* Indian Agricultural Research Institute, New Delhi, India *E-mail: madanpal@yahoo.com Abstract High temperature (HT) is a major constraint for crop productivity under changing climate. Mitigation of the adverse impacts of HT is requisite for sustainable agricultural productivity. External application of some signalling 2+ molecules like salicylic acid (SA) and calcium (Ca ) have been shown to acclimatize plants for abiotic stress environment. This study reports the mitigation of HT (~10°C higher than the optimum) through foliar application of SA (50 µM) and CaCl2 (10 mM) in two kabuli chickpea genotypes raised under controlled environment in growth chambers. HT exposure enhanced generation of reactive oxygen species (ROS) and membrane lipid peroxidation significantly and caused substantial loss in membrane stability (36 to 42%). However, low generation of ROS and peroxidation of lipids noted in the plants pre-treated with SA and CaCl2. Activities of antioxidant enzymes viz. superoxide dismutase (SOD), ascorbate peroxidase (APOX) and guaiacol peroxidase (GPOX) significantly increased under HT as well as SA and CaCl 2 application per se, though, additive effects on enzyme activities were observed in plants pre-treated with SA or CaCl2 followed by HT exposure. Soluble protein profile indicated different patterns in SA, CaCl2 and HT treated plants compared with control. Besides significant reductions in growth rate and fresh mass under HT, both SA and CaCl2 treatments reverted above losses significantly. The findings reveal that acclimation of plants with salicylic acid or calcium chloride may impart HT tolerance in chickpea, but warrants further studies to confirm the role of these signalling molecules for mitigation of high temperature stress effects. VI ICLGG - Posters In silico identification and validation of selected drought responsive genes in pigeonpea (Cajanus cajan) Sinha P, Azam S, Saxena RK, Krishnamurthy L, Saxena KB, Varshney RK P-ABS35 * International Crop Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India * E-mail: r.k.varshney@cgiar.org Abstract One of the most unique features of pigeonpea is its tolerance to drought stress compared to other legume crops. In this context, analysis of 511 universal drought-responsive protein sequences from the Viridiplantatein the pigeonpea genome identified 111 homologous sequences. Further analysis of these homologous sequences for the universally stressed protein (USP) domain that has shown to confer resistance for environment stress, such as drought in many species at higher stringency using the Hidden Markov Models (HMM) resulted in identification of 49 genes. Primer pairs have been designed for all the 49 genes for monitoring gene expression and validation in drought responsive genotypes (ICPL 151, ICPL 227 and ICPL 8755).Slow drought (dry down) stress has been imposed on above mentioned genotypes and RNAs has been isolated from stress challenged and control root tissues. qRT-PCR for these genes will be presented in the conference. In summary, this study is expected to provide the validated drought responsive genes that could be useful for enhancing drought tolerance in drought sensitive pigeonpea varieties as well as in other related legume crops. VI ICLGG - Posters P-ABS36 Reverse Northern: A novel tool for differential expression analysis of drought stress induced-related genes in chickpea (Cicer arietinum L.) Soren KR*, Patil PG, Alok D, Chaturvedi SK, Nadarajan N Indian Institute of Pulses Research (IIPR), Kanpur, India *E-mail: sorenars@gmail.com Abstract The wild and cultivated chickpea species are useful resources to identify and isolate the genes conferring drought tolerance. An enriched cDNA library for chickpea genes expressed specifically in responses to water stress was constructed using C.arietinum (RSG-88) and C.judacum (ICC-17150). Water stress was imparted through different means like withholding water supply (5070%, RWC), induction with polyethylene glycol (PEG-800) and abscisic acid (ABA) treatment (6-8hr). Total RNA was isolated from water-stressed induced wild and cultivated chickpea leaves at 21 days after sowing (DAS) by using Plant RNeasy kit (Qiagen). Total RNA (10µg) pools from these three treatments were used for the synthesis cDNA library and the expression patterns of novel sequences were characterised to verify their differential expressed gene during water stress conditions. The sequence homology showed 26% genes regulated under (water stress), 7% (regulatory gene), 5% (defence responsive genes), 33% (photosynthetic gene) and 29% others. Furthermore, reverse northern hybridization (RNH), demonstrated that most of the up-regulated genes belonged to dehydration induced like protein of Medicago ab1Di19 super family, salt stress responsive PLN00046 super family of Medicago, zip transcription factor, cell cycle regulatory genes etc which are differentially expressed in response to water stress in chickpea. VI ICLGG - Posters Genetic studies for saline and alkaline tolerances in soybean 1,* Xu D , Tuyen DC 1, 2 , Hamweish A 1, 3 1, 4 , Lal KS , Zhang H P-ABS37 1, 5 1 Japan International Research Center for Agricultural Sciences (JIRCAS), Tsukuba, Japan 2 Cuulong Delta Rice Research Institute (CLRRI), Thoilai, Vietnam 3 International Center for Agricultural Research in the Dry Areas (ICARDA),Aleppo, Syria 4 Indian Agricultural Research Institute, New Delhi, India 5 Institute of Vegetable Crops, Jiangsu Academy of Agricultural Sciences, Nanjing Jiangsu, China *E-mail: xudh@jircas.affrc.go.jp Abstract Salt-affected soils are generally classified into two main categories: saline and alkaline (sodic). Both kinds of salt stresses were reported to inhibit soybean (Glycine max (L.) Merr) plant growth, nodule formation, and seed yield. Developing of soybean cultivars with high salt tolerance is an effective way of maintaining sustainable production in areas where soybean growth is threatened by salt stress. Several soybean genotypes with high salinity tolerance were identified in our recent studies. QTL analysis using different mapping populations constantly detected a major salinity tolerant QTL on chromosome 3 flanking by SSR markers Sat_91, Satt255, and Satt339. This QTL accounted for 44-64.0% of the total variance for saline tolerance in the populations. In addition, several near isogenic lines (NILs) for the QTL were developed using DNA marker-assisted selection (MAS). These NILs are currently used for fine-mapping of the QTL and identification of candidate gene that contributes to saline tolerance. In the case of alkaline tolerance, a wild soybean (Glycine soja Sieb. & Zucc.) accession from Japan with high alkaline tolerance was identified, and a major QTL for alkaline tolerance was detected on chromosome 17. Fine-mapping analysis narrowed down the QTL to a 3.3cM interval region between the markers GM17-11.6 and Satt447 with a physical map length of approximately 771 kb. These studies indicated that saline and alkaline tolerances were controlled by different genes in soybean. DNA markers closely associated with these QTLs can be used for MAS to pyramid tolerance genes in soybean for both saline and alkaline stresses. VI ICLGG - Posters Theme Pathogenesis and Disease Resitance P-PAD05 Breeding for rust (Uromyces viciae-fabae) resistance in faba bean (Vicia faba L.) in Australia 1, 2 1 Adhikari KN *, van Leur J , Sadeque A , Trethowan R 1 1 The University of Sydney, IA Watson Grains Research Centre, Narrabri, Australia 2 Tamworth Agricultural Institute, Department of Primary Industries, Calala, Australia *E-mail: kedar.adhikari@sydney.edu.au Abstract Faba bean is one of the oldest grain legumes grown in many countries for both human consumption and animal feed. Rust, caused by Uromyces viciaefabae, is a serious disease of faba beans in sub-tropical New South Wales and southern Queensland. Although faba bean rust is a major disease in this region, limited information is available on the genetic basis of the rust resistance found in the germplasm. Except for the recently released variety Doza, all faba bean varieties are susceptible to rust. The aims of this study were to assess the variation of rust resistance in the germplasm and enhance the level of resistance in the breeding program. Various lines were obtained from the Australian Temperate Field Crop Collection and tested for rust resistance. A wide range in resistance was found, but only few lines remained rust-free under high disease pressure. Crosses were made between the best resistant germplasm available in the breeding program and agronomically suitable parents. Their progenies were advanced to F 4/F5 stage and they were screened against rust at Narrabri under artificially created disease pressure. Many lines showed a higher level of resistance than shown by current cultivars including Doza. The level of resistance was significantly higher in lines at early stage of development than those of advanced lines showing a significant improvement in the breeding program. The level of resistance displayed by new lines is high enough to grow reliable crop without any fungicide protection. VI ICLGG - Posters Towards identification of single nucleotide polymorphisms (SNPs) linked to disease resistance genes in pigeonpea {Cajanus cajan (L.) Millsp.} Agbagwa IO 1,2* 2 , Patil P , Nadarajan N P-PAD06 2 1 Department of Plant Science & Biotechnology, University of Port Harcourt, Nigeria 2 Indian Institute of Pulses Research (IIPR), Kanpur, India *E-mail: ikechukwu.agbagwa@gmail.com Abstract Sequences of cloned resistance genes from a wide range of plants taxa reveal significant similarities in sequence homology and structural motifs. In this study, oligonucleotide primers designed from conserved nucleotide binding site (NBS) domain were used to amplify related sequences from two pigeonpea genotypes (ICPL 87119 and UPAS 120), differing in disease reaction against Fusarium wilt (FW) and Sterility Mosaic disease (SMD). Sequencing of amplified products and homology search implicates association with cell death process linked to hypersensitive response (HR). Based on consensus sequence of resistance gene analogues, five new sets of primers were designed and specific open reading frames (OFRs) amplified in a set of 8 selected pigeonpea genotypes. Sequence alignment of the amplified products revealed several single nucleotide polymorphisms (SNPs) within the coding region. Dendrogram constructed using similarity matrices of the 40 sequences revealed phylogenetic relationship among the genotypes based on pedigree. The sequence variation (SNPs) observed shall be helpful in developing CAPS markers and identifying candidate resistance genes associated with FW and SMD. VI ICLGG - Posters P-PAD07 Intra-field sequence diversity in NP gene of Groundnut bud necrosis virus population infecting wild Vigna in India Akram M*, Naimuddin, Pratap A, Malviya N, Yadav P Indian Institute of Pulses Research (IIPR), Kanpur, India *E-mail: akram23859@gmail.com Abstract Tospoviruses have wide host range and are emerging as serious pathogens affecting several crops worldwide. Groundnut bud necrosis virus (GBNV) is identified as most prevalent tospovirus in India. NP-gene diversity in GBNV population in a field was studied. During 2011, twelve accessions of wild species/sub-species of Vigna (V. umbellata, V. glabrescens, V. hainiana, V. mungo var. mungo, V. radiata var. radiata and V. radiata var. sublobata) grown at the IIPR, Kanpur, India, exhibited chlorotic and necrotic spots on leaves, stems and petioles, symptoms resembling those induced by GBNV. Disease incidence ranged 2-20% in different accessions. Identity of the causal virus was confirmed on the basis of symptoms on the diagnostic host and the NP-gene sequence identities. Isolates of GBNV studied were designated as [Vmm1-Knp], [Vmm2-Knp], [Vmm3-Knp], [Vmm4-Knp], [Vum1-Knp], [Vum2Knp], [Vum3-Knp], [Vrr1-Knp], [Vrr2-Knp], [Vha-Knp], [Vgl-Knp] and [Vrs-Knp] and their NP-gene sequences were deposited at NCBI GenBank under the accessions JQ406578, JQ406579, JQ406580, JQ406581, JQ406582, JQ406583, JQ406584, JQ347266, JQ347265, JQ406577, JQ406576 and JQ347264, respectively. Analysis of NP-gene sequences of GBNV isolates revealed 2-4% variation in nucleotide. The variation in amino acids was 0-3%. Common type of symptoms produced on indicator host and the level of variation present in NP-gene sequences indicated that the GBNV isolates from a field constitute the same population, though these isolates were collected from different wild species and sub-species of Vigna. To the best of our knowledge this is the first report of Groundnut bud necrosis virus infection on all the wild accessions of Vigna studied. VI ICLGG - Posters Differential expression of Lathyrus cicera and Lathyrus transcriptomes in response to rust (Uromyces pisi) infection 1, 1 2 2 sativus 3 Almeida NF *, Leitão ST , Rotter B , Winter P , Rubiales D , Vaz Patto MC P-PAD08 1 1 Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal 2 GenXPro, D-60438 Frankfurt am Main, Germany 3 Institute for Sustainable Agriculture, Córdoba, Spain *E-mail: nalmeida@itqb.unl.pt Abstract Chickling pea (Lathyrus cicera L.) and grass pea (Lathyrus sativus L.) have great potential among grain legumes due to their good adaptability to inauspicious environments, high protein content and resistance to serious diseases. Due to the need of increasing sustainability of agricultural production systems the interest in the conservation and utilization of Lathyrus genetic resources is rising. Nevertheless, due to its past underuse, just a few molecular breeding tools were developed specifically for Lathyrus species. Research on more important economically related legumes can to a certain extend contribute to surpass this limitation. We have compared the leaf transcriptome profiles of two contrasting accessions each of L. cicera and L. sativus in rust (Uromyces pisi) infected and non infected conditions. For L. cicera a normalized cDNA library was created using Illumina sequencing. For L. sativus a semi-quantitative RNA-seq method was chosen in order to detect also variation in expression level. These new data will allow bridging Lathyrus genomic information with model species and better studied related legume crops in order to develop molecular tools that will allow breeding these ancient crops interesting for modern farmers. VI ICLGG - Posters P-PAD09 Characterization of Uromyces appendiculatus isolates collected from snap bean growing areas in Kenya 1, 1 1 Arunga EE *, Ochuodho JO , Kinyua MG , Owuoche JO 2 1 Chepkoilel University College, Eldoret, Kenya Egerton University, Egerton, Kenya 2 * E-mail: eea08ke@hotmail.com Abstract Bean rust (Uromyces appendiculatus (Pers.:Pers.) Unger var. appendiculatus) is one of the most devastating and variable pathogens of common bean (Phaseolus vulgaris L.) worldwide that can cause total crop loss. Characterization of bean rust races can help in screening resistant materials during gene pyramiding. The aims of the present research were first, to characterize bean rust isolates collected from snap bean growing areas in Kenya. Secondly, to identify which of the available rust resistance genes in common bean differentials are most effective to control rust in those areas. Snap bean leaf tissues with rust pustules were collected from different farms in eight locations in Central and Western Kenya during the years 2010 and 2011. Forty seven single pustule isolates were obtained and inoculated on 12 bean rust differential cultivars. For consistent results, the inoculation was repeated twice. The new international classification system and the binary nomenclature grouped the 47 single pustule isolates of Uromyces appendiculatus into nine different races most of this affected the Andean gene pool. Hierarchical cluster analysis grouped the races into two major clusters depending on the virulence of the races on the host differential cultivars. The most resistant genes for pyramiding in Kenya were identified as Ur-5, Ur-11 and Ur-CNC. An important output of this study was the identification of races with potential use during gene pyramiding process. . VI ICLGG - Posters Fusarium virguliforme FvTox1 toxin causes foliar sudden death syndrome presumably through interaction with a carbonic anhydrase in soybean (Glycine max L. Merr.) P-PAD10 Brar HK, Pudake RN, Swaminathan S, Baumbach JL, Sahu BB, Brooke C, Bhattacharyya MK* Department of Agronomy, Iowa State University, Ames, USA *E-mail: mbhattac@iastate.edu Abstract Soybean is one of the world’s most valuable crops. In 2010, the U.S. produced 35% of the world soybean crop followed by Brazil (27%) and Argentina (19%). Soybean suffers yield suppression from various biotic stresses. In 2010, 14.4% of total yield valued $5.59 billion was suppressed by pathogenic diseases caused by microbes and nematodes. Soybean sudden death syndrome (SDS) is a major threat to soybean production in the U.S. Nationwide, the estimated soybean yield suppression from SDS in 2010 was valued at $0.82 billion. SDS is caused by Fusarium virguliforme. Foliar SDS symptoms are caused by toxins. A proteinacious toxin, FvTox1, produces foliar SDS-like symptoms only in presence of light. Expression of a single chain variable fragment antibody against the toxin molecule enhanced the foliar SDS resistance in transgenic soybean plants. Analyses of five independent F. virguliforme fvtox1 mutants created by a homologous recombination approach revealed that FvTox1 is a virulence factor required for foliar SDS development. FvTox1 accumulates in chloroplasts and binds to a soybean carbonic anhydrase isoform that transports carbon dioxide (CO2) in the form of carbonic acid into chloroplasts for photosynthesis. Expression of the maize homologue of the protein lacking affinity to FvTox1 provides enhanced foliar SDS resistance in transgenic soybean plants. Thus, most likely FvTox1 produces foliar SDS symptoms through its interaction with a carbonic anhydrase isoform. We hypothesize that following binding of FvTox1 to carbonic anhydrase the CO 2 transport is impaired leading to formation of highly toxic free radicals which initiate foliar SDS development. VI ICLGG - Posters P-PAD11 Screening the Vietnamese mungbean germplasm collection for resistance to mungbean Yellow Mosaic disease 1,* 2 3 3 Bui H , Nair R , Keryon L , Ebert A 1 Plant Resouces Center, Hanoi, Vietnam AVRDC - Regional Center for South Asia (RCSA), ICRISAT Campus, Hyderabad, India 3 AVRDC - The World Vegetable Center, Headquarters, Tainan, Taiwan 2 * E-mail: Bui_thu_huyen84@yahoo.com Abtract Mungbean (Vigna radiata (L.) Wilczek.) is a traditional legume food crop which is consumed in several different ways in Vietnam. One of the most serious threats to mungbean cultivation, particularly in the arid central and southern parts of Vietnam, is mungbean yellow mosaic disease (MYMD). In order to identify sources of resistance against mungbean yellow mosaic desease, 50 lines of mungbean were evaluated under field conditions in two sites in centralsouthern Vietnam in Binh Dinh (from July to September 2011) and in Phu Yen (from March to May 2012). Out of the 50 mungbean accessions screened, three were found to be highly resistant (HR), one was resistant (R), 10 were moderately resistant (MR) and 34 accessions were found to be susceptible (S) to highly susceptible (HS) to the disease. This is the first time resistance against MYMD in mungbean has been reported in Vietnam. The identification of sources of resistance against the Vietnamese strains of the disease is very important for the future development of mungbean in Vietnam. VI ICLGG - Posters Repeat motifs putatively linked to leaf crinkle disease resistance in cowpea (Vigna unguiculata (L.) Walp.) P-PAD12 Dhanasekar P*, Reddy KS Bhabha Atomic Research Centre, Mumbai, India * E-mail: sekar@barc.gov.in Abstract Cowpea (Vigna unguiculata (L.) Walp.) an arid legume and protein rich pulse crop, construed as an orphan crop until now, is being perceived as a prospective crop for its hardiness in the present perspective of climate change. The low productivity of cowpea could be attributed to its susceptibility to biotic stresses like diseases. Leaf crinkle caused by Urdbean leaf crinkle virus is one such disease gaining importance leading to heavy yield losses. With a view to identifying resistance source, 47 elite genotypes including varieties, mutants and advanced breeding lines were screened for two successive seasons under high incidence of the disease. Of the eight resistant lines identified, the mutant derivative TC1-6-10 along with supposedly genetically similar susceptible parent EC394763 were used in the parental screening with 66 SSR, 99 ISSR and 38 RAPD primers to identify putative markers linked to resistance trait. These markers were successful in discerning 17.2 percent polymorphism between TC1-6-10 and EC394763. Two SSR and 25 ISSR polymorphic markers were identified that were present in resistant and absent in susceptible parents. After confirming the reproducibility, these markers were validated in the rest of the seven resistant lines. One SSR (VM28 300) and one ISSR (UBC 8911200) showed positive amplification in six and five resistant lines respectively, enhancing the credibility of these markers for possible linkage to the resistance loci. This however, needs to be confirmed using segregating population/RILs for its further use in marker assisted breeding and to determine the linkage distance. VI ICLGG - Posters P-PAD13 Varietal screening of horse gram genotypes to pulse beetle, Callosobruchus chinensis (Coleoptera: Bruchidae) 1, 2 Durga KK *, Rajasri M 1 Seed Research and Technology Center, Acharya NG Ranga Agricultural University, Hyderabad, India 2 Farmers Call Center, Secunderbad, India Abstract Studies on the relative susceptibility of Horse gram (Macrotyloma uniflorum) genotypes to pulse beetle (Callosobruchus chinensis) was undertaken at Seed Research and Technology Centre, Acharya N.G. Ranga Agricultural University, Rajendranagar, Hyderabad, Andhra pradesh, India during 2011-12. 23 horse gram genotypes were screened for their relative response to the attack by C. chinensis with artificial infestation and were classified based on ovipositional preference, per cent adult emergence, weight loss % of seed and per cent insect damage. Significant differences were observed between genotypes with regard to their relative susceptibility to ovipositional preference, bruchid infestation and % weight loss. HG 24 (3.0) followed by HG 14 (4.0), HG 54 (6.7) and HG 18 (9.3) recorded less number of eggs/100 seeds and were less preferred by the pulse beetle for oviposition, while Palem 1 (42.3), HG 52 (40.0), AK 38 (34.3) and HG 38 (33.3) were most preferred for egg laying. Least progeny production was noticed in HG 75 (2.3), HG 24 (3.7), HG 11 (4.3), HG 59 (8.3), HG 54 (8.7) while high adult progeny production was noticed with HG 38 (162.3) followed by HG 41 (77.3), AK 38 (59.7), Palem 1 (59.0) and HG 52 (51.0). Among the genotypes, low insect damage was recorded with HG 14 (0.36) followed by HG 75 (0.51) and HG 24 (0.81) While maximum % insect damage was noticed with HG 32 (6.22) and HG 17 (4.35). The horse gram genotypes viz., HG 54, HG 59, HG 63, HG 14 and HG 11 were found to be resistant to pulse beetle which recorded less insect damage and adult emergence with no weight loss after 45 days of storage where as Palem 1, HG 38 and AK 38 were found to be susceptible with high insect damage and adult emergence. The seed colour of HG 54 was black indicating the morphological trait for resistance which is less preferred by the pulse beetle than the straw coloured seeds. Correlation analysis between fecundity, adult emergence, % weight loss and % insect damage indicated the positive relationship between fecundity with adult emergence (r = 0.6971) and % weight loss (0.6067). Similarly adult progeny production was positively correlated with % weight loss (r = 0.40301). VI ICLGG - Posters Viral diseases infecting legume crops in Azerbaijan P-PAD14 Sultanova N, Mammadov A, Huseynova I* Institute of Botany, Azerbaijan National Academy of Sciences, Baku, Azerbaijan *E-mail: i_guseinova@mail.ru Abstract Viral diseases are an important limiting factor in many crop production systems, legumes affected to a wide ranges virus diseases that can cause yield and quality losses under unfavorable environmental conditions. In this purpose field surveys were conducted in Azerbaijan during 2009-2011 growing seasons to identify of viruses showing mosaic, stunting, yellowing, wilting, shortening of the internodes, and phloem discoloration symptoms on chickpea, faba bean, lentil, pea, alfa-alfa crops. All collected samples were tested for the presence of viruses using the DAS-ELISA method. At the results of different legume samples investigation, we were determined following viruses- Bean leaf roll virus (Luteovirus, BLRV), Bean common mosaic virus and Bean yellow mosaic virus (Potyviruses, BCMV, BYMV), Alfa-alfa mosaic virus (Bromovirus, AMV) and Nanoviruses. Legume samples relevant positive results for nanoviruses were also analyzed by RCA and PCR methods. DNA of two chickpea (Cicer arietinum L.) and two lentil (Lens culinaris L.) plants was amplified by RCA, restricted by endonucleases AatII or HindIII and analyzed on 1, 5 % agarose gel. Restriction by AatII was produced predominant 1-kb fragment. This fragment was a strongly indication of the presence of a nanovirus infection in these samples. The template DNA of chickpea and lentil samples was also amplified by using specific primer pairs F103/R101 and C5F/C5R for nanoviruses. These samples yielded PCR products of the expected size 770 bp and 660 bp, respectively and this confirms the presence of viruses. As more research is carried out, additional pathogens can be expected on these crops. VI ICLGG - Posters P-PAD15 Cross talk between signaling cascades and redox balance regulation in soybean plant reaction to biotic stress mediators Iakovenko OM*, Kalachova TA, Kretynin SV, Kravets VS Institute of Bioorganic Chemistry and Petrochemistry, Kyiv, Ukraine *E-mail: yon@bpci.kiev.ua Abstract Exceptional research interest is dedicated to lipid signaling cascades involved in metabolic responses regulation under biotic stress. The present study investigates the lipid derived second messengers, reactive oxygen species formation in the signal transduction of salicylic acid (SA). Significant increase in phosphatidic acid content in SA-treated soybean seedlings represents the evidence of lipid metabolism enzymes activation. The addition of 1-butanol to the incubation medium led to a more than two fold increase of phosphatydylbutanol participation of formation after 30 and 60 minutes of salicylic acid treatment and suggests inclusion of phospholipase D to the elicitor-induced reaction cascade. Antioxidant system balance changes were registered under salicylic influence and correlated with endogenous peroxide accumulation in legumes tissues with maximum effect observed under the joint phytohormone action. Displayed superoxide dismutase activity increase was attended by decreased catalase activity that maintains peroxide content at the level required for protective responses development. Thereby our findings suggest the involvement of lipid signaling and reactive oxygen species generation and detoxification enzymes to plant defense reactions. We also indicated that exogenous SA application causes an increase of total reducing activity in leaf tissues, which correlates with the accumulation of phenolic compounds. Furthermore SA pre-treatment of soybean plants at the early flowering stage increased α-tocopherol content in mature seeds tissues. The findings suggest activation of compensatory mechanisms of ROS production and detoxification regulation by antioxidant enzymes and lipophilic antioxidants during plant adaptive reactions formation. VI ICLGG - Posters Genetic studies on chlorophyll content in chickpea (Cicer arietinum L.) P-PAD16 Jayalakshmi V*, Yamini BR, Ronald RG Regional Agricultural Research Station, Nandyal, India *E-mail: veera.jayalakshmi@gmail.com Abstract Integration of physiological selection tools with conventional breeding methods is essential to develop photosynthetically efficient and stress tolerant genotypes of crop plants. Seven chickpea genotypes and their 21 half diallel F 1 crosses were evaluated for leaf chlorophyll content and SPAD chlorophyll meter reading (SCMR, a trait related to nitrogen acquisition capability/ water use efficiency) during Rabi 2011-12 at Regional Agricultural Research Station, Nandyal, Andhra Pradesh, India. Analysis of variance indicated highly significant differences among the genotypes for chlorophyll ‘a’, chlorophyll ‘b’, total chlorophyll and SCMR. NBeG 3, JG 11 and Vihar were identified as high performing genotypes for chlorophyll content and SCMR. Heterotic effects related to mean of parents were positive and significant in 13,14,13 and 12 F1s and that due to better parent were in desirable direction in 12,8,8 and 8 F 1s respectively for chlorophyll ‘a’, chlorophyll ‘b’, total chlorophyll and SCMR. Apart from high per se performance, crosses NBeG 3 x MNK 1, ICCV 05106 x MNK 1, ICCV 05106 x KAK 2, MNK1 x KAK 2, MNK 1 x Vihar, ICCV 95333 x KAK 2 and ICCV 05106 x Vihar displayed desirable heterosis for total chlorophyll content and SCMR. A positive and significant association between chlorophyll ‘a’, chlorophyll ‘b’ total chlorophyll and SCMR and seed yield of chickpea was observed. Chlorophyll ‘a’, chlorophyll ‘b’, total chlorophyll and SCMR exhibited mutual association among themselves and also with root length and shoot biomass of chickpea. Promising parental genotypes and crosses with desirable chlorophyll content and SCMR identified in the study can be utilized further for selection and breeding for improved yield in chickpea through improved nitrogen acquirement. VI ICLGG - Posters P-PAD17 Infectivity studies of the mastrevirus, Chickpea chlorotic dwarf Pakistan virus (CpCDPKV) causing chickpea stunt disease in India. 1 2 3 1 Kanakala S , Akshay S , Verma HN , Malathi VG * 1 Division of Plant Pathology, Indian Agricultural Research Institute, New Delhi, India 2 Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi, India 3 Jaipur National University, Jaipur, India *E-mail: vgmalathi@rediffmail.com Abstract Chickpea stunt disease (CSD) was identified as an important viral diseases causing 75-95% yield loss in chickpea growing region of India as early as 1993. The disease was also reported to occur in Africa, Middle East and Asia. Though the disease showing stunting and smalling of leaf was recorded first in India, little was known about the pathogen causing the disease. Here we have determined the complete nucleotide sequence of the mastrevirus associated with CSD in Delhi region. Genomic component of the virus was cloned by Rolling circle amplification method and the virus isolate was found to show 99% sequence identity with Chickpea chlorotic dwarf Pakistan virus (CpCDPKV). Phylogenetic analysis of all the chickpea infecting mastreviruses world wide revealed the dichotomy between the viruses originating from Africa, Middle East, Asia on one clade and viruses originating from Australia on another clade. Agroinoculation of complete tandem dimeric construct of the cloned genome of the virus on chickpea (Cicer arietinum) plants induced severe stunting, smalling of leaves, drying and eventual death of plant. The virus was highly infectious on Nicotiana benthamiana, N.tabacum, N. glutinosa and Solanum lycopersicum. Ability of the CpCDPKV to interact with alpha and betasatellites, normally associated with begomoviruses was tested by coinoculating CpCDPKV with alphasatellite, betasatellite and Tomato leaf curl New Delhi virus. The mechanism of induction of severe stunting during viral pathogenesis was studied, which revealed changes in host-gene expression. . VI ICLGG - Posters Growth of collar rot (Aspergillus niger van Tieghem) and stem rot (Sclerotium rolfsii Saac.) fungi under artificial epiphytotic conditions and its implications for disease management in groundnut 1 2 1 P-PAD18 1, Rathnakumar AL , Kumar V , Bagwan NB , Kumar N * 1 Directorate of Groundnut Research, Junagadh, India National Research Centre for Litchi, Muzaffarpur, India 2 * E-mail: narendrapb09@gmail.com Abstract Collar rot caused by the Aspergillus niger and stem rot caused by Sclerotium rolfsii are major threats to groundnut production worldwide. To identify genotypes with desired levels of resistance, screening of a large number of germplasm under artificial epiphytotic conditions (sick plots) is essential. The inoculums of virulent isolates were mass multiplied and mixed with FYM and applied separately to each of the micro plots of sick plot at Directorate of Groundnut Research, Junagadh, India. The growth and multiplication of A. niger and S. rolfsii in the soil was monitored periodically at monthly intervals during kharif 2011. The study revealed that the soil population of A. niger had 3 steadily increased from 1.8 to 23.4 X 10 cfu during June to October in spite of the heavy rainfall (1613 mm in 66 rainy days). While it was converse in case of 3 Sclerotium rolfsii where the soil population decreased drastically (24.4 x 10 3 cfu) from the initial level (30.2 x 10 cfu) and subsequently got increased in the 3 3 months of September (21.6 x 10 cfu)) and October (24.4 x 10 cfu) when the rainfall decreased (312 mm in 14 rainy days and 55 mm in 2 rainy days in September and October month respectively). Thus the study clearly indicated that the population built up of A. niger was unaffected due to heavy rainfall and once the rain ceases and optimal conditions restored, there will be a sudden spurt in the incidence of the collar rot disease. While in case of stem rot, the incidence of the disease will be low following a heavy rainfall. Thus, chemical control is possible for stem rot while genetic resistance is the suitable option in case of collar rot. VI ICLGG - Posters P-PAD19 Analysis of chickpea transcriptome in response to Ascochyta rabiei infection using Roche/454 sequencing Kumar K, Mehta P, Purayannur S, Verma PK* National Institute of Plant Genome Research (NIPGR), New Delhi, India *E-mail: praveen_verma@nipgr.res.in Abstract An important legume crop, chickpea (Cicer arietinum) ranks third in world legume crop production. Its productivity is hampered by several biotic factors viz. Ascochyta blight, Helicoverpa and Fusarium. So far the chickpea transcriptome has been analysed using the next-generation sequencing technologies under the normal plant growth conditions and abiotic stress conditions. To elucidate the chickpea transcriptome in compatible interaction with Ascochyta rabiei on a large scale, we sequenced the transcriptome using Roche 454 pyrosequencing platform. Six cDNA libraries of chickpea (3 control and 3 fungal spore inoculated) were generated using the RNA samples of 1, 3 and 6 dpi. After 454 FLX sequencing 4,78,780 raw reads were obtained. The quality reads (3,78,863) were pooled together for hybrid de novo assembly of transcriptome. These reads were assembled into 39,192 contigs with average length of 644 bp. The quality of assembly was validated by blastx against the protein databases of Medicago truncatula, Glycine max and Arabidopsis thaliana which revealed about 75-84% matches. Digital expression analysis showed differential expression of several genes at different time intervals. Transcriptome analysis revealed many gene families of chickpea that are involved in signaling, antioxidant response and transcriptional regulation. Many fungal genes were also isolated in this work. Validation of selected chickpea genes for differential expression was carried out by real-time PCR analysis. VI ICLGG - Posters Characterization of Fusarium solani, the causal agent of sudden death disease in soybean Lestari P 1,2 1,3 1 , Sun S , Lee Y-H , Lee S-H 1,4, P-PAD20 * 1 Department of Plant Science and Research Institute for Agriculture and Life Sciences, Seoul National University, Seoul, South Korea 2 Indonesian Center for Agricultural Biotechnology and Genetic Resources Research and Development, Bogor, Indonesia 3 Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing, China 4 Plant Genomics and Breeding Institute, Seoul National University, Seoul, South Korea *E-mail: sukhalee@snu.ac.kr Abstract Sudden death disease (SDS) is one of important disease in soybean and can cause severe losses in major soybean production areas. Fusarium solani indigenous from South Korea, the causal agent of SDS, has not been characterized well. This study aimed to identify and characterize morphology, anatomy and pathogenicity of the fungal pathogen on host plant soybean. Total about 50 fungi isolated from soybean plants tissues were found to majorly belong to Fusarium species based on inter transcribed spacer (ITS) sequence. The fungal isolates were classified as Fusarium sp., F. moniliformis, F. equiseti, F. solani, and Epicoccum singurm. Ten isolates identified as Fusarium solani were strongly classified as F. solani species complex according to translation elongation factor-α (EF-α). Macroscopic and microscopic morphology suggested the specific properties of F. solani. Both stem and root of soybean infected by F. solani altered to be damaged and patchy. Xylem seemed to be affected higher than phloem, indicating an effective zone of SDS pathogen infection. Virulence varied among F. solani isolates on SDS reactions on resistant and moderately resistant soybean cultivars in growth chamber test. This interaction between SDS and F. solani pathogen is important for evaluation of soybean germplasm that may provide a source of resistance needed to develop new SDS-resistant soybean breeding lines or cultivars. VI ICLGG - Posters P-PAD21 Pathogenomics of the Didymella spp. causal agents of Ascochyta blights in legumes 1,* 2,3 2,3 2,3 Lichtenzveig J , Williams AH , Kessie F , Mohd-Shah R , Oliver RP 1,2 1 Department of Environment and Agriculture, Curtin University, Western Australia 2 Australian Centre for Necrotrophic Fungal Pathogens, Curtin University, Western Australia 3 Murdoch University, Western Australia *E-mail: judith.lichtenzveig@curtin.edu.au Abstract The Didymellaceae family includes some of the most important pathogens of legume crops: the causal agents of Ascochyta blight in chickpea, pea, lentil, and faba beans among others (Aveskamp et al 2010). Despite their substantial economic impact, little is known about the molecular aspects of pathogenicity in these closely related species. At this meeting, the first genome assemblies of Ascochyta rabiei (pathogen of chickpea), Peyronellaea pinodes (syn. Mycosphaerella pinodes; pathogen of field pea), and Phoma medicaginis (pathogen of the model legume Medicago truncatula) will be presented and discussed. Untrained in silico annotation resulted in the identification of 9,00012,000 genes per species, of which 91-97% are complete gene models. The sequence data is serving to: pinpoint areas of synteny and clusters of genes associated with reproduction and adaptation; identify pathogenicity-related gene-candidates through proteomic, transcriptomic and in silico comparative analyses; and design DNA makers for diagnostics and studies in population structure. Among the pathogenicity related genes, we have identified potential necrotrophic effectors that seem to operate similarly to those found other fungal pathogens of the order of the Pleosporales. Detailed understanding of the molecular mechanism involved in fungal adaptation in general, and pathogenicity in particular, is facilitating the development of novel tools and strategies in crop protection. VI ICLGG - Posters Characterization of Fusarium udum isolates of southern Telangana zone of Andhra Pradesh causing wilt in pigeonpea (Cajanus cajan L. Millsp) by RAPD analysis. 1,* 1 2 Suresh M , SameerKumar CV , Jagadeeshwar R , Rajeswari R P-PAD22 1 1 ARS, ANGRAU, Tandur, India ARI, ANGRAU, Hyderabad, India 2 *E-mail: sureshiari@yahoo.co.in Abstract Genetic variability among 17 isolates of Fusarium udum causing wilt in Pigeonpea (Cajanus cajan L. Millsp.) collected from the Pigeonpea growing areas of Rangareddy and Mahaboobnagar districts under Southern Telangana Zone (STZ) of Andhra Pradesh, India were assessed using RAPD markers. A total of 120 markers of Operon Technologies were used in the study. Though the markers detected high levels of polymorphism among the fungal pathogen isolates, the degree of polymorphism varied depending on the marker selected. OPK and OPI series markers detected high level of polymorphism among the Fusarium udum isolates. NTSYSpc (version 2.0) cluster analysis based on Jaccard’s similarity co-efficient grouped all the isolates in to two major distinct groups with a minimum of 15% similarity index, suggesting the existence of a minimum of 2 specific races of the pathogen prevailing in the pigeonpea growing areas of STZ . The maximum variation was seen among the pathogen isolates collected from Rangareddy compared to others. Fu11 and Fu12 isolates of Fusarium udum showed 100% similarity with most of the RAPD markers. M3 and M4 isolates of Mahaboobnagar district were grouped separately from rest of the isolates of Rangareddy district (at 20% similarity index) with around 60% similarity among the two isolates, thus suggesting high levels of variability among the isolates collected from the same agro-climatic region of Andhra Pradesh. VI ICLGG - Posters P-PAD23 A soybean cyst nematode resistance gene points to a new mechanism of plant resistance to pathogens 1† 2† 3 2 2 2 4 Liu S , Kandoth PK , Warren SD , Yeckel G ,Heinz R , Alden J ,Yang C , 1 1 4 4 5 Jamai A ,El-Mellouki T , Juvale PS , Hill J4,Baum TJ , Cianzio S ,Whitham 4 3 2,* 1* SA , Korkin D , Mitchum MG , Meksem K 1 Department of Plant, Soil and Agricultural Systems, Southern Illinois University, Carbondale, USA 2 Division of Plant Sciences, Christopher S. Bond Life Sciences Center and Interdisciplinary Plant Group, University of Missouri, Columbia, USA 3 Department of Computer Science, Christopher S. Bond Life Sciences Center and Informatics Institute, University of Missouri, Columbia, USA 4 Department of Plant Pathology and Microbiology, Iowa State University, Ames, USA 5 Department of Agronomy, Iowa State University, Ames, USA *E-mail: meksemk@siu.edu Abstract Soybean cyst nematode (SCN), Heterodera glycines Ichinohe, causes more than a billion dollar in yield losses annually in the United States, it is indeed the most economically important pathogen on soybean. Moreover, virulent populations are overcoming most known resistance sources, therefore, the urgent need to identify, isolate and deploy new genes for resistance to SCN. Genes for resistance to SCN has been identified and mapped to several regions of the soybean genome by both, classical and molecular genetics using a variety of soybean germplasm. The Rhg1 locus on chromosomes 18 and the Rhg4 locus on chromosome 8 were identified by several research teams as two major resistances QTL to SCN. An integrated approach, combining positional cloning with newly developed functional genomics tools in soybean (VIGS, RNAi, Hairy root complementation and TILLING), allowed the isolation and confirmation of a metabolic gene at the Rhg4 locus in resistance to SCN. The discovery is a major breakthrough in the field of plant disease resistance that will impact greatly our understanding of soybean's resistance against SCN and will allow the development of soybean lines with durable resistance to soybean cyst nematode. VI ICLGG - Posters Molecular mechanism of whitefly transmission of yellow mosaic viruses infecting grain legumes Jyothsna P, Archana K, Malathi VG P-PAD24 * Advanced Centre for Plant Virology, Division of Plant Pathology, Indian Agricultural Research Institute, New Delhi, India *E-mail:jyothsnap177@gmail.com Abstract Yellow mosaic disease in legumes is caused by six different legumoviruses in India, of which Mungbean yellow mosaic India virus (MYMIV) is the most devastating agent for leguminous crops in Northern regions of the country. The virus characterized as a bipartite begomovirus encodes eight functional proteins both in viral and complementary strands of both the genomic components (DNA A & DNA B) and the coat protein is found to be indispensable for virion assembly and insect transmissibility. Seven site directed mutations (S129K, K130Q, T135Q, K139F, C141V, F147Y and D152E) in CP gene were generated using MYMIV (Acc No: AY049772) are infectious on French bean (100%), mungbean (90-96%) and cowpea (80-90%). Systemic spread and symptom expression were seen in all the mutants. The mutants T135Q, K139F and C141V showed sparse yellow mosaic unlike other four mutants where the latter showed severe yellow mosaic as seen in wild. Unlike wild type the percent infectivity was less in all the mutants in all the three hosts tested. The mutants T135Q, K139F and D152E were found non-transmissible. Retention kinetics and tracking of the virions inside the whitefly were analyzed by dissecting the body parts (stylet, filter chamber, midgut, hindgut, whole digestive system and salivary gland) of an individual whitefly fed on the infected plants. Results indicate that in the mutants T135Q and K139F transport of the virus to salivary gland from hoemocoel is hindered. The study is expected to reveal the domain in CP which is required for acquiring and transmission of YMV affecting grain legumes is discussed. VI ICLGG - Posters P-PAD25 Deciphering molecular facet of MYMIV-induced pathogenesis in Vigna mungo employing genomic and proteomic techniques Pal A*, Kundu A, Kundu S Bose Institute, Kolkata, India *E-mail: amita@bic.boseinst.ernet.in Abstract Mungbean yellow mosaic India virus (MYMIV) causes yellow mosaic disease in Vigna mungo. Successful disease establishment resulted in irregular, chlorotic, yellow patches on the leaves, the characteristic phenotype of MYMIV-infected susceptible plants. Substantial crop-yield-penalty is a consequence of patho-inhibition in the function of photosystem II induced by MYMIV with an associated decline in quantum photosynthetic efficiency of diseased plants. Severe yield penalty occurs when MYMIV infects susceptible plants at the juvenile stage of development. To understand the molecular basis of compatible interaction comprehensively during MYMIV-induced pathogenesis in V. mungo, genomic and proteomic approaches have been undertaken in our laboratory. Four suppression subtractive hybridization libraries were generated and 113 unigenes with 90 singletons were identified during compatible interaction. Differentially expressed ESTs were functionally categorized into nine subgroups; induced vs. repressed were 47 and 53%, respectively. ESTs that were extensively modulated during disease establishment were associated mainly with primary metabolism and photosynthesis/energy. Successful evasion of host’s surveillance system and pathogen’s effector proteins counteracting the effects of various defense and signal transduction-related proteins already present or produced by the host plants during the pathogen attack have been comprehensively analyzed by proteomic approach. Hundred and nine proteins were identified and characterized, of which 37 were mostly down-regulated during pathogenesis. Surrender of these genes/proteins functionally involved in the photosynthetic machinery and a shift in primary metabolisms of the host plant during MYMIV invasion is the decisive factor for the establishment of pathogenesis. Perhaps during the compatible interaction MYMIV target these physiological processes of the plant to their advantage. VI ICLGG - Posters How resistant are the Portuguese common bean traditional landraces to powdery mildew and rust fungi? 1,* 1 1 2 2 Leitão ST , Dinis M , Almeida NF , Moral A , Rubiales D , Vaz Patto MC P-PAD26 1 1 Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal 2 Institute for Sustainable Agriculture, Córdoba, Spain *E-mail: sleitao@itqb.unl.pt Abstract Common bean (Phaseolus vulgaris L.) was introduced in Portugal more than 5 centuries ago, becoming a popular crop throughout the country due to its high organoleptic and nutritional properties. Nowadays it represents 80% of the National grain legume income (INE). However, common bean is vulnerable to several diseases, such as rust and powdery mildew, which leads to significant yield losses, limiting its cultivation. The very diverse bean landraces that are still grown at farmers’ fields represent unique and valuable genetic resource for breeding and disease resistance improvement. From the Portuguese Germplasm Bank (BPGV) common bean collection, 89 traditional landraces were evaluated for resistance to powdery mildew and 92 to rust fungi in seedling tests, under controlled conditions. Powdery mildew disease severity (DS) was assessed 7 and 15 days after inoculation (DAI) as the percentage of leaf tissue covered by mildew colonies. Many landraces showed intermediate levels of DS (6-25%) when compared to the susceptible check, 3 had no visible symptoms and 12 showed low levels of DS (0-5%) with hypersensitive resistant reaction. Rust infection type (IT) was analyzed 15 DAI using a 0-4 scale and a range of patterns of disease reaction was observed in the collection. DS was also estimated, with 42 landraces showing low values (DS<5%) and IT=4, indicative of partial resistance. The high levels of resistance here described and the associated high genetic diversity detected in these landraces in previous studies anticipate great potential of the traditional national germplasm for the enhancement of this important crop. VI ICLGG - Posters P-PAD27 Anthracnose resistance in lupin – increasing yield stability and promoting growth of homegrown legumes 1, 1 2 1 Ruge-Wehling B *, Fischer K , Dieterich R , Wehling P 1 Julius Kühn-Institut, Federal Research Centre for Cultivated Plants, Groß Lüsewitz, Germany 2 Saatzucht Steinach GmbH, Bornhof, Germany *E-mail: brigitte.ruge-wehling@jki.bund.de Abstract Anthracnose, caused by the fungus Colletotrichum lupini, is among the most important lupin diseases. For the time being, fungicide application and seed treatment are necessary to prevent C. lupini infection in the field. Stability of lupin seed production shall be improved by the development of diseaseresistant lupin cultivars. Wild accessions and cultivated varieties of L. angustifolius and L. luteus were tested to select novel resistance resources to anthracnose. Resistance tests under controlled greenhouse conditions revealed one breeding line of L. angustifolius and two cultivars of L. luteus with high resistance. Field tests confirmed the strong resistance in all cases. From the potential resistance resources, segregating mapping populations are being developed for genetic and molecular marker analysis. Segregation data from a mapping population (N= 150) indicated a dominant inheritance of the resistance detected in narrow-leafed lupin. The resistance phenotype observed under field conditions was different from that of the Australian cv. 'Tanjil'. This and the differing sources of resistance suggest that the anthracnose resistance in our narrow-leafed lupin material is different from the one effective in cv. 'Tanjil'. For mapping purposes, anchor markers provided by M. Nelson (University of Western Australia, Perth) are used. Additionally, sequences differentially expressed in the resistant vs. susceptable parents of the mapping population are used to achieve closely linked markers which can be used in resistance breeding. VI ICLGG - Posters Rhizosphere microbes in community alleviates biotic stress in chickpea through enhanced antioxidant and phenylpropanoid activities 1 2, 1 Singh A , Sarma BK *, Upadhyay RS , Singh HB P-PAD28 2 1 Department of Botany, Banaras Hindu University, Varanasi, India Department of Mycology and Plant Pathology, Banaras Hindu University, Varanasi, India 2 *E-mail: birinchi_ks@yahoo.com Abstract We examined the efficacy of a rhizospheric microbial consortium comprising of a fluorescent Pseudomonas (PHU094), Trichoderma (THU0816) and Rhizobium (RL091) strain in stimulating the physiological defense responses in chickpea against the collar rot pathogen Sclerotium rolfsii. Results of individual microbes were compared with dual and triple strain mixtures with reduced microbial load (1/2 and 1/3rd, respectively, of individual microbial load compared to single microbe application) in the mixtures. Periodical studies revealed maximum activities of phenylalanine ammonia lyase [E.C. 4.1.3.5] and polyphenol oxidase [E.C. 1.14.18.1] and accumulation of total phenolics in chickpea in the triple microbe consortium treated plants challenged with the pathogen compared to the single microbe and dual microbial consortia. Similarly, the expression of the antioxidant enzymes superoxide dismutase [E.C.1.15.1.1] and peroxidase [E.C.1.11.1.7] was also highest in the triple microbial consortium which was correlated with lesser lipid peroxidation in chickpea under the biotic stress. Histochemical staining clearly demonstrated higher and uniform lignification in vascular bundles of chickpea stems treated with the triple microbes. The physiological responses were directly correlated with the mortality rate as least plant mortality was recorded in the triple microbe consortium treated plants. 2-D gel electrophoresis analysis also revealed expression of 30 differential proteins induced in the treatments comprising mostly from the photosynthetic, defence and antioxidant pathways further confirming the previous results. The results thus suggest an augmented elicitation of stress response in chickpea under S. rolfsii stress by the triple microbial consortium in a synergistic manner under reduced microbial load. VI ICLGG - Posters P-PAD29 Existence of the second gene (er2) for powdery mildew resistance in pea (Pisum sativum) not confirmed by complementation test Sharma B* Division of Genetics, Indian Agricultural Research Institute, Delhi, India *E-mail: drbrsh@yahoo.co.in Abstract Several reports have suggested about two-gene control of powdery mildew resistance in pea. After the first report of Harland (1948) about one recessive gene causing resistance against Erusiphe pisi, the second published report of Heringa et al. appeared in 1969 in which a second gene for powdery mildew resistance in pea was claimed to exist, which was assigned gene symbol er2. However, the use of Heringa’s pea strains in hybridization did not confirm the possibility of the second gene. In fact, these strains were demonstrated to carry the same gene which was earlier reported by Harland and was named as er1. Several studies subsequently reported the er2 gene to exist. However, these studies also revealed some deficiency in the methodology of experiment or interpretation of results. The discussion about monogenic or digenic control of powdery mildew resistance continues till today, and the dispute has not been resolved even after 43 years. Considering the importance of powdery mildew in pea all over the world and the extent of crop losses due to this disease, which is under the control of Mendelian gene(s), it is absolutely essential to understand the genetic nature of the disease controlling mechanism. With the simplicity of major gene control of powdery mildew in pea, it should have been possible to confirm the situation through the simple complementation test by crossing the confirmed sources of the er1 allele and the putative donors of er2. Surprisingly, this was never done while claims have been made about molecular tagging of er2 on chromosome 3. Location of er1 on chromosome 6 was established long ago. An attempt has been made to make crosses between the known donors of er1 (variety Stratagem, or JI 2302 in the John Innes Institute collection) and the disputed allele er2 (Svp 951 or JI 2480). The results so far suggest that the two genes, one real and the other hypothetical, do not complement and the F1 plants remain resistant. This is a very strong argument against the existence of er2 gene even though it is claimed to be non-linked with er2. Further experiments to demonstrate linkage of er2 with marker genes for other visible traits on chromosomes 3 and 6 are in progress. VI ICLGG - Posters Identification of candidate defence-related genes to Sclerotinia sclerotiorum in soybean; functional analysis in soybean, Arabidopsis, and Nicotiana benthamiana 1,* 1 1 2 P-PAD30 1 Simmonds DH , Blahut-Beatty L , Koziol L , Buchwaldt L , Zhang Y , Calla 3 3 3 B , Neece DJ , Clough SJ 1 Agriculture and Agri-Food Canada, Ottawa, Canada Agriculture and Agri-Food Canada, Saskatoon, Canada 3 United States Department of Agriculture and University of Illinois, USA 2 * E-mail: daina.simmonds@agr.gc.ca Abstract In soybean, the wheat germin gene (gf-2.8) confers a high degree of resistance to Sclerotinia sclerotiorum, a necrotrophic fungus. The transgene product, oxalate oxidase (OxO), catalyzes oxalic acid (OA) to produce H 2O2. To identify genes with a role in defence, microarray studies have been used to examine the changes in soybean gene expression in response to infection of the transgenic (resistant) and parental (susceptible) lines. In addition, the effect of OA, a major virulence factor of S. sclerotiorum, was evaluated by leaf infiltration with OA. Thousands of genes were found to be significantly differentially expressed in each of the two studies. To identify genes related to defence, genes were classified functionally, based on the annotation of their best sequence match in public databases. Cluster analyses identified many defence-related genes that were induced across the studies, including genes annotated as GSTs, P450s, MMPs, PR proteins, WRKYs and genes of the phenylpropanoid pathway. Verification of a defence-related role of the candidate genes by over-expression or silencing is being assessed in soybean, Arabidopsis and in Nicotiana benthamiana. The goal of this study is to identify genes with the most relevant role in defence, either to aid molecular breeding or for transgenic modification to improve disease resistance. VI ICLGG - Posters P-PAD31 Screening of mungbean [Vigna radiata (L.) Wilczek] genotypes for resistance to mungbean yellow mosaic virus (MYMV) Suman S*, Kishore C, Sharma VK, Kumar R, Kumar H, Shahi VK Rajendra Agricultural University, Pusa (Samastipur), India *E-mail: sugandhsuman@gmail.com Abstract Pulses have been grown since millennia as a vital ingredient of the human diet. India is the largest producer of pulses, accounting for about 25 per cent of the global share. The important Asiatic species of genus Vigna, Vigna radiata (L.) Wilczek (mungbean, green gram) is widely cultivated pulse crop globally and in India. The yield of this crop is low due to its susceptibility to diseases including Mungbean Yellow Mosaic disease caused by mungbean yellow mosaic virus (MYMV), the most devastating pathogen transmitted through whitefly, Bemesia tabaci.Thirty-five genotypes of mungbean were screened for their resistance to MYMV on the basis of agro-morphological traits under natural conditions in summer, 2012. The genotypes were scored for MYMV disease and grouped into different categories from highly resistant to highly susceptible based on 1 to 9 rating scale. None was found to be highly resistant (HR), 2 were resistant (R), 5 moderately resistant (MR), 16 moderately susceptible (MR), 11 susceptible (S) and 1 highly susceptible (HS) among the 35 genotypes including one as a susceptible check. The highly resistant genotypes were IPM 2-14 and PDM 139, while the highly susceptible genotype was LGG 450. The resistance and susceptibility against MYMV varies with the environmental conditions under which these genotypes were grown and evaluated VI ICLGG - Posters Differential gene expression between tolerant and susceptible reactions in pea to Aphanomyces euteiches 1 P-PAD32 2, Saha G , Vandemark G * 1 Washington State University, Pullman, USA United States Department of Agriculture-Agricultural Research Service, Pullman, USA 2 *E-mail: george.vandemark@ars.usda.gov Abstract Aphanomyces root rot, caused by the soil borne oomycete Aphanomyces euteiches, is possibly the most globally destructive disease of pea (Pisum sativum L.). Commercial varieties at best have limited tolerance to this disease and losses can approach 100% in highly infested fields. Tolerance to disease is quantitatively inherited. Associating distinct disease reactions with differential expression of specific genes would provide robust evidence of gene function. Our objective was to examine levels of expression of selected genes in tolerant (90-2079) and susceptible (Dark Skin Perfection) pea genotypes in response to inoculation by A. euteiches. A preliminary analysis of expression patterns of 36 selected genes was performed with real-time PCR (Sybr Green). Based on these results, a set of 15 genes were selected for further study in repeated experiments. Expression profiles were examined 1, 2, and 3 days after inoculation and additional experiments were performed to examine expression profiles at 5 and 30 min after inoculation. Several genes were differentially expressed in tolerant and susceptible reactions. Significantly higher levels of expression of β-1,3 glucanase- 2 were observed in inoculated tolerant plants than in inoculated susceptible plants, for which expression of this gene could not be detected within 1 day after inoculation. A significant up-regulation of genes for disease resistant response protein-206 and ethylene receptor protein-5 were observed at 5 and 30 min after inoculation. Results will be presented for several other genes that showed differences in expression levels between tolerant and susceptible pea genotypes. VI ICLGG - Posters P-PAD33 Role of quarantine in biosecuring India from plant viruses: The case of legume germplasm imported during 2000-2012 Chalam VC*, Parakh DB, Maurya AK, Singh S, Khetarpal RK, Agarwal PC National Bureau of Plant Genetic Resources, New Delhi, India *E-mail: mailcelia@gmail.com, celia@nbpgr.ernet.in Abstract About 131 viruses are transmitted through true seeds, of which ~50% occur in legumes alone and can be introduced through import of seeds. This calls for stringent quarantine processing of imports. During 2000 to 2012, a total of 6,228 accessions of different legume crops imported from different countries and CGIAR Centres were subjected to quarantine processing by National Bureau of Plant Genetic Resources, New Delhi, which is empowered for the same. The accessions were grown in post-entry quarantine greenhouses and the plants showing virus-like symptoms were further subjected to a combination of more than one virus detection technique viz., infectivity test, electron microscopy, variants of ELISA and RT-PCR. Thirty viruses have so far been intercepted, which includes 8 viruses not yet reported from India viz., Bean pod mottle virus on soybean, Broad bean stain virus on faba bean and pea; Cherry leaf roll virus on French bean, green gram and soybean; Cowpea mottle virus on Bambara groundnut; Cowpea severe mosaic virus on cowpea, green gram and soybean; Pea enation mosaic virus on faba bean and pea; Raspberry ringspot virus and Tomato ringspot virus on soybean. Besides, 14 viruses not known to occur on particular host(s) in India have been intercepted. It may be noted that vectors of these viruses are present in India. The infected plants were incinerated and harvest from only virus-free plants was released to the indenters. The risk of introduction of 30 seed-transmitted viruses or their strains into India was thus eliminated. Greater risk is associated with bulk commercial imports that need to be addressed through proper sampling procedures with a group testing approach and detection techniques. The importance of quarantine has increased manifold due to liberalization of international trade and adopting not only the appropriate technique but also the right strategy for virus detection would go a long way in ensuring biosecurity of the Indian agriculture from plant viruses. VI ICLGG - Posters Characterization and pathogenicity of geminiviruses infecting pulse crops in India P-PAD34 * Malathi VG , Jyothsna P, Archana K, Kanakala S Advanced Centre for Plant Virology, Division of Plant Pathology, Indian Agricultural Research Institute, New Delhi, India *E-mail:vgmalathi@rediffmail.com Abstract Yellow mosaic disease of grain legumes and stunt disease of Chickpea are the major constraints in improving their productivity. The disease is caused by virus members belonging to the family Geminiviridae. They have circular single stranded DNA genome encapsidated in a characteristic geminate particle. YMD is caused by the viruses belonging to the genus Begomovirus, and the virus infecting chickpea belongs to the genus Mastrevirus. Molecular characterization has been completed for YMV infecting grain legumes, blackgram, mungbean, cowpea, soybean and dolichos from different states of India. The study revealed that four begomovirus species, MYMV, MYMIV, HgYMV and DoYMV cause YMD; they have unique identity and are not phylogenetically related to begomoviruses infecting other crops. Agroinoculation of cloned components led to systemic symptom expression in all the hosts tested. Among MYMIV isolates sequence identity is high (9498%). However, genetic complementation between DNA A and DNA B components of blackgram and cowpea isolates do not occur. One interesting feature of YMV is occurrence of DNA B variants. DNA B components can be differentiated into three groups, two groups typically associated with MYMV or MYMIV, the third group comprising DNA B variants that share 92% identity between them and may have intergenic region derived either from MYMV or MYMIV. The virus causing chickpea stunt disease has been identified as Chickpea chlorotic dwarf Pakistan virus. Comparison of nucleotide sequence of all chickpea infecting mastreviruses revealed that viruses occurring in Asia, Africa and Middle East are closely related to each other distinct from those occurring in Australia. Importance of results of our study on phylogenetic relationship, viral gene functions and host gene expression will be discussed. VI ICLGG - Posters Theme: Genomic Resources and Trait Mapping P-GRTM07 QTL mapping of floral scent compounds in cowpea (Vigna unguiculata) Andargie M 1,3,4 5 1, 2 , Knudsen JT , Pasquet RS 4 3,* , Muluvi GM , Timko MP 1 International Center of Insect Physiology & Ecology, Nairobi, Kenya Institut de Recherché pour le Développement, Montpellier, France 3 University of Virginia, Charlottesville, USA 4 Kenyatta University, Nairobi, Kenya 5 Lund University, Sweden 2 *E-mail: mpt9g@cms.mail.virginia.edu Abstract The development of genetically modified crop plants has aroused the biosafety concern associated with their release and the potential transgene escape from a GM crop to its non-GM crop counterparts or wild relatives. Identifying the different volatile compounds and tagging the genes using the closely linked molecular markers in order to suppress the scent of cowpea flowers is an important but complex breeding goal. The aim of this study was to use a molecular linkage map of cowpea to identify QTLs that are responsible for the scent compounds. The cowpea genetic map was constructed using the two parental lines, ‘524B’ and ‘219-01’ and the 159 F7 recombinant inbred lines by the help of SSR markers. ‘524B’ and ‘219-01’ showed very different volatile patterns with ‘219-01’ having higher volatile concentrations than ‘524B’. About 63 putative QTLs for a total of 22 different cowpea flower volatiles were detected through composite interval mapping by using the genotypic data of 159 F7 RILs of the mapping population together with phenotypic data obtained by GS-MS. QTLs for volatile compounds putatively involved in cowpea flower scent were found on 9 out of the 11 cowpea chromosomes, but they were not evenly distributed. QTLs were mainly clustered on linkage groups 1, 2 and 4. This new procedure is the first step for eventually blocking pollen flow from domesticated to wild cowpea and thus preventing the risk of transgene escape into wild cowpea gene pool. VI ICLGG - Posters Association of univalent frequency with gametic nonreduction and restoration of reductional meiotic cell division in interspecific hybrids of the genus Trifolium: A molecular cytogenetic analysis P-GRTM08 Ansari HA*, Ellison NW, Verry IM, Williams WM AgResearch Ltd, Grasslands Research Centre Palmerston North, New Zealand *E-mail: helal.ansari@agresearch.co.nz Abstract A near-sterile F1 hybrid (2n=4x=32) between 6x Trifolium ambiguum and 2x T. occidentale was formed with three genomes of T. ambiguum and one of T. occidentale. Because the functional female gametes from this F 1 were unreduced (2n), we undertook a molecular cytogenetic analysis to study the chromosomal processes leading to 2n gametes. In PMCs, at metaphase I (MI), 10-14 univalents failed to align at the equatorial plate, indicating delayed spindle-kinetochore attachment. At anaphase I (AI), the properly aligned bivalents segregated reductionally, indicating normal syntelic kinetochore attachment. Univalents invariably displayed equational division (precocious separation of sister chromatids), indicating amphitelic attachments of sister kinetochores. Laggards comprising individual and incompletely separated sister chromatids were frequent. Almost 35% of PMCs displayed minimal poleward chromosome movement. This was consistent with spindle failure caused by the large number of univalents, each generating spindle fibre tension because amphitelic sister chromatid separation was restrained by kinetochore cohesion. At AII, the cells with undivided nuclei produced dyads with unreduced chromosome number. Where AI produced proximally located daughter nuclei, there was either failure of poleward chromosomal movement at AII or reorientation to form parallel or triangular spindles leading to dyads and triads. By contrast with the F1 hybrid, two open-pollinated progeny plants (derived from unreduced female gametes) had restored reductional meiosis. At MI, both progeny exhibited only 2-4 univalents among 51-56 chromosomes, apparently insufficient to cause major spindle disruption. Results suggest that a high proportion of amphitelic univalents at AI can cause spindle disruption, leading to the formation of 2n gametes. VI ICLGG - Posters P-GRTM09 Development, characterization and cross-species transferability of ESTderived microsatellite markers for mungbean (Vigna radiata (L.) Wilczek). Bansal R*, Gupta SK, Vaidya UJ, Gopalakrishna T Bhabha Atomic Research Centre, Mumbai, India *E-mail: ravindra.barc@gmail.com Abstract Microsatellite or simple sequence repeat (SSR) markers, which detect sequence variation in the hypervariable regions of tandem repeats of 2 to 6 base pairs, are a powerful tool for genome analysis because of their codominant nature, locus specificity and high reproducibility. The large EST databases available in public domain provide a valuable resource for the identification and development of SSR markers. EST-SSRs, being part of the genes, are more useful as genetic markers. This study reports the development and characterization of EST-based microsatellite markers in mungbean (Vigna radiata (L.) Wilczek). A total of 889 EST sequences of mungbean from NCBI were searched for the presence of microsatellite repeats and 33 EST sequences with microsatellite repeats were identified. A total of 20 microsatellite markers were developed and were evaluated for the polymorphism on 22 mungbean genotypes. Ten microsatellite markers (50%) were found to be polymorphic and the number of alleles ranged from 2 to 3 with an average of 2.3 alleles per locus. Polymorphic information content of the microsatellite markers ranged from 0.09 to 0.59 with an average of 0.31. Clustering analyses using microsatellite data distributed the 22 mungbean genotypes into 5 main clusters with Jaccard’s similarity coefficient ranging from 0.22 to 1.0, indicating that moderate genetic variability is present in the mungbean genotypes. These microsatellite markers were also showing high transferability (90%) to other Vigna species. This study demonstrated that mungbean EST-derived microsatellite markers are highly polymorphic and will be useful for linkage mapping, gene tagging and genetic diversity studies in mungbean and other related Vigna species. VI ICLGG - Posters Molecular diversity and association of simple sequence repeat markers to peanut bud necrosis (PBND) in cultivated groundnut (Arachis hypogaea L.) Kamdar JH, Goswami BR, Bera SK P-GRTM10 * Directorate of Groundnut Research, Junagadh, India *E-mail:berask67@yahoo.co.in Abstract Molecular markers are useful tools for assaying genetic variation and provide an efficient means for early and reliable selection of genotypes having resistance to peanut bud necrosis disease (PBND) in groundnut breeding programs. Molecular diversity and association of simple sequence repeat (SSR) markers with resistance to PBND was detected in 24 groundnut genotypes differing in degree of resistance to PBND. Forty-eight primer pairs yielded 568 bands of which 380 were polymorphic, with an average of 7.91 polymorphic bands per primer. Polymorphism ranged from zero to 100% with an average of 65.87%. Cluster analysis (UPGMA) revealed two main clusters separated at 76% Jaccard’s similarity coefficient based on resistance to PBND. All 14 susceptible genotypes were grouped in one cluster, while 11 resistant genotypes grouped into separate cluster. AMOVA among 24 genotypes detected 35% (P<0.001) of total variation associated with resistance to PBND. Kruska-Wallis ANOVA detected the significant association of 10 primers with resistance to PBND. Seven out of ten primers explained more than 10% of phenotypic variation due to resistance to PBND. These loci are associated with the resistance to PBND 2 in groundnut and seemed to be major QTLs with regression coefficient value (r ) ranging from 15 to 63%. Besides, both loci PM145140 and PM1590 differentiated the DNA bulked of resistant genotypes from the DNA bulked of susceptible genotypes. VI ICLGG - Posters P-GRTM11 The identification of QTLs for disease resistance and agronomic traits in a population derived from a cultivated x synthetic cross, and introgression of disease resistance into cultivated peanut. 1, 2 3 2 Leal-Bertioli SCM *, Shirasawa K , Ozias-Akins P , Isobe S , Guimarães 1 4 5,6 1 PM , Araújo AE , Bertioli DJ , Moretzsohn MC 1 EMBRAPA, Genetic Resources and Biotechnology, Brasília, Brazil Kazusa DNA Research InstituteChiba, Japan 3 University of Georgia, Tifton, USA 4 EMBRAPA Cotton, Brazil 5 University of Brasília, Campus Darcy Ribeiro, DF, Brazil 6 Catholic University of Brasília, Brasília, Brazil 2 *E-mail: soraya.bertioli@embrapa.br Abstract Cultivated peanut has an exceptionally narrow genetic base and low DNA polymorphism. This imposes certain limitations on genetic improvement, in particular with regard to disease resistance and has hindered molecular breeding. The use of wild species has the potential to bring much needed genetic diversity to breeding programs and introduce DNA polymorphism to facilitate the construction of genetic maps, and molecular breeding. We developed a recombinant inbred mapping population from a cross of cultivated peanut with a synthetic amphidiploid derived from the two most probable wild diploid ancestors. A genetic map was constructed using microsatellite, MITE and SNP markers, the population phenotyped for resistance to late leaf spot, plant architecture, and productivity. QTLs were identified. In parallel, a simple proof-of-principle introgression of wild genes was conducted. Forty BC 1s were developed and BC1F2 families field evaluated. Twenty-four BC1F3 families were selected based on evaluations for disease resistance, plant architecture and productivity. Further selection was done and 12 families of BC 1F4 were planted in the following year. When compared to the recurrent cultivated parental, a considerable number of lines exhibited superior resistance to leaf diseases, had agronomically adapted architecture and similar levels of productivity. These lines were genotyped at low density, and an analysis of the introgressed wild segments carried out. In summary, we identified QTLs for disease resistance and agronomic characteristics in a synthetic amphidiploid, and showed that disease resistance can be improved, and a surprisingly rapid recovery of agronomic traits made, through a simple back-crossing scheme and selection. VI ICLGG - Posters The first consensus genetic map for cultivated pigeonpea [Cajanus cajan (L.) Millsp.] based on SSR markers 1,2 1 3 1 P-GRTM12 3 Bohra A , Saxena RK , Gnanesh BN , Saxena KB , Byregowda M , Rathore 1 4 5 1,6,* A , Kavi PBK , Cook DR , Varshney RK 1 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 2 Present address: Indian Institute of Pulses Research (IIPR), Kanpur, India 3 University of Agricultural Sciences, Bengaluru, India 4 Osmania University, Hyderabad, India 5 University of California Davis, Davis, USA 6 CGIAR Generation Challenge Programme (GCP), CIMMYT, Mexico DF, Mexico *E-mail: r.k.varshney@cgiar.org Abstract Pigeonpea [Cajanus cajan (L.) Millsp] occupies an important place among the food legume crops of tropical regions and accounts for 5.7 percent of the worldwide pulses production from 6.5 percent of the total pulses area. Though sincere efforts have been made to improve pigeonpea productivity using traditional breeding, appreciable success has not been witnessed so far. Therefore, necessitating integration of molecular tools in regular breeding programme to experience the anticipated progress. However in pigeonpea inadequate supply of DNA polymorphisms and dearth of genetic maps/QTLs have posed major obstacles in undertaking molecular breeding for pigeonpea improvement. Although recently some simple sequence repeat (SSR) based genetic maps have been reported in cultivated pigeonpea, these genetic maps could provide map positions to approximately 70-120 SSR loci only. With the objective of leveraging the pigeonpea genomics repertoire, here we report development of four new intra-specific genetic maps and synthesis of one integrated genetic map merging segregation data from six different intraspecific F2 mapping populations. The consensus genetic map comprised of a total of 339 SSR loci and covered a map length of 1,059 cM. To make the consensus map more informative, the map was divided into several bins of 10 cM each and all the mapped SSR markers were assigned polymorphism information content (PIC) values which would help geneticists and breeders to select a good set of SSR markers representing the genome best as well as displaying higher level of DNA polymorphism. In summary, the developed consensus genetic map would act as reference map for cultivated pigeonpea and would facilitate precise selection of markers for future linkage/association analyses, genetic diversity studies and practising foreground and background selection via marker-assisted back crossing (MABC) for pigeonpea genetic improvement. VI ICLGG - Posters P-GRTM13 Mapping of QTLs for biological yield and days to flowering in red clover (Trifolium pratense) Chahota RK, Sharma TR, Verma B, Sharma S, Gupta M* Department of Agricultural Biotechnology, CSK Himachal Pradesh Agricultural University, Palampur, India *E-mail: rkchahota@yahoo.com Abstract Red clover (Trifolium pratense) is one of the most important components of permanent pastures and meadows in temperate parts of the world. During the last decades many improved red clover cultivars have been released but they often suffer from diseases and susceptibility to water stress resulting into poor yield. Days to flowering and biological yield are the important traits determining the fodder yield and duration of vegetative phase. These two complex traits are influenced by the many factors and are negatively correlated with each other. Molecular markers may help to improve these traits rapidly as in case of MAS. Therefore, the present study was undertaken to identify markers linked to quantitative traits loci (QTLs) for their use in the marker assisted selection. A genetic linkage map was constructed using RIL population of 94 lines and 195 PCR-based markers were mapped for QTL analysis of these traits. The genetic linkage map covered a total length of 421.5 cM with an average distance of 1.2 cM between loci. A total of 5 QTLs were identified for these traits, of these three QTLs were found associated with biological yield explaining 35.1% variation and two QTLs with days to flowering explaining 14.4% of the total variation. These QTLs rich genome regions represent the valuable candidate regions for further characterization and development of markers closely linked to these traits. VI ICLGG - Posters Identification of QTL for grain yield and other traits in chickpea Anuradha Ch 1,2,* 2 2 , Gaur PM , Ganesh M , Kumar J P-GRTM14 1,3 1 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 2 Acharya NG Ranga Agricultural University, Hyderabad, India 3 Hendrick Beans-for-Health Research Foundation, Inkerman, Canada *E-mail: anu_dna@rediffmail.com Abstract Quantitative trait loci (QTL) were identified for grain yield and several phenological (days to first flower, days to maturity) and morphological traits (number of nodes below the first flower, plant height, plant width, number of primary branches per plant, number of pods per plant, number of seeds per plant, number of seeds per pod, 100-seed weight) in chickpea (Cicer arietinum L.) based on the genotypic and phenotypic evaluation of ICCV 2 x JG 62 RILs. QTL that explained more than 10% of the phenotypic variation were identified for grain yield, number of nodes below the first flower, plant height, plant width, number of primary branches per plant and number of seeds per pod. One QTL that explained 15.2% of the phenotypic variation for grain yield was identified on LG6A at LOD 3.08 with flanking markers TA44X and TR1s. One QTL was identified each for number of nodes below the first flower and plant height on LG 3 accounting for 10.7 and 15.1% of the phenotypic variation, respectively. Three QTL, together accounted for 22.4% of the phenotypic variation, were identified for plant width and mapped on LG6A, LG3 and LG9. Seven QTL, explaining 25.6% of the phenotypic variation, were identified for number of seeds per pod. Genomic regions were identified which contained QTL for multiple traits. For example, LG 3, LG 4 and LG 9 each carried six QTL for various traits. Further studies are needed on validation of these QTL for use in chickpea improvement program. VI ICLGG - Posters P-GRTM15 EST-SSRs from a Pisum sativum transcriptome 1 2 3 4, 2 2 McGee RJ , Cheng C-H , Mockaitis K , Coyne CJ *, Jung S , Zheng P , 1 1 2 Vandemark GJ , Chen W , Main D 1 USDA-ARS, Grain Legume Genetics and Physiology Research Unit, Pullman, USA 2 Department of Horticulture, Washington State University, Pullman, USA 3 Indiana University Center for Genomics and Bioinformatics, Bloomington, USA 4 USDA-ARS, Western Regional Plant Introduction Station, Pullman, USA *E-mail clarice.coyne@ars.usda.gov Abstract We used the next generation sequencing technology to develop a public pea transcriptome resource, mined the assembled pea unigenes for SSRs. RNA pools representing seven tissue types from pea cv ‘Aragorn’ produced 3.084 million high quality reads, which averaged 1044 bp in length after cleaning. Assembly of the cleaned reads with NEWBLER and MIRA resulted in 37,455 unigenes of which approximately 80% had homology to previously identified proteins in the TAIR, TrEMBL and Swiss-Prot databases. We used a tool developed for the Genome Database for Rosaceae to identify microsatellites. This tool will allow you to upload a batch of sequences in FASTA format and return information on those sequences that contain microsatellites based on the motif frequency that you specify. Having submitted your sequences for processing you will be sent an email after the job has completed that will direct you to a website with links to files you can download. These files will contain an EXCEL spreadsheet with information on the type of motif, frequency, location of SSR in the sequence, sequence ORF information and whether the repeat in the ORF. If you have selected "generate primers" you will also be provided with the optimal primers generated from PRIMER3 and the expected product size. A separate output file gives you all alternate primers suggested by PRIMER3. A fasta file of the sequences containing the SSRs is also generated as is the original file you uploaded. From a total of 37,455 pea sequences 9,346 contained microsatellites with a total of 537 unique motifs. A summary of the analysis is provided in the tabbed EXCEL file available from the Cool Season Food Legume Genome Database (http://www.coolseasonfoodlegume.org). This database is being developed to assist in translating genomics into crop improvement and facilitate cool season food legume breeding and research by serving as a genomic, genetic and breeding data resource. The database is built using Tripal which provides simplified site development by merging the power of Drupal, a popular web Content Management System (CMS), with that of Chado, a community derived database schema for storage of genomic, genetic and other biological data. VI ICLGG - Posters Identification of groundnut SSR markers suitable for multiple resistances QTL mapping in African germplasm 1, 2 2 2 1 P-GRTM16 3 De Villiers SM *, He G , Prakash C , Zhao Y , Njunge VM , Monyo ES , 4 4 4 Mweetwa AM , Kaimoyo E , Siambi M 1 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Nairobi, Kenya 2 Tuskegee University, Alabama, USA 3 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Lilongwe, Malawi 4 University of Zambia, Lusaka, Zambia E-mail: s.devilliers@cgiar.org Abstract Groundnut is an important legume and contributes to household food security and nutrition in many countries in Africa. Good quality groundnuts from Malawi and Zambia in southern Africa earn valuable incomes from exports to Europe but yields suffer from diseases such as groundnut rosette disease (GRD), early leaf spots (ELS), rust and aflatoxin contamination. Sources for resistance or tolerance have been identified and the next step is to introgress this resistance into farmer- and market preferred varieties through QTL mapping and markerassisted back-crossing (MAB). As a first step, this study identified SSR markers that are well distributed across the genome and showed good polymorphism amongst the resistance sources and high yielding, market preferred varieties. We developed a comprehensive database of published polymorphic SSR markers in groundnut by analyzing information on these markers from publications, genome databases and laboratory screening. We then screened 18 varieties, including the sources for resistance to GRD (ICG 12991, ICGV-SM 90704), ELS (ICG 7878, ICGV-SM 95714), rust (ICGV-SM 94114, ICGV-SM 95342) and aflatoxin tolerance (55-47) as well as market preferred varieties (CG7, Chalimbana, Fleur 11, ICGV 86124, ICGVSM 99557, JL24, MGV5, Nyota, Pendo, Red Mwitunde and 47_10) with more than 1200 SSRs markers. Results showed that more than 500 SSR markers were polymorphic, ranging from 120 to 410 for different combinations of two varieties. Thus, we were able to compile a toolkit indicating the polymorphic markers across all 18 varieties that can be used to select the best combinations for bi-parental mapping population development and/or markerassisted introgression of the target disease or aflatoxin resistance. VI ICLGG - Posters P-GRTM17 Consensus genetic map and QTLs for drought tolerance traits in cultivated groundnut (Arachis hypogaea L.) 1,2 1 1 1 1 Gautami B , Pandey MK , Vadez V , Nigam SN , Ratnakumar P , 1 3 4 2 Krishnamurthy L , Radhakrishnan T , Gowda MVC , Narasu ML , Hoisington 1 5 1, 6,* DA , Knapp SJ , Varshney RK 1 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 2 Institute of Science and Technology, Jawaharlal Nehru Technological University, Hyderabad, India 3 Directorate of Groundnut Research (DGR), Junagadh, India 4 University of Agricultural Sciences, Dharwad, India 5 The University of Georgia, Athens, USA 6 Generation Challenge Programme (GCP), CIMMYT, Mexico DF, Mexico * E-mail: r.k.varshney@cgiar.org Abstract Drought stress is one of the major production constraints in semi-arid tropics (SAT) regions of the world in groundnut (Arachis hypogaea L.), an important food and cash crop. With an objective to use genomics tools together with conventional approaches for improving drought tolerance, efforts were made to identify QTLs and markers using two RIL populations namely ICGS 44 × ICGS 76 (RIL-2) and ICGS 76 × CSMG 84-1 (RIL-3). Genetic maps were constructed with 82 (RIL-2) and 119 (RIL-3) SSR loci after screening of 3,215 SSR markers on the parental genotypes of these populations. Together with these two maps and the already available reference map with 191 SSR loci based on TAG 24 × ICGV 86031 (RIL-1), a consensus map was constructed with 293 SSR loci distributed over 20 linkage groups, spanning 2,840.8 cM. As all these three populations segregate for drought tolerance related traits, a comprehensive QTL analysis identified 153 main effect QTLs (M-QTLs) and 25 epistatic QTLs (E-QTLs) for drought tolerance related traits. Localization of these QTLs on the consensus map provided 16 genomic regions that contained 137 QTLs. Few key genomic regions were selected on the basis of QTLs identified in each regions and their expected role for drought adaptation discussed. Given that no major single QTL for drought adaptation was identified, novel breeding approaches like marker-assisted recurrent selection (MARS) and genomic selection (GS) approaches are likely to be the preferred approaches for introgression of a larger number of QTLs to breed drought tolerant groundnut genotypes. . VI ICLGG - Posters Cross species transferability of simple sequence repeat markers in blackgram (Vigna mungo L. Hepper) 1,* 1,2 2 3 P-GRTM18 2 Gupta S , Anjum TK , Pratap A , Kumar J , Gupta DS 1 AICRP on MULLARP, Indian Institute of Pulses research (IIPR), Kanpur, India Division of Crop Improvement, Indian Institute of Pulses research (IIPR), Kanpur, India 2 *E-mail: saniipr@rediffmail.com Abstract Blackgram (Vigna mungo L. Hepper) is an important food legume species in South-east Asia, particularly Indian sub-continent. Use of transferable SSR markers in linkage maps of blackgram was limited and only 47 SSR loci were assigned to the 11 linkage groups Transferability of simple sequence repeat (SSR) markers was studied to enrich genomic resources in blackgram. Three hundred sixty three (361) simple sequence repeat markers developed in the past in other food legumes were used in present study for this purpose. Out of these, 247 SSR markers (68.04%) amplified though only 39 (15.8%) were highly polymorphic among 24 diverse genotypes of blackgram. Each of these SSR marker amplified for 2 to 3 alleles. The polymorphism information content ranged between 0.08 to 0.66 with an average of 0.48. Cluster analysis based on UPGMA grouped the studied genotypes into three major clusters. Jaccard’s similarity coefficient was ranged from 0.13 to 0.90. It was observed that transferability of SSR markers containing AG motif was more frequent as compared to other motifs in blackgram. A few SSRs also generated genotype specific allele that could be useful in maintaining the genetic purity of respective genotypes. These transferred SSR markers can be an important source for enriching available genomic resources of blackgram for mapping and tagging of the gene(s) of agronomic importance. VI ICLGG - Posters P-GRTM19 Towards fine mapping of drought tolerance related QTL region in chickpea (Cicer arietinum L.) Jaganathan D, Thudi M, Tuteja R, Gaur PM, Varshney RK* International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India *E-mail: r.k.varshney@cgiar.org Abstract Terminal drought is one of major constraints that lead to considerable yield losses (~50%) in chickpea. By using linkage mapping approach on ICC 4958 × ICC 1882 population, a genomic region (~35cM) harbouring several QTLs for drought tolerance related traits was identified on linkage group 4 (LG 4). This region contained only four markers (TAA170, ICCM0249, STMS11 and GA24). In order to increase marker density in this region, two approaches were adopted: (a) targeted mapping from the corresponding region from published maps in chickpea, and (b) Medicago-Cicer synteny approach. In the former approach, a set of 43 SSR markers were chosen from the corresponding region on the published genetic maps and genotyping data were for generated 19 polymorphic markers. In the later approach, syntenic regions on Medicago chromosome 1 were identified by aligning the tentative orthologous (TOG) markers present in the corresponding genomic region on the interspecific map (ICC 4958 × PI 489777). In summary, a set 591 primer pairs were designed from this syntenic region, of which 144 primers produced successful amplification in the intraspecific mapping population. A total of 75 SNPs were identified between the parental genotypes through allele specific sequencing. Only 32 SNPs were found CAPS convertible and 3 markers were polymorphic between the parental lines. By combining these two approaches, one SSR marker loci and three DArT loci were mapped in the QTL region for the intraspecific mapping population. At present, efforts are underway to saturate this region by using RAD-sequencing approach on the mapping population. VI ICLGG - Posters Transcriptome sequencing of chickpea for discovery of novel genes and genetic variations P-GRTM20 Jain M* National Institute of Plant Genome Research (NIPGR), New Delhi, India *E-mail: mjain@nipgr.res.in Abstract Chickpea is an important food legume because of its high nutritional value and ability to fix atmospheric nitrogen. Very low levels of genetic diversity within cultivated chickpea species has been an important constraint in chickpea improvement. The importance of wild species having important agronomic traits in improvement of chickpea has been realized. Thus, sequencing the transcriptome of wild species will lead to the identification of genetic variations of agronomic use. We sequenced the transcriptomes of cultivated desi (Cicer arietinum) and wild (Cicer reticulatum) chickpea using next generation sequencing technologies. Various softwares and approaches were used to obtain the best optimized assembly. A total of 34,760 and 37,265 transcripts were generated for desi and wild chickpea, respectively. The comprehensive functional annotation identified transcripts involved in various cellular processes. More than 4,000 simple sequence repeats (SSRs) were identified in the transcriptome sequences. At least 561 SSRs were found to be polymorphic between desi and wild chickpea. In addition, we identified 36,446 single nucleotide polymorphisms (SNPs) between the two chickpea types. Many of these SSRs and SNPs were found to be present in the tissue-specific and transcription factor encoding transcripts. The experimental validation of few polymorphic SSRs and SNPs showed very good correlation with the in silico prediction. The results of this study will help to accelerate research in various areas of genomics and provide a rich resource for exploiting genetic variations for breeding programmes in chickpea. VI ICLGG - Posters P-GRTM21 Enriching genomic resources to localize multiple disease resistance genes in field pea (Pisum staium L.) 1 2 1 3 4 1* Jain S , Weeden N , Miranda AL , Porter L , Eigenbrode S , McPhee K 1 North Dakota State University, Fargo, USA Montana State University, Bozeman, USA 3 USDA-ARS, Prosser 4 University of Idaho, Moscow, Russia 2 *E-mail: kevin.mcphee@ndsu.edu Abstract Pea is affected by many pests and pathogens that reduce seed yield and quality. A pea recombinant inbred line mapping population (PRIL-5) derived from the cross ‘Lifter’/‘Radley’ was developed to study the many desirable agronomic attributes, including resistance to various fungal and viral pathogens segregating in the population. PRIL-5 is comprised of 393 F7derived recombinant inbred lines advanced by single seed descent. Lifter has resistance to diseases e.g. pea enation mosaic virus, bean leaf roll virus and Fusarium wilt race 1 while Radley is resistant to Fusarium wilt race 2 and has partial resistance to Ascochyta blight. Gene-based markers were developed using the Intron Targeted Amplified Polymorphism (ITAP) approach based on synteny with Medicago truncatula. Pea expressed sequence tags (ESTs) were aligned with the M. truncatula genome sequence to identify conserved exon sequences and flanking one or more introns. Conserved primers were designed to exploit length variation or single nucleotide polymorphism within the intron and 22% of the markers were polymorphic in PRIL-5. Target Region Amplified Polymorphism (TRAP) markers were also developed using one conserved primer from a pea EST and one arbitrary primer. A genetic map of PRIL-5 has been developed with 179 loci including SSR, RAPD, CAPS, ITAP, TRAP and one morphological marker including 48 novel genic markers distributed on seven linkage groups. Disease screening for the pathogens is currently underway in greenhouse and field conditions. This genetic map will provide additional insights to localize various disease resistance genes/QTLs and has implications for further crop improvement. VI ICLGG - Posters Genomic translocation in Medicago truncatula 1,2, 1 1 1 P-GRTM22 1 1 1 Kiss GB *, Kaló P , Szabó Z , Balogh M , Miró K , Iliescu C , Harsányi I , 2 3 4 Lustyik G , Cook DR , Ellis THN 1 Agricultural Biotechnology Center, Gödöllő, Hungary Soft Flow Hungary Ltd, Pécs, Hungary 3 University of California Davis, Davis, California, USA 4 Aberystwyth University, Aberystwyth, UK 2 *E-mail: gbkiss@abc.hu Abstract Medicago truncatula (Mt) genetic map using an F2 segregating population from the cross of Mt A17 x Mt A20 was published by Choi et al, in 2004. During the course of this genetic work, the genotypes of some markers displaying high degree of genetic linkage (> 5 cM) showed unusual (cruciform) linkage characteristics with other linked markers analyzed (G.B. Kiss and D. Cook, unpublished). With the help of the genetic map of M. sativa (Kaló et al., 2000) Mt markers were sorted according to the M. sativa genetic map supposing genetic co-linearity between the two species. Later, Kamphuis et al. (2007) carried out pollen viability and linkage analysis using different Mt accessions and suggested that „The Medicago truncatula reference accession A17 has an aberrant chromosomal configuration”. The results of genetic mapping of Mt, M. sativa and pea (Pisum sativum), in addition to BAC contig constructions and BAC sequencing of the aberrant region on LG4 and LG8 will be shown and discussed. VI ICLGG - Posters P-GRTM23 Conserved synteny between the narrow-leafed lupin and sequenced legumes: a case study of selected gene-rich regions 1,* 1 1 1 1 Ksiazkiewicz M , Wyrwa K , Szczepaniak A , Rychel S , Przysiecka L , 2 2 1 1 Zielezinski A , Karlowski WM , Naganowska B , Wolko B 1 Institute of Plant Genetics, Polish Academy of Sciences, Poznan, Poland Institute of Molecular Biology and Biotechnology, Adam Mickiewicz University, Poznan, Poland 2 *E-mail: mksi@igr.poznan.pl Abstract Lupinus angustifolius is a legume crop widely cultivated across several continents from America through Europe to Asia, Africa and Australia. It has become an object of molecular studies, since the development of genomic resources including sequence defined markers, linkage maps and bacterial artificial chromosome (BAC) libraries. Here, we exploited these resources to analyze the structure and organization of narrow-leafed lupin gene-rich regions and their patterns of synteny to already sequenced legumes. Molecular probes based on marker sequences containing particular short sequence repeats (SSRs) were used for hybridization with BAC library. Clones selected by this approach have undergone restriction fingerprinting and physical mapping. Contigs of overlapping clones were constructed and representatives of contigs and singletons were analyzed by fluorescent in situ hybridization (BAC-FISH) on metaphase chromosomes. BAC-FISH was applied as a supporting tool to select BAC clones for sequencing and functional annotation. The process of annotation included de novo detection of specific signals located on genomic sequence as well as a comparative analysis. All clones which showed single locus signals in BAC-FISH turned out to be rich in gene encoding sequences and completely or partially free of transposable elements. Cross species genomic analysis revealed very well conserved synteny between these generich regions and homologous Glycine max, Medicago truncatula and Lotus japonicus sequences. On the contrary, BACs which displayed dispersed BACFISH signals on numerous chromosomes contained large amounts of repetitive content, occupying about 45% of sequences. The synteny patterns in such regions of genome were retracted by DNA rearrangements. VI ICLGG - Posters Large-scale development of cost-effective SNP marker assays for diversity assessment and genetic mapping in chickpea and comparative mapping in legumes 1,2 1,3 4 5 P-GRTM24 6 Hiremath PJ , Kumar A , Penmetsa RV , Farmer A , Schlueter JA , 1 6 4 1 Chamarthi SK , Whaley AM , Carrasquilla-Garcia N , Gaur PM , Upadhyaya 1 2 1 4 1,7,* HD , Kavi PBK , Shah TM , Cook DR , Varshney RK 1 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), India 2 Osmania University, Hyderabad, India 3 College of Agriculture, Jawahar Lal Nehru Krishi Vishwa Vidyalaya (JNKVV), Jabalpur, India 4 University of California, Davis, USA 5 National Center for Genome Resources (NCGR), Santa Fe, USA 6 University of North Carolina, Charlotte, USA 7 Generation Challenge Program (GCP), Mexico DF, Mexico *E-mail: r.k.varshney@cgiar.org Abstract A set of 2486 single nucleotide polymorphisms (SNPs) were compiled in chickpea using four approaches, namely (i) Solexa ⁄ Illumina sequencing (1409), (ii) amplicon sequencing of tentative orthologous genes (TOGs) (604), (iii) mining of expressed sequence tags (ESTs) (286) and (iv) sequencing of candidate genes (187). Conversion of these SNPs to the cost-effective and flexible throughput Competitive Allele Specific PCR (KASPar) assays generated successful assays for 2005 SNPs. These marker assays have been designated as Chickpea KASPar Assay Markers (CKAMs). Screening of 70 genotypes including 58 diverse chickpea accessions and 12 BC 3F2 lines showed 1341 CKAMs as being polymorphic. Genetic analysis of these data clustered chickpea accessions based on geographical origin. Genotyping data generated for 671 CKAMs on the reference mapping population (Cicer arietinum ICC 4958 × Cicer reticulatum PI 489777) were compiled with 317 unpublished TOG-SNPs and 396 published markers for developing the genetic map. As a result, a second-generation genetic map comprising 1328 marker loci including novel 625 CKAMs, 314 TOG-SNPs and 389 published marker loci with an average inter-marker distance of 0.59 cM was constructed. Detailed analyses of 1064 mapped loci of this second-generation chickpea genetic map showed a higher degree of synteny with genome of Medicago truncatula, followed by Glycine max, Lotus japonicus and least with Vigna unguiculata. Development of these cost-effective CKAMs for SNP genotyping will be useful not only for genetics research and breeding applications in chickpea, but also for utilizing genome information from other sequenced or model legumes. VI ICLGG - Posters P-GRTM25 Identification and mapping of QTLs conferring resistance to Ascochyta blight in chickpea (Cicer arietinum L.) Vashist U, Kumar P*, Sangwan MS, Dhillon S, Boora KS Chaudhary Charan Singh Haryana Agricultural University, Hisar, India *E-mail: pardeep4yadav@gmail.com Abstract Ascochyta blight, caused by Ascochyta rabiei (Pass.) Lab. is a devastating disease of chickpea (Cicer arietinum L.) worldwide. The disease affects stems, leaflets, pods and seeds causing necrotic lesions and breakage of stems and petioles. Ascochyta blight infections may cause 100% yield loss. Resistant germplasm has been identified and the genetics of resistance has been the subject of numerous studies. The objective of this study was to identify and map the QTLs for ascochyta blight resistance genes in chickpea using F 2:3 populations, as well as to quantify the contribution of each QTL to observed phenotypic variation. F2:3 populations were derived from cross HC-5 (ascochyta blight resistance) x HC-3 (susceptible). Disease reactions of the parents and F2:3 populations were scored using 1 to 9 scale. The 102 SSR primers were used to discriminate the parental chickpea genotypes. By using the primers which discriminated the parental genotypes for ascochyta blight disease, three major QTLs: H2B202, H1A19 and P78 were identified and mapped on linkage group 5 (LG 5) of chickpea linkage map. The major QTL (H2B202) contributed 46 % of the total phenotypic variation for ascochyta blight resistance along with two QTLs, P78 and H1A19 having phenotypic variation 17% and 9% respectively. These three QTLs explained 72% of the phenotypic variation in the population suggesting the involvement of these three segments having several minor genes with robust effects on the trait. The QTLs identified for resistance to ascochyta blight can be helpful in crop improvement programmes and resistance breeding in chickpea. VI ICLGG - Posters Comparative analysis of functional and anonymous SNP diversity in Legumes and conversion of SNPs into PCR based markers 1,* 2 2 2 1 P-GRTM26 2 Lohithaswa HC , Shilpa HB , Kumar KS , Jyothi K , Bi K , Hittalmani S 1 Department of Genetics and Plant Breeding, College of Agriculture V.C. Farm, Mandya India 2 Department of Genetics and Plant Breeding, University of Agricultural Sciences, Bangalore, India *E-mail: lohithhc@rediffmail.com Abstract The legumes are the third largest family (Leguminosae) among angiosperms with more than 650 genera and 19000 species. Many agronomically and nutritionally important legumes are considered orphan due to limited resources in the GenBank. The comparative genome analysis to assess synteny facilitates use of genomic resources between different legume species. With the progress in sequencing efforts in model plant species like M. truncatula, it has become possible to characterize syntemic relationships between model and other economically useful members of the legume family. Conserved Intron Scanning Primers (CISPs) thus developed can provide large numbers of genomic tools suitable for linking genomics research in crops of critical nutritional and economic importance that lack appreciable sequence information, to growing knowledge in botanical models and better studied crops. A total of 1024 CISP pairs from 67,429 Medicago-Common Bean, 1231 CISP pairs from 62,591 Medicago-Cowpea and 278 CISP pairs from 25,132 Medicago-Cajanus cajan alignments were designed. Out of these, 384 Cowpea-Medicago truncatula, 192 – Pigeonpea-Medicago truncatula and 288 Commonbean-Medicago truncatula CISP sets predicted to amplify nonredundant Medicago truncatula loci were synthesized and employed for PCR amplification in ten legumes. The CISPs worked in individual taxa for which DNA sequence information was not considered in primer design (chickpea, blackgram, soybean, groundnut and horsegram) which proved that CISPs are an effective means to explore poorly characterized genomes and also provide anchor points for comparative genomics across a diverse range of species. A total of 56, 76 and 147 single nucleotide polymorphisms were detected in chickpea, pigeonpea and cowpea, respectively. Sufficiently good number of single nucleotide polymorphisms was converted into dCAPS. VI ICLGG - Posters P-GRTM27 Preliminary Investigation of the genetic linkage of winter hardiness and photoperiodic response in lentil (Lens culinaris Medik.) 1 2,* 3 4 Sakar D , McGee RJ , Coyne CJ , Kahraman A , Muehlbauer FJ 2 1 Dicle University, Department of Field Crops, Diyarbakir, Turkey USDA-ARS, Grain Legume Genetics and Physiology Research Unit, Pullman, USA 3 USDA-ARS, Western Regional Plant Introduction Station, Pullman, USA 4 Harran University, Department of Field Crops, Sanliurfa, Turkey 2 *E-mail: rebecca.mcgee@ars.usda.gov Abstract Lentils (Lens culinaris Medic.) are an integral component of cereal-based cropping systems. They contribute to agricultural sustainability by fixing atmospheric N, breaking disease and weed cycles, improving the cereals grain and protein yields, and providing diet diversification. Autumn-sown lentils have higher yield potential, improved efficiency of field operations and rotational and agronomic benefits in farming systems. However, they must have adequate levels of cold tolerance, tolerate harsh winter conditions and delay flowering until after the main winter freezing periods have passed. The objectives of this study were 1) determine the response to photoperiod within a population of F6 derived recombinant inbred lines (RILs) from a cross of ‘Precoz’ x WA86499090; 2) determine whether photoperiod response is related to winter hardiness and 3)determine the genetic map position of genes for photoperiod response. The population was previously screened for winter hardiness. Here we will report data collected when we screened the RILs for response of flowering to photoperiod in growth cabinets maintained at 8, 10, 12 and 16 o hour day length and constant temperature (20 C). We identified a photoperiod response on gene linkage group 3 that co-localizes with a previously identified QTL for frost tolerance. VI ICLGG - Posters A new QTL controlling growth habit in chickpea (Cicer arietinum L.) 1 2 3 3 4 1 Ali L , Rubio J , Madrid E , Cobos MJ , Rameau C , Gil J , Millan T P-GRTM28 1,* 1 Dept of Genetics, University of Cordoba, Cordoba, Spain Area de Mejora y Biotecnología, IFAPA, Centro ‘Alameda del Obispo’, Córdoba, Spain 3 Institute for Sustainable Agriculture, CSIC, Cordoba, Spain 4 Station de Génétique et d’Amélioration des Plantes, Institut National de la Recherche Agronomique,Versailles Cedex, France 2 *E-mail: teresa.millan@uco.es Abstract Growth habit is a morphological trait affecting adaptation ability of genotypes to a particular environment. The growth habit in chickpea plants can be erect, semi-erect (bushy) or prostrate. Erect or bushy types are present in cultivated Cicer arietinum while prostrate is present in annual wild Cicer sp. The aim of this study was to develop pairs of Near-isogenic lines (NILs) for erect vs prostrate growth habit using residual heterocigosity detected in a RIL (Recombinant Inbred Line) population derived from an interspecific cross (ILC72xC. reticulatum Cr5-10). The screening of 47 microsatellite markers distributed across all linkage groups (LG) in the chickpea genetic map showed that the NILs were different in LG 1 and 3. So, we could hypothesize that the genes involved in this trait could be located in either of these two LGs. Evaluation of growth habit in plant mature stage and genotyping for STMS markers located in LG1 and LG3 in two RIL populations, ILC72xCr5-10 (RIP-1) and WR315xILC3279 (RIP-2), revealed a QTL located in LG1 of chickpea genetic map. In RIP-1, the major QTL peak lay in RAPD (Random Amplified Polymorphic DNA) marker AD091053 accounting 18.8 and 17% of the phenotypic variation obtained in two different environments. This QTL was 2 confirmed in RIP-2 (intraspecific) being associated with STMS TA8 (R = 21.7%). The sequence CaDet, mapped in LG3 that were homologous to the gene controlling determinate vs non-determinate growth habit in Pisum sativun (PsTFL1) resulted non-related with growth habit in chickpea. VI ICLGG - Posters P-GRTM29 Phenotypic and genotypic diversity of selected pigeonpea [Cajanus cajan (L.) Millsp.] parental lines and hybrids Bharathi M 1, 2,* 1 1 1 , Vales MI , Saxena RK , Varshney RK , Saxena KB 1 1 International Crop Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 2 Jawaharlal Nehru Technological University Hyderabad, Hyderabad, India *E-mail: bharati_bth@yahoo.com Abstract Forty seven pigeonpea [Cajanus cajan (L.) Millsp.] Genotypes including 24 CMS (cytoplasmic nuclear male-sterile) based hybrids and 23 parental lines were used to study the phenotypic and genotypic variability for assessing the relationship between morphological characters and molecular markers which could be useful in future crop improvement programs. Principal component analysis (PCA) of the phenotypic data revealed the formation of two distinct clusters that were based on maturity (short vs. medium). Analysis of variance indicated significant differences among all the pigeonpea genotypes for days to flowering, days to maturity, plant height, 100 seed weight and grain yield. Genotypic analysis was carried out using 24 simple sequence repeat (SSR) markers. The dendrogram based on genetic distance (1-percent of shared alleles) revealed two distinct clusters of short duration and three clusters of medium duration genotypes. The polymorphic information content (PIC) values ranged from 0.04 to 0.90. A total of 192 SSR fragments were amplified with an average of eight alleles per marker. Genetic distance between the parental lines did not show its association with the yield of the respective hybrid. The SSR fingerprints generated could be used for identification of pure lines and for confirmation of hybrids. The study demonstrated that the clustering of the pigeonpea genotypes using both phenotypic and molecular approaches was mainly based on maturity. VI ICLGG - Posters Lupinus angustifolius genome mapping: rDNA and centromeres P-GRTM30 Wyrwa K, Szczepaniak A, Ksiazkiewicz M, Wolko B, Naganowska B* Institute of Plant Genetics of Polish Academy of Sciences, Poznań, Poland E-mail: bnag@igr.poznan.pl Abstract Narrow leafed lupin (Lupinus angustifolius L., 2n-40), a cultivated lupin species, has been a subject of our genomic studies based on the nuclear genome BAC library and the linkage map. A range of cytogenetic and genetic markers have been generated and used for assigning linkage groups to chromosomes via an approach combining BAC-FISH and genetic mapping. Here, we present current results: mapping of rRNA genes and centromere sequences in L. angustifolius chromosomes. Two BAC clones containing the sequences of two rRNA genes (26S and 5S) were selected from the L. angustifolius nuclear genome BAC library. These clones revealed single-locus BAC-FISH signals and thus served as cytogenetic markers for rRNA genes. Co-localization of these BACs with conserved sequences 5S and 25S from reference species confirmed the presence of one 25S and one 5S locus in the narrow-leafed lupin genome, in two distinct chromosome pairs. This corroborates with results published earlier. However, we localized genetically rDNA loci in lupin for the first time. The presence of particular rRNA genes in these clones was confirmed by BAC-end sequencing. The sequences of BAC-ends served as anchors for development of molecular markers for genetic mapping. 25S and 5S loci were localized in linkage groups NLL-16 and NLL-02, respectively. Furthermore, during the comprehensive screening of BAC library, a clone revealing clear BAC-FISH signals in the centromeres of all 20 chromosome pairs was identified. Comparative cytogenetic mapping of that BAC in several other species showed that this clone contains centromere sequences specific for lupins. VI ICLGG - Posters P-GRTM31 Improved simple sequence repeat panel for high-throughput genotyping in soybean Sayama T*, Sasama H, Ishimoto M National Institute of Agrobiological Science (NIAS), Ibaraki, Japan *E-mail: tsayama@affrc.go.jp Abstract Among commonly used molecular markers, simple sequence repeats (SSRs) have the advantages of a high level of polymorphism and codominant inheritance at individual loci. To facilitate rapid, systematic genetic mapping of soybean, we constructed a genotyping panel of 304 SSR markers selected for allelic diversity and chromosomal location so as to provide wide coverage of the entire genome (Sayama et al. 2011, DNA Res. 18: 107-115). Most primer pairs for the markers were redesigned to yield amplicons of 80–600 bp in multiplex polymerase chain reaction (PCR) and fluorescence-based sequencer analysis, and they were labeled with one of four fluorescent dyes. Multiplex PCR with sets of six to eight primer pairs per reaction allowed genotyping of multiple loci at the same time, which corresponded to loci previously determined. The SSR panel enabled us to construct genetic linkage maps for 11 populations composed of 1791 lines in two years. The average polymorphic ratio among the analyzed markers was 61.0%, and 21 markers showed no polymorphisms. Several gaps of >20 cM between markers remained. We upgraded the SSR panel to increase the polymorphism and even out the marker distribution by replacing the 21 nonpolymorphic markers with 65 new SSR markers. We used the new panel to genetically map 8 populations composed of 1369 lines in one year. It gave an average polymorphism ratio of 70.9%. This new SSR panel will promote the genetic analysis and improvement of soybean. VI ICLGG - Posters Mapping of QTLs for flowering time and seed number in lentil 1 1 2 Pote TD , Chahota R , Muehlbauer FJ , Sharma TR P-GRTM32 1* 1 Department of Agricultural Biotechnology, CSK Himachal Pradesh Agricultural University, Palampur, India 2 Department of Agriculture, Washington State University, Pullman, USA *E-mail: sharmat88@yahoo.com Abstract Our objectives was to identify quantitative trait loci (QTLs) for early flowering and seed number in a recombinant line population derived from lentil (Lens culinaris ssp culinaris Medikus) cross WA8649090 x Precoz. The WA8649090 parent was late maturing and had higher seed number, while Precoz was early maturing with less seeds per plant. QTL analysis was performed by composite interval mapping and the analysis revealed one major QTL on linkage group 3 which explained 28.60% of the phenotypic variation for early flowering. The QTL for seed number was mapped in the same region, however, showed no significant association with genomic region harbouring genes for earliness. The QTLs detected may provide useful information for improving the seed yield and developing early maturing lentils using marker-assisted selection. VI ICLGG - Posters P-GRTM33 An integrated genetic linkage map in Arachis spp. with diploids, tetraploids, and artificial amphidiploids mapping populations. 1, 1 2 3 4 Shirasawa K *, Hirakawa H , Bertioli DJ , Varshney RK , Moretzsohn MC , 4 3 3 5 5 Leal Bertioli SCM , Thudi M , Pandey MK , Rami JF , Foncéka D , Gowda 6 7 8 9 9 1 1 MVC , Qin H , Guo B , Hong Y , Liang X , Tabata S , Isobe S 1 Kazusa DNA Research Institute, Chiba, Japan University of Brasília, Brasília, Brazil 3 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 4 Embrapa Genetic Resources and Biotechnology, Brasília, Brazil 5 Centre de coopération Internationale en Recherche Agronomique pour le Développement (CIRAD), Montpellier, France 6 University of Agricultural Sciences (UAS-D), Dharwad India 7 Hubei Academy of Agricultural Sciences (HAAS), Wuhan, China 8 USDA-Agricultural Research Service, Tifton, USA 9 Guangdong Academy of Agricultural Sciences (GAAS), Guangzhou, China 2 *E-mail: shirasaw@kazusa.or.jp Abstract Peanut (Arachis hypogaea) is an autogamous allotetraploid legume (2n=4x=40), comprised of A and B genomes, which derived from two diploids, most likely A. duranensis (A genome) and A. ipaënsis (B genome). In the past decade, >10,000 molecular markers have been developed and used for construction of genetic linkage maps in cultivated peanut and its wild relatives. Integration of linkage maps of Arachis spp. has a potential to be a standard tool for genetics, genomics, and breeding in peanut. In this study, we mapped large number of genomic/EST-SSR and transpospn markers on the three RIL populations in Arachis spp. derived from crosses between: 1) A. duraensis and A. stenosperma for A genome (Moretzsohn et al. 2005); 2) A. ipaënsis and A. magna for B genome (Moretzsohn et al. 2009); and 3) (A. ipaensis x A. x4 duranensis) and A. hypogaea for AB genome, and the two F2 populations in peanut (Shirasawa et al. 2012). The developed maps were integrated with the published maps of cultivated peanuts (Varshney et al. 2009, Hong et al. 2010, Khedikar et al. 2010, Ravi et al. 2011, Sarvamangala et al. 2011, Gautami et al. 2012, Sujay et al. 2012, Qin et al. 2012) and artificial amphidiploids (Foncéka et al. 2009) to construct an integrated Arachis linkage map. The resultant map consisted of 20 linkage groups covering a total length of 2,651 cM with 3,693 marker loci, and revealed chromosome structures of A and B genomes in peanut. Comparative analysis of the consensus maps with the genomes of Lotus, Medicago, Glycine, and Cajanus identified several conserved chromosomal segments among them. VI ICLGG - Posters Development of expressed sequence tags based microsatellite markers for pigeonpea from wild relative Cajanus scarabaeoides. P-GRTM34 Singh P*, Mahfooz S, Patil PG, Chaudhary AK, Agbagwa IO, Chaturvedi SK , Nadarajan N, Datta S Indian Institute of Pulses Research (IIPR), Kanpur, India *E-mail: pallavi_pbi@rediffmail.com Abstract Pigeonpea (Cajanus cajan L. Millspaugh) is one of the major pulse crops of tropics and subtropics and occupies the second position in terms of area and production in India after chickpea. Wild relatives of any crop are important sources of resistance to biotic and abiotic constraints. Cajanus scarabaeoides an Indian origin wild species of Pigeonpea, has many desirable characters and is cross compatible with cultivated pigeonpea, with an objective to develop and utilize genomic tools for cultivated pigeonpea, a total of 350 EST clones from the root tissue of this wild relative were developed and sequenced. RNA was isolated and converted into cDNA, which was then ligated to positive selection vector pJET 1.2. Recombinant plasmids were transformed in competent E. coli XL-1Blue strain. Plasmid DNA from 100 putative positive clones was isolated and sequenced. Full length cDNA sequences ranging from 280 to 895 bp were obtained. Total sixty three EST sequences were submitted to NCBI dbEST. The microsatellite motifs in each sequence were identified and each sequence was compared against legume sequences in NCBI database in order to identify homologous clones. The initial analysis of sequencing data from 100 clones revealed sequence homology with gene sequences of many legumes such as Glycine max, Medicago truncatula, Lotus japonica, Cicer arietinum, Astragalus canadensis and Phaseolus vulgaris. Twenty six primer pairs were also designed based on sequences flanking the microsatellite motifs using the PRIMER 3.0 software. The primer pairs were validated on forty five pigeonpea genotypes (cultivar and wild) for their ability to amplify and polymorphic primers were identified. These new markers will add to the pool of available markers and accelerate molecular breeding for pigeonpea improvement. VI ICLGG - Posters P-GRTM35 Association mapping for abiotic stress tolerance in chickpea 1 1 1 1 1 Thudi M , Upadhyaya HD , Rathore A , Gaur PM , Krishnamurthy L , 1,2 1,3 4 5 6 Roorkiwal M , Nayak S , Cheturvedi S , Gangarao NVPR Fikre A , 7 2 1,8,* Kimurto P , Sharma P , Varshney RK 1 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 2 Guru Gobind Singh Indraprastha University, Delhi, India 3 University of Florida, Gainville, USA 4 Indian Institute of Pulse Research (IIPR), Kanpur, India 5 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 6 Ethiopian Institute of Agricultural Research (EIAR), Addis Abeba, Ethiopia 7 Egerton University, Egerton, Kenya 8 Generation Challenge Program (GCP), Mexico DF, Mexico *E-mail: r.k.varshney@cgiar.org Abstract Abiotic stresses like drought and heat are major constraints to chickpea production in arid and semi-arid regions of the world, where chickpea is grown extensively. To understand the genetic basis of these complex traits association mapping approach was employed in the present study. Phenotypic data generated on reference set (300 genotypes) for drought tolerance related root traits (three seasons at Patancheru), heat tolerance (one season at Patancheru and Kanpur), yield and yield component traits (three seasons at Patancheru under rainfed environment) and drought tolerance phenotyping data (two seasons at three locations Debre Zeit, Nairobi and Koibatek) was assembled. In addition seven season drought tolerance phenotyping data on mini-core collection (210 genotypes) was also used in the present study. Large phenotypic variation for different traits was observed with in the reference set. DArT markers equally distributed across chickpea genome were used to determine population structure as results three sub-populations were identified in the reference set using admixture model in STRUCTURE. Genome-wide and candidate gene-based association studies were conducted using 1872 markers (1072 DArTs, 651 SNPs, 113 gene-based SNPs and 36 SSRs) and phenotyping data generated on the reference set employing mixed linear model (MLM) analysis with compression method and kinship matrix. As a result a total of 583 significant marker-traits associations (MTAs) were identified and maximum number of MTAs was identified for 100-seed weight (113) and the loci identified through association studies explained ~38.5% phenotypic variation. Among 113 candidate gene-based SNP markers 6 SNPs in ASR, 2 SNPs in chickpea apetala, 3 SNPs in dehydrin and 3 SNPs in DREB were found to have significant associations with different traits like 100-seed weight, delta carbon, plant height, root surface area, root dry weight, pods per plant and yield. This study provides significant marker trait associations for drought and heat tolerance in chickpea and enables increasing the mapping resolution. Further the marker trait associations can used for enhancing drought and heat tolerance through molecular breeding. VI ICLGG - Posters Development of genomic resources in lentil (Lens culinaris) for the generation of linkage map and genetic diversity analysis P-GRTM36 Verma P , Shah N, Bhatia S* National Institute of Plant Genome Research, New Delhi, India *E-mail: sabhyata_bhatia@nipgr.res.in Abstract Genomic resources such as ESTs, molecular markers and linkage maps are still limited in lentil (L. culinaris Medik.). Hence their large scale development is reported in this study. Genomic SSR markers were generated through construction of microsatellite enriched library and more than 500 gSSRs were developed. These markers were used for genotyping in the RIL population (Precoz X L830) to construct a SSR based linkage map of lentil. Further transcriptome analysis using Illumina Genome Analyzer II platform resulted ~120 million reads. The de novo assembly was done using different assembly programs at various parameters which resulted in a number of non-redundant high quality transcripts. The average length of transcripts, N50 value, average number of reads per transcript, total coverage was also identified. Similarity search with the unigenes of other plant genomes was carried out using BLASTX and tBLASTX and identification of transcription factor families were also performed. Functional annotation using Gene Ontology (GO) terms distributed the sequences in the GO functional categories of biological process, cellular components and molecular function. SSR identification resulted in a number of sequences containing microsatellite motifs. Some of these markers were validated and utilized for analysis of genetic diversity across lentil genotypes. This study, which provides a large transcript dataset as well as gene based and genomic SSR markers, would serve as a foundation for various applications in lentil breeding. VI ICLGG - Posters .P-GRTM37 Identification of microsatellite markers linked to bacterial leaf blight resistance in cowpea (Vigna unguiculata (L.) Walp) 1, 2 1 1 Viswanatha KP *, Khan H , Amita K , Pallavi MS 1 University of Agricultural Sciences, GKVK, Bangalore, India University of Agricultural Sciences, Raichur, India 2 *E mail: viswanathakp55@gmail.com Abstract Cowpea bacterial leaf blight, caused by Xanthomonas campestris pv. vignicola, is a worldwide major disease of cowpea. Among different strategies to manage the disease including cultural practices, intercropping, application of chemicals, and sowing pathogen-free seeds, cowpea genotypes with resistance to the pathogen would be the most prudent option to the resource poor farmers in Asia and sub-Saharan Africa. Breeding for resistant cultivars would be facilitated by marker assisted selection. A set of 196 cowpea genotypes were screened for resistance to bacterial blight both under natural and controlled conditions to identify resistant source. Two genotypes, C-152 (susceptible parent) and V-16 (resistant parent) were identified and used to develop the F2 mapping population. Twelve of 42 SSR primers which were able to differentiate the parents of the mapping population were subjected to BSA to identify putative linked markers for bacterial leaf blight resistance. Among these 12 SSR primers, three were found to be polymorphic between the resistant and susceptible F2 bulks, indicating that these markers are putatively linked to BLB resistant gene. Band information of three SSR markers that showed Mendelian segregation ratio was compared with disease reaction. The data from individual marker analysis on F 2 population along with disease reaction were subjected to Single Marker Analysis. From the ANOVA of single factor analysis of these three markers, only marker CP 641/642 showed the significant difference at 5 % level of significance and hence it can be predicted that there is a linkage between the marker and the BLB resistance gene. VI ICLGG - Posters Fine mapping of and candidate genes for classical mutations on (Pisum sativum) linkage group III P-GRTM38 Weeden NF*, Biswas P Montana State University, Bozeman, USA *E-mail: nweeden@montana.edu Abstract Linkage group III of pea (LG III) represents one of the larger chromosomes in the standard pea karyotype and appears to be a combination of all of chromosome 3 and a portion of chromosome 2 of Medicago truncatula. The region of pea LG III linking one end of M. truncatula chromosome 3 with chromosome 2 does not display as strong a homology with the M. truncatula gene order as is observed in many other regions of the respective genomes. This region also contains the breakpoint of a translocation in pea and appears to be a fragile site. Using both the medicago and soybean genomic sequences we have assembled a fine-scale map for the lower arm (as classically drawn) of pea LG III consisting primarily of gene-based markers. A number of classical mutations in pea, including reduced stipule (st), apulvinic (apu), purple pod (Pu), clariroseus (b), Neoplasm (Np), Serratus (Ser), burnt leaf (bulf), tolerance to powdery mildew (er2), and resistance to fusarium wilt race I (Fw) have been placed on this map, and candidate genes identified for several of these. The gene order on lentil LG III appears to be highly conserved to that in pea. VI ICLGG - Posters P-GRTM39 Molecular mapping of QTLs for resistance to fusarium wilt in chickpea (Cicer arietinum L.) Vashist U, Kumar P, Sangwan MS, Dhillon S , Boora KS* Chaudhary Charan Singh Haryana Agricultural University, Hisar, India *E-mail: kboora@gmail.com Abstract Chickpea wilt caused by Fusarium oxysporum f. sp. ciceris, is one of the major yield limiting factors in chickpea. The disease causes 10–90% yield losses annually in chickpea. Eight physiological races of the pathogen (0, 1A, 1B/ C, 2, 3, 4, 5 and 6) are reported so far whereas additional races are suspected from India. The distribution pattern of these races in different parts of the world indicates regional specificity for their occurrence leading to the perception that F. oxysporum f. sp. ciceris evolved independently in different regions. Among these races, race 4 is prominent in northern part of India. For identification and mapping of QTL(s) resistance to fusarium wilt foc-4, two cultivars of chickpea HC-5 (fusarium wilt resistantace) and ICCV96030 (susceptible) along with the 111 recombinant inbred lines (RILs) derived by their cross were used. The screening for fusarium wilt was done in wilt sick plot and by sand culture method having Fusarium oxysporum f. sp. ciceris race 4. By using the primers which discriminated the parental genotypes for fusarium wilt, three major QTLs, H3A052, H4H07 and P79 were identified and mapped on the same chromosome covering 46.7cM. The interval QTL mapping for fusarium wilt showed 10.0 per cent of the resistance trait mapped in the region of three loci H3A052, H4H07 and P79 covering 46.7 cM with 95% level of confidence. Identification and further evaluation of disease resistance gene clusters would improve our understanding of wilt resistance in chickpea and facilitate the transfer of resistance genes to new cultivars. VI ICLGG - Posters PRIL_A, PRIL_B and PRIL_C: Pigeonpea [Cajanus cajan (L) Millsp] Recombinant Inbred Line mapping populations segregating for resistance to Fusarium wilt (FW) and sterility mosaic disease (SMD) P-GRTM40 Vales MI, Saxena KB, Saxena RK, Patil SB, Sawargaonkar SL, Sultana R, * Sharma M, Pande S, Varshney RK International Crops Research (ICRISAT),Hyderabad, India Institute for the Semi-Arid Tropics * E-mail: r.k.varshney@cgiar.org Abstract Pigeonpea bi-parental populations segregating for various traits of interest are being developed. The three most advanced populations, named PRIL_A, PRIL_B and PRIL_C (Pigeonpea Recombinant Inbred Line, population A, B and C) have reached F6 generation. PRIL_A: derived from the cross ICPB 2049 x ICPL 99050 segregates for FW, 329 lines. PRIL_B: derived from the cross ICPL 20096 x ICPL 332 segregates for FW and SMD, 342 lines. PRIL_C: derived from the cross ICPL 20097 x ICP 8863 segregates for SMD, 266 lines. Marker genotyping of the parental lines, however, showed low level of genetic variation. After screening over 4,616 (3,000 simple sequence repeats (SSRs) and 1,616 single nucleotide polymorphism (SNPs)) markers on parental genotypes of each mapping population, a total of 159 (104 SSRs and 55 SNPs), 80 (52 SSRs and 28 SNPs) and 157 (143 SSRs and 14 SNPs) markers were found polymorphic for ICPB 2049 vs ICPL 99050, ICPL 20096 vs ICPL 332 and ICPL 20097 vs ICP 8863, respectively. The polymorphic markers will be used for constructing the genetic linkage maps. The populations will be screened for FW and SMD in 2012-13. Marker-trait association analysis will be conducted in order to understand the genetic basis of resistance to FW and SMD. In addition, a parallel effort was initiated in 2010 to select lines with high agronomic potential from the above indicated mapping populations. Further selection during 2011-12 resulted in twenty-eight lines with high yield (up to 2.4 t/ha) and disease resistance. VI ICLGG - Posters Theme Genomics – Assisted Breeding P-GAB07 Marker-assisted selection for low phytic acid in soybean [Glycine max (L.) Merrill] * Narmadha , Jency PJ, Mohan S, Shadakshari TV, Pande R, Kalaimagal T Tamil Nadu Agricultural University, Coimbatore, India *E-mail: manosandhiya_87@yahoo.co.in Abstract Phytic acid, otherwise known as myo-inositol (1,2,3,4,5,6)- hexakisphosphate, is the major storage form of phosphorus in soybean [Glycine max (L.) Merr.] which comprises 75% of total seed phosphorus. It decreases the availability of some essential elements (Iron, Zinc, Calcium, Potassium and Magnesium) via bonding between the negatively charged phytic acid and the positively charged elements. Thus, diets high in phytate may lead to nutrient deficiencies. So, identification of lines with low phytic acid is of paramount importance. In the present study, 50 soybean accessions were analyzed for phytic acid following Davis and Reed method. The genotype NSO 15 recorded the lowest phytic acid value of 0.17 mg/100g. About thirty three genotypes recorded significantly higher values than the grand mean of 3.60 mg/100g. The genotypes with low phytic acid content are Co 1, VLS 41, CSB 0808, NSO 15, CSB 0903, LU 90, NSO 111 and JS 56. Two SSR markers namely, Satt416 and Satt168 which are closely linked to lpa loci were used to screen the germplasm. The eight lines which recorded lowest phytic acid were amplified through these markers. Hence, it is confirmed that the low phytate trait was inherited in concert with Satt416 and Satt168 markers. So, the above genotypes can be considered as good donors to transfer target gene of lpa into improved varieties in future. In summary, the markers Satt416 and Satt168 proved to be effective for dual marker assisted selection for gene transfer of two recessive genes which collectively confer the low phytate trait, as evidenced by chemical analysis of seed phosphorous in selected accessions. VI ICLGG - Posters Genomic approaches towards yield resilience in chickpea 1, 1 1 2 3 P-GAB08 3 Bharadwaj C *, Kumar J , Tripathi S , Hegde V , Jain PK , Srinivasan , Gaur 4 4 1 1 1 1 PM , Varshney RK , Yadava S , Kumar T , Faruqui A , Chauhan SK , Kumar 1 5 6 1 1 1 A , Chaturvedi SK , Sabhyta B , Santosh HB , Srivastava R , Sarika K 1 Division of Genetics, Indian Agricultural Research Institute, New Delhi, India Dharwar Centre, IARI, India 3 National Research Centre for Plant Biotechnology, New Delhi, India 4 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 5 Indian Institute of Pulses research, Kanpur, India 6 National Plant Genetic Resources, New Delhi, India 2 *E-mail: drchbharadwaj@gmail.com Abstract Chickpea (Cicer arietinum L.) is the most important pulse crop of India contributing to over 40% of country’s total pulse production. Though India is the largest chickpea producing country with a share of about 67% in global chickpea production, the production is not adequate to meet the domestic demand. Because of warm environments in Indian peninsula, the crop is challenged by fusarium wilt, a major yield reducing disease. Recent advances in the area of biotechnology have offered the tools in the form of molecular markers to assist the breeding practices. Use of MABC and MARS to develop high yielding wilt resistant lines in chickpea is being adopted to develop cultivars with higher yield as well as pyramided to wilt resistance genes. Validation of a core set of markers reported to be tagged to the foc genes in chickpea identified. Phenotyping and genotyping 175 RILs was carried out with one hundred and twenty polymorphic STMS primers. In comparison to allelic distortion observed by Winter et al. (2000), allelic distortion in the RILs from the intraspecific cross that we used was minimal. Using single point marker analysis, four markers (TR 56, TA78, TR 29, TA 146) were significantly associated with yield related traits i.e 100-seed weight and seeds per plant. These associations were determined at a LRS threshold of 12.7 and explained over 70% of total trait variation. The QTL (QTL1) for seed size individually explained between 19% of the total phenotypic variance. MABC approach is being deployed to introgress wilt resistance in high yielding chickpea genotypes using these markers in Pusa 256 x WR 315 and Pusa 372 x WR 315 and MARS is being followed in elite x elite crosses. VI ICLGG - Posters P-GAB09 Incorporation of resistance to angular leaf spot and bean common mosaic necrosis virus diseases in adapted common bean (Phaseolus vulgaris, L) genotype * Chilagane LA , Msolla SN, Kusolwa PM Sokoine University of Agriculture, Morogoro, Tanzania *E-mail: lusekochilagane@yahoo.co.uk Abstract A study was conducted to incorporate resistant genes for ALS and BCMNV diseases into adapted bean genotype using Marker assisted selection. The parents Mexico 54 and UBR(25)95 donor of Phg-2 and bc-3 genes for ALS and BCMNV respectively were used and the recipient being Kablanketi. SCAR markers SNO2 and ROC11 linked to Phg-2 and bc-3 respectively were used to assist selection. Kablanketi was crossed to both Mexico 54 and UBR (25)95 and their F1 crossed to make a double cross F1. The F1, F2 and backcrosses from single crosses were characterized. The Chi square values for ALS were 0.081 (P<0.776) and 0.017 (P<0.896) and for BCMNV were 1.609 (P<0.205) and 1.2 (P<0.273) for molecular and phenotypic screening respectively. The resistance to ALS and BCMNV was found to be monogenic and the genes involved are dominant and recessive respectively. The heritability of ALS was found to be high (0.772) implying that selection for ALS can be done early in segregating populations. High correlation values, r = 0.741 and 0.624 for ALS and BCMNV were obtained between phenotypic and molecular data, indicating high reliability for markers. In selection SW13 marker linked to I gene for BCMV resistance was included. Among the 20 F1 double cross hybrids, four showed resistance to ALS, BCMV and BCMNV, five to ALS and BCMNV, four to ALS and BCMV, one to BCMV/BCMNV, three to ALS, two to BCMNV and one had none of the three genes. This work signifies the use of MAS for multiple gene screening. VI ICLGG - Posters Introgression of drought tolerance trait in ICCV 10, a mid-duration desi variety of chickpea employing MAS 1,2 1 1 1 1 P-GAB10 1 SongokS , Thudi M , Khera P , Srinivasan S , Sivakumar C , Gaur PM , 2 1 2 1,3* Kimurto P , Krishnamurthy L , Mulwa R , Varshney RK 1 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 2 Egerton University, Njoro, Kenya 3 CGIAR Generation Challenge Programme (GCP), CIMMYT, Mexico DF, Mexico *E-mail: r.k.varshney@cgiar.org Abstract Chickpea (Cicer arietinum L.) is third most important grain legume cultivated in semi-arid regions of South Asia and sub Saharan Africa. Until recently, breeding efforts to improve drought tolerance have been hindered due to its quantitative genetic basis and poor understanding of the physiological basis of yield in water-limited conditions. Therefore efforts are being made to deploy marker-assisted back crossing (MABC) approach for enhancing drought tolerance in chickpea. For MABC, a chromosomal region harbouring several QTLs for drought tolerance related traits contributing >30% phenotypic variation identified in the Phase I of the Tropical Legume (TLI) of Generation Challenge Programme/Bill and Melinda Gates Foundation has been targeted. Based on marker polymorphism, ICCV 10, a medium-duration desi variety was chosen as recurrent parent and three cycles of MABC has been completed using ICC 4958 as donor. SSR markers (TAA170, ICCM0249, STMS11 and GA24) in the QTL region were used for foreground selection and 80 SSR markers distributed across chickpea genome were employed for background selection. A set of 20 backcross lines homozygous for QTL allele was identified. Selected homozygous plants are grown in fields in the main crop season 2010- 2011 for seed multiplication. These selected lines BC3F4will be eventually tested in field conditions in Kenya and India during 2012-13 crop season to identify the best performing lines for their possible release as drought tolerant varieties. VI ICLGG - Posters P-GAB11 Application of marker-assisted breeding to improve drought tolerance in elite Kenyan chickpea genotypes 1 1,* 1 1 2 Songok S , Kimurto PK , Mulwa RK , Towett BK , Gangarao NVPR , Silim 2 3 3 3 3 S , Gaur PM , Sivakumar C , Thudi M , Varshney RK 1 Egerton University, Egerton, Kenya International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Nairobi, Kenya 3 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 2 *E-mail: kimurtopk@gmail.com Abstract Chickpea (Cicer arietinum L.) is a good alternative food-security legume crop for semi-arid and dry highlands of Eastern Africa, since it can act as a relay crop in cereal-legume cropping systems. However terminal drought stress causes significant yield losses since moisture is depleted in upper profile at critical stages of grain filling and podding. To enhance drought tolerance through improving water uptake through root traits, ICRISAT has mapped root traits QTL in two donor parental genotypes ICC 4958 and ICC 8261 which can contribute significantly (33%-36%) to drought tolerance. This study therefore introgressed these root traits into 2 Kenyan Elite lines (ICCV 95423 and ICCV 10) using Marker Assisted Backcross breeding scheme using 4 SSR markers including TAA 170 and ICCM 0249 flanking the genomic regions. Backcross breeding and DNA extraction was done at Egerton University while DNA analysis was outsourced from ICRISAT India until BC3F3, when seed was multiplied. BC3F4 progenies developed from these two crosses will be evaluated with both parents and ICC 4958 as donor parent in one location in India (ICRISAT, Patancheru) and two locations in Kenya (ATC Koibatek and KARI Marigat) during short rains 2012. Preliminary observation at Patancheru showed some promising progenies that are expected to do better than both parents ICCV 95423 and ICCV 10 and donor parent ICC 4958 in relation to root length density (RLD), root biomass, depth and consequently better yield under drought. The results from this study are very encouraging and demonstrate that application of molecular breeding in breeding programs in developing countries is possible as it accelerates the process of varietal development especially for complex traits like drought.. VI ICLGG - Posters Precision breeding in chickpea (Cicer aerietinum L.) resistance to wilt through MAS 1 2 Mannur DM , Jailaxmi SK , Mahiboobsa M 4 5 Sivakumar C , Bharathi S 3, Gaur PM 4, P-GAB012 4 Varshney RK , 1 Dept. of Genetics and Plant Breeding, ARS, Gulbarga, India Dept. of Plant Pathology, ARS, Gulbarga, India 3 Dept. of GPB, ARS, Gulbarga, India 4 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 5 ARS, Gulbarga, India 2 *E-mail: dmmannur@rediffmail.com Abstarct Chickpea (Cicer arietinum L.) is a major source of protein in human and animal diet and the world’s most important pulse crop after beans (Phaseolus vulgaris L.) and peas (Pisum sativum L.). The present investigation was conducted at Agricultural Research Station, Gulbarga, U.A.S; Raichur, India with prime objective to study and transfer of wilt resistance to Annigeri-1 (A-1) a popular chickpea variety through back crossing programme by using “Marker Assisted Selection” (MAS) .The source of resistance to wilt in this study for race 4 is WR-315. Using a set of 384 SSR markers to check polymorphism on parental genotypes, Annigeri-1 and WR 315, a total of 87 markers were identified as polymorphic and 9 multiplexes were prepared with 40 markers which represents all 8 LG’s of chickpea. Through DNA sampling of 55 F1 plants of cross A-1 x WR-315 three were found true F1 plants .These three F1 plants were used for backcrossing with A-1 [(A-1 x WR-315) x A-1]. 311 F2 (A1 x WR315) seeds obtained were sown in wilt sick garden at ARS Gulbarga to screen for resistance to wilt. Total of 169 plants were found resistant. These 169 progenies will be evaluated along with both the parents. Based on per-se performance the progenies will be selected for large scale evaluation and further selection process which will yield the precise result. Simultaneously BC1F1 seeds obtained during the same season were subjected for DNA screening through MAS using same lab and true BC1F1’s were back crossed by A-1 and produced BC2F1 seeds for further study till the genome recovery of candidate variety A-1 with wilt resistant race 4 (WR-315). VI ICLGG - Posters P-GAB13 Validation of late leaf spot resistance QTLs in groundnut (Arachis hypogaea L.) 1,* 1 1 1 1 Manivannan N , Pavithradevi S , Priyadharhshini M , Anitha BK , Shoba D , 1 1 2 2 2 Vindhiyavarman P , Ganesamurthy K , Janila P , Pandey MK , Varshney RK 1 Tamil Nadu Agricultural University, Coimbatore, India International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 2 *Email: nmvannan@gmail.com Abstract Groundnut is affected by many biotic and abiotic stresses. Among these, late leaf spot (LLS) is one of the major yield limiting factor. Markers assisted breeding helps the breeder to target the traits efficiently. Molecular markers associated with QTL regions for LLS resistance were already reported viz., IPHAM103, GM2079, GM2301 and GM1536 (Sujay et al., 2012); PGP08D09, GM1009 and GM1573 (Sujay et al., 2012); PMc588, PM384, Ah4-26 (Shoba, 2010) and PM377, PM137 and PM3 (Shoba, 2010). These markers were tested in a mapping population derived from the cross TMV 2 x COG 0437. The parents TMV 2 and COG 0437 are susceptible and resistant to LLS respectively. A total of 140 SSRs were screened for parental polymorphism including the markers reported to be associated with LLS QTLs. Among the markers linked to LLS QTLs, GM 1536, GM 1573 and GM 2079 were monomorphic for these parents. Thirty seven primers were found as polymorphic and used to survey 94 RILs of the mapping population TMV 2 X COG 0437. Phenotypic data were recorded during post rainy season of 2010. LLS incidence was observed in the range of 1-9 scale among parents and RILs. Linkage and QTL analyses were performed using the software QTL IciMapping (Version 3.2). Two linkage groups were formed with 8 and 2 markers respectively. The total length covered by these markers is 81.62 cM with an average of 8.16 cM. The results revealed two QTLs for LLS with 12.3 13.4 per cent phenotypic variation explained. Both QTLs were found in the same linkage group. The nearest marker for each QTL are PM 384 and PGP08D09. As the marker PM 384 and PGP08D09 were already reported for LLS resistance, these markers could be considered as potential markers in marker assisted breeding programme for LLS resistance. VI ICLGG - Posters Towards marker assisted backcross breeding to enhance drought tolerance in Kenyan chickpea (Cicer arietinum L.) germplasm 1 1,* 1 1 P-GAB14 1 Oyier MO , Kimurto PK , Mulwa RK , Towett BK , Cheruiyot EK , Gangarao 2 2 3 3 R , Silim S , Varshney RK , Gaur PM 1 Egerton University, Egerton Kenya International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Nairobi, Kenya 3 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Patancheru, India 2 *E-mail: pkimurto@yahoo.co.uk Abstract Terminal drought is globally the number one constraint for production of chickpea and other crops as well. In the past, breeding efforts to improve terminal drought tolerance have been hindered due to its quantitative genetic basis and poor understanding of the physiological basis of yield in waterlimited conditions. This is because quantitative traits are controlled by multiple genes whose inheritance patterns are difficult to predict. In chickpea, recent research efforts, has enabled the development of a large number of molecular markers, expressed sequence tags (ESTs), dense genetic maps and molecular markers associated with QTL/gene conferring resistance to drought become available. Among the cultivars identified as sources of resistance with QTL for drought traits are ICCV 4958 (Desi) and ICCV 8261 (Kabuli). These linked markers have been validated and deployed in chickpea breeding. With an aim of enhancing the drought tolerance in Kenyan cultivars through marker assisted introgression, ICCV97105 was selected. ICCV 97105 exhibited high polymorphism with the SSR markers and was chosen as recurrent parent among the ten agronomically superior elite cultivars. The improvement has been done to BC3F1 with seeds multiplied for BC2F2. With identification of suitable recurrent parent highly polymorphic with the markers, crosses were made between ICCV 97105 and ICC 4958 to introgress the drought QTL traits to obtain F1. The F1 seeds were planted, DNA of the plants were extracted and used for foreground selection. True heterozygozity was tested as well. The heterozygotes were used as donors and backcrossed to the recurrent parent to obtain BC1F1 seeds. The seeds were planted and the process repeated to obtain BC2F1 and finally BC3F1. The results indicated that it is possible to transfer QTL confers drought tolerance using marker assisted selection. The two sets of seeds viz: BC3F1 and BC2F2 were planted in the field, increased and preserved for further research. VI ICLGG - Posters P-GAB15 Validation of STS markers linked with bruchid resistance in greengram (Vigna radiata L. Wilczek) Jayamani P*, Sowmini K, Surya M Tamil Nadu Agricultural University, Coimbatore, India *E-mail: jayamani1108@gmail.com Abstract Greengram or mungbean (Vigna radiata L. Wilczek) is an important grain legume crop of India, where diet of the majority of the people predominantly follow cereal based diet. Post harvest damage to the greengram seeds caused by a group of insect pest of the genus Callasobruchus often referred to as been weevil or bruchid is quite extensive in India. A RFLP marker Mgm 213 is known to be closely linked (1.3 cM) to a major locus controlling bruchid resistance in ACC 41, Vigna radiata var. sublobata from Australia. Later two PCR based locus specific markers namely STSbr1 and STSbr2 were developed and validated in ACC41 (resistant), Berken (susceptible) and RILs derived from the above parents. Both the markers were co- segregated with the initial RFLP maker and also linked with the bruchid resistance. In the present study, the two STS makers namely STSbr1 and STSbr2 were validated in Indian originated V. radiata var. sublobata accession resistant to burchid damage. Analysis of V. radiata var. sublobata/1 (resistant) and VBN (Gg) 2 (susceptible) and their hybrids showed co – segregation of resistant locus with polymorphic fragments. STSbr1 behaved as dominant marker and produced approximately 225 bp band only in resistant parent and in hybrids and absent in susceptible parent. STSbr2 produced an allele with a size of approximately 470 bp and found to be monomorphic. Hence, STSbr1 could be used in the germplasm screening to identify new source for bruchid resistance and in marker assisted selection to develop bruchid resistant varieties in greengram. VI ICLGG - Posters Disease screening and yield assessment of introgression lines developed through marker-assisted backcrossing for rust resistance in cultivated groundnut (Arachis hypogaea L.) 1 1 1 2 P-GAB16 1 Pandey MK , Janila P , Nigam SN , Gowda MVC , Manohar SS , Sriswathi 1 1,3* M , Varshney RK 1 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 2 University of Agricultural Sciences (UAS), Dharwad, India 3 CGIAR Generation Challenge Programme (GCP), c/o CIMMYT, Mexico DF, Mexico *E-mail: r.k.varshney@cgiar.org Abstract Rust, caused by Puccinia arachidis, is widespread in most of the tropical countries and severely affecting the yield and quality of the produce in groundnut (Arachis hypogaea L.). By using linakge mapping approach on a RIL population (TAG 24 × GPBD 4), a major QTL (QTL rust01) associated with four SSR markers (IPAHM103, GM2079, GM2301, GM1536) has been identified that contributes upto 82.96% phenotypic variation for rust resistance. This major QTL region and associated markers were further identified in another RIL population (TG 26 × GPBD 4). Subsequently these markers were validated on a range of diverse germplasm. The validated markers were deployed in marker-assisted backcrossing (MABC) programme to improve three elite groundnut varieties (ICGV 91114, JL 24 and TAG 24) for rust resistance using GPBD 4 as a donor genotype. After undertaking 2-3 rounds of backcrossing, a total of 76 homozygous BC 3F2 and 158 BC2F3 introgression lines developed were subjected to field screening for rust resistance in the rainy 2011 season. Several promising lines with remarkable reduction in disease spread and other desirable agronomic traits have been selected for further multiplication and generation advancement. At present, a total of 200 introgression lines in advanced generation (117 BC2F5 and 83 BC3F5) are grown in replication for yield assessment and selection of superior lines for multilocation evaluation. The promising lines with desirable yield and higher resistance to leaf rust could be released an improved varieties. VI ICLGG - Posters P-GAB17 Hybridity testing of inter-specific lentil and chickpea crosses using ISSR markers * Rana MK , Singh M, Kumar K, Singh S, Singh AK, Deo S National Bureau of Plant Genetic Resources (NBPGR), New Delhi, India *E-mail: mkrana@nbpgr.ernet.in Abstract Pulses are important in India not only as a vegetarian diet but also for their ability to fix nitrogen, provide complementary protein in combination with cereals and as source of some micronutrients essential for human consumption. In the past, variability in pulse crops has mostly been generated using hybridizations and mutations and has been used to attain the desired levels of production. However, new variability needs to be created in order to attain further strides in pulse production. Lentil and chickpea are the most important grain legumes grown in the arid and semi-arid regions of India. The narrowing genetic base in the cultivated varieties of these crops is the main concern for realizing further improvement in the production. In this regard, prebreeding efforts for genetic enhancement of the narrowing genetic base are underway at NBPGR. Thirteen inter-specific crosses in lentil and seven in case of chickpea had been attempted and for confirming their hybridity, SSR (27) and ISSR (100) primers were used. After screening of these primers in all the hybrids and their respective parents of chickpea and lentil crosses, only male parent specific diagnostic dominant ISSR markers were identified. True hybrid nature of nine lentil and two chickpea inter-specific crosses was confirmed using the diagnostic markers. These confirmed crosses would further be raised for generation advancement. VI ICLGG - Posters Detection and/or validation of QTL for resistance to Ascochyta blight and for Fusarium wilt (race 1) in chickpea (Cicer arietinum L.) 1 1 1 1 P-GAB18 1,2 Sabbavarapu MM , Sharma M , Nayakoti S , Bairapaka VK , Chamarthi SK , 1 1 1 3 1 1,4,* Rathore A , Samineni S , Gaur PM , Singh S , Pande S ,Varshney RK 1 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 2 Mc Gill University, Quebec, Canada 3 Punjab Agricultural University (PAU), Ludhiana, India 4 CGIAR Generation Challenge Programme (GCP), CIMMYT, Mexico DF, Mexico *E-mail: r.k.varshney@cgiar.org Abstract The aim of the present study is to validate and/or to identify new quantitative trait loci (QTL) for resistance to Ascochyta blight (AB) caused by Ascochyta rabiei and Fusarium wilt (FW) by Fusarium oxysporum f. sp. ciceris. For this purpose, two mapping populations (188 F2:3 families each) derived from different crosses (C 214 × ILC 3279 and C 214 × WR 315 for AB and FW respectively) were developed and phenotyped for resistance to seedling resistance and adult plant resistance for AB and to FW (race 1). A linkage map with a length of 386.3 cM using 58 SSR loci for AB cross and, a linkage map spanning 1119.0 cM with 57 SSRs for the FW cross were developed. QTL analysis identified a total of 6 main effects QTL (M-QTL) explaining up to 31.97% of phenotypic variation and an epistatic QTL (E-QTL) in the AB population. In the case of FW, two M-QTLs explaining 11.0% and 17.0% of phenotypic variation respectively were detected. The QTL for AB resistance explaining 31.97% phenotypic variation was detected in both field and controlled conditions in this study and, the same was reported previously using different genetic backgrounds and using resistant parent (ILC 3279) used in present study. Hence, markers associated with this QTL could be used in marker-assisted breeding to improve AB resistance. QTLs for FW resistance detected in the present study are not reported so far, hence, these QTLs could be new and, need to be confirmed in other genetic backgrounds. VI ICLGG - Posters P-GAB19 Marker assisted recurrent selection (MARS) for enhancing drought tolerance in chickpea 1 1,2 1 3 4 Gaur PM , Kumar A , Rathore A , Kimurto PK , Asnake Fikre , Gangarao 5 1 1 1,6* NVPR , Sama VSAK , Thudi M , Varshney RK 1 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 2 Jawahar Lal Nehru Krishi Vishwa Vidyalaya (JNKVV), Jabalpur, India 3 Egerton University, Egerton, Kenya 4 Ethiopian Institute of Agricultural Research (EIAR), Addis Abeba, Ethiopia 5 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Nairobi, Kenya 6 CGIAR Generation Challenge Programme (GCP), Mexico DF, Mexico *E-mail: r.k.varshney@cgiar.org Abstract Terminal drought is considered as the major constraint to chickpea production in arid and semi-arid regions of the world. Marker-assisted recurrent selection (MARS), a modern breeding approach was employed for enhancing drought tolerance in chickpea, which enables increasing frequency of several beneficial alleles having additive effect and small individual effects in recurrent crosses. For this purpose, four superior desi genotypes based on their performance have been selected ICCV 04112, ICCV 05107, ICCV 93954 (released as JG 11 in India) and ICCV 94954 (released as JG 130 in India) and two crosses were made by using elite and elite lines (a. JG 11 × ICCV 04112 and b. JG 130 × ICCV 05107). The F3:5 progenies (188) from the two crosses developed were evaluated at three locations (Ethiopia, Kenya and India) under rainfed and irrigated conditions, in an alpha-lattice design with two replications. Genotyping data was generated on F3 plants for 70 and 92 SSR markers on JG 11 × ICCV 04112 and JG 130 × ICCV 05107 respectively. A total of 65 SSRs were mapped in case of JG 11 × ICCV 04112 and 55 SSRs on JG 130 × ICCV 05107. To pyramid, superior alleles of the favorable QTLs were identified for both crosses using QTLCartographer. A set of eight lines were selected for each cross using OptiMAS 1.0. It is anticipated that at the end of the project, RC3F4 progenies will be available for evaluation at multi-locations. These efforts will lead to development of superior lines for drought tolerance. . VI ICLGG - Posters GDMS: genotyping data management system implementation for tropical legumes 1 1 1 1 1 P-GAB20 1 Shah T , Pottluri SK , Peteti P , Reddy PT , Thudi M , Pandey MK , 1 2 2 2 1,2,* BhanuPrakash A , Sawkins M , Portugal A , McLaren G , Varshney RK 1 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 2 Generation Challenge Program (GCP), Mexico DF, Mexico *E-mail: r.k.varshney@cgiar.org Abstract The critical component of any Marker Assisted Breeding program is the effective management of genotyping data with its associated germplasm and phenotyping data. The Genotyping Data Management System (GDMS) focuses on handling the low to high throughput genotyping data from different marker systems and platforms (SSRs, DArTs and SNPs). In addition to the marker, genotyping and fingerprinting information the system also handles maps, QTLs and allows for exchange of data with the analytical tools. The GDMS has been implemented in the Tropical Legumes project of the Generation Challenge Program, currently for chickpea and groundnut, and is available at http://220.227.242.214:8085/GDMS. The system provides the functionality to query for polymorphic markers, screened markers, germplasm and select markers based on distribution on the map or regions of interest. It allows for export of the data in a variety of formats including the Flapjack format for visualization. The system is also linked to CMap for visualizing comparative maps, and the Crop Ontology database for the detailed traitinformation. The GDMS can be installed locally too, and will allow users to upload and manage their genotyping and marker information through predefined templates. The GDMS has been developed in Java as a browser based system with a MySQL database back end. The database is integrated within the Integrated Breeding Database (IBDB) and is therefore tightly coupled with the management of phenotyping and germplasm information. VI ICLGG - Posters P-GAB21 Validation of DNA markers linked to bruchid resistance gene(s) in urdbean (Vigna mungo (L.) Hepper) Souframanien J*, D’Souza SF Bhabha Atomic Research Centre (BARC), Mumbai, India *E-mail: souf@barc.gov.in Abstract Urdbean and other legume crops are severely affected by bruchid species of the genus Callosobruchus (Coleoptera: Bruchidae). The wild relative V. mungo var. silvestris is a potential source of bruchid resistance in urdbean breeding programme. Bruchid resistance in V. mungo var. silvestris has been mapped using a saturated linkage map constructed with 428 markers. Eight regions associated with adult emergence and developmental period have been previously identified. For successful implementation of markers in breeding programme, the available linked markers needs to be validated in diverse genetic backgrounds. Hence, in the present study, reported markers associated with bruchid resistance from urdbean, mungbean and common bean were validated using diverse black gram genotypes and recombinant inbred lines known for their bruchid resistance. Of the ninteen primers (5 ISSR, 3 RAPD and 11 SSR) tested, one ISSR (ISSR8551357) and two RAPD markers (OPL11500 and OPO-061400) amplified bands specific to resistant lines, that were absent in susceptible RILs and other diverse genotypes studied. The identification, mapping and validation of the bruchid resistance genes and QTLs should facilitate the development of resistant cultivars of urdbean and closely related mungbean. It will also be useful for the pyramiding of bruchid resistance with other traits, particularly through the use of marker- assisted selection. VI ICLGG - Posters Application of marker-assisted selection for Striga resistance in cowpea (Vigna unguiculata (L.) WALP) 1 2 1 2 P-GAB22 3 Tignegre JBS , Satoru M , Ouedraogo JT, Drabo I , Boukar O , Moutari N , 4 4 4 Timko MP , Southern E , Koebner R 1 INERA, Burkina Faso IITA, Nigeria 3 INRAN, Niger 4 Kirkhouse Trust Foundation, UK 2 *E-mail: racinetignegre@yahoo.com Abstract Striga gesnerioides is a noxious root parasite that can cause complete yield loss if only susceptible varieties are involved. The research being initiated in that project aims at applying available molecular markers for selecting Striga gesnerioides-resistant sources in cowpea. To achieve this, the most prevailing two Striga races in Burkina Faso (race 1), Niger and Nigeria (race 3) were targeted as damaging constraints to address. Participatory variety selection and rural appraisal, including farmers, other end-users, researchers, and developers were conducted. Adapted cowpea resistant lines were implemented in Burkina Faso and Nigeria in 2007 and 2008. Farmer’s preferred cowpea cultivars were selected through field-screening and then involved in a backcross selection associated with MAS to combine both Striga resistance and farmer’s desirable traits in a single variety. An attempt to validate sets of AFLP and SSR markers shows some polymorphisms specific to the crosses. No marker showed polymorphisms to all combinations of crosses. A SSR marker (SSR1) showing polymorphism for wide Striga resistance in Burkina Faso was identified at 150 bp was validated and is being exploited in MAS selection for the control of Striga in Burkina Faso. VI ICLGG - Posters Theme: Nutrition and Quality P-NAQ05 Consumption pattern of pulses and legumes among tribal population in India Arlappa N*, Balakrishna N, Hrikumar R, Meshram II, Rao MK, Galreddy Ch , Kumar S, Ravindranath M, Laxmaiah A National Institute of Nutrition, Hyderabad, India *E-mail: arlappan@yahoo.com Abstract Pulses and legumes is the very important food group and is the richest source of protein especially for the traditional vegetarians and low socio-economic groups. The objective of this communication is to assess the consumption pattern of pulses and legumes among tribal population in India. A community based cross-sectional study was, carried out by National Nutrition Monitoring Bureau (NNMB) in 9 states of India to study the dietary pattern of tribals using 24-hr re-call method of diet survey in about 10,000 households (HHs).The tribal population was consuming 13 types of pulses & legumes. Of them, consumption of red gram (16.8g/day) was high, followed by green gram (3.4 g/day), Bengal gram/ chickpea and black gram (2.7g/day each), lentil (2.3g/day), cow pea (1.6g/day) and horse gram (1.4g/day). Red gram is the most common pulse consumed in all states except in West Bengal. Highest consumption of red gram was reported in Maharashtra followed by Gujarat and Tamil Nadu. In general, the mean intake of pulses & legumes was 30.1 g/day (SD: 36.7), and only Gujarat and Maharashtra were meeting recommended dietary intakes (RDI) of 40g. The mean consumption ranges from a high 53.5g (SD: 64.6) in Gujarat to a low 10.1g (SD: 15.9) in West Bengal. Similarly, about 49% of HHs was not consuming even 50% of RDI and only 30% of HHs were meeting RDI and the proportion ranges from a high 57.5g in Maharashtra to a low 6.2g in West Bengal. The consumption was significantly (p<0.01) different with different socio-economic status. VI ICLGG - Posters Potential of Vigna unguiculata as a phytoremediation plant in the remediation of Zn from contaminated soil P-NAQ06 * Basak J , Singh J Visva-Bharati, Santiniketan, India * E-mail: jolly.basak@visva-bharati.ac.in Abstract Due to global industrialization and the constant increase in the human population in the last few decades, heavy metal contamination of air, water and soil has created diverse fatal effects on humans and also on the stability of the ecosystem. Heavy metals are not biologically degradable, and therefore can remain in the ecosystem for a long time. Heavy metal toxicity and the threat associated with their bioaccumulation in food chains represent one of the major environmental and health problems of our present day society. Plants that take up heavy metals from the soil offer an alternative and less expensive method to strip heavy metals directly from the soil. The use of such plants to cleanup soil and water contaminated with pollutants, a technique known as phytoremediation, is emerging as a new tool for in-situ remediation. Zinc is a potential environmental hazard. In the present work, the uptake of Zn by Vigna unguiculata plants has been studied. The experiment was continued for a period of 15 days. The Zn content in plant tissues was quantified using Atomic Absorption Spectroscopy. The phytoremediation of Zn using V. unguiculata plants in hydroponic solution shows that, during the period of the experiment (i.e. 15 days), the plants could tolerate and remove up to 41.2µg/ml zinc from the environment and so can be considered as good accumulators of zinc. VI ICLGG - Posters P-NAQ07 Phenotypic expression and inheritance of determinate habit (deh) mutation in pea (Pisum sativum L.) Belyakova AS, Sinjushin AA* M.V.Lomonosov Moscow State University, Moscow, Russian Federation *E-mail: asinjushin@mail.ru Abstract Increase of productivity of agricultural plants is available via breeding of cultivars with improved characteristics, often novel morphotype. One of the serious needs for garden pea (Pisum sativum L.) is growing limitation, i.e. simultaneous production of pods without further stem proliferation. Hence, forms with determinate growth are of special interest in pea breeding. The mutation determinate habit (deh) was firstly described in 1989 and few cultivars with such type of determinacy were bred out in Russia (Flagman, Batrak, Orlovchanin 2 etc.). However, peculiarities of phenotypic manifestation of this mutation together with its inheritance remain obscure. Our work was aimed at investigation of developmental characteristics of deh mutants (cv. Flagman) compared with control (cv. Nemchinovskii 766). The mutant plants are characterized with early apical arrest (apical meristem finishes proliferation) together with reduction of stipules’ area. Stipules exhibit strong asymmetry, one or both of them remaining underdeveloped beginning before flowering. Number of productive nodes decreases but connection between incomplete development of stipules and cessation of apical growth seems unclear. Cells of stipules’ epidermis are smaller, with less incurved anticlinal walls. Comparison with another mutant having reduced stipules, st, showed that only cell size decreases in deh plant, not number. Hybridological analysis of deh×DEH and DEH×deh crosses showed that heterozygous plants exhibit somewhat intermediate phenotype, while F 2 plants usually deviate from 3:1 ratio evidencing for existence of additional genes which influence deh expression. It may complicate breeding of deh cultivars. VI ICLGG - Posters A novel effector CtNUDIX secreted by the anthracnose pathogen Colletotrichum truncatum contributes to the transition from biotrophy to necrotrophy in hemibiotrophic fungal pathogens 2 2, 2 3 P-NAQ08 1, Bhadauria V , Banniza S *, Vandenberg A , Selvaraj G , Wei Y * 1 Department of Biology, University of Saskatchewan, Saskatoon, Canada Crop Development Centre, University of Saskatchewan, Saskatoon, Canada 3 Plant Biotechnology Institute, National Research Council of Canada, Saskatoon, Canada 2 *E-mail: sabine.banniza@usask.ca & yangdou.wei@usask.ca Abstract The in planta transition from biotrophy to necrotrophy known as the biotrophynecrotrophy switch (BNS) in hemibiotrophic fungal and fungus-like phytopathogens is critical in disease development. We herein report a novel effector gene CtNUDIX from Colletotrichum truncatum, the causal agent of anthracnose on pulse crops that exclusively expresses precisely before the BNS and elicits severe hypersensitive response (HR)-like cell death in tobacco leaves transiently expressing the effector. CtNUDIX containing a signal peptide and a Nudix hydrolase domain is unique to hemibiotrophic phytopathogens. Nudix hydrolases are known to hydrolyze a range of nucleotide and nonnucleotide derivatives like ADP-ribose, diadenosine polyphosphate, inositol pyrophosphates. Cell death triggered by CtNUDIX requires its accumulation at the host cell plasma membrane, indicating that the effector may cause perturbation in cell surface dynamics. Overexpression of CtNUDIX in C. truncatum and the agronomically important rice blast pathogen, Magnaporthe oryzae, blocked the BNS and resulted in incompatibility with hosts lentil and barley, respectively. A HR-like response was also observed in infected host cells associated with the biotrophic invasive hyphae. These results provide compelling evidence that hemibiotrophic fungal phytopathogens deliver Nudix effectors to the host cell plasma membrane to promote pathogenesis by causing massive cell death during the in planta differentiation of necrotrophic hyphae from biotrophic hyphae. VI ICLGG - Posters P-NAQ09 Development of a yellow seeded mung bean line with good nutritional quality Biswas SC * BRAC Agriculture Research & Development Centre (BARDC), Gazipur, Bangladesh *E-mail: siteshbiswas@rediffmail.com Abstract Mung bean (Vigna radiata L.) is an excellent source of high quality vegetable protein in the diets of Asian countries. In Bangladesh mung bean is being grown traditionally from the very ancient times as scented pulse crop for its delicious taste and easy fitting in the various cropping system. Due to its good nutritional quality, soil amelioration properties and also for its huge consumption by the local confectionaries for making value added high quality nutritious confectionary products viz, Dal Bhaja, Papar etc. mung bean has become an excellent agricultural commodity in Bangladesh in recent times. Besides during the harvesting period a huge no. of poor women are employed to collect the fruits of mung bean and hence mung bean cultivation generates income opportunities as well particularly for the underprivileged section of women in Bangladesh. Therefore, mung bean cultivation not only providing scope for generating income opportunities to the poor people it is also playing a significant role in strengthening the national economy of Bangladesh. Undoubtedly recently developed short duration mung bean varieties by the national research institutes with the help of AVRDC has made possible this tremendous achievement in the mung bean cultivation in Bangladesh. These short duration but high yielding mung bean varieties can fit easily in the existing cropping pattern although asynchronous ripening of fruits discouraging the mung bean growers for its cultivation in few cases. Keeping view of these above facts a relatively synchronously matured yellow seeded short duration advance line (YS-1) having moderate seed size has been developed through selection from a local green seeded variety recently at BRAC Agricultural Research and Development Centre (BARDC) which can be harvested only 57-58 days. It is a low fat line and it has got relatively high level of minerals than the check popular varieties BARImoog-5 and BINAmoog-7 released by the national research institute. This high yielding line can be grown in the local agro climatic condition both in the Kharif-1 & Kharif-2 season. It is relatively tolerant to MYMV disease and gave 37% increased yield than BARImoog-5 and near about 70% higher yield than BINAmoog-7. Hopefully due to its attractive golden yellow colour, better taste and moderate size seeds farmers also expressed their willingness to grow this high value crop and this advance line will be released very soon for its commercial cultivation. Since it fits well in the existing cropping pattern this nutritious mung bean line will not only provide better returns to the farmers in a shorter period but will also help in providing sustainability of the productivity of the cereal system. VI ICLGG - Posters Validation of QTLs for pod and kernel traits in groundnut 1 1 1 P-NAQ10 1 Anitha BK , Pavithradevi S , Zaheer Hussain VM , Selvamuthu K , Saranya 1 1 1 2 ER, Manivannan N *, Anandakumar CR , Ganesamurthy K , Janila P , Pandey 2 2 MK and Varshney RK 1 Tamil Nadu Agricultural University, Coimbatore, India International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 2 *E-mail: nmvannan@gmail.com Abstract Marker assisted breeding is getting momentum in groundnut due to the identification of potential QTLs for important traits. One of the important steps in marker assisted breeding is to validate the identified QTLs. QTLs associated with pod and kernel traits of groundnut were reported by many authors. In the present study, a set of 64 genotypes were selected to validate the reported markers associated with QTLs of pod and kernel traits viz., PM36, PGS19D09 and IPAHM103 for oil content; PM375 for 100-pod weight, pod length and kernel length; PM137, PGS19D09 and PM384 for 100-kernel weight. Genotype data were generated using 64 genotypes and the six markers associated with various traits. These 64 genotypes were evaluated for pod and kernel traits during post rainy season, 2011-12 at Oilseeds Farm, Tamil Nadu Agricultural University, Coimbatore. Single marker analysis was performed to ascertain the relationship between the marker and traits. The results indicated that markers PM36, IPAHM103 and PGS19D09 for oil content; PM375 for 100 pod weight, pod length and kernel length; PM137 and PM384 for 100-kernel weight explained more than 10 per cent of phenotypic variation. In addition, these markers also showed association with other traits. Hence these markers could be considered as a potential tool in the marker assisted breeding in groundnut. VI ICLGG - Posters P-NAQ11 Functional characterization of PvMRP1 and PvMRP2, two putative phytic acid transporters * Cominelli E , Panzeri D, Laterza AR, Palmieri G, Bollini R, Sparvoli F Institute of Agricultural Biology and Biotechnology, Milano, Italy *E-mail: cominelli@ibba.cnr.it Abstract Phytic acid is the main storage form for phosphate in plant seeds. It decreases the nutritional value of the seeds by trapping phosphate and chelating nutritionally important minerals, such as iron, zinc, magnesium, and calcium. Therefore, the isolation of low phytic acid (lpa) mutants is considered a highly desirable objective in the genetic improvement of the nutritional quality of grain crops. The lpa1(280-10) mutant isolated in Phaseolus vulgaris confers a 90% reduction in seed phytic acid content, a 25% reduction in raffinosaccharide content, sevenfold increase of free iron cations in the seeds and does not display pleiotropic effects and agronomic defects. The mutated gene corresponds to PvMRP1, coding for an ATP-Binding Cassette transporter. The Arabidopsis AtMRP5 homolog of PvMRP1 has been characterized by reverse genetics approach: the atmrp5-1 insertion mutant displays lpa seeds, modified root architecture, reduced sensitivity to abscisic acid in seed germination and in guard cells, improved drought tolerance and water use efficiency. Recently it was reported the first direct evidence that AtMRP5 is a vacuolar phytic acid transporter. In Phaseolus there is also PvMRP2, paralog of PvMRP1, while in Arabidopsis there is only one gene. PvMRP2 is probably able to complement the lpa1 (280-10) phenotype in other tissues than the seed, thus explaining the lack of pleiotropic effects in the bean mutant. The aim of this work is the functional characterization of PvMRP1 and PvMRP2 through expression analysis, subcellular localization and complementation of the Arabidopsis atmrp5 mutant. Here we show preliminary data and our working model. VI ICLGG - Posters Fatty acid profiling for pod fragrance and impact on pod borers in field bean ( Dolichos lablab L.). P-NAQ12 Doss DD*, Rao KS, Gowda MB. Department of Plant Biotechnology, University of Agricultural Sciences, Bangalore, India *E-mail: dds3a1win@gmail.com Abstract Field Bean ( Dolichos lablab L.) is an important leguminous pod vegetable used mostly in Asian countries and the consumers showed much perference for the varieties with fragrance. Present invetigation on biochemistry of fatty acid profiling of pod fragrance in Field Bean was attempted for the first time by selecting High- fragrant and Low-fragrant germ lines. Gas chromatographic analysis of pod exudates revealed the fatty acid profile dominating 2-DoDecenoic acid, Lauric acid, Myristic acid and Palmitic acid in sequence. Fattyacid, 2-Do-Decenoic acid was the major constituent (30.34%) in exudates of high fragrant germ lines (GL-16, 17, 18, 21 and 26) and was responcible for the high frgrance in field bean. In contrast, Luaric acid (17.90%) dominates in low fragrant germ lines (GL-6,22,24,32 and 34). To correlate the fatty acid profile with the infestation of pod borers, Helicoverpa armigera and Adisura atkinsoni, a mean per cent inhibition study was done during the crop growth under the field condition. The high frgerance content was postively corellated with the per cent pod wall damage ( 29.00%) by pod borer H. armigera. Present investigation also confirmed the presence of other fatty acids like Stearic acid, Oleic acid, Linoeic acid and Linolenic acid in comparatively less per cent and were completely absent in low fragrant germ lines. VI ICLGG - Posters P-NAQ13 Cytological and biochemical analysis of large seed mutants of mungbean induced by gamma rays * Dhole VJ , Reddy KS Nuclear Agriculture and Biotechnology Division, Bhabha Centre, Mumbai, India Atomic Research *E-mail: vjdhole@yahoo.co.in Abstract Mungbean is important short duration pulse crop cultivated for seeds. Demand for large seed mungbean is increasing because of better milling quality and higher dhal recovery. Mungbean cultivars with large seed size and MYMV resistance are limited. Samrat is high yielding and MYMV resistant variety but having small seed size. Four large seed mutants (LSM) of Samrat were developed through gamma rays (450 Gy). The present study was undertaken to find out the cytological and nutritionally important biochemical changes in LSM’s. LSM’s were evaluated in different seasons for seed weight and yield. Observations were recorded on cell size, starch granule size, shape and number/cell, protein, amylose and starch content. The LSM-1, LSM-2, LSM-3, LSM-4 and Samrat showed 6.44, 6.19, 5.80 5.20 and 3.90 g mean 100 seed weights respectively, which were 32.99 to 64.96 % more over Samrat. The cell size, starch granule size and number of starch granules/cell increased in all four LSMs (17-20/cell) than Samrat (12/cell). The starch granule was found oval shape (1.13 to 1.22 Length: Breadth ratio) in LSM’s, while elongated in Samrat (1.45 Length: Breadth ratio). Protein and amylose content were at par in LSM-1 (25.46% and 19.9%), LSM-2 (25.40% and 21.08%), LSM-3 (25.16% and 21.89%) and LSM-4 (26.24% and 20.11%) with Samrat (25.85% and 22.05%) respectively. Starch content was increased significantly in LSM’s (41.4 to 48.27%) as compared to Samrat (35.80%). Therefore, the increased seed size was attributed to increased cell size, starch granule size, number of starch granules/cell and starch content. VI ICLGG - Posters Studies on antinutritional factors in lentil (Lens culinaris Medic) 1,* P-NAQ14 2 Gaikwad NB , Kothekar VS 1 Shivaji University, Kolhapur, India Dr. B. A. M. University, Aurangabad, India 2 *E-mail: nbgaikwadsu@gmail.com Abstract Lentil (Lens culinaris Medic) is a nutritional food legume which has a relatively higher content of proteins, carbohydrates and calories and it is the most desired crop in lentil growing regions because of its high average protein content and fast cooking characteristic. Presence of antinutritional factors such as trypsin inhibitor (TI), polyphenol and lectin content inhibits the nutritional quality of lentil. Forty –nine lentil genotypes were analyzed for trypsin inhibitor, polyphenol and lectin content. Among the 49 genotypes, lowest TI content (140.25±0.17 TIU/Min/g seed meal) was recorded in IC-201695genotype, while the IC-208326 genotype revealed the highest TI content (830.66±0.20 TIU/Min/g seed meal). Highest polyphenol content (409.3±0.64mg/100g seed meal) was recorded in IC-208326 genotype, whereas L-4642 genotype showed lowest (159.0±0.40 mg/100g seed meal) polyphenol content. Lectin activity was expressed in terms of specific activity and it was observed that L332 genotype demonstrate lowest (3.571 HAU/mg protein), whereas L-4611 genotype reported highest (23.188 HAU/mg protein) specific activity. All 49 lentil genotypes exhibit the wide range of variation for presence of TI, polyphenol and lectin content, from this information one can select a genotype with least amount of antinutritional factors for further breeding program. VI ICLGG - Posters P-NAQ15 The indirect effect of leaf-feeding insects on soybean quality improvement Gu H, Chen H, Chen X*, Yuan X, Zhang H, Peng C, Cui X Institute of Vegetable Crops, Jiangsu Academy of Agricultural Sciences, Nanjing, China *E-mail: ghp@jaas.ac.cn Abstract The hazards of 640 soybean germplasm were evaluated from leaf feeding insects under the natural environment in Nanjing from 2006 to 2008. Insect species were divided into two kinds of all leaf-eating insects (mainly incloud Spodoptera litura, silver agnata, Great bridge-building worm and soybean leaf borer) and soybean leaf borer; there is a linear relationship among the leaves victims, and the soybean seed protein content and taste of fresh soybean seeds statistically. The indirect selection effects on seed quality of the hybrid offspring of two combinations were analyzed under two types of insects (all insect species and soybean leaf borer) pressure. The results showed that: the integrated pest species against the soybean grain protein content and taste evaluation of the correlation coefficient were 0.792 and 0.648 in the evaluation of three consecutive years of germplasm resources, respectively. Correlation coefficient of soybean leaf borer corresponding to protein content and taste evaluation were 0.656 and 0.499. In the F4 generation of the two combinations, soybean integrated pest hazard and the protein content of 0.781 and 0.579, very significant; and taste evaluation value of the correlation coefficient is 0.765 and 0.39, the former significantly, the latter was not significant. Soybean leaf borer and protein, taste evaluation value of the correlation coefficient is 0.471 and 0.848, respectively, are very significant. F2F3 two generations of leaf-eating insect species to select a combination (58161 × Haimen chihuangdou) grain protein content were increased by 3.0% and 2.5%, the second combo 3.2 % and 2.2%; combination of taste evaluation value increased by 5% and 4%, 4.1% and 3.1% in the second combo. Preliminary view that: the hazards of leaf-eating insects on the leaves can be used as indirect selection indicators for seed quality improvement. VI ICLGG - Posters Construction and use of saturated genetic map for identification of QTLs associated with disease resistance, oil quality, morphological descriptors, and yield components in cultivated peanut (Arachis hypogaea L.) 1,2,3 2,4 2 5 5 P-NAQ16 6 Pandey MK , Feng S , Culbreadth A , Wang M , Barkley N , Holbrook CC , 7 4 3 1,* Zhang XY , Wu YT , Varshney RK , Guo B 1 USDA-ARS, Crop Protection and Management Research Unit, Tifton, USA Department of Plant Pathology, the University of Georgia, Tifton, USA 3 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 4 Qiongzhou University, Sanya, China 5 USDA-ARS, Plant Genetics Resources Conservation Unit, Griffin, USA 6 USDA-ARS, Crop Genetics and Breeding Research Unit, Tifton, USA 7 Henan Academy of Agricultural Sciences, Zhengzhou, China. 2 Abstract Peanut production and consumer acceptability are adversely affected by several biotic/abiotic stresses and poor oil quality. In order to deal with these concerns, the developed genetic map from one peanut population derived from SunOleic 97R × NC94022 by Qin et al. (2012) using 190 subset with 172 markers was further advanced with additional 77 polymorphic marker loci on all 353 lines. The saturated genetic map possesses 249 loci distributed on 23 linkage groups with total map distance of 3,433.5 cM. Multiple phenotyping data of all lines on several economically important traits was used for quantitative trait loci (QTL) analysis, resulting in identification of a total of 155 QTLs with phenotypic variance (PVE) ranged from 1.48 to 65.2% and the logof-odds (LOD) ranged from 2.51 to 83.68. One major QTL (16.7% PVE) for Tomato spotted wilt virus (TSWV) resistance, three major QTLs for late leaf spot (12.42 -20.59%), two major QTLs for growth habit (11.90 - 14.05% PVE) while one major QTL each for leaf color (10.43% PVE), pod shape (11.55% PVE) and pod reticulation (14.01% PVE), and four QTLs for seed color (10.1218.45% PVE) were identified. QTL analysis also revealed that FAD2B gene contributes up to 74.03% PVE while FAD2A gene could contribute only 11.13% PVE for high oleic/linoleic (O/L) ratio. Interestingly, the A- and Bgenome mutations of FAD2 were also found to contribute to other fatty acids, such as palmitic acid (PVE 21.26 and 27.77%) and gadoleic acid (PVE 13.73 and 10.32%), respectively. The markers linked to major QTLs could be utilized for peanut genetic improvement through molecular marker-assisted selection in breeding. VI ICLGG - Posters P-NAQ17 Marker development and alleles distribution of both flavonoid 3'hydroxylase and flavonoid 3',5'-hydroxylase genes in soybean (Glycine max) Guo Y, Qiu L-J * The National Key Facility for Crop Gene Resources and Genetic Improvement (NFCRI), Institute of Crop Science, Beijing, China * E-mail: qiu_lijuan@263.net Abstract Color is one of the most widely used phenotypic markers to study genetic, molecular and biochemical process for their easy recognizability. Hydroxylation pattern of B-ring in flavonoids plays an important role in coloration. Two P450dependent monooxygenases, flavonoid 3'-hydroxylase and flavonoid 3', 5'hydroxylase, catalyze the hydroxylation of the B-ring in flavonoids. Previous study showed that soybean F3’H and F3’5’H co-segregated with the T and W1 loci respectively, which belonged to genetic loci controlling seed coat, flower and pubescence color. However, whether these two genes have other alleles and the distribution of these alleles in germplasm still remains to be revealed. In this study, coding sequences of GmF3’H and GmF3’5’H were identified and deduced amino acid sequences were analyzed with sequences from other organisms. Four different alleles of GmF3’H and three alleles of GmF3’5’H were elucidated on the basis of sequence variation from 30 different soybean materials, including modern elite cultivars, local landraces and annual wild soybeans. A set of gene-tagged markers were developed based on the sequence diversity of these alleles and verified. In addition, the distribution of these alleles in germplasm were also analyzed by using about 250 germplasm including cultivars, landraces and wild soybeans. The results revealed that different alleles were dominantly distributed in cultivars or wild soybeans. The genotype of GmF3’H was correlated with pubescence color and that of GmF3’5’H was correlated with flower color. Furthermore, combination of these two genes could explain more than 90% variety of yellow seed coat color from the materials in our study. . . VI ICLGG - Posters Biochemical characterisation of rajamash germplasm collected from Jammu hills P-NAQ18 Gupta M, Mallick SA, Mir RA, Guleria S, Singh V Division of Biochemistry & Plant Physiology SKUAST-J, Jammu, India *E-mail: sachinmoni@rediffmail.com Abstract Biochemical composition of red kidney beans locally called as Rajmash collected from hills of Doda district of Jammu Division were analysed to identify the nutritionally rich germplasm having the potential of being used for pulse breeding programme. A lot of variation was observed in the chemical composition of seeds collected from different villages giving an idea of the variability among the samples. Among the parameters evaluated, protein contents were found to vary from18.9 to 23.2 per cent. The minimum protein solubility of isolated proteins from all seeds was at around pH of 4-5. The seed samples collected from Chirala village exhibited the highest carbohydrate content of 59.9%, while the highest total starch, total soluble sugar and reducing sugar was found highest in the samples from Manthal. The highest trypsin inhibitory activity was found in bean collected from Chatroo (15.3 TUI/mg dw). However, highest carotenoid contents, phenols, tannins were found in red beans collected from Kansar. Essential limiting amino acid content i.e Methionine content was found to range from19.4 to 24.8 mg/gdw in the analysed samples. SDS-PAGE profiling of seed protein is being carried out. VI ICLGG - Posters P-NAQ19 Comparative transcription analysis of high seed protein alleles on chromosome 20 of soybean [Glycine max (l.) Merr.] using near isogenic lines 1 1 1 Jang YE , Kim MY , Van K , Lee S-H 1,2 1 Department of Plant Science and Research Institute for Agriculture and Life Sciences, Seoul National University, Seoul, Korea 2 Plant Genomics and Breeding institute, Seoul National University, Seoul, Korea *E-mail: sukhalee@snu.ac.kr Abstract The major sources of protein for human consumption are plants and cultivated soybean is known as one of the most efficient protein sources. Increasing seed protein content of soybean is important for supply of protein, however, trait of increasing protein content is related with other agronomic traits such as delayed seed maturity. The purposes of this research are understanding gene expression and inferring the process of controlling seed protein content. We had been investigated the major quantitative trait locus for high seed protein content on Chr 20 of soybean cultivar, Danbaekkong, which has about 480 g -1 kg of seed protein content. Simple sequence repeat (SSR) markers on Chr 20, Satt239 and Satt496, were used to select near isogenic line (NIL) from two recombinant inbred line (RIL) populations from Sinpaldalkong 2 x Danbaekkong (SD) and Daewonkong x Danbaekkong (DD) crosses. Residual heterozygous line (RHL) strategy was efficient to reduce time for selecting NIL pairs from each RIL population. SSR marker genotyping that inherited from Danbaekkong was corresponded with high seed protein content in all NIL pairs. To investigate transcription and translation patterns during seed development, immature seeds were harvested at 1, 2, 3 and 4 week after flowering (WAF). Transcriptomic change during seed development at 1, 2, 3 and 4 WAF of a NIL pair from DD RIL population was investigated using nextgeneration sequencing. Proteomic changes in 2, 3 and 4 WAF seed of two NIL pairs from SD and DD populations were examined by protein 2-dimension electrophoresis technologies. VI ICLGG - Posters Lentil (Lens culinaris L.): a whole food rich in prebiotic carbohydrates P-NAQ20 * Johnson CR, Thavarajah D , Thavarajah P North Dakota State University, Fargo, USA *E-mail: dilrukshi.thavarajah@ndsu.edu Abstract Prebiotics, which are important in maintaining healthy hindgut microflora, reduce factors associated with obesity and metabolic syndrome. Biofortification of these prebiotic compounds in pulses through traditional plant breeding could be a sustainable strategy to reduce obesity-related metabolic disorders. Several groups of prebiotic carbohydrates are contained in lentils (Lens culinaris L.). The present study was carried out to determine the prebiotic carbohydrates present in 10 lentil genotypes grown in Ward and McLean Counties, North Dakota, USA in 2010 and 2011. Significant variations were observed in lentil prebiotic carbohydrates: sugar alcohols varied with both genotype and locations, raffinose-family oligosaccharides (RFO) varied with genotype, and resistant starch (RS) varied with location. Sugar alcohols, fructooligosacharides (FOS), RFO, and RS mean concentrations were 1.4 g, -1 0.06 g, 4.1 g, and 7.5 g 100 g dry matter, respectively. These results show that a quarter-cup serving of cooked lentils (30 g lentils by dry weight) could provide approximately 3.6 g of prebiotic carbohydrates. Study results also show that lentils contain nutritionally significant amounts of prebiotic carbohydrates. Moreover, variation in prebiotics with genotype and location suggests potential to enhance their concentrations through biofortification and locational sourcing. VI ICLGG - Posters P-NAQ21 Antinutritional factors in moth bean (Vigna aconitifolia (Jacq.) Marechal): effect of heat treatment and germination 1, Khadke SG *, Kothekar VS 2 1 Department of Botany, Smt. Kasturbai Walchand College, Sangli. India Department of Botany, Dr. Babasaheb Ambedkar Marathwada University, Aurangabad,India 2 *E-mail: shivramkhadke@gmail.com Abstract Moth bean (Vigna aconitifolia (Jacq.) Marechal) is one of the important pulse species of India. It is widely grown in the arid and semiarid regions of Rajashtan, Gujarat, Maharashtra, Utter Pradesh and Haryana. Moth bean possesses a great amount of antinutritional factor like trypsin inhibitors. Trypsin inhibitor assays were carried out to find out the trypsin inhibitor activity among seven viable mutants and 78 micromutants of moth bean which were developed by induced mutation breeding. The pertinent studies of seven viable mutants and 78 micromutants of moth bean have indicated considerable variation regarding TI level. The lowest TI (180.00 TIU/min/gm meal) could be recorded in 6/IV-mutant, while the 7/IV-mutant revealed the highest TI content (326.16 TIU/min/gm meal). The electrophoretic profiles of trypsin inhibitors on X-rays film revealed 3 to 7 isoinhibitors in different viable mutants and micromutants of moth bean. The mutants showing reduced trypsin inhibitor bands were quantitatively estimated. The lowest TI activity (187.98 TIU) was observed in B6 mutant, which was evolved by the 0.15% EMS treatment. To nullify trypsin inhibitor activity in moth bean seeds, so as to use it in diet, the heat treatment and germination studies have been tried in some of its mutant lines. The processing method involving heat treatment almost eliminated trypsin inhibitor activity while soaking and germination partly removed the activity VI ICLGG - Posters Utilizing next generation sequencing in soybean product development P-NAQ22 Hyten D, Woodward J, Kuhlman L, Massman J, King M*, Zhang Y, Daines B, Sebastian S, Deschamps S, Shendelman J, Mendez E, Kalvig A, Hayes K DuPont Pioneer, Johnston, USA *E-mail: matt.king@pioneer.com Abstract Pioneer has had tremendous success in utilizing molecular markers throughout the soybean product development breeding scheme to increase productivity through the Accelerated Yield Technology (AYT(TM)) system and the utilization of native traits. Next Generation sequencing has created a revolution in the world of genomics that is enabling Pioneer to look past the use of molecular markers to a fuller integration of genomics within product development. Through the use of this technology we have resequenced a set of soybean lines which include the lines that make up soybean’s ancestral base, several key lines throughout Pioneer’s history and many elite lines which are being used as parents in our current product development. This resequencing dataset has enabled us to discover a large proportion of the variation that is present within our germplasm for soybean improvement and allows us to identify regions of the genome where diversity has become extremely low. This data is also being used to increase our ability to identify and delimit regions of the genome that contribute to important native traits enabling discovery and better marker development. In addition, this resequencing data allows an unprecidented view into the inheritance of progeny lines within breeding populations. Genomics technology such as next generation sequencing has allowed a further integration of the use of genomics into the product development strategy which will result in higher performing products within farmer’s specific growing environments. VI ICLGG - Posters P-NAQ23 Studies on high O/L ratio in cultivated peanut by marker assisted backcross selection (MABS) 1 2 2 1 1# 1 Koilkonda P , Kuwata C , Fukami M , Shirasawa K , Aoki K , Tabata S , 2 2 2 1 1 Hasegawa M , Kiyoshima H , Suzuki S , Sasamoto S , Kurabayashi A , 1 1 1, Tsuruoka H , Wada T , Isobe S * 1 Kazusa DNA Research Institute, Kisarazu Chiba, Japan Chiba Prefectural Agriculture and Forestry Research Center, Chiba, Japan 2 *E-mail:sisobe@kazusa.or.jp Abstract Peanut (Arachis hypogaea L.), allotetraploid (AABB; 2n=4X = 40), is a major food legume, oil seed crop covering a large area with high production annually throughout the world. Cultivated peanut is highly important because of its applications in various aspects which is raised by breeding in most of the public sectors. The present research mostly project on the use of molecular strategies like candidate gene approach ie., SNP (ahFAD2A) and transposon (ahFAD2B) markers and SSRs in backcross selection (upto BC cycle 2, BC cycle 3 in progress) to generate high O/L ratio (determinant of oil quality) in peanut germplasm. The two parental lines ‘Nakateyutaka’ (Normal O/L ratio; seed and backcross parent) and ‘YI-0311’ (High O/L ratio; pollen and donor parent) were used in the breeding scheme. Simultaneously construction of a consensus linkage map was also carried out using Genome Wide (GW) polymorphic markers on 186 F2 lines to increase the number of markers. By Genotype Matrix Mapping (GMM) analysis, it was found that high O/L was associated with the combination recessive alleles of ahFAD2A and ahFAD2B. The result also suggested limited diversity in the mutation alleles on the two FAD2 genes and the markers developed can be more effective on multiple donor lines showing high O/L ratios. Finally nine BC2F2 plants showed homozygous ‘YI-0311’ genotypes (High O/L ratio) on the FAD 2 loci, designated for use in the next BC Cycle 3. Backcrossing and selfing were performed alternately to suffice the problem of less temperature during off season in the greenhouse. Conclusively our breeding programme is cost effective in terms of labour, time and money. Higher efficiency of MABS can be achieved by using more number of GW polymorphic markers and increasing the amount of breeding material used in the scheme. VI ICLGG - Posters Genetics of cotyledon colour in lentil (Lens culinaris Medik.) 1,* 2 3 4 5 P-NAQ24 5 Kumar Y , Sarker A , Akhtar J , Singh PK , Mahto CS , Singh MM , Sharma 6 B 1 Central Rainfed Upland Rice Research Station (CRURRS), Hazaribagh, India South Asia Regional Program (ICARDA), New Delhi, India 3 Division of Quarantine, NBPGR, New Delhi, India 4 Department of Entomology, BAU, Ranchi, India 5 Department of Plant Breeding & Genetics, BAU, Ranchi, India 6 Division of Genetics, IARI, New Delhi, India 2 *E-mail: dryogeshtiwari70@gmail.com Abstract The genetics of cotyledon colour was studied using F 1, F2 and F3 generations de from 36 cross combinations in lentil. A monogenic inheritance pattern was obta when analyzed F2 seeds from the crosses between orange vs yellow (12), orang brown (2), orange vs dark green (2) and yellow vs light green (5) seeded parents F1 seeds always showed orange cotyledon and F 2 seeds segregated into 3:1 rat all combinations (3 orange: 1 yellow; 3 orange: 1 brown; 3 orange: 1 dark green, 2 respectively) with non-significant values. The digenic inheritance of cotyl colour and its epistatic interaction were confirmed from the crosses between or vs light green (3) and yellow vs brown cotyledon (3) parents, which gave a dig ratio of 9 (orange): 3 (brown): 3 (yellow): 1 (light green) cotyledon. The orange c of cotyledon was demonstrated to be a digenic trait, in which two genes (Y and B yellow and brownish (called brown) jointly cause orange and light green cotyledo dominant and recessive homozygous conditions, respectively. Digenic segreg with recessive epistasis (9:3:4) was obtained when the genes Y or B were combi with dg (third gene) through hybridization, while independent assortment without kind of epistasis (9:3:3:1) resulted when the gene dg was excluded. The trige inheritance pattern was obtained when two crosses were made with light green dark green cotyledons, which brought all three genes (Y, B, Dg). Although parents in such crosses had green (light or dark) cotyledons, the F1 seed developed orange cotyledons and F2 seeds segregated into all five pheno classes: orange, yellow, brown, light green and dark green with a good fit to trigenic ratio of 27:9:9:3:16. A detailed study has confirmed the hypothesis on basis of segregation patterns in all possible gene combinations. VI ICLGG - Posters P-NAQ25 Rice bean (Vigna umbellata (Thunb.) Ohwi and Ohashi) for nutritional security and conservation of natural resources in north-western Himalayas Mehta H*, Srimali SS, Ghosh BN, Singh C, Joshi BP, Chaturvedi OP Central Soil and Water Conservation Research and Training Institute, Dehradun, India *E-mail: harshmehta41@rediffmail.com Abstract Pulses grown in the hilly regions of north-western Himalayas are the integral components of human diet as they provide quality proteins for tribal and hilly people. Kharif pulses viz. rice bean (Jhilangi): Vigna umbellata; kidney bean (Chhimi) : Phaseolus vulgaris, horse gram (Kulthi): Dolichos uniflorus and soybean (Kalibhat) : Glycine max are the prominent crops usually grown under mixed cropping with other staple crops like maize and minor millets. Deep root system of pulses complements well with shallow root system of cereals in mixed and inter cropping systems. Pulses are known to improve the soil fertility by fixing atmospheric nitrogen and provide effective canopy cover over land, thereby preventing runoff and soil loss in fragile undulating topography of Himalayas. Rice bean (Vigna umbellata (Thunb.) Ohwi and Ohashi) is a traditional pulse crop generally gown in marginal lands but its soil conservation value is not studied. Studies were conducted for five years, initiated in 2004-05 to utilize the existing waterways at the farm for conserving natural resources and productive purposes in scientifically planned waterways under rice bean- gobhi sarson (Brassica napus) cropping sequence ( first segment ,0.9% slope), finger millet - lentil cropping sequence (second segment 1.1% slope) and guinea grass (1.3% slope). Productivity levels of rice bean, lentil, gobhi sarson finger millet and guinea grass were assessed alongwith their role in conserving natural resources and improving soil productivity in degraded lands of nearly 1 KM long waterways with the catchment area of 12.0 ha under rainfed situation. The average biomass -1 productivity of rice bean was recorded at 3.6 t ha maturing in 147 days over -1 five years , while the average grain yield was 0.72 t ha . Rice bean provided a very good canopy cover over the soil, covering 80% of the area within 40 days. The crop has low HI at 0.25 and high biomass production levels and hence suitable for fodder purposes as well. Cultivation of finger millet and guinea grass in adjoining segments of the water way with rice bean in the first segment resulted in moderation of peaks flows of run off water in the waterways during the rainy seasons. The outflows from the downstream pond of the waterway were reduced from the average of seven in past to four or five. The treatments of rice bean, finger millet and guinea grass in successive plots of the waterway were quite effective in arresting soil loss. Treatments trapped soil sediments and their net deposition was to the tune of 15.41 cum and 25.40 cum in segment II and segment III respectively which otherwise would have resulted in reduction of capacity of downstream pond. Integration of rice bean in farming systems is advocated not only in improving biomass productivity for nutritional security but also for conserving natural resources, utilizing wastelands and improving soil fertility for environmental security and sustainable productivity. VI ICLGG - Posters The effect of moisture deficiency on protein content of legumes P-NAQ26 * Narits L , Keppart L Jõgeva Plant Breeding Institute, Estonia * E-mail: lea.narits@jpbi.ee Abstract The purpose of this experiment was to determine the impact of the moisture deficiency to the seed protein content of different legumes. Experiment was carried out at the Jõgeva Plant Breeding Institute in 2010-2011. The following legumes were tested in the experiment: common vetch, field bean, leafy and semi-leafless pea, and soybean. Precipitation rates varied significantly, depending on the length of the growing period. Pea cultivars of the shortest growing period suffered most by the moisture deficiency. Precipitation for leafy pea was 70% of the Long Term Average (LTA) (1964-2009) in 2010 and 52% of LTA in 2011. Seed protein content of leafy pea decreased by 2.23% in 2010 and 1.19% in 2011. Precipitation of the growing period of semi-leafless pea was 70% of LTA in 2010 and 54% of LTA in 2011. Seed protein content of the varieties of semi-leafless pea decreased the least compared to the other species: 0.21% and 0.39%, respectively. In 2010, soybeans got only 10.6% less precipitation compared to LTA but the seed protein content decreased to 3.48%, while in 2011 of the precipitation deficit of 35% of LTA, the protein content decreased only 0.78%. Protein content of field bean - increased by 0.11% in 2010 (moisture deficiency 18% of LTA), but decreased by 0.97% in 2011 (deficiency 33%). Protein content of common vetch decreased both years: 1.14% and 2.41% respectively (deficiency 17 and 37%). Correlation analysis proofed significant correlation between precipitation and the seed protein content (r = 0.73 ***). VI ICLGG - Posters P-NAQ27 Effect of nutrient sprays on yield and seedling quality parameters of chickpea (Cicer arietinum L.) Parimala K*, Anitha G, Reddy VA Acharya N.G. Ranga Agricultural University, Hyderabad, India *Email: pari_mala123@rediffmail.com Abstract Chickpea (Cicer arietinum L) is one of the important pulse crops in rainfed areas of Andhra Pradesh. The crop requires higher levels of Nitrogen and Phosphorus and is often susceptible to deficiencies of Iron, Sulphur and Zinc. A field experiment was conducted at Seed Research and Technology Centre, Rajendranagar, Hyderabad to study the effect of macro and micronutrients on yield and seedling quality parameters of chickpea variety JG-11. Foliar applications of DAP (2%), Urea (2%), Zn (0.5%), FeSO4 (0.5%), KCl (1%) and KNO3 (1%) were done twice at pre flowering and flowering stage. DAP (2%) and Urea (2%) sprays recorded maximum number of branches per plant which suggests that overall biomass could be increased by DAP and Urea sprays. Zn (0.5%) enhanced the number of pods/plant and led to maximum seed yield/plant compared to all other treatments. Application of Zinc enhanced the root growth, nodulation and nitrogen content of nodules indirectly contributing to the increased yield. KCl (1%) sprays had a positive effect on root and shoot growth recording maximum seedling root length, shoot length, fresh weight, dry weight and vigour index. However, the nutrient sprays had no effect on plant height and germination. VI ICLGG - Posters Marker-assisted introgression of high oleate trait to improve oil quality in elite genotypes of groundnut (Arachis hypogaea L.) 1* 1 1 1 P-NAQ-28 1 Janila P , Pandey MK , Nagesh MP , Manohar SS , Roorkiwal M , Sriswathi 1 2 3 4 5 2 M , Radhakrishnan T , Manivannan N , Vasanthi R , Dobariya KL , Bera SK , 2 1 1,6,* Mishra JB , Nigam SN , Varshney RK 1 c International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 2 Directorate of Groundnut Research (DGR), Junagadh, India 3 Tamil Nadu Agricultural University (TNAU), Coimbatore, India 4 Regional Agricultural Research Station (RARS), ANGRAU, Tirupathi, India 5 Junagadh Agricultural University (JAU), Junagadh, India 6 CGIAR Generation Challenge Programme (GCP), CIMMYT, Mexico DF, *E-mail: p.janila@cgiar.org; r.k.varshney@cgiar.org Abstract Groundnut kernel contains 48-50% oil and stability (shelf- life) of the oil depends on oleic to linoleic acid ratio (O/L ratio). The O/L ratio in normal groundnut genotypes varies from 1-4, however, in case of mutants it can go >20 due to mutations in FAD (fatty acid desaturase) genes in both the genomes. Realizing the potential of this trait towards oil quality improvement and availability of associated markers, marker-assisted backcrossing (MABC) approach has been initiated to improve this trait in elite genotypes with high oil content (>50%). Cleaved amplified polymorphic sequence (CAPS) and allelespecific PCR-based markers associated with high O/L ratio were optimized and validated on a set of 22 genotypes. Allele specific PCR-based markers differentiated the genotypes possessing mutations in A and B genomes, while CAPS markers could not show consistency due to complex technical genotyping. Crosses were effected between six selected recurrent parents (RP) and SunOleic 95R, donor carrying both mutant alleles (high O/L ratio). Allele-specific markers were used for confirmation of F1 plants as well as for foreground selection to select heterozygotes among backcrossed BC 1F1 plants. It is planned to undertake three backcrosses followed by selfing to develop MABC products with enhanced O/L ratio and oil content. VI ICLGG - Posters P-NAQ29 Assessment of quality parameters in greengram seeds treated with newer insecticides against the pulse beetle (Callosobruchus chinensis L.) Raheem A*, Sridevi D, JhansiRani K, Rajasri M ANGRAU, College of Agriculture, Hyderabad, India *E-mail: amieraheem04@gmail.com Abstract Mungbean [Vigna radiata (L.) Wilczek] is damaged to a great extent during storage by C. chinensis both in terms of quality and quantity. The uninterrupted use of malathion i.e., a commonly used insecticide resulted in the development of high degree of resistance invariably requiring suitable alternative insecticide with low mammalian toxicity. Hence, a study was undertaken to evaluate the residual toxicity of newer insecticides on green gram seeds and also to assess their effect on seed quality parameters. Seven insecticides viz., abamectin, emamectin benzoate, spinosad, novaluron, lufenuron, deltamethrin and neem were studied. The seeds were treated @ 2, 4, 2, 2, 5, 1 and 75 ppm, respectively and 5 pairs of adults were released into each jar containing the treated seeds in three replications. The mortality was assessed each day upto 14 days and thereafter the adults were removed. After 40 days the residual toxicity in terms of per cent mortality and adult emergence of C. chinensis, per cent seed germination, seed vigour, per cent moisture content and per cent insect damage was recorded. The results showed that abamectin caused 100 per cent mortality at 5 days after treatment (DAT), emamectin benzoate at 6 DAT, while spinosad and deltamethrin took more than 7 days, whereas, novaluron, lufenuron and neem recorded relatively low mortality rates. After 40 days spinosad, abamectin, emamectin benzoate and deltamethrin recorded lower adult emergence of 16, 17, 32 and 28, respectively. The spinosad treated seeds showed lower moisture content (8.0%), per cent seed damage (0.0%), oviposition (5.6 eggs) and higher vigour index (2362) and germination (93.33%) than all other treatments. Thus, it was found that for a short period of storage the seed can be stored by using spinosad as a protectant, as it is comparatively efficient in maintaining seed quality by reducing the damage. VI ICLGG - Posters Comparative and evolutionary analysis of major peanut allergen gene families 1 1 1 1 1 1 P-NAQ30 1 Ratnaparkhe MB , Lee TH , Tan X , Wang X , Li J , Kim C , Rainville LK , 1 1 1 2 3 1,* Lemke C , Compton RO , Robertson J , Gallo M , Bertioli D , Paterson AH 1 The University of Georgia, Athens, Georgia, USA University of Florida, Gainesville, Florida, USA 3 Universidade Católica de Brasília, Brasília, DF, Brazil 2 *E-mail: paterson@plantbio.uga.edu Abstract Peanut (Arachis hypogaea) allergy is one of the most serious food allergies. Peanut seed proteins, Arah1, Arah2 and Arah3, are considered to be one of the most important peanut allergens. We sequenced ~ 617 kb from cultivated peanut and ~ 215 kb from wild species (A genome) to gain insights into genome organization and evolution of Arah1, Arah2 and Arah3 allergen genes. To assign polarity to differences between homoeologous regions in peanut, we used as outgroups the single orthologous regions in Medicago, Lotus and pigeonpea which diverged from peanut approximately 50 million years ago (mya) and have not undergone subsequent polyploidy. We also compared these regions with orthologs in soybean, grape, tomato, poplar and Arabidopsis, to help clarify the timing of evolutionary events. The peanut allergen genes are interspersed with low-copy genes and transposable elements. Phylogenetic analyses revealed lineage-specific expansion and loss of low-copy genes between species and homoeologs. We infer that two rounds of segmental duplication and tandem duplications led to the establishment of the Arah3 gene family. Our analysis indicates differences in conserved motifs in both Arah1 and Arah3 proteins and in the promoter regions. Phylogenetic analysis and genomic organization studies provide new insights into evolution of major peanut allergen genes. VI ICLGG - Posters P-NAQ31 Suppression of soybean storage proteins results in altered changes in seed proteome that may be caused or resulted from genome methylation Schmidt MA*, Herman EM University of Arizona, Tucson, USA *E-mail: monicaschmidt@email.arizona.edu Abstract Soybean (Glycine max L.) has numerous uses that largely revolve around its high oil and protein content. We are interested in how the seed content is determined and, in particular, if the content can be altered for specific end uses. We have engineered soybean via RNAi technology to have a seedspecific suppression of both its storage proteins (RNAiSP), glycinin and betaconglycinin. Together these proteins constitute over 80% of soybeans near 40% seed protein content. The resultant transgenic plants are phenotypically comparable to the nontransgenic counterparts and the seed composition in the transgenics are still ~40% protein/ 18% oil as in wildtype but the proteins making up the proteome have been altered. We have also engineered soybean seeds via RNAi to suppress only the beta-conglycicin storage protein (RNAiBC), and like the RNAiSP seeds they also contain wildtype levels of oil and protein and their seed protein content composition also has changed to now contain increasing levels of alternate compensating proteins. Interestingly, although the RNAiSP plants have no visible phenotypes, the RNAiBC plants display a number of altered phenotypes compared to wildtype plants, namely dark hilium, white-to-purple flower color, change in leaf morphology and dwarf stature. These plant phenotypes displayed in RNAiBC, but not RNAiSP, plants can most readily be explained by epigenetic events. Methylation analysis of the RNAiBC and RNAiSP genomes compared to wildtype revealed a marked, 3-fold, increase in global methylation. The comparison of the maps of methylation in the genomes of RNAiBC and RNAiSP compared to wildtype will give insights to the mechanism of protein compensation in the seeds and to the RNAiBC plant phenotypes that cannot be explained by the introduced transgene. VI ICLGG - Posters Effect of elevated atmospheric CO2 concentration on nutrient quality of black gram (Vigna mungo L. hepper) P-NAQ32 Kobaku SS*, Vanaja M, Venkateswarulu B Central Research Institute for Dryland Agriculture (CRIDA), Hyderabad, India *E-mail: k.sreedevishanker@crida.in. Abstract Elevated atmospheric CO2 plays an important role in determining many nutritional aspects of food crops and atmospheric CO 2 concentration is unknown to specifically alter the concentrations of food crop constituents related to human health. A pot experiment was taken up in open top chambers (OTCs) tied with transparent PVC sheet each having 3m×3m with 3m height, where elevated CO2 was maintained for 24h per day until maturity of Black gram crop (cv.T9). The three chambered CO2 treatments comprised ambient i.e., chamber control and elevated CO2 treatments of 550 and 700ppm. Food crop nutrient quality analysis of black gram was carried out to determine protein, crude fibre, total ash and mineral content in edible parts of Black gram crop. The result showed an increased content to elevated CO 2 levels in terms of protein and was observed that the response to enhanced CO 2 levels was higher at 700ppm compared to other two treatments. The total ash content of black gram crop enriched with elevated CO2 concentration was significantly higher at 550ppm level compared to ambient CO 2 level. Crude fibre content of black gram was found to be increased with increased CO 2 concentration i.e. 550 and 700ppm. Magnesium content recorded significant decrease with enriched CO2 levels at 550ppm. The response of manganese content had lower effect due to enriched CO2 concentration from 550 to 700ppm and the trends was also repeated in the case of iron and copper content, in black gram crop. VI ICLGG - Posters P-NAQ33 Efficience and assimilation of phosphorus in common bean (Phaseolus vulgaris) Silva DA*, Messias U, Chiorato AF, Esteves JAF, Carbonell SAM The Agronomic Institute (IAC), Campinas, Brazil *E-mail: daiagrouel2002@hotmail.com Abstract The common bean is the most important legume from Brazilian consumption, being part of the daily diet and the principal source of protein and iron. This crop is often cultivated in soils of low phosphorus availability, causing themain limitation of the crop production.The objective was to evaluatethe morphophysiologicalresponse for P absorption and its recovery of20 common beans genotypesunder differentconcentrations.The experiment was conducted in a hydroponic greenhouse located at theInstitute of Agronomic of Campinas (IAC). The experiment was in split plot design with four concentration of P(35, 17.5, 8.75 e 2 ppm) in the nutrient solution, with six replicates for genotype. Three replicates were collected at the flowering phase to evaluatethe leaf area, fresh and dry weight, length of plant, P level on leavesand chlorophyllcontent; the roots traits evaluatedwere superficial area, length and volume, fresh and dry weight. Three replicates of each genotype were measured for yield.The th symptoms of deprivation of P started around the 30 day at the 2ppm concentrationplot, presenting reduced plant size andearly leaf abscission.Allconcentrations of P present significant differences on grow of plantsbetween plots and between genotypes for most of the traits evaluated in P deprivation condition.Theconcentrations used were efficient to characterize these genotypes to select for P efficiency. VI ICLGG - Posters Evaluation of elite lentil (Lens culinaris subsp culinaris) lines for iron and zinc concentration and their molecular characterization using SSR markers 1 1,* 1 1 P-NAQ34 1 Kumar H , Singh Akanksha , Dikshit HK , Singh D1, Jain N , Prabhu KV , 2 Sarkar A 1 Division of Genetics, Indian Agricultural Research Institute, New Delhi, India ICARDA South Asia Office, New Delhi, India 2 *E-mail: a_singh1388@yahoo.in Abstract Forty one IVT entries of lentil were evaluated at IARI,New Delhi,GBPUAT Pantnagar and PAU Ludhiana during 2010-11 cool season for estimation of Fe and Zn concentration,GxE interaction studies and analysis of genetic diversity using SSR markers. Both micronutrients are influenced by environment, however Fe more sensitive. Analysis of stability based on Russel Eberhart model revealed that IPC320 and L4704 were most stable for Zn concentration and LH 07-26 and LH848. AMMI biplot revealed that all three locations discriminated Zn concentration; IARI was most discriminative of Fe concentration. The studied lines were characterized using thirty four SSR markers. The genetic similarity based on Jaccard’s coefficient observed during the investigation ranged from 9 to 66% which indicated the existence of high degree of variability among the studied lines. The studied genotypes were grouped in two clusters and genotypes originating at different locations were grouped in same cluster. Cluster I comprised of seventeen genotypes which were further sub grouped into two sub clusters and Cluster II included twenty four genotypes which were further sub grouped into two sub clusters. No association between clustering pattern eco-geographical distribution/ origin of genotypes was recorded. The results obtained could benefit lentil breeders in selection of suitable parents for hybridization programme, study of genetics and development of mapping population for these traits. VI ICLGG - Posters P-NAQ35 Breeding mungbean for bio-fortification of iron and zinc content 1, 1 1 1 Singh V *, Yadav RK , Yadav R , Yadav NR , Singh J 2 1 CCS Haryana Agricultural University, Hisar, India Central Soil Salinity Research Institute, Karnal, India 2 *E-mail: vijayatasingh.gpb@gmail.com Abstract Micronutrient deficiency leads to malnutrition or “hidden hunger”. There is an international effort to breed staple food crops for enhanced micronutrient content through CGIAR. However, mungbean is not included in their priority crop list even it being consumed on large scale by Indian population. Iron and Zinc are two important micronutrients that required for maintaining metabolic regulation and organ function. Thus, there is a need to improve mungbean varieties with high iron and zinc content. An experiment was conducted during Kharif 2011 to assess predictability of performance through stability parameters for seed yield/ plant, protein content and iron and zinc content of 12 mungbean genotypes tested at two locations in 14 environments. Pooled analysis of variance indicated highly significant differences for variation components. The partitioning of G x E interaction into linear and non-linear components indicated that both predictable and unpredictable components shared the interaction. Deviation from regression was highly significant revealing unpredictable response of these genotypes and better suitability with 2 better environments. The basis of stability parameters (Mean, b and S di) best three genotypes; ML 776, MH 565 and SML 668 with higher performance for seed yield, protein, iron and zinc content, may be recommended for better environment. Consequently, RILs have been developed using ML776 and Sattya with contrasting micronutrient content having per 100g dry weight of seed Fe (10.45mg and 4.02mg, respectively) and Zn (2.64mg and 1.51 mg, respectively) for gene/QTL tagging associated with high Fe and Zn content and for further mungbean breeding programmes. VI ICLGG - Posters Genetic variation in storage proteins and isozymes among Vicia faba L. cultivars of Bihar 1 Pradhan A , Sinha SK P-NAQ36 1, 2* 1 Faculty of Basic Sciences & Humanities, Rajendra Agricultural University, Pusa, India 2 National Research Centre on Plant Biotechnology, IARI campus, New Delhi, India * E-mail: subsinha@gmail.com Abstract Legumes are an important source of proteins and minerals in human and animal nutrition. People of developing countries largely depend on seed proteins for their protein requirement. Faba bean is the traditional legume crop in Bihar which has a long history of cultivation and wider distribution. Characterization of the genetic variation in the different germplasms being grown is important for its further genetic improvement. In present study, genetic variation among ten different genotypes of faba bean (Vicia faba L.) collected from all different agro-climatic zones of Bihar along with Vikrant as reference variety of this pulse crop, have been analysed using electrophoretic profiles of total seed storage proteins, globulins and albumins. Attempt was also made to establish the diversity among these germplasms on the basis of the expression profile of two reactive oxygen species scavenging isozymes viz. superoxide dismutase (SOD) and catalase. Electropherogram of total proteins, albumins and globulins shows slight variation and some polymorphic bands were observed. Number of bands in total proteins, albumins and globulins ranges from 17-21, 17-20 and 16-20, respectively. Random selection of 100 seeds shows variation in seed weight also. There is negative correlation between seed globulin content and both seed weight (-0.488) and seed albumin (-0.319) while it shows positive correlation (0.633) with seed albumin concentration. The similarity coefficient of the total seed proteins, globulins and albumins diversity of all 10 genotypes of V. faba including Vikrant has been revealed by SDS-PAGE which varies from 0.83-1.0, 0.89-1.0 and 0.75-1.0 respectively. Zymogram of super oxide dismutase showed no variation whereas polymorphism was observed in case of catalase. VI ICLGG - Posters P-NAQ37 Iron biofortification in bean: is the lpa1 mutant a good candidate? 1 1 1 2 3 4 Panzeri D , Cominelli E , Bollini R , Campion B , Nielsen E , Glahn R , 1* Sparvoli F 1 Institute of Agricultural Biology and Biotechnology, Milan, Italy CRA, Montanaso Lombardo, Lodi, Italy 3 University of Pavia, Pavia, Italy 4 USDA-ARS, Ithaca, USA 2 *E-mail: sparvoli@ibba.cnr.it Abstract Iron deficiency is the most prevalent micronutrient deficiency affecting more than 2 billion people throughout of the world. Due to its worldwide consumption and nutritional qualities, common bean has been identified as a strategic target crop for iron biofortification. To this purpose we adopted a strategy aimed to reduce seed phytic acid (PA) and polyphenols (PP) content, being these strong inhibitors of iron absorption. In bean, low PP genotypes can be easily identified, conversely, natural variability in PA content is not very high and the best way to gain significant PA reductions is by obtaining low phytic acid (lpa) mutants. PA reduction is often associated with negative agronomic traits, such as lower seed viability and emergence, reduced plant growth rate and grain yield. These traits may limit the use of lpa mutants for iron biofortification, since acceptable agronomic performance should be guaranteed to small farmers and poor populations that would benefit from biofortified crops. Recently, we have identified a bean lpa mutant and showed it carries a defective MRP type ABC transporter (Pvmrp1) necessary for phytic acid transport to the vacuole. lpa seeds have a 90% PA reduction, seven fold higher free iron and 25% less raffinosaccharides. Assessment of iron bioavailability in lpa lines, using an in vitro Caco-2 cell model, showed a twelve times increase in low PP lpa seeds, while non significant increases were detected in high PP lpa seeds. Agronomic analyses of the original lpa mutant and of derived lpa lines have shown that seedling emergence, seed yield and plant growth were not affected. Contrary to other species, bean possesses a MRP paralog (PvMRP2) that is poorly expressed in seeds but not in other plant parts. This gene may complement the effects of the defective Pvmrp1 copy in vegetative tissues and thus explain the absence of negative phenotypes. VI ICLGG - Posters Marker-assisted selection for low phytic acid in Chickpea (Cicer arietinum) P-NAQ38 * Kalaimagal T , Narmadha B, Mohan S, Jency PJ, Shadakshari TV, Pande R Tamil Nadu Agricultural University, Coimbatore, India *E-mail: kalaimagal.t@gmail.com Abstract Phytic acid (Myo-inositol phosphates) is an anti-nutritional compound found in the seeds of cereals and legumes. Dietary phytate has a negative effect on the bioavailability of minerals like Iron, Zinc, Magnesium, Calcium and Potassium. Development of low phytate crops provides enhanced nutrition and metabolism for monogastric consumers such as human beings, poultry and swine. The trait also is favorable to the environment by reducing phosphorous loads to agricultural lands and surface waters. Chickpea being nutritionally rich crop, contains about 51.2% phytate phosphorus in its seeds. The major inositol Phosphorous in chickpea is IP6. In this study, about 50 germplasm accessions of chickpea were screened for low phytic acid using the soybean SSR markers viz., Satt416 and Satt168, Satt237, Satt561 that are tightly linked to lpa loci. Since the trait is governed by recessive genes, screening using co-dominant markers like SSR is reliable. Among fifty genotypes, none of them were amplified by the markers Satt416, Satt168 and Satt561. The genotypes RC 102-2, ICHRN 1 and ICC 9677 were amplified by the marker Satt237 which is linked to a major locus associated with seed phytate. This indicates that these genotypes are low in phytic acid and they may be used in breeding program to reduce the phytic acid content of agronomically superior lines. VI ICLGG - Posters P-NAQ39 Lentils (Lens culinaris L) micronutrients and prebiotic carbohydrates rich whole food source: future pulse biofortification opportunities 1,* 1 1 Thavarajah D , Thavarajah P , Johnson CR , Combs Jr. GF 2 1 School of Food System, North Dakota State University, Fargo, USA Grand Forks Human Nutrition Research Centre, Grand Forks, USA 2 *E-mail: dilrukshi.thavarajah@ndsu.edu Abstract Micronutrient malnutrition, mainly selenium (Se), iron (Fe), and zinc (Zn), and diet related obesity are major global health issues. Micronutrient deficiencies affect more than two billion people worldwide, especially women and preschool children in Asia, Africa, and Latin America. The World Health Organization estimated 1.5 billion adults (age 20 and over) were overweight in 2008, and more than one of every ten adults worldwide were obese. Previous attempts to prevent micronutrient malnutrition and through food fortification, supplementation, and enrichment of staple crops with these micronutrients had limited success. Diets rich in refined carbohydrates increased calorie intakes and individual weight gains; however, whole food rich with complex and prebiotics carbohydrates could reverse this trend. Our current research effort is to develop whole food grain legumes as a possible food source to provide adequate daily Se, Fe, Zn, and prebiotic carbohydrate requirements through a 50g of single serving. Lentils (Lens culinaris L) are rich in micronutrients: Se -1 -1 -1 (425 – 673 µg kg ), Fe (73 – 90 mg kg ), Zn (44 – 54 mg kg ), and with very -1 low concentration of phytic acid (2.5 – 4.4 mg g ). The major prebiotic carbohydrates in pulses are fructans; a quarter-cup serving of lentils (30 g lentils by dry weight) could provide approximately 3.6 g of prebiotic carbohydrates. In addition to minerals and prebiotics, lentils are also rich -1 source of beta carotene (1 – 5 mg kg ), bio-beneficial phenolics, and numerous folates. This paper provides an overview of the pulse’s promise and biofortification opportunities, particularly lentil, as a whole food strategy to combat global micronutrient malnutrition and obesity. Lentil’s genetic potential towards superior mineral bioavailability and diverse prebiotic carbohydrate profiles could be further exploited in developing whole food solutions to the global micronutrient malnutrition and obesity epidemics. VI ICLGG - Posters Identification of mineral related genes in the legumes using high throughput sequencing P-NAQ40 Santos C, Silva AI, Serrão I, Carvalho AL, Vasconcelos MW* CBQF – Centro de Biotecnologia e Química Fina, Escola Superior de Biotecnologia, Centro Regional do Porto da Universidade Católica Portuguesa, Rua Dr. António Bernardino Almeida, 4200-072 Porto, Portugal *E-mail: mvasconcelos@porto.ucp.pt Abstract Humans and other animals require several essential micronutrients in their diet for proper maintenance of health. Iron and zinc deficiencies affect about 30% of the world population. Legumes, such as soybean (Glycine max L.) and common bean (Phaseolus vulgaris), are important sources of micronutrients in the Mediterranean diet. Our goal is to unravel the mechanisms that will enable us to address two major challenges concerning the legumes: susceptibility to iron deficiency in the soil (affecting productivity and associated economics); and nutrient biofortification for iron and zinc. In order devise programs of biofortification that aim at enhancing the levels of micronutrients in legume seeds, it is important to clearly identify the principal molecular players involved in mineral uptake, transport and deposition. Using High throughput sequencing (Illumina Hiseq2000) we have identified specific genes that are associated with plant mineral metabolism in G. max, P. vulgaris and M. truncatula (a model plant for the legumes). Plants were grown hydroponically in iron sufficient and deficient conditions and shoot and root RNA from Fe-reducing and Fe-non reducing regions were extracted. We obtained a library with 43246 transcripts, among which we were able to find iron-related genes, namely, YSL (Yellow Stripe Like) in M. truncatula, IRT (iron-regulated transporter) in P. vulgaris and ZIP, sigG (spolllG) and ZnuA genes in G. max. VI ICLGG - Posters P-NAQ41 Genetic potential of lentils (Lens culinaris) for selenium biofortification * Viall EW, Thavarajah D , Thavarajah P School of Food Systems, North Dakota State University, Fargo, USA *E-mail: dilrukshi.thavarajah@ndsu.edu Abstract Selenium (Se) is an essential nutrient for proper function of the human body. Deficiencies in Se can create severe developmental problems such as Keshan and Kashin-Beck disease. These diseases are endemic to areas with naturally low soil selenium levels. Biofortification of staple crops provides nutrient rich foods to affected populations. Agronomic biofortification utilizes fertilization to increase micronutrient content of staple food crops. Implementing agronomic biofortification programs may provide an important source of essential micronutrients while breeders develop biofortified varieties. No studies have shown conclusive evidence that Se is an essential nutrient in plants. However, research suggests that Se provides plants with increased oxidative stress resistance. The objectives of this study were to determine 1) genetic and environmental variation of Se uptake in lentil genotypes grown in ND, USA and 2) the effect of Se fertilization on antioxidant activity and yield of commonly grown commercial lentil varieties. Three replicates of ten lentil genotypes were planted in three North Dakota locations for three years (2010, 2011, and 2012). Seeds were analyzed for total Se using ICP. Work investigating antioxidant protection after Se-fertilization is in progress. Preliminary results indicate that lentil seed Se concentrations can be enriched via Se-fertilization. Conventional lentil plant breeding will enable selection of genetic materials for higher seed Se accumulation and increased grain yields. Therefore, Se biofortification of lentils may be a sustainable solution to address global Se deficiency. VI ICLGG - Posters Theme: Translational Genomics P-TLG07 Groundnut breeding at ICRISAT: from tools of genetics to genomics 1* 1, 2 Janila P , Varshney RK 1 Nigam SN 1 1 1 1 Pandey MK , Vadez V , Sudini HK , Patne N , 1 International Crops Research Institute for Semi-Arid Tropics (ICRISAT), Hyderabad, India 2 CGIAR Generation Challenge Programme (GCP), Mexico *E-mail: p.janila@cgiar.org Abstract The breeding program for genetic enhancement of groundnut has begun in 1976 at International Crops Research Institute for the Semi-Arid Tropics (ICRISAT). So far over 10,000 crosses were made using germplasm, breeding lines and interspecific derivatives. Since 1986, a total of 138 improved varieties of groundnut, bred at ICRISAT were released by national partners in 36 different countries of Asia and Africa. The adoption of improved varieties resulted in increased yields and production and thus improved the livelihoods of small families in semi-arid tropical regions, engaged in groundnut cultivation. At ICRISAT’s Asia program, traditional breeding procedures based on the principles of genetics along with the physiological tools and screening for pest and diseases were largely used in the development of improved varieties of groundnut. Significant progress has been made to breed varieties with high yield potential, short maturity duration, resistance to foliar fungal diseases, drought tolerance, enhanced oil content and traits for food uses. Since both, empirical and trait-based approaches were equally efficient in breeding for drought tolerance; a combination of these approaches was adopted. With the development of molecular tools in groundnut during the last decade, it is now possible for integrated marker assisted selection (MAS) in groundnut breeding for improved efficiency and optimal utilization of time and resources. The introgression of QTLs for rust and late leaf spot resistance and mutant FAD alleles (for enhanced oil quality) are under progress. Further, initiatives have been taken for the development of genomic tools for improvement/selection of traits such as, drought tolerance, high oil content, aflatoxin resistance and fresh seed dormancy. Is the trehalose pathway involved in the response of Phaseolus vulgaris L. toward water deficit? 1 1 2 Carvalho T , Morgado AR , Paiva JAP , Fevereiro P 1,3 , Araújo SS 1,2* 1 Instituto de Tecnologia Química e Biológica-UNL (ITQB-UNL), Oeiras, Oeiras, Portugal 2 Instituto de Investigação Científica Tropical (IICT), Lisboa, Lisboa, Portugal 3 Faculdade de Ciências da Universidade de Lisboa (FCUL), Lisboa, Lisboa, Portugal *E-mail:saraujo@itqb.unl.pt Abstract Trehalose metabolism plays a major role in plant, growth, development and stress response signaling. In a previous work, we addressed the impact of water deficit (WD) on trehalose metabolism in the model M. truncatula by qPCR and the results supported the assumption that trehalose metabolism is involved in legume WD and WD recovery responses. Among grain legumes, the common bean (Phaseolus vulgaris L.) is a major food crop in Latin America and Sub-Saharan Africa, where drought is a significant problem. In this study, we addressed the impact of WD on trehalose metabolism of this crop, using two bean lines differing in adaptation to drought. Plants were submitted to a moderate (MWD) and severe (SWD) stress by suppressing pot irrigation. Their physiological performance was compared to a well watered (WW) group, irrespective to the line studied. Using a qPCR approach, we quantified the mRNA expression levels of the three enzymes of the trehalose pathway (TPS- trehalose-6-phosphate synthase; TPP- trehalose phosphate phosphatase and TRE-trehalase) in leaf and root tissue samples across the treatments imposed. In leaves under SWD, the drought resistant line presented TPP levels identical to the WW ones but TPP levels were upregulated in drought susceptible line. This indicates indicating a differential behavior of each studied line suggesting a regulation of T6P levels through extensive regulation of TPP expression under WD. Interestingly, data indicated that a differential regulation of the trehalose pathway might occurs between aerial and root part of the plants. Possible implications of these results will be discussed. P-TLG07 P-TLG08 A spontaneous mutation in a semi-leafless pea cultivar restores leaflet formation and improves photosynthetic function 1, 1 2 Avercheva O *, Sinjushin A , Zelenov A 1 M.V. Lomonosov Moscow State University, Moscow, Russia The All-Russia Research Institute of Legumes and Groat Crops, Orel, Russia 2 *E-mail: olga.avercheva@gmail.com Abstract Photosynthesis is one of the main parameters that determine productivity in crop plants. The development of the photosynthetic apparatus (PSA) is closely linked to the development of the leaf as a whole. Many pea (Pisum sativum L.) cultivars used in agriculture are semileafless. This type of leaf structure has many agricultural benefits. However, tendrils have an impaired photosynthetic function and need to compensate with higher tendril biomass to support high productivity. We still do not understand how the tendril development program is linked with the formation of the PSA. We compared the physiological state of the PSA in two pea cultivars: cultivar ‘Batrak’ (semi-leafless, bred at the All-Russia Research Institute of Legumes and Groat Crops) and cultivar ‘Ras’ (which appeared from ‘Batrak’ due to spontaneous mutation, in which some of the tendrils transformed back into leaflets). We studied photosynthetic pigment content and chlorophyll fluorescence parameters in leaflets and tendrils of both pea cultivars. Most parameters studied in leaflets of ‘Ras’ were higher than in ‘Batrak’ tendrils and were close to the parameters of normal leaflets in other pea cultivars. This shows that the spontaneous mutation occurred in a gene or group of genes that determine not only leaflet/tendril development but also the photosynthetic function of the organ. Further study is needed to determine the gene(s) affected by the mutation and uncover their functions. Transgenic peanut (Arachis Hypogaea L.) expressing a rice chitinase cDNA shows varied levels of resistance against attack by fungal pathogen Phaeoisariopsis Personata (Berk. & Curt.) V. Arx. Beena MR* CTCRI, Thiruvananthapuram, India Email: beenamr@gmail.com Abstract Cultivated peanut (Arachis hypogaea L.), an allo-tetraploid (2n=40), is an annual legume crop that traces its origin to Southern Bolivia/ North-West Argentina in South America and presently cultivated in 108 countries of the world. Peanut is one among those several crop plants that suffer severe losses in productivity due to fungal attack. Most common among the diseases caused by fungi in peanut is late leaf spot disease caused by Phaeoisariopsis personata (Berk. & Curt.) V. Arx. Transgenic peanut lines possessing fungal resistance genes offer an alternative to traditional resistance and fungicide application in managing fungal diseases. Chitin is a linear homopolymer of β (1-4) linked 2-deoxy-2-acetamido-D-glucopyranosyl residues, and is a major component of fungal cell walls, which could be broken down by the enzyme chitinase. Hence, chitinase is mostly related to resistance against fungal pathogens and renders general resistance in plants. Expression of rice chitinase - cDNA via Agrobacterium – mediated transformation of peanut provided varied levels of resistance against the attack of this pathogen. P-TLG09 P-TLG10 Genetics, genomics and biotechnology of the biofuel feedstock tree legume Pongamia pinnata 1,* 1 1 2 1 Biswas B , Jiang Q , Kazakoff S , Edwards D , Gresshoff PM , Scott PT 1 1 ARC Centre of Excellence for Integrative Legume Research, Australia The University of Queensland, Brisbane, Australia 2 * E-mail:b.biswas@uq.edu.au Abstract The legume tree Pongamia pinnata is rapidly gaining acceptance as a biofuel feedstock. It is fast-growing, high-yielding, stresstolerant and adaptable to various environments including winterfrosted areas of Queensland1. It produces profuse quantities of oil-rich seeds (35-45%) with high oleic acid (C18:1) triglycerides, making it suitable for biodiesel and aviation fuel production 2,3. It is an outcrossing tropical/sub-tropical tree, native to India, southeast Asia and northern Australia. Pollination is by insects, principally bees. ISSR markers showed high genetic diversity of trees, supporting the proposed outcrossing reproduction of Pongamia and confirming phenotypic data4. Seedlings derived from a single parent tree were less diverse than those sourced from different parent trees. Whole plant regeneration from immature cotyledons was achieved5; regenerated plants from the same tissue were clonal as determined by ISSR markers. For further improvement genetic transformation protocols, mediated by both Agrobacterium tumefaciens and Agrobacterium rhizogenes are being developed. Genes to be targeted such as those involved in the seed oil biosynthesis are being characterized and their levels of gene expression during throughout seed development have been determined. Deep DNA sequencing (Illumina®) has enabled a partial characterization of the Pongamia genome and orthologues of nodulation-associated genes to be identified. Illumina® reads were used to assemble and annotate for the first time the chloroplast (153 kb) and mitochondrial (426 kb) genomes of Pongamia. Towards plant-parasitic nematodes control using RNAi technology in fieldpea (Pisum sativum L.) 1,* 1 1 1 2 P-TLG11 1 Das A , Nandeesha P , Datta S , Singh B , Subramanium K , Chaturvedi SK , 1 Nadarajan N 1 Indian Institute of Pulses Research, Kanpur, India BSBE, Indian Institute of Technology, Kanpur, India 2 *Email: adas@icar.org.in Abstract Plant-parasite nematodes (PPNs) are a menace to crop species, and the yield loss range from 10-40%. Despite this enormous impact, biology of these nematodes is currently poorly understood. Owing this parasitic nature, it has not been possible to culture them in laboratory and hence, forward genetic and biochemical approaches could not be applied. RNA interference (RNAi) technology is a promising reverse genetic tool that has to potential to specifically down regulate gene expression in organism. The oesophageal gland secretory proteins are nematodes arsenal for entry into the root zone. We designed a double stranded RNAi constructs in a binary vector pBI121for salivary gland gene (Accession number AF531160). An in vitro regeneration system (direct organogenesis) using thin embryonic slices of axes of immature seeds; amenable to genetic transformation was developed. Genetic transformation was attempted in the susceptible field pea cultivar HUDP15 using Agrobacterium tumefaciens stain LBA4404. Homozygous fieldpea lines have been identified and nematode bio-assay (Meloidogyne incognita and M javanica) indicated significant reduction (56.0 – 88.6 %) in average gall formation and reduced percent conversion (13.34 – 30.00 %) to egg mass. Thus, RNAi technology can be used as an important component of Integrated Nematode Management Programme. P-TLG12 RNAi approach for the development of resistance in grain legumes against Mungbean yellow mosaic India virus using IR and Rep gene Archana K, Singh VB, Malathi VG * Indian Agricultural Research Institute, New Delhi, India * E-mail:archana_dayal2001@yahoo.com Abstract Mungbean yellow mosaic India virus (MYMIV) belonging to the family Geminiviridae is a bipartite legume virus, infecting grain legumes. They are transmitted by whitefly, Bemisia tabaci and cause yield loss to the tune of 69%. RNAi a homology-dependant gene silencing mechanism in which down regulation of the expression of specific gene occurs is widely used for developing transgenic resistance in plants against viruses. Transforming plants with hairpin RNAi (hpRNAi) construct has been used to discover or validate the functions of number of genes. In the present study, functional validation of RNAi construct targeting two different regions of the genome of MYMIV attempted: The hp RNAi constructs were made targeting 500 bp sequence in Replication associated protein (Rep) and 190 bp sequence in Intergenic region (IR) of MYMIV .The sequences were amplified and cloned in sense and antisense orientation respectively interspersed with an intron in a cloning vector pHannibal. The cassettes of Rep and IR which carry 35S promoter, sense sequences followed by intron then antisense target sequences and OCS terminator, were subcloned in a binary vector pCambia 1301 to express the hp RNAi transcript in the plant system. The resulting construct was mobilized into Agrobacterium tumefaciens. The constructs were used for Agroinfiltration on plants of Cowpea (Vigna unguiculata) cv. Pusa Komal. The hp RNAi constructs were co-agroinoculated along with infectious constructs of MYMIV. The infectious construct of MYMIV was also agroinoculated to the plants to serve as control. The transient assay of both the hp RNAi constructs revealed that it prevents the viral pathogenesis and the Realtime quantitative PCR analysis identified the low-level accumulation of viral DNA in the inoculated plants. The viral siRNA level in both hp RNAi inoculated plants are being looked into. . RNA interference highlights the role of SymRK in symbiosome development in an aeschynomeneae legume Arachis hypogaea 1 1 Dutta A , Saha S , Sinharoy S 1, 2 , Gupta MD 1,* 1 University of Calcutta, Kolkata, India Samuel Roberts Noble Foundation, Oklahoma, USA 2 * E-mail: maitrayee_d@hotmail.com Abstract Among the prime candidates that are responsible for adaptations specific to root nodule symbiosis is SymRK (Symbiosis receptor kinase), a member of the ‘Sym pathway’, since there is a correlation between the occurrence of the fulllength SymRK version in the nodulating clade and the potential of this version, as opposed to the shorter ones present outside the clade, to support functional endosymbiosis with bacteria. SymRK is demonstrated to have a role in release of the bacteria from infection threads towards formation of symbiosomes in Lotus japonicus and Sesbania rostrata. Our objective was to investigate the role of SymRK in an aeschynomeneae legume Arachis hypogaea, where rhizobial invasion bypass the complex intracellular infection-thread mediated processes adapted by most legumes. Instead, the invasion occurs through epidermal cracks and the rhizobia are endocytosed directly by a cortical cell. RNA interference of SymRK in Arachis reveals that there was >80% reduction in nodulation which is consistent with its role in inception of these nodules. Infected zone of the RNAi nodules was mostly empty and scattered with pockets of cells containing filamentous rhizobia without any sign of development of spherical symbiosomes. This is in contrast to our previous results on RNA interference of CCamK where also we observed scattered pockets of infected cells but the rod shaped bacteria were all developed into spherical symbiosomes. The possible roles of SymRK and CcaMK in Arachis nodule development would be discussed. P-TLG13 P-TLG14 Genomic characterization of a fast neutron-induced soybean mutant 1 1 1 1 1 1 Hwang WJ , Kim MY , Kang YJ , Shim SR , Lee JE , Lestari P , Suk-Ha 1,2* Lee 1 Department of Plant Science and Research Institute for Agriculture and Life Sciences, Seoul National University, Seoul, Korea 2 Plant Genomics and Breeding Institute, Seoul National University, Seoul, Korea *E-mail: sukhalee@snu.ac.kr Abstract Mutagenesis approach with whole genome sequencing has become an important role in genetic and molecular biology study and breeding of crop plants. In this study, we screened the fast neutron M4 soybean mutant plants based on morphological phenotypes and characterized the mutant of interest using resequencing. Screened mutants showed abnormal phenotypes in plant heights, number and color of leaves, maturity and number of branches etc. Among them, the mutant displaying short plant height and bush type of growth habit was selected to perform resequencing for identification of the altered genomic regions. Mutant sequence reads generated by paired-end shotgun library were mapped on a draft soybean reference soybean (Glycine max cv. Williams 82). Among the predicted deletion sites, a total of 3 deletion regions were confirmed by PCR. Glyma03g02390 gene and Glyma03g03560 gene were involved in the deletion regions. Glyma03g02390 gene was related to AMP binding, cofactor binding and metabolic process of cell growth and Glyma03g03560 gene was concerned to oxygen binding and especially process of indole acetic acid (IAA) biosynthesis. These genes detected in this mutant will be studied their molecular function in stunted phenotype. In planta transformation of pigeonpea (Cajanus cajan (L.) Millsp.) with cry1Ac gene for resistance against its major pest Helicoverpa armigera Jain M*, Khatodia S, Kharb P, Batra P, Chowdhury VK Haryana Agricultural University, Hisar, India *E-mail: meenakshi110@gmail.com Abstract Pigeonpea [Cajanus cajan (L) Millsp.] is an important protein rich legume crop grown in many semi arid tropical region of the world. Development of transgenics in pigeonpea remains dogged by poor plant regeneration in vitro from transformed tissue and low frequency transformation protocol. Transgenic pigeonpea plants of variety Manak expressing cry1Ac gene were generated by tissue culture independent , in planta transformation method using Agrobacterium tumefaciens strain LBA4404, harboring pBIN-35Scry1Ac. Putative transformants were screened at early seedling stage by subjecting small leaf pieces to PCR amplification using plant direct PCR kit and transformation frequency of 45.8% was obtained. Quantitative ELISA of T0 transgenic plants showed expression of CRY protein ranging from 10-54 ng -1 mg of total soluble protein in fresh leaves. Of the 374 seeds harvested from all the T0 plants, 324 were sown to obtain progeny (T1) plants and 123 of these were randomly analysed to determine their transgenic nature. Sixty of the 123 T1 plants amplified cry1Ac gene, substantiated the integration and transmission of the gene. The protocol ensured generation of pigeon pea transgenic plants with considerable ease in a short time and offers immense potential as a supplemental or an alternative protocol for generating transgenic plants of difficult-to-regenerate pigeon pea. Further, the protocol offers the option of doing away with a selection step in the procedure and so facilitates marker free transformation. P-TLG15 P-TLG16 New advances in plant growth promoting Rhizobacteria for stress alleviation- role of bacterial ACC deaminase 1 2 1 1 Dhanushkodi R , Govindasamy G , Lakkineni V , Bose P , Sharma V, 1,* Kannepalli A 1 Indian Agricultural Research Institute, New Delhi, India National Institute of Abiotic Stress Management, Pune, India 2 *E-mail: annapurna96@yahoo.co.in Abstract Plant growth promoting rhizobacteria (PGPR) colonize rhizosphere of many plant species and confer beneficial effects, through direct and indirect mechanisms. Direct stimulation includes providing plants with compounds such as fixed nitrogen, phytohormones, or solubilized iron from the soil. Many PGPR contain enzyme 1-aminocyclopropane-1-carboxylate (ACC) deaminase which can cleave the ethylene precursor ACC to α-ketobutyrate and ammonia thereby lower the level of ethylene in developing or stressed plants and plays a key role in plant-microbe associations. In our study we screened rhizobia of chickpea, pea, soybean, fenugreek, lentil and frenchbean for the presence of acdS gene; the structural gene for ACC deaminase through PCR. Multiple alignment of the acdS gene sequences showed diversity in the nucleotide sequences. acdS was more prevalent in pea nodulating rhizobia than in rhizobia of other legumes. Legume inoculations with ACC deaminase positive rhizobia helped the plants survive heavy metal, salinity and water stress. Following five days in gonobiotic growth pouches, pea roots produced from seeds treated with ACC positive rhizobia and E. coli DH5α clones with acdS gene showed a significant increase in root length when compared to negative control. SDS PAGE profiles of the positive clones showed a 41kD protein. Under Cadmium stress plant ethylene levels were found low in wheat seedlings treated with ACC positive bacteria. Treated seedlings were protected from the deleterious effects of ethylene. The data is in agreement with the model in which ACC deaminase-containing plant growth promoting bacteria reduce plant stress and induce root elongation and proliferation in plants, largely by lowering ethylene levels. Novel tissue culture independent protocol for transformation of chickpea (Cicer arietinum L.) with cry1Aa3 for resistance against its major pest, Helicoverpa armigera 1,* 1 1 2 Kharb P , Khatodia S , Batra P , Kumar PA , Chowdhury VK 1 1 Haryana Agricultural University, Hisar, India Indian Agricultural Research Institute, New Delhi, India 2 *E-mail: pkharbhau@gmail.com Abstract Chickpea (Cicer arietinum L.) is the third most important food legume, cultivated in arid and semi-arid regions of the world. The transgenic plants of the chickpea cv. C-235 expressing cry1Aa3 gene were generated using a novel tissue culture independent in-planta transformation method (Patent No. 252590) using Agrobacterium tumefaciens strain EHA105 harboring pBIN35Scry1Aa3. PCR analysis indicated the putative transgenic nature of the T 0 plants. Seeds of T0 plants allowed to continue into the next generation amplified gene of interest in most of the plants tested. The expression level of CRY protein in T0 and T1 transgenic chickpea plants was achieved maximum -1 up to 17 ng mg of total soluble protein, which efficiently causes 90% mortality to first instar larvae of Helicoverpa armigera as analyzed by insect mortality bioassay. Single copy of transgene was detected in the transgenic chickpea lines using real time PCR assay. These results substantiated the integration and expression of crylAa3 gene in the transgenic chickpea plants developed for resistance against Helicoverpa armigera with considerable ease in a short time. Further, the protocol offers the option of doing away with a selection step in the procedure and so facilitates transformation, which is free of marker genes. The study demonstrated potentialities of developing insect resistant transgenic chickpea and provides an opportunity for further test and advancement to reach the field utility level. P-TLG17 P-TLG18 A rapid and efficient transformation system of chickpea (Cicer arietinum L.) with high frequency using tissue culture independent protocol * Khatodia S , Kharb P, Batra P, Chowdhury VK Chaudhary Charan Singh Haryana Agricultural University, Hisar, India *E-mail: khatodia99@gmail.com Abstract The transgenic plants of the chickpea (Cicer arietinum L.) cv. C-235 and HC-1 expressing cry1Ac gene were generated using a tissue culture independent inplanta transformation method using Agrobacterium tumefaciens strain LBA4404, harboring pBIN-35Scry1Ac. The putative transgenic events in the early stage of seedlings identified by subjecting small leaf piece to PCR amplification using Plant Direct PCR kit, representing transformation frequency of 13.4% and 41% in cv. C-235 and HC-1 respectively. Quantitative assessment of Bt Cry toxins by ELISA of T 0 transgenic chickpea plants showed -1 expression of toxin ranging from 8.8 to 17.6 ng mg of total soluble protein in leaves of six of each cv. C-235 and HC-1 transgenic plants. Bioassays against major pest of the chickpea, Helicoverpa armigera revealed, T0 plants perform well against the larvae. The cry1Ac gene was transmitted to the next generation as analyzed by PCR of T 1 transgenic plants. Our results demonstrate a rapid, simple and efficient transformation system of chickpea with high frequency which does not require the complex steps for selection of the transgenic events. These findings will certainly accelerate the development of chickpea plants with novel traits with the possibility of developing marker free transgenic events, which minimizes the risk of introducing unwanted genetic changes and allows stacking of multiple genes. . . An integrated systems approach to identify key pathways and metabolites affected by water-deficit stress during seed development in peanut 1* 1 1 2 Kottapalli KR , Zabet-Moghaddam M , Francisco SS , Puppala N , Paxton 3 Payton P 1 Texas Tech University, USA New Mexico State University, USA 3 United States Department of Agriculture Cropping Systems Research Laboratory, Texas, USA 2 * E-mail:rao.kottapalli@ttu.edu Abstract Legume seeds and peanuts, in particular, are an inexpensive source of plant proteins and edible oil. Peanuts are primarily grown on marginal soils with limited irrigation under environmentally challenging conditions. These conditions have a predominant effect on the developing seed and reduce the seed oil and protein quantity and quality. A comprehensive understanding of seed development and the effects of water-deficit stress on the incorporation of the main storage reserves in seeds, such as proteins, fatty acids, starch, and secondary metabolites will enhance our ability to improve seed quality and yield through molecular breeding programs. In the present study we employed a combined transcriptomics, proteomics, and metabolomics approach to study the primary transcriptional networks, functional proteins, and secondary metabolites altered in the developing peanut seed during water-deficit stress. We observed a decline in total fatty acid content coupled with significant change in oleate and linoleate composition due to drought in the developing seed. These changes corresponded to the reduction in the gene transcripts and proteins related to fatty acid metabolism. Interestingly, the integrated pathway analysis revealed, that the nitrogen is shunted into developing seeds possibly to avoid nitrogen toxicity during drought stress. Additionally, the presentation will focus on the integration of transcript, protein and metabolite data to understand key metabolic pathways altered during stress. P-TLG19 P-TLG20 Over-expression of Tfgd2:Rs-AFP2 defensin gene increases resistances to early leaf spot, late leaf spot and stem rot in transgenic groundnut (Arachis hypogaea L.) * Madhubala, Kumar A , Dobaria JR, Radhakrishnan T Directorate of Groundnut Research, Junagadh, India * E-mail: abhayiari@gmail.com Abstract Groundnut is an important oilseed crop of the Indian subcontinent. Fungal diseases, especially the soil borne, take a heavy toll on the production of peanut. As a possible alternative to the very heavy use of pesticides in chemical controls, we have developed peanut transgenics (cv. GG 20) carrying a fusion gene of Tfgd2 (Trigonella foenum-graecum defensin2) and Rs-AFP2 (Raphanus sativus antifungal protein2) through Agrobacterium- mediated genetic transformation. The putative transgenics were tested for transgene integration by PCR, dot blot and Southern blotting. Transgene expression was confirmed in T0 and T2 stage using Real-Time PCR. Fourteen independent transgenic events were evaluated for tolerance to early and late leaf spots under artificial conditions in T 2 generation. The transgenics showed not only reduction in the number of spots but also the onset of disease was delayed on artificially screening with Cercospora arachidicola and Phaeoisariopsis personata inoculums. The average number of days to defoliation was found to be less in transgenic plants compared to the controls. Constitutive overexpression of the defensin genes in transgenic groundnut also showed enhanced resistance against the soil borne pathogen, Sclerotium rolfsii, causing stem rot disease. Seeds of the transgenic lines germinated and grew to quadrifoliate seedling even in the presence of fungal mycelia on the seeds while the wild type seed (GG-20) and the resistant control (CS-19) failed to germinate, in artificially inoculated germination media. More elaborate screening may confirm the observations and the selected events can be used for breeding programmes on fungal disease tolerance. Molecular cloning and characterization of phytochelatin synthase (LlPCS) and metallothionein (LlMT) genes from Leucaena leucocephala Sunita MSL 1, 2, *, Prashant S 1, 2 1 2 , Renuka S , Narasu ML , Kavi PBK 1 1 Osmania University, Hyderabad India Jawaharlal Nehru Technological University, Hyderabad, India 2 *E-mail: m.s.l.sunita@gmail.com Abstract Leucaena leucocephala, a leguminous tree with high biomass, has metal tolerance and survival ability in arid, infertile and metal-contaminated areas. But so far, no reports are available on identification and functional characterization of the metal-tolerance genes in this species. In the present study, we report the isolation and characterization of phytochelatin synthase (LlPCS) and metallothionein (LlMT) genes from this tree species. The predicted 3D structure of these proteins exhibited potential possible binding sites for the metals like Cd, Zn, Cu, Fe and Mn. The role of LlPCS and LlMT genes in metal tolerance and accumulation was characterized in E. coli. Both LlPCS and LlMT genes were sub-cloned individually into pET32a expression vector and bacterial transformation was performed. SDS-PAGE analysis of the total protein isolated from the recombinant E. coli revealed the over expression of LlPCS and LlMT proteins along with His-tag. When grown in toxic metal enriched media, recombinant E. coli exhibited enhanced tolerance to broad range of metals like arsenic, cadmium, cobalt, copper, mercury, tin and zinc compared to controls. Further, these recombinant E. coli cells could also accumulate several folds increased concentrations of toxic metals like arsenite, copper and arsenate compared to controls. Thus the present study suggests that LlPCS and LlMT genes confer enhanced toxic metal tolerance and accumulation in E. coli and might play crucial role in toxic metal detoxification in Leucaena leucocephala. This study can help us in better understanding the toxic metal tolerance ability of L. leucocephala for its successful usage in phytoremediation technologies. P-TLG21 P-TLG22 Isolation and characterization of carbonic anhydrase gene from cowpea (Vigna unguiculata) * Mondal TK , Ravi NR, Rana MK National Bureau of Plant Genetic Resources, New Delhi, India * E-mail: mondaltk@yahoo.com Abstract Around 50% of Indian soil is zinc (Zn) deficient that ultimately affects severely human and cattle health. Although, carbonic anhydrase is well-known as zinc deficiency responsive gene, yet it is not cloned from cowpea plant, which is an important legume of India. We have cloned the coding sequence of the gene from young leaf of cowpea cv. PUSA Kamal (Genbank: JQ429799) and deduced its structural organization in the genome. It was also found that expression of this gene was up-regulated under dehydration but downregulated under Zn-deficient conditions as revealed by RT-PCR analysis. However, under normal Zn conditions, the expression was highest in young leaf and lowest in root. In silico analysis of the DNA as well as deduced protein sequence was carried out to understand domain structure, sequences logos of the domain, phylogenetic relationship of this gene among the various legume species, predicted secondary structure of amino acid sequence and 3D structure by homology modeling of the protein. Additionally, to find out the allelic variation, CA gene from 6 cowpea genotypes (IC202801, IC588972, IC20574, IC588976, IC93782, BKS/YR/KAP P-1) of diverse origin were cloned, sequenced which registered 5 non-synonymous SNPs. These genespecific SNPs may be a potential marker for the use for identifying Zn use efficient cowpea cultivar in future. Could an extra shot of TPS confer better tolerance to water deficit stress in Medicago truncatula? 1 1 1,2 Morgado R , Branco D , Fevereiro PS , Araújo SS 1,3,* 1 Instituto de Tecnologia Química e Biológica-UNL (ITQB-UNL), Oeiras, Portugal 2 Faculdade de Ciências da Universidade de Lisboa (FCUL), Lisboa, Portugal 3 Instituto de Investigação Científica Tropical (IICT), Lisboa, Portugal *E-mail:saraujo@itqb.unl.pt Abstract Drought is the major environmental constraint affecting plant growth and production yield, and considering the pronounced global environmental changes, water deficit will be evermore a challenge to be overcome. Consequently, it becomes urgent to develop crops showing a better tolerance to water deficit stress (WDS). Studies from our group highlighted that the accumulation of trehalose6-phospate synthase (TPS) may improve plant tolerance to water stress. The goal of this study was to investigate the effect of the ectopic expression of the functional A. thaliana TPS in the response to water stress of the model legume Medicago truncatula. To accomplish that goal, we transformed M9-10a embryogenic line with the constitutive driven AtTPS cDNA using the Agrobacterium-mediated transformation and the resulting transgenic lines were characterized by measuring its expression by qRT-PCR. The results showed that we were able to obtain lines with different AtTPS expression, being grouped concerning their expression level: low, medium and high. This gave us means to proceed with further studies about how different levels of the transgene can have an impact on the response of M. truncatula to WDS conditions. In this context, experiments are being performed to study the response of these different transgenic and wild type plants submitted to moderate and severe WDS and also to recovery from it. The physiological and molecular studies will help us to understand how the alteration of the trehalose metabolism can interfere with the response of plants to water deficit. P-TLG23 P-TLG24 Text mining to link clinical and laboratory research for translational genomics research Natarajan J* Bharathiar University, Coimbatore, India *E-mail: n.jeyakumar@yahoo.co.in Abstract Translational research is the process of translating basic scientific discoveries into clinical applications such as new diagnostics and treatments. Translational research serves as a bridge between lab bench discoveries and the patient bedside. Information collected at the patient bedside can circle back to the laboratory to fuel additional discoveries. So, it necessary to link the clinical data and laboratory research data which will be the key issue in translational research. This presentation high lights the application of text mining in translational genomics research. Text mining, sometimes alternately referred to as text data mining refers to the process of deriving high-quality information from text. The application of biomedical text mining includes named entity tagging (e.g. genes, proteins, enzymes, drugs etc.), entity concept relation extraction (e.g. protein-protein interactions, drug-disease relations etc.) and mining new facts (e.g. biomedical pathways and functions). Integrating the above text mining results to a data warehouse related to a broad range of human diseases will make these data usable/available at the point-of-need for a translational research community. This presentation will highlight our vision of text mining in translational genomic research. This will show how a Data Warehouse for Translational Research can be created using text mining. Clinical information is available in clinical records and on the other hand the laboratory research outputs are available online in the form of abstracts (e.g. PubMed/MEDLINE) and full text research articles (e.g. PubMED Central, BioMed Central). However, the growth in clinical records and research publications is exponential and at this rate of publication, it is difficult or impossible for biologists to keep up with the underlying hypotheses. Text mining methods will help automatically mine the knowledge from both resources and create Data Warehouse for Translational Genomic Research. Exploration and in vitro propagation of Desmodium gangeticum (L.) – a medicinal species of leguminosae/fabaceae 1,* 2 Puhan P , Rath SP 1 Institute of Biotechnology, Acharya NG Ranga Agricultural University, Hyderabad, India 2 Koraput Central University, Koraput, India *E-mail: puspapuhan@gmail.com Abstract Desmodium gangeticum (L.) commonly known as Salaparni or Prishniparni (Hindi & Sanskrit) belonging to the family Fabaceae/Leguminosae has multiple uses in the Indian Ayurveda system of medicine. As a single drug its effect has been described on pregnancy with reference to abortions. It contains indole alkaloids in its green parts and three ptero-carpenoids namely gangetin, gangetinin and desmodin. The plant is commonly used in conditions, which are due to vata-disorders. In the present study nodal explants and explants from in vitro grown seedlings were cultured on MS basal medium with Kn or BA at different concentrations for multiplication, organogenesis and somatic embryogenesis. Nodal explants showed axillary shoot multiplication in 0.5 mg/l conc. of BA. Different in vitro grown explants cultured on MS medium containing different concentrations of BA (0.1-1.0mg/l) produced friable nonembryogenic callus (brownish white) with many small, smooth, rounded, discreet and greenish unit of embryogenic callus within the first week of inoculation. For induction of somatic embryogenesis, these friable embryonic callus were transferred to MS medium supplemented with various growth regulators like BA, Kn, 2,4-D, NAA, CH and BA. On regeneration medium most of the somatic embryos germinated like normal embryos and some produced greenish sprouted embryoids like structures with only roots or shoots or secondary somatic embryos. Regenerated shoots gave 100% rooting response with IBA treatment at 1.0 mg/l. The medium containing BA 0.5 mg/l and the cotyledon is proved to be best explants over all. Somatic embryogenesis derived plantlets of D. gangeticum showed 60 % of survival while axillary bud multiplied plants gave 80% survival. Thus the in vitro propagation protocol standardized can be highly useful in raising quality planting materials of Desmodium gangeticum for commercial plantation programmes and secondary metabolite production. P-TLG25 P-TLG26 Emergence of new virus isolate with changed genomic constituent: a challenge for durable resistance against yellow mosaic disease in black gram germplasm Roy A*, Bag MK, Gautam NK, Prasad TV, Dutta M, Bansal KC National Bureau of Plant Genetic Resources (NBPGR), New Delhi, India *E-mail: anirbanroy75@yahoo.com Abstract Yellow mosaic disease, one of the most limiting factors for cultivation of grain legumes, is caused by four different begomoviruses in India. Among these begomoviruses, Mungbean yellow mosaic India virus (MYMIV) and Mungbean yellow mosaic virus (MYMV), are most important. MYMIV is more predominant in northern, central and eastern regions of India and MYMV in southern and western regions. Black gram germplasm (344 accessions) were evaluated under natural field condition against yellow mosaic disease. Two accessions were found without any symptom, 11 accessions highly resistant and 19 accessions resistant based on co-efficient of infection. For detection and characterization of virus(es), rolling circle rase. Digestion of RCA product with HindIII generated virus specific 2.7 kb full genomic length fragment which was purified and cloned. Two distinct clones (MF1 and MF2) with different restriction profiles were sequenced. Sequence analyses showed that MF1 (DNA B) and MF2 (DNA A) shared highest sequence identity (92.8% and 88.2%) with two different MYMV isolates reported from Cambodia and Pakistan, respectively but had low sequence identity with MYMIV strains reported to be associated with the disease in northern India. In phylogenetic analysis both these components clustered in separate phylogenetic clad. This virus appeared to be a new isolate of MYMV and is the first report of MYMV in black gram from northern India. Further gene wise sequence analysis and recombination analysis indicated that this virus probably evolved due to recombination. Emergence of such new isolates of the virus and their spread to newer geographical locations could be a potential reason for breakdown of host resistance and hence, pose a serious threat to germplasm screening against virus diseases. Assessing the genetic and agronomic performance of the biofuel feedstock tree legume Pongamia pinnata in northern Australia 1,* 1, 1 1 1 1 Scott PT , Biswas B , Jiang Q , Samuel S , Kazakoff SH , Idris Z , Dietzgen 2 1 R , Gresshoff PM 1 ARC Centre of Excellence for Integrative Legume Research, School of Agriculture and Food Sciences, Australia 2 The University of Queensland, Brisbane, Australia E-mail: p.scott@uq.edu.au Abstract It is widely accepted that the foreseeable demand for liquid fuels will be sourced, in part at least, from biological feedstocks (i.e., biofuels). The future success of any proposed biofuel feedstock will rely on harnessing the genetic potential of biofuel production and simultaneous sustainable agronomic performance. Pongamia pinnata has been identified as a future feedstock for biodiesel and aviation fuel because of its abundant production of oil-rich (~40% v/v) seeds, its ability to grow on poor quality soils and under conditions of stress (i.e., drought and salinity), and importantly because as a legume it is capable of biological nitrogen fixation. In addition to its future as a biofuel feedstock we are assessing the potential of Pongamia for rehabilitation of degraded landscapes and as a crop for “carbon farming”. Field trial sites have been established in different geo-climatic regions of Queensland, Australia. The performance of Pongamia at these sites will be presented, with particular reference to growth, flowering and seed production. With respect to flowering, seed production and current threats from insect pests, preliminary findings suggest that Pongamia may be more suited to the dry tropics than the wet tropics. Biochemical assays demonstrated greater variation in protein and oil production in seeds from different trees than with seeds from the same parent tree, supporting the molecular assays (ISSRs) and the outcrossing reproductive nature of Pongamia. As expected, preliminary experiments show that total protein and oil content increase with seed mass and relative maturity. Interestingly, oil content as a percentage of seed content is maximal at stage of development prior to maturity, while protein content increases throughout seed development. Nodulation in Pongamia is not species or strain-specific with ontogeny resembling determinate development. Autoregulation of nodulation (AON) has been observed in Pongamia with parallel studies to determine the nodulation gene network currently in progress. P-TLG27 P-TLG28 Overexpression of ALDRXV4 gene from a resurrection plant enhances tolerance to multiple environmental stresses in blackgram * Singh P , Sarin NB Jawaharlal Nehru University, New Delhi, India *E-mail:preetimku@gmail.com Abstract Aldose reductases belong to the well-conserved aldo–keto reductase super family of enzymes and play numerous roles in the metabolism of steroids, sugars, and other carbonyls in plants and animals.We used the aldose reductase (ALDRXV4) gene isolated from Xerophyta viscose for engineering abiotic stress tolerance in an important legume; Vigna mungo (blackgram). Molecular analyses of independent transgenic lines performed by PCR, Southern, RT-PCR and Western blot revealed the stable integration and expression of the transgene in the progeny. These Southern positive transgenic plants showed high aldose reductase enzyme activity as compared to the untransformed control plants. The leaf discs from transgenic plants were able to tolerate upto 800mM of mannitol, 20% of polyethylene glycol (PEG) and 10mM of methylglyoxal stress as revealed by retention of more chlorophyll and delayed senescence as compared to the untransformed control plants. The transgenic plants were tolerant to higher and toxic levels of methylglyoxal (MG). These could also resist a long period of water deficiency and salt stress and exhibited improved recovery after rehydration. The health of the transgenic plants was assessed by reduction in lipid peroxidation-derived reactive aldehydes and better photosynthetic efficiency under drought and salt stress. Together, these results indicate that ALDRXV4 plays a key role in drought and salinity tolerance via ROS detoxification. Over-expression of bacterial mtlD gene confers enhanced tolerance to salt and water-deficit stress in groundnut (Arachis hypogaea L.) Tengale BD, Kumar A, Dobaria JR, Radhakrishnan T * Directorate of Groundnut Research, Junagadh, India *E-mail: radhakrishnan.nrcg@gmail.com Abstract Groundnut (Arachis hypogaea L.), is an economically important oil seed crop. Water deficit is one of the major production constraints of the crop worldwide. To develop transgenic resistance to abiotic stress, in the present study, bacterial mtlD gene encoding mannitol-1-phosphate dehydrogenase was introduced into groundnut by Agrobacterium-mediated genetic transformation. The integration of the transgene was verified by PCR, dot blot and Southern blotting. The transformation efficiency obtained was 5.29%. Homozygous transgenic lines in T2 generations were evaluated for tolerance to NaCl-induced salt-stress and polyethylene glycol (PEG)-induced water-deficit-stress under in vitro and in vivo conditions. Expression of the transgene resulted in accumulation of mannitol which was confirmed in T 0 and T2 generations using RT-PCR and HPLC. The transgenics showed better shoot length, root length and root volume as compared to the wild type and there was no retardation in growth under stress. Increased salt tolerance of transgenic plants was observed both in vitro and in hydroponic culture; where transgenics could withstand the stress condition of 200 mm NaCl or 20% PEG. Under the same NaCl and PEG stress, relative water content, chlorophyll content and proline content were higher in transgenic plants, while the cell membrane leakage was lower. The manitol accumulation in transgenics ranged from 5.8-13.8 μg.g 1 -1 fr.wt. of tissue at 200Mm NaCl and 4.5-5.6 μg.g fr.wt. of tissue at 20% PEG whereas mannitol was not detected in wild type. The study demonstrated that mtlD gene was expressed in transgenic groundnut plants, resulting in mannitol accumulation and improved tolerance to salt and water-deficit-stress. P-TLG29 P-TLG30 Transgenic resistance to groundnut bud necrosis virus (GBNV) in peanut (Arachis hypogaea L.) plants expressing nucleocapsid protein gene of GBNV 1,* 2 1 1 1 Yadav R , Jain RK , Dobaria JR , Kumar A , Thirumalaisamy PP , 1 Radhakrishnan T 1 Directorate of Groundnut Research, Junagadh, India Indian Agricultural Research Institute, New Delhi, India 2 *E-mail: reena.siya@gmail.com Abstract Peanut bud necrosis caused by Groundnut Bud Necrosis Virus (GBNV) is a very serious problem in peanut (Arachis hypogaea L.) production. Since confirmed sources of genetic resistance are not available to use in the conventional breeding programmes, we have made an attempt to develop transgenic peanut lines with resistance to GBNV, using Nucleocapsid Protein gene of GBNV, through Agrobacterium mediated transformation in the popular cultivar K6. Stable integration of the gene was confirmed by molecular methods in 5 independent transgenic events (T 0) and in 60 plants in T 1 and T2 generations. In Real-time PCR, three transgenic lines viz. PBN.1041.21, 656.1.02 and 417.48.05, had 132.8 and 93 fold; and PBN.887.04, 443.15.01 and PBN.689.01.02 had more than 150 folds the expression as compared to the calibrator having the lowest level of expression. Transgenic peanut progenies expressing the nucleocapsid protein of GBNV were subjected to challenge infection with the virus under glasshouse conditions. The inoculated leaves of the transgenic and non-transgenic controls showed necrotic lesions within 5 to 10 days post inoculation and they tested positive to GBNV and DAC-ELISA analysis indicated that expression of the GBNV gene was higher in transgenic T3 plants. Significantly lower incidence of bud necrosis was observed in the transgenic progeny in comparison to the non-transgenic checks under controlled environmental conditions. These transgenic events could be tested further and may find a potential for use in conventional breeding programs to enhance host resistance. Genome wide Isolation and characterization of functional genes relevant to bio/abio-stresses employing High-density microchip in Arachis hypogaea 1, 1 1 1 1 1 Zhuang WJ *, Zhang C , Cai TC , Chen H , Zhuang CH , Deng Y , Cook, 2 1 1 3 DR , Chen SZ , Lin ZC , Zheng YX 1 Fujian Province Key Lab of Plant Molecular and Cell Biology, Fujian Agriculture and Forestry University, Fuzhou, China 2 University of California Davis, Davis, USA 3 Zhongkai University of Agriculture and Engineering, Guangzhou, China E-mail: weijianz1@163.com Abstract Peanut is an important oil and protein crop worldwide and it often suffers from many abio / bio -stresses, such as drought, low temperature, low calcium in soil, bacterial wilt and aflotoxin contamination, etc, causing a great deal of loses in peanut yield or to human health each year. Understanding the molecular mechanism underlined the stresses would be benefit to alleviate the constraint. Employing established high density microchip with 100,000 unigene probes in peanut, we carried out a widely screening of functional genes relevant to the above stresses as well as treatments with growth regulators such as SA, ABA, ETH, paclobutrazol (PAC). A great amount of differential expression genes were isolated and characterized with high throughput. Nine thousand nine hundred seventy seven genes of pods >3 folds up/down regulated induced by A. flavus infection, 1739 genes from root induced by R. solanacearum, 4982 and 3823 genes from leaf and pod carp, respectively induced by drought, 7313 genes of leaves in duced by low temperature, 7132, 8462, 8577 and 11305 genes from leaves over 3 folds up/down regulated expression induced by SA, ABA, ETH and PAC treatments, respectively, were isolated. There were 392 NBS-LRR genes were characterized by microchip. Among them, 4 NBS-LRR genes were found to be relevant to R. solanacearum and 47 NBS-LRR genes relevant to A. flavus were isolated and characterized. These indicate that the heritance of peanut resistance to R. solanacearum is regulated by few genes, while the resistance to A. flavus appears to be quantitative inheritance. These results established functional genomics resource for further study on peanut resistance network and the molecular mechanism of resistance to environment stresses. P-TLG31 List of Participants LIST OF PARTICIPANTS Adhikari, Kedar The University of Sydney Australia email: kedar.adhikari@sydney.edu.au Almeida, Nuno Instituto de Tecnologia Quimica e Biologia Portugal email: nalmeida@itqb.unl.pt Adivi, Sowmya ICRISAT India email: sowmya_sona75@yahoo.com Anbazhagan, Krithika ICRISAT India email: krithika.anbazhagan@gmail.com Agbagwa, Ikechukwu O IIPR India email: ikechukwu.agbagwa@gmail.com Andargie, Mebeaselassie Haramaya University Ethiopia email: mebhel@yahoo.com Agrama, Hesham International Institute of Tropical Agriculture Zambia email: agrama.hesham@gmail.com Anitha, BK Tamil Nadu Agricultural University India email: bkani22@gmail.com Akila, Chandra Sekhar Yogi Vemana University India email: acsekhar@yogivemanauniversity.ac.in Anjum, TK IIPR India email: k_tuba786@yahoo.co.in Akram, M IIPR India email: akram23859@gmail.com Ansari, Helal AgResearch Limited New Zealand email: helal.ansari@agresearch.co.nz Ali, Latifeh University of Cordoba Spain email: alatifa23bhr@yahoo.com Araujo, Susana Instituto de Tecnologia Quimica e Biologia Portugal email: ssaraujo@gmail.com Ali, Masood IIPR India emai: masoodali53@rediffmail.com Arlappa, N NIN India email: arlappan@yahoo.com Alizadeh, Khoshnod Islamic Azad University Iran email: khoshnod2000@yahoo.com Arunga, Esther Chepkoilel University College Kenya email: eea08ke@hotmail.com VI ICLGG – List of Participants Aski, Muraleedhar IIPR India email: murali2416@gmail.com Bansal, Ravindra BARC India email: ravindra.barc@gmail.com Avercheva, Olga Lomonosov Moscow State University Russia email: olga.avercheva@gmail.com Barkley, Noelle USDA-ARS USA email: elle.barkley@ars.usda.gov Azam, Sarwar ICRISAT India email: s.azam@cgiar.org Basak, Jolly Visva-Bharati Siksha Bhavana India email: jolly.basak@visva-bharati.ac.in Babbar, Anitha JNKVV-Jabalpur India email: anitababbarjnkvv@gmail.com Basamma, B UAS-Dharwad India email: basammapatil09@gmail.com Bae, Ahra Seoul National University Korea email: ar35335@snu.ac.kr Batley, Jacqueline The University of Queensland Australia email: j.batley@uq.edu.au Bag, Manas K NBPGR India email: manas.bag@gmail.com Beena, MR Central Tuber Crops Research Institute India email: beenamr@gmail.com Baga, Monica University of Saskatchewan Canada email: monica.baga@usask.ca Belamkar, Vikas Iowa State University USA email: belamkar@iastate.edu Bandara, Manjula Alberta Agricultural University Canada email: manjula.bandara@gov.ab.ca Belyakova, Alexandra Lomonosov Moscow State University Russia email: alixletter@yandex.ru Bansal, Kailash C NBPGR India email: kcbansal27@gmail.com Bentota, Amitha P GLORC Sri Lanka email: amithabentota@gmail.com Bansal, Mangla CCSHAU India email: monabansal30@gmail.com Bera, Biswajit Tea Board of India India email: biswajittra@yahoo.co.in VI ICLGG – List of Participants Bera, SK Directorate of Groundnut Research India email: berask67@yahoo.co.in Bharathi, S UAS-Raichur India email: bharathi_bup@rediffmail.com Bergvinson, David Bill & Melinda Gates Foundation USA email: david.bergvinson@gatesfoundation.org Bhat, Ramesh UAS-Dharwad India email: bhatramesh12@gmail.com Berhanu, Fenta University of Pretoria South Africa email: berhanufenta@yahoo.com Bhatia, Sabhyata NIPGR India email: sabhyata_bhatia@nipgr.res.in Bertioli, David University of Brasília Brazil email: djbertioli@gmail.com Bhattacharyya, Avisek University of Calcutta India email: helpmeavi.83@gmail.com Bertioli, Soraya EMBRAPA Brazil email: soraya.bertioli@embrapa.br Bhattacharyya, Madan Iowa State University USA email: mbhattac@iastate.edu Bhadauria, Vijai University of Saskatchewan Canada email: bhadauria.vijai@gmail.com Bhimana, Gautami ICRISAT India email: gautaminaidu@gmail.com Bhalla, Prem L The University of Melbourne Australia email: premlb@unimelb.edu.au Bird, David North Carolina State University USA email: bird@ncsu.edu Bhar, Kaushik University of Calcutta India email: kaushik_chembio@yahoo.co.in Biswas, Bandana The University of Queensland Australia email: b.biswas@uq.edu.au Bharadwaj, C IARI India email: drchbharadwaj@gmail.com Bohra, Abhishek IIPR India email: abhi.omics@gmail.com Bharti, Arvind NCGR USA email: akb@ncgr.org Boora, Khazan S CCSHAU India email: kboora@gmail.com VI ICLGG – List of Participants Boukar, Ousmane IITA Nigeria email: o.boukar@cgiar.org Calderini, Ornella University of Perugia Italy email: ornella.calderini@igv.cnr.it Brasileiro, Ana EMBRAPA Brazil email: ana.brasileiro@embrapa.br Cannon, Steven B USDA-ARS USA email: steven.cannon@ars.usda.gov Brear, Ella The University of Sydney Australia email: ella.brear@sydney.edu.au Chahota, Rakesh K CSK Himachal Pradesh Agricultural University India email: rkchahota@yahoo.com Briñez Rodriguez, Boris State University of Campinas Brazil email: borisbrinez@hotmail.com Chakraborty, Dipjyoti Banasthali Vidyapith India email: dciitkgp@gmail.com Bruggemann, Edward DuPont Pioneer USA email: edward.bruggemann@pioneer.com Chan, Crystal Saskatchewan Pulse Growers Canada email: cchan@saskpulse.com Bui, Huyen Thi Thu Vietnam Academy of Agricultural Science Vietnam email: huyenbuong@gmail.com Chandra Reddy, AS ANGRAU India email: chanduforbiotech@gmail.com Buirchell, Bevan The University of Western Australia Australia email: bevan.buirchell@agric.wa.gov.au Chaturvedi, SK IIPR India email: sushilk.chaturvedi@gmail.com Burstin, Judith INRA-Dijon France email: burstin@dijon.inra.fr Chaudhary, RG IIPR India email: rgc_1234@yahoo.com Bustos-Sanmamed, Pilar CNRS France email: bustos@isv.cnrs-gif.fr Chen, Charles Auburn University USA email: cyc0002@auburn.edu Butler, Larry Generation Challenge Programme Mexico email: l.butler@cgiar.org Chen, Xiao Ping Guangdong Academy of Agricultural Sciences China email: xpchen1011@gmail.com VI ICLGG – List of Participants Chetukuri, Anuradha ANGRAU India email: anu_dna@rediffmail.com Crespi, Martin ISV-CNRS France email: crespi@isv.cnrs-gif.fr Chibbar, Ravindra University of Saskatchewan Canada email: ravi.chibbar@usask.ca da Silva , Daiana Alves, Agronomic Institute of Campinas Brazil email: daiagrouel2002@hotmail.com Chilagane, Luseko Sokoine University of Agriculture Tanzania email: lusekochilagane@yahoo.co.uk Datta, Swapan K ICAR India email: swpndatta@yahoo.com Cisse, Ndiaga Institute Senegalais de Recherche Agricole Senegal email: ncisse@refer.sn Das, Alok IIPR India email: alokbio@gmail.com Close, Timothy UC-Riverside USA email: timothy.close@ucr.edu Das, Debapriya R University of Calcutta India email: debapriyarajlakshmi@gmail.com Cnops, Gerda Ghent University Belgium email: gerda.cnops@ilvo.vlaanderen.be Das, Mousumi University of Calcutta India email: mou.19may@gmail.com Cominelli, Eleonora Università degli Studi di Milano Italy email: cominelli@ibba.cnr.it Das, Sampa Bose Institute India email: sampa@bic.boseinst.ernet.in Cook, Douglas R UC-Davis USA email: drcook@ucdavis.edu Dayal, Archana IARI India email: archana_dayal2001@yahoo.com Cordeiro, Matilde UC-Davis USA email: mcordeiro@ucdavis.edu Debelle, Frederic INRA-Toulouse France email: frederic.debelle@toulouse.inra.fr Coyne, Clarice J USDA-ARS, USA email: clarice.coyne@ars.usda.gov Desclaux, Dominique INRA France email: dominique.desclaux@supagro.inra.fr VI ICLGG – List of Participants Deshmukh, Milind Mahatma Phule Krishi Vidyapeeth India email: drmpdeshmukh@gmail.com Domergue, Odile INRA France email: domergue@supagro.inra.fr Devi, Pavithra Tamil Nadu Agricultural University India email: pavithra613@gmail.com Domoney, Claire John Innes Centre UK email: claire.domoney@jic.ac.uk de Villers, Santie ICRISAT Kenya email: s.devilliers@cgiar.org Doss, Dayal D UAS-Banglore India email: dds3a1win@yahoo.com Dhanasekar, P BARC India email: sekar@barc.gov.in Doss, Parimala K ANGRAU India email: pari_mala123@rediffmail.com Dhar, Shri IARI India email: shridhar@iari.res.in Doyle, Jane Cornell University USA email: jjd5@cornell.edu Dharmaraj, PS ARS-Gulbarga India email: psdharmaraj@yahoo.com Doyle, Jeffrey Cornell University USA email: jjd5@cornell.edu Dhole, VJ BARC India email: vjdhole@yahoo.co.in Drevon, Jean-Jacques INRA Eco&Sols France email: drevonjj@supagro.inra.fr Diop, Ndeye Ndack Generation Challenge Programme Mexico email: nn.diop@cgiar.org Dudhe, Mangesh Directorate of Oilseeds Research India email: mangeshdudhe@gmail.com Dixon, Richard The Samuel Roberts Noble Foundation USA email: radixon@noble.org Dutta, Ayan University of Calcutta India email: a.dutta2010@gmail.com Djordjevic, Michael Australian National University Australia email: michael.djordjevic@anu.edu.au Duvvurumuni, Rajasekhara Reddy Monsanto Research Centre India email: rduvv@monsanto.com VI ICLGG – List of Participants Easdown, Warwick AVRDC India email: warwick.easdown@worldveg.org Fischer, Kristin Julius Kühn-Institut (JKI) Germany email: kristin.fischer@jki.bund.de Edwards, Dave The University of Queensland Australia email: dave.edwards@uq.edu.au Fonceka, Daniel CIRAD France email: daniel.fonceka@cirad.fr Ehlers, Jeffrey Bill & Melinda Gates Foundation USA email: jeff.ehlers@gatesfoundation.org Friesen, Maren University of Southern California USA email: maren.l.friesen@gmail.com El-Rodeny, Walid ARC Egypt email: wrodeny@yahoo.com Froenicke, Lutz UC-Davis USA email: lfroenicke@ucdavis.edu Ellis, Noel Aberystwyth University UK email: noe2@aber.ac.uk Frugoli, Julia Clemson University USA email: jfrugol@clemson.edu Eshete, Million EIAR Ethiopia email: million102001@yahoo.com Fukai, Eigo National Institute of Agrobiological Sciences Japan email: efu@affrc.go.jp Eyles, Rod The Australian National University Australia email: rod.eyles@anu.edu.au Gaikwad, Nikhil Shivaji University India email: nbgaikwadsu@gmail.com Fedoruk, Michael University of Saskatchewan Canada email: fedorukmichael@gmail.com Ganga Rao, NVPR ICRISAT Kenya email: n.gangarao@cgiar.org Fevereiro, Pedro Instituto de Tecnologia Quimica e Biologia Portugal email: psalema@itqb.unl.pt Gangisetty, Nagaraju NIPGR India email: nagagangi08@gmail.com Fikre, Asnake EIAR Ethiopia email: fikreasnake@yahoo.com Garg, Neera Punjab University India email: gargneera@gmail.com VI ICLGG – List of Participants Garg, Rohini NIPGR India email: rohini@nipgr.res.in Gorntla, Annapurna ICRISAT India email: anu5299@gmail.com Gaur, Pooran M ICRISAT India email: p.gaur@cgiar.org Gowda, CLL ICRISAT India email: c.gowda@cgiar.org Geffory, Valérie INRA France email: valerie.geffroy@u-psud.fr Grunz, Fabian Univeristy of Frankfurt Germany email: fgrunz@stud.uni-frankfurt.de Ghanta, Anuradha ANGRAU India email: saps_61@yahoo.com Gu, Jenny USAID USA email: jgu@usaid.gov Ghoulam, Cherki Guéliz Marrakech Morocco email: ghoulamcherki@yahoo.fr Gu, Heping Jiangsu Academy of Agricultural Sciences China email: ghp@jaas.ac.cn Ghosh, Bikramaditya University of Calcutta India email: ghosh.bikramaditya10@gmail.com Guha, Sohini University of Calcutta India email: phagealpha@gmail.com Giri, Ashok P National Chemcial Laboratory India email: ap.giri@ncl.res.in Guimaraes, Patricia EMBRAPA Brazil email: patricia.guimaraes@embrapa.br Gnanasambandam, Annathurai DPI- Horsham Australia email: annathurai.gnanasambandam@dpi.vic.gov.au Guo, Baozhu University of Georgia USA email: baozhu.guo@ars.usda.gov Gopalakrishnan, R Genotypic Technology (P) Ltd India email: gopalakrishna.r@genotypic.co.in Guo, Yong Chinese Academy of Agricultural Sciences China email: guoyongbj@gmail.com Gopala Krishna, M Agri Biotech Foundation India email: bt_gopal@rediffmail.com Gupta, Debjyoti S IIPR India email: debgpb@gmail.com VI ICLGG – List of Participants Gupta, Moni SKUAST-Jammu India email: sachinmoni@rediffmail.com Hien, Edmond CAMES Burkina Faso email: edmond.hien@ird.fr Gupta, Priyanka IARI India email: priyanka13biotech@gmail.com Hernandez, Georgina Universidad Nacional Autónoma de México Mexico email: gina@ccg.unam.mx Gupta, Sanjeev IIPR India email: saniipr@rediffmail.com Higgins, Thomas J CSIRO Australia email: tj.higgins@csiro.au Gupta, Shefali NIPGR India email: shefali_1403@yahoo.co.in Himabindu, K ICRISAT India email: k.himabindu@cgiar.org Gupta, Vidya National Chemical Laboratory India email: vs.gupta@ncl.res.in Hingane, Anupama ICRISAT India email: h.anupama@cgiar.org Guseinova, Irada Azerbaijan National Academy of Sciences Azerbaijan email: i_guseinova@mail.ru Hofer, Julie Aberystwyth University UK email: jmh18@aber.ac.uk Han, Kwang-soo Seoul National University Korea email: overmax@snu.ac.kr Hoisington, David ICRISAT India email: d.hoisington@cgiar.org Harry, David TerViva BioEnergy, Inc USA email: deharry@peak.org Hong, Yanbin Guangdong Academy of Agricultural Sciences China email: hongyanbin1979@yahoo.com.cn Hatira, Abdessattar Faculté des sciences de Tunis Tunisia email: absat.hatira@gmail.com Hu, Jinguo USDA-ARS USA email: jinguo.hu@ars.usda.gov Hay, Alison ACPFG Australia email: alison.hay@acpfg.com.au Hwang, Wonjoo Seoul National University Korea email: hwj450@naver.com VI ICLGG – List of Participants Ibrahim, Hatem Université de Tunise Tunisia email: brahim_hatem@yahoo.fr Jang, Young E Seoul National University Korea email: j.easttree@gmail.com Iakovenko, Oskana Inst. Bioorganic Chem &Petrochem Ukraine email: lottin@rambler.ru Janila, P ICRISAT India email: p.janila@cgiar.org Isobe, Sachiko Kazusa DNA Research Institute Japan email: sisobe@kazusa.or.jp Jarso, Mussa EIAR Ethiopia email: mjarso@yahoo.com Iwata, Aiko University of Georgia USA email: aiwata@uga.edu Javadi, Firouzeh Kyushu University Japan email: fjavascb@kyushu-u.org Jackson, Scott University of Georgia USA email: sjackson@uga.edu Jayalakshmi, Veera RARS-Nandyal India email: veera.jayalakshmi@gmail.com Jaganathan, Deepa ICRISAT India email: deebiotech@gmail.com Jhansi Rani, K ANGRAU India email: jhansisagar@yahoo.com Jain, Meenakshi CCSHAU India email: meenakshi110@gmail.com Johnson, Casey North Dakota State University USA email: dilrukshi.thavarajah@ndsu.edu Jain, Mukesh NIPGR India email: mukesh_nipgr@yahoo.com Joshi, Priyanka RAK College of Agriculture India email: priyanka.joshi95@yahoo.com Jain, Shalu North Dakota State University USA email: Sujain_jnu@yahoo.com Kaashyap, Mayank ICRISAT India email: mayankkaashyap@yahoo.com Jang, Hyunju Seoul National University Korea email: janghyunju12@gmail.com Kadoo, Narendra National Chemical Labrotary India email: ny.kadoo@ncl.res.in VI ICLGG – List of Participants Kahl, Guenter University of Frankfurtam Main Germany email: kahl@em.uni-frankfurt.de Kaur, Livinder Punjab Agricultural University India email: livinderk@rediffmail.com Kakilaru, Kanaka Durga ANGRAU India email: kana@yahoo.com Kawashima, Cintia The Sainsbury Laboratory UK email: cintia.kawashima@tsl.ac.uk Kaiser, Brent Norman The University of Adelaide Australia email: brent.kaiser@adelaide.edu.au Kereszt, Attila Hungarian Academy of Sciences Hungary email: kereszta@baygen.hu Kalaimagal, T Tamil Nadu Agricultural University India email: kalaimagal.t@gmail.com Khadke, Shivram Smt. Kasturbabai Walchand College of, Arts and Science, India email: shivramkhadke@gmail.com Kamphuis, Lars The University of Western Australia Australia email: lars.kamphuis@csiro.au Khandelwal, Neha National Chemical Laboratory India email: neha.biogene@gmail.com Kanakala, Surapathrudu IARI India email: kanakalavit@gmail.com Kharb, Pushpa CCSHAU India email: pkharbhau@gmail.com Kang, Yang J Seoul National University Korea email: kangyangjae@gmail.com Khan, Aamir W ICRISAT India email: a.khan@cgiar.org Kannepalli, Annapurna IARI India email: annapurna96@yahoo.co.in Khatodia, Surender CCSHAU India email: khatodia99@gmail.com Kant, Chandra NIPGR India email: ckantop@gmail.com Khazaei, Hamid University of Helsinki Finland email: hamid.khazaei@helsinki.fi Kaur, Jagmeet Punjab Agricultural University India email: jagskaur@gmail.com Khera, Pawan ICRISAT India email: k.pawan@cgiar.org VI ICLGG – List of Participants Kileo, Robert LZARDI Tanzania email: kileorobert@yahoo.com Krishna Shilpa, P ICRISAT India email: krishnashilpa.11@gmail.com Kim, Sue Seoul National University Korea email: suekim83@snu.ac.kr Krishnamurthy, L ICRISAT India email: l.krishnamurthy@cgiar.org Kim, Dong Hyun ICRISAT India email: d.kim@cgiar.org Ksiazkiewicz, Michal Institute of Plant Genetics of PAS Poland email: mksi@igr.poznan.pl Kimurto, Paul Egerton University Kenya email: kimurtopk@gmail.com Kuchanur, Prakash H UAS-Raichur India email: prakashkuchanur@yahoo.co.in King, Matthew DuPoint Pioneer USA email: matt.king@pioneer.com Kulwal, Pawan Mahatma Phule Agricultural University India email: pawankulwal@gmail.com Kiss, Gyorgy B Agricultural Biotechnology Centre Hungary email: gbkiss@abc.hu Kumar, Abhay Directorate of Groundnut Research India email: abhayiari@gmail.com Kobaku, Shankar S CRIDA India email: sreedevikobaku@gmail.com Kumar, Ashish JNKVV-Rewa India Email: ashishashish2612@gmail.com Koilkonda, Padmalatha Vanasthalipuram India email: padmalathakk@gmail.com Kumar, Ashutosh Birla Institute of Technology India email: kr.ashutosh@yahoo.com Kosgei, Alice Egerton University Kenya email: alicesongio@yahoo.com Kumar, Jitendra IIPR India email: jitendra73@gmail.com Kottapalli, Kameswara R Texas Tech University USA email: rao.kottapalli@ttu.edu Kumar, Kamal NIPGR India email: kamal_kumar@nipgr.res.in VI ICLGG – List of Participants Kumar, Narendra Directorate of Groundnut Research India email: narendrapb09@gmail.com Lazali, Mohamed ENSA Algérie email: m.lazali@yahoo.fr Kumar, Yogesh Central Rainfed Upland Rice Research Station India email: dryogeshtiwari70@gmail.com Lee, Jay E Seoul National University Korea email: leejayern@gmail.com Kumari, Archana Birla Institute of Technology India email: archana.bioinfo87@gmail.com Lee, Suk-ha Seoul National University Korea email: sukhalee@snu.ac.kr Kundagrami, Sabyasachi University of Calcutta India email: skundagrami@gmail.com Lee, Taeyoung Seoul National University Korea email: alima9002@gmail.com Kundu, Anirban University of Calcutta India email: discoveranirban@yahoo.co.in Lee, Yeongho Seoul National University Korea email: yhl413@hanmail.net Kundu, Anindya University of Calcutta India email: anindya.kundu65@gmail.com Lestari, Puji Seoul National University Korea email: lestari_71@yahoo.com Kute , Nand Kumar Mahatma Phule Krishi Vidyapeeth India nskute2004@rediffmail.com Li, Ying-Hui Chinese Academy of Agricultural Sciences China email: dadousoybean@yahoo.com.cn Lal, Chuni Directorate of Groundnut Research India email: chunilal@nrcg.res.in Liang, Xuanqiang Guangdong Academy of Agricultural Sciences China email: liang804@yahoo.com Laouar, Meriem INRA Algeria email: laouar_m@yahoo.fr Lichtenzveig, Judith Curtin University Australia email: judith.lichtenzveig@curtin.edu.au Latati, Mourad ENSA Algerie email: m.latati@yahoo.com Lightfoot, David Southern Illinois University USA email: ga4082@siu.edu VI ICLGG – List of Participants Liu, Chuang ShanDong Shofine Seed Technology Co Ltd China email: lichunyan@shofine.com Markmann, Katharina University of Aarhus Denmark email: kama@mb.au.dk Lohithaswa, HC UAS-Dharwad India email: lohithhc@rediffmail.com Marla, Soma S NBPGR India email: soma.marla@nbpgr.ernet.in Ma, Jianxin Purdue University USA email: maj@purdue.edu Martin, Xavier Queensland Department of Agriculture Fisheries and Forestry (DAFF), Australia email: martinx@northnet.com.au Maougal, Rim ENSA Algérie email: rym.maougal@gmail.com Mashamba, Philippo Naliendele Agricultural Research Institute Tanzania email: phillipomas@yahoo.com Malathi, VG IARI India email: vgmalathi@rediffmail.com Massawe, Festo University of Nottingham Malaysia Campus Malaysia Festo.Massawe@nottingham.edu.my Mallikarjuna, Nalini ICRISAT India email: n.mallikarjuna@cgiar.org McGee, Rebecca USDA-ARS USA email: rebecca.mcgee@ars.usda.gov Malviya, Nupur IIPR India email: malviya.nupur@gmail.com Meena, Mukesh K NIPGR India email: mukesh_nipgr@yahoo.com Manivannan, N Tamil Nadu Agricultural University India email: nmvannan@gmail.com Mehta, Harsh Central Soil and Water Conservation India email: harshmehta41@rediffmail.com Mannur, Durdundappa ARS-Gulbarga India email: dmmannur@rediffmail.com Meksem, Khalid Southern Illinois University Carbondale USA email: meksemk@siu.edu Mantri, Nitin RMIT University Australia email: nitin.mantri@rmit.edu.au Merla, Srilakshmi S JNTU India email: m.s.l.sunita@gmail.com VI ICLGG – List of Participants Mfaume, Juma Naliendele Agricultural Research Institute Tanzania email: mfaumer2000@yahoo.co.uk Morgado, Ana R Instituto de Tecnologia Quimica e Biologia Portugal email: rmorgado@itqb.unl.pt Millan, Teresa University of Cordoba Spain email: teresa.millan@uco.es Mudaraddi, Bharati ICRISAT India email: bharati_bth@yahoo.com Millawithanachchi, MC GLORC Sri Lanka email: withanamcm@yahoo.com Mukherje, Shreyartha Iowa State University USA email: shreyartha@gmail.com Mini, ML Bharathidasan University India email: minitnau@gmail.com Mulwa, Richard Egerton University Kenya email: rmulwa@egerton.ac.ke Mir, Reyazul R SKUAST-Jammu India email: imrouf2006@gmail.com Mundree, Sagadevan G Queensland University of Technology Australia sagadevan.mundree@qut.edu.au Misra, Hari O CIMAP India email: ho.misra@cimap.res.in Muniswamy, S ARS-Gulbarga India email: munis01@rediffmail.com Misra, Neelu IIPR India email: neelumishra111@gmail.com Muralidharan, Rajalakshmi Department of Biotechnology (DBT) India email: m.rajalakshmi@nic.in Mondal, Suvendu BARC India email: suvenduhere@yahoo.co.in Nadarajan, N IIPR India email: n.nadarajan@gmail.com Mondal, Tapan K NBPGR India email: mondaltk@yahoo.com Naganowska, Barbara Institute of Plant Genetics of PAS Poland email: bnag@igr.poznan.pl Monyo, Emmanuel S ICRISAT Malawi email: e.monyo2@cgiar.org Nagesh, PN ICRISAT India email: patnenagesh@gmail.com VI ICLGG – List of Participants Naik, KSS ARS-Kadiri India email: drkssnaik@yahoo.com Niebel, Andreas CNRS-LIPM France email: andreas.niebel@toulouse.inra.fr Naimuddin IIPR India email: naimk@rediffmail.com Nouhou, Rekia ICRISAT Niger email: rekadave2002@yahoo.fr Nair, Ramakrishnan M AVRDC India email: r.nair@cgiar.org Ntare, Bonny ICRISAT Mali email: b.ntare@cgiar.org Namdeo, Ghanawat Yashwantrao Chavan Institute of Science India email: namdeoghanawat249@gmail.com Nwosu, Victor C MARS Chocolate USA email: victor.nwosu@effem.com Narits, Lea Jõgeva Plant Breeding Institute Estonia email: Lea@jpbi.ee O’Sullivan, Donal National Institute of Agricultural Botany UK email: donal.osullivan@niab.com Narmadha, B Tamil Nadu Agricultural University India email: manosandhiya_87@yahoo.co.in Obarley, Yue Macrogen Millennium Genomics Co. Ltd. China email: yuyue@macrogencn.com Natarajan, Jeyakumar Bharathiar University India email: n.jeyakumar@yahoo.co.in Oldroyd, Giles John Innes Centre UK email: giles.oldroyd@jic.ac.uk Nayakoti, Swapna ICRISAT India email: nswapnaforu@gmail.com Olson, Mark Pacific Agri-Food Research Centre Canada email: mark.olson@gov.ab.ca Nelson, Matthew The University of Western Australia Australia email: matthew.nelson@uwa.edu.au Omoigui, Lucky University of Agriculture Nigeria email: lomoigui@yahoo.co.uk Nguyen, Henry University of Missouri USA email: nguyenhenry@missouri.edu Ontagodi, Timmanna UAS-Dharwad India email: timmu07uas@gmail.com VI ICLGG – List of Participants Oyier, Moses O Egerton University Kenya email: oyierkilli2@yahoo.com Parker, Jeff Saskatchewan Pulse Growers Canada email: jparker@saskpulse.com Ozias-Akins, Peggy University of Georgia USA email: pozias@uga.edu Patel, Anju Bose Institute India email: anjupatel2009@gmail.com Jyothsna, P IARI India email: jyothsnap177@gmail.com Patil, BR UAS-Dharwad India email: patilbhuvaneshwara@gmail.com Pal, Amita Bose Institute India email: amita@bic.boseinst.ernet.in Patil, BS IARI's Center for Pulse Improvement India email: bs_patil2000@yahoo.com Pansu, Marc Supagro France email: marc.pansu@ird.fr Patil, Prakash G IIPR India email: patilbt@gmail.com Palaniappan, Jayamani Tamil Nadu Agricultural University India email: jayamani1108@hotmail.com Patil, Suyash B ICRISAT India email: P.Suyash@cgiar.org Palavalasa, Hima K Osmania University India email: phimakumari@gmail.com Patil, Jaiprakash UAS-Raichur India email: jrpatil_uas@yahoo.com Panchangam, Sameera S ICRISAT India email: sameera.panchangam@gmail.com Patil, Somnath Nirmal Seeds Pvt Ltd India email: srpatil@nirmalseedsindia.com Pandey, Dev M Birla Institute of Technology India email: dmpandey@bitmesra.ac.in Patto, Maria CV Instituto de Tecnologia Quimica e Biologia Portugal email: cpatto@itqb.unl.pt Pandey, Manish K ICRISAT India email: m.pandey@cgiar.org Paul, Anindita University of Calcutta India email: bitinfo_anin@yahoo.co.in VI ICLGG – List of Participants Paul, Pronob J Sam Higginbotom Institute of Agriculture, Technology & Sciences, India email: pronobpaul@gmail.com Pushpavalli, R ICRISAT India email: siva_sasi11@yahoo.co.in Penmetsa, R Varma UC-Davis USA email: rvpenmetsa@ucdavis.edu Putterill, Jo The University of Auckland New Zealand email: j.putterill@auckland.ac.nz Pouryousef, Mahmood Islamic Azad University Iran email: pooryousefm@yahoo.com Radhakrishnan, T Directorate of Groundnut Research India email: radhakrishnan.nrcg@gmail.com Pradhan, Seema NIPGR India email: seemapradhan3@gmail.com Rahayu, Sherly Center for Application of Isotopes and Radiation Technology, Indonesia email: sherlyrahayu@yahoo.com Prajapati, Gopal K Birla Institute of Technology India email: gk.prajapati5@gmail.com Raheem, Amtul ANGRAU India email: amieraheem04@gmail.com Prasad, Akhouri Center for Bioinformatics-Rachi India email: abprasad35@yahoo.com Rajan, Neha IIPR India email: neha_rajan96@yahoo.com Pratap, Aditya IIPR India email: adityapratapgarg@gmail.com Raju, Dhandapani Directorate of Seed Research India email: dandyman2k3@yahoo.co.in Puhan, Puspasree ANGRAU India email: puspapuhan@gmail.com Ramírez Yáñez, Mario Universidad Nacional Autónoma de México Mexico email: mario@ccg.unam.mx Purayannur, Savithri NIPGR India email: savithri@nipgr.res.in Rana, Jai C NBPGR India email: ranajc2003@yahoo.com Purushothaman, R ICRISAT India email: purushothmurthy@gmail.com Rana, MK NBPGR India email: mkrana_nbpgr@yahoo.co.in VI ICLGG – List of Participants Rane, Jagadish National Institute of Abiotic Stress Management India email: jagrane@hotmail.com Rotter, Bjoern GenxPro Germany email: rotter@genxpro.de Rao, AR Indian Agricultural Statistics Research Institute India email: arrao@iasri.res.in Roy, Anirban NBPGR India email: anirbanroy75@yahoo.com Rastogi, Khushboo Seoul National University India email: khushboorastogi5@gmail.com Roy Choudhury, Debashree University of Calcutta India email: drc.gpb@gmail.com Rathore, Abhishek ICRISAT India email: a.rathore@cgiar.org Rueda, Janice USA Dry Pea & Lentil Council USA email: jrueda@pea-lentil.com Ratnaparkhe, Milind Directorate of Soybean Research India email: milind.ratnaparkhe@gmail.com Ruge-Wehling, Brigitte Julius Kuehn Institute Germany email: brigitte.ruge-wehling@jki.bund.de Reddy, Chandra Obul Yogi Vemana University India email: coreddy@yogivemanauniversity.ac.in Sabbavarapu, Murali Mohan ICRISAT India email: m.sabbavarapu@cgiar.org Reynoso, Mauricio A Instituto de Biotecnología y Biología Molecular UNLP-CONICET. Argentina email: mauricio_a_reynoso@yahoo.com.ar Saha, Sudip University of Calcutta India email: saha4sudip@gmail.com Ribaut, Jean Marcel Generation Challenge Programme Mexico email: j.ribaut@cgiar.org Salimath, PM UAS-Shimoga India email: salimathpm@gmail.com Roopa Lavanya, G Sam Higginbotom Institute of Agriculture, Technology & Sciences, India email: lavanya.roopa@gmail.com Sama, VSAK ICRISAT India email: a.sama@cgiar.org Roorkiwal, Manish ICRISAT India email: m.roorkiwal@cgiar.org Samaddar, Sandip University of Calcutta India email: samsan700@gmail.com VI ICLGG – List of Participants Sameerkumar, CV ICRISAT India email: c.sameerkumar@cgiar.org Saxena, KB ICRISAT India email: k.saxena@cgiar.org Sanchez, Vanessa Florida International University USA email: vanessa.sanchez.velez@gmail.com Sayama, Takashi National Institute of Agrobiological Science Japan email: tsayama@affrc.go.jp Sanyal, Abhijit Pioneer DuPont India email: abhijit.sanyal@pioneer.com Schafleitner, Roland AVRDC Taiwan email: roland.schafleitner@worldveg.org Sarin, Neera B JNU India email: neerasarin@rediffmail.com Schmidt, Monica University of Arizona USA email: monicaschmidt@email.arizona.edu Sarkar, Monolina University of Calcutta India email: monolinasarkar@gmail.com Scholz, Todd USA Dry Pea & Lentil Council USA email: scholz@pea-lentil.com Sarker, Ashutosh ICARDA Syria email: a.sarker@cgiar.org Scott, Paul The University of Queensland Australia email: p.scott@uq.edu.au Sarma, BK BHU India email: birinchi_ks@yahoo.com Shah, Divya IARI India email: divya_shah1984@yahoo.co.in Sarma, Shivali ICRISAT India email: s.sarma@cgiar.org Shah, Niraj NIPGR India email: n_shah_321@yahoo.co.in Satyanarayana, N Sikkim University India email: sathyan_dixit@yahoo.in Shah, Trushar ICRISAT India email: tm.shah@cgiar.org Saxena, Rachit K ICRISAT India email: r.saxena@cgiar.org Sharma, Balram IARI India email: drbrsh@yahoo.co.in VI ICLGG – List of Participants Sharma, Tilak CSK Himachal Pradesh Agricultural University India email: sharmat88@yahoo.com Singam, Prashant JNTU India email: prashantsingam@gmail.com Shasidhar, Y ICRISAT India email: bioshasi@gmail.com Singh, Ainmisha IIPR India email: ainmishasingh1@gmail.com Shetty, Prateek Sir M Visvesvaraya Institute of Technology India email: prateekshettys@gmail.com Singh, Akanksha BHU India email: bhuaks29@gmail.com Shridevi, BM UAS-Dharwad India email: bmshree.4795@gmail.com Singh, Bikram SKUAST-Jammu India email: bikramsgh@rediffmail.com Shim, Sangrea Seoul National University Korea email: kev8305@gmail.com Singh, Karam The University of Western Australia Australia email: Karam.Singh@csiro.au Shirasawa, Kenta Kazusa DNA Research Institute Japan email: shirasaw@kazusa.or.jp Singh, Madan Pal IARI India email: madanpal@yahoo.com Shirkanth, Sandhya ICRISAT India email: sandhya.thudumu@gmail.com Singh, Mahendranarain BHU India email: mnsbhu@yahoo.co.in Siddhanta, Anirban Univesity of Calcutta India email: asiddhanto@yahoo.com Singh, Pallavi IIPR India email: pallavi_pbi@rediffmail.com Simmonds, Daina Agriculture and Agri-Food Canada Canada email: daina.simmonds@agr.gc.ca Singh, Preeti JNU India email: preetimku@gmail.com Sine, Bassirou CERAAS/ISRA Senegal email: bsinefr@yahoo.fr Singh, Sarvjeet Punjab Agricultural Univesity India email: sarvjeetm@rediffmail.com VI ICLGG – List of Participants Singh, Vinay K BHU India email: vinaysingh@bhu.ac.in Somanaboina, Anil K Osmania University India email: anilkumarou@gmail.com Singh, Harikesh B BHU India email: hbs1@rediffmail.com Soren, KR IIPR India email: sorenars@gmail.com Singh, Vijayata Central Soil Salinity Research Institute India email: vijayatasingh.gpb@gmail.com Souframanien, J BARC India email: souf@scientist.com Sinha, Pallavi ICRISAT India email: p.sinha@cgiar.org Sparvoli, Francesca Institute of Agricultural Biology and Biotechnology ,Italy email: sparvoli@ibba.cnr.it Sinha, Subodh K NRCPB India email: subsinha@gmail.com Sriswathi, M ICRISAT India email: sriswathi.17@gmail.com Sinharoy, Senjuti The Samuel Roberts Noble Foundation USA email: ssinharoy@noble.org Srivastava, Ekta IIPR India email: ekta29nov@gmail.com Sinjushin, Andrey Lomonosov Moscow State University Russia email: asinjushin@mail.ru Stephens, Brian University of Houston-Clear Lake USA email: stephensb@uhcl.edu Smith, Penelope University of Sydney Australia email: penny.smith@sydney.edu.au Stougaard, Jens Aarhus University Denmark email: stougaard@mb.au.dk Sneha, Sonam Jayoti Vidyapeeth Women's University India email: sonam.sneha2@gmail.com Stupar, Milanko Department of Animal Science Serbia email: milanko.stupar@laposte.net Sofi, Parvaze SKUAST-K India email: parvazesofi@gmail.com Stupar, Robert University of Minnesota USA email: stup0004@umn.edu VI ICLGG – List of Participants Sudini, Hari Kishan ICRISAT India email: h.sudini@cgiar.org Taran, Bunyamin University of Saskatchewan Canada email: bunyamin.taran@usask.ca Sunkar, Ramanjulu Oklahoma State University USA email: ramanjulu.sunkar@okstate.edu Taunk, Jyoti CCSHAU India email: jyotibiotech86@gmail.com Surapaneni, Koteswara Rao JNKVV- Rewa India email: skrao_jnau@yahoo.co.in Tebkew, Damte EIAR Ethiopia email: tebkew@yahoo.com Sugandh Suman Rajendra Agricultural University India email: sugandhsuman@gmail.com Thakare, Dipali P ICRISAT India email: dipalithakare02@gmail.com Suresh, M ARS-Tandur India email: sureshiari@yahoo.co.in Thavarajah, Dil North Dakota State University USA email: dilrukshi.thavarajah@ndsu.edu Sutton, Tim ACPFG Australia email: tim.sutton@acpfg.com.au Thompson, Courtney University of Saskatchewan Canada email: clt788@mail.usask.ca Swarnalakshmi, K IARI India email: swarna_bga@yahoo.com Thompson, Richard INRA France email: thompson@dijon.inra.fr Swathi, P ICRISAT India email: parupalliswathi_17@yahoo.com Thudi, Mahendar ICRISAT India email: t.mahendar@cgiar.org Szczyglowski, Krzysztof Agriculture and Agri-Food Canada Canada email: Krzysztof.Szczyglowski@agr.gc.ca Tian, Jing Hebei Academy of Agricultural and Forestry Sciences, China email: nkytianjing@163.com Tadege, Million The Samuel Roberts Noble Foundation USA email: million.tadege@okstate.edu Tignegre, Jean B CREAF de Kamboinse Burkina Faso email: racinetignegre@yahoo.com VI ICLGG – List of Participants Tiwari, Gunjan GB Pant University of Agriculture & Technology, India email: gnjntiwari1@gmail.com Van, Kyujung Seoul National University Korea email: kvan@snu.ac.kr Torres, Ana M IFAPA Spain email: anam.torress.romero@juntadcandatucia.es Vandemark, George USDA-ARS USA email: george.vandemark@ars.usda.gov Tripathi, Shailesh IARI India email: shaitri@rediffmail.com Vandenberg, Albert University of Saskatchewan Canada email: bert.vandenberg@usask.ca Udayakumar, M UAS-Bangalore India email: udayakumar_m@yahoo.com Varshney, Rajeev K ICRISAT India email: r.k.varshney@cgiar.org Udvardi, Michael The Samuel Roberts Noble Foundation USA email: mudvardi@noble.org Vasanthi, RP RARS-Tirupati India email: vasanthi.rrs@gmail.cm Upadhyaya, Hari ICRISAT India email: h.upadhyaya@cgiar.org Vasconcelos, Marta W Universidade Católica Portuguesa Portugal email: mvasconcelos@porto.ucp.pt Uppalapati, Srinivasa R The Samuel Roberts Noble Foundation USA email: srinivasaraou@yahoo.com Vasimalla, Celia C NBPGR India email: mailcelia@gmail.com Vadez, Vincent ICRISAT India email: v.vadez@cgiar.org Verdier, Jerome The Samuel Roberts Noble Foundation USA email: javerdier@noble.org Valentine, Howard The Peanut Foundation USA email: hvalentine@peanutsusa.com Verma, Priyanka NIPGR India email: priyankabiochem29@gmail.com Valliyodan, Babu University of Missouri USA email: valliyodanb@missouri.edu Viall, Eric North Dakota State University USA email: Eric.viall@ndsu.edu VI ICLGG – List of Participants Vijayakumar, AG UAS-Dharwad India email: vijay_gpbd@yahoo.co.in Williams, Jonathan University of Georgia USA email: crspgrf@griffin.uga.edu Vinoth, R Tamil Nadu Agriculture University India email: rvinothagri@gmail.com Wilson, Richard The Peanut Foundation USA email: rfwilson@mindspring.com Viswanatha, KP UAS-Bangalore India email: viswanathakp55@gmail.com Wolko, Bogdan Institute of Plant Genetics of PAS Poland email: bwol@igr.poznan.pl von Wettberg, Eric Florida International University USA email: evonwettberg@fairchildgarden.org Xu, Donghe JIRCAS Japan email: xudh@jircas.affrc.go.jp Vu, Wendy University of Southern California USA email: wvu@usc.edu Xuzhen, Cheng Chinese Academy of Agricultural Sciences China email: chengxz@caas.net.cn Wang, Jianping University of Florida USA email: wangjp@ufl.edu Yadav, Aneeta GB Pant University of Agriculture & Technology India email: yadav.aneeta@gmail.com Wany, Aakanksha Birla Institute of Technology India email: aakanksha.wany@yahoo.com Yadav, Pardeep CCSHAU India email: pardeep4yadav@gmail.com Warkentin, Tom University of Saskatchewan Canada email: tom.warkentin@usask.ca Yadav, Reena JNU India email: reena.siya@gmail.com Weeden, Norman F Montana State University USA email: nweeden@montana.edu Yamini, KN ANGRAU India email: yaminikn@yahoo.com Widders, Irvin E Michigan State University USA email: widders@anr.msu.edu Yang, Shi Ying The University of Adelaide Australia email: shi.y.yang@adelaide.edu.au VI ICLGG – List of Participants Yasin, Mohammad RVSKVV India email: myasin23@gmail.com Yoon, Min Seoul National University Korea email: rainfan1013@snu.ac.kr Zargar, Sajad SKUAST-Jammu India email: smzargar@rediffmail.com Zhang, Xinyou Henan Academy of Agricultural Sciences China email: haasz@sohu.com Zhuang, Weijian Fujian Agriculture and Forestry University China email: weijianz1@163.com Zulhedi, Yuliasti Center for Application of Isotopes and Radiation Technology, Indonesia email: upikyuliasti@yahoo.com VI ICLGG – List of Participants Thanks to...... 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