Project: Biotechnology tools for improving disease resistance and seed quality in legumes

At present, where sustainable agriculture and a reduced utilisation of intrants is needed, protein legumes (i.e. large-seeded, protein-rich legumes) have an important role to play, due to their capacity to fixate atmospheric nitrogen, requiring little fertilization. However, widening of their cultivation is handicapped by the reduced stability of yields from one year to the next and a lack of genotypes capable of resisting various biotic and/or abiotic stresses, while their acceptability by the cattle feed industry is limited by their reduced protein content/quality compared to cereals. These factors result in a limited availability of options based on protein legumes for agriculture. An analysis of the present economical situation for legumes in Europe and worldwide, produced during the recent FP6 GLIP project is annexed to this form._x000D_Both UMRLEG and AGRITEC, carry out research in protein legumes aimed at resolving these problems since several years, both being interested in developing in vitro biotechnology approaches to widen their genetic variability, be it by interspecific hybridisations, through induction of somaclonal variations followed by selection in vitro, generating doubled haploid populations to fixate genes and for gene mapping, or still by gene transfer for validation of function of genes identified. _x000D_This led us to identify genetic transformation for gene function validation and seed and embryo morphogenesis and development as the CO areas of common interest for both Ps. We have been collaborating, formally and informally on these for a number of years. _x000D_The CO goal of this collaboration is hence to add our complementary expertise to exploit in vitro biotechnology for a widening of the genetic variability in grain legumes._x000D_Within the context of gene transfer, legumes have been quite recalcitrant so far, with responses that were strongly dependent on species and, even, on genotype studied. _x000D_AGRITEC has a long experience in pea tissue culture, from establishment of callus and cell suspensions to regeneration and isoenzymatic characterization of plants produced, and is already successfully using gene transfer to improve disease and insect resistance in pea. _x000D_At INRA, we also have a long experience in legume biotechnology, having produced transformants in the model legume Medicago truncatula with various constructs of Agrobacterium tumefaciens (both with reporter and interesting genes), and we carry out research on interspecific hybridisation, on control of recalcitrance for conversion of somatic embryos into plants, on detailed characterization of in vitro regenerants and for a comprehension of the fundamental physiological, cyto-histological and genetic mechanisms underlying embryo development and seed filling in legumes. _x000D_Experiments undertaken in this collaboration between Agritec and INRA UMRLEG will focus on the exploitation of methods for gene transfer in pea and M. truncatula, via plant regeneration from leaf explants co-cultured with Agrobacterium tumefaciens constructs carrying genes of interest for new disease resistance traits, and for the control of embryo development and seed filling, important on yield. _x000D_Work will concern in parallel the two species and, for pea, both Czech and French varieties willl be used. All plants obtained will be characterized in terms of phenotype (in planta, in situ cyto-histology), by biochemical methods (isoenzymes), by molecular (RAPD, RFLP) and cytomic approaches (flow cytometry). Particular attention will be paid to the analysis of the stable incorporation and expression of the transgenes in the primary transformants and also their progeny, assessed up to T2-T3 generations. For this, molecular (Southern Blot, PCR) and flow cytometry analyses in particular will be applied, and generation cycles will be accelerated in vitro in order to save time. All experiments involving GMO materials will be conducted with full respect of the governmental regulations in force in both countries. _x000D__x000D_During the nearly three years of this collaboration, both Ps will exchange their respective methods. It is expected that, by the end of this collaboration results of interest will have been produced at three levels :_x000D_- Economic : by the production of novel genotypes of pea with interesting characters that may be susceptible of being used as the baseline genitors for new cultivars or even per se) ;_x000D_- Fundamental : with the acquisition of a better knowledge of the function of key genes in the resistance of legume plants to disease and on the control of seed filling, of capital importance for yield ; and,_x000D_- European : through the formalisation of a collaboration between a private company and a public institute, belonging to two different countries, working on the same genotypes, sharing their expertise and thereby contributing to the breeding of better legumes for a more sustainable agriculture in Europe._x000D_

Acronym	PEASTAR (Reference Number: 4770)
Duration	01/04/2009 - 31/12/2011
Project Topic	The content of this project is the development of transgenic pea lines with improved resistance to fungal pathogens and insect pests, and increased seed yield. Resulting transgenic breeding lines will have novel characters demanding by breeders, and COtaining wide market applications.
Project Results (after finalisation)	The CO results of the project are represented by: _x000D_(I) commercially exploitable plant/seed materials of pea , _x000D_(II) new biotechnological protocols adopted specifically for pea (with a possibility of their transfer/modification to other grain legume crops), _x000D_(III) new theoretical knowledge in the field of genetics and physiology of legumes. _x000D_Commercially exploitable results will be primarily finalized within the project consortium, new protocols - when published - as well as new theoretical knowledge will be on disposal for the broad scientific community. The CO results were represented namely by:_x000D_(1) Development of transgenic lines of pea (and Medicago truncatula) via Agrobacterium tumefaciens-mediated transformation protocol with constructs carrying genes for improved resistance to pests and pathogens and for improved yield. _x000D_(2) Study of key genes controlling biosynthesis and deposition of seed storage substances and thus, yield parameters in Pisum sativum and M. truncatula. _x000D_(3) Publication of results in scientific journals, presentations at international congresses as well as at the meetings of Czech/Moravian pea breeders, seed producers, growers and processors (annual general assembly of CAAS - Czech Academy of Agricultural Sciences, CMSSA - Czech Seed Trade Association and AZLP - Association of Pulse Growers and Processors CR).
Network	Eurostars
Call	Eurostars Cut-Off 2

Project partner

Number	Name	Role	Country
2	AGRITEC, Research, Breeding and Services, Ltd.	Coordinator	Czech Republic
2	Unité Mixte de Recherches en Génétique et Ecophysiologie des Légumineuses à Graine - INRA C.R. Dijon	Partner	France