URGV, INRA,2 Rue Gaston CrémieuxCP 5708, 91 057 Evry, France Mini-biographyI obtained my Master degree in plant science from the university of Paris XI, France and Ph.D. from the university of East Anglia, Norwich, UK. Currently I m heading the Crop Functional Genomics team, URGV, Evry. The major goal of CFG team is to develop and apply modern genomics approaches to study crop plants. The most significant achievement of the last three years is the development of a HTP pea TILLING platform and the construction of a pea BAC library of more than 400 000 clones. These resources are being used to isolate genes of agronomic importance from pea. I am also currently involved in GL-TTP through my commitment to the GLIP and the desire to make the results of the pea TILLING platform available for exploitation by the broad user community. As member of GL-TTP Council, I am strongly supporting the development of the GL-TTP in forging strong links between research and industry.
TILLING to validate gene function and generate new genetic resources for pea
Abdel Bendahmane
URGV, 2 Rue Gaston Crémieux, CP 5708, 91 057 EVRY Cedex, FRANCE
bendahm@evry.inra.frA limiting step in the characterisation of DNA sequences derived from genome sequencing projects as well as genes identified in transcriptome projects is the ability to study the function of these genes in a cellular context to relate the gene sequence to a phenotype. Such studies cannot be carried out only in model plants in which some agronomic characters cannot be characterised.
To bridge the gap between genomics and functional genomics in pea, we developed a high throughput gene knockout system as a reverse genetics tool. This system is based on the TILLING concept. This strategy has many advantages; first it permits the identification of mutations in target genes without the production of genetically modified organisms that cause public concern. Second, it can be automated in a high throughput system. Third, it is an efficient way to identify mutants in a specific gene with a leaky phenotype and consequently with a higher potential agronomic value.
Three distinct research areas were at the core of our project: (1) the production and management of large collections of chemically mutagenised populations, (2) the development of HTP tools for rapid and systematic identification of mutations in target sequences and (3) the creation of interactive and evolving databases.
The pea-TILLING platform, PEATIL, exploit a mutant population of more than 8000 M2 families derived from cultivar Terese and 4700 M2 families derived from cultivar Cameor. The phenotypes segregating in the mutant population were recorded and stored in a public database that will be soon accessible by the web. Concerning the screening methods, a main improvement was achieved in the sensitivity of the detection system. One allele in pools of thirty was reproducibly detected without any loss in sensitivity.
As a proof of concept a list of agronomic traits were selected for TILLING. These traits have an impact on functional properties of the harvested plants for use in food, animal feed, or industrial products. Examples of these agronomic traits includes allergenicity and indigestibility of pea proteins, increase in biomass, enhanced disease resistance, amino acids compositions, flowering time, cold tolerance, plant architecture, etc. The PEATIL platform is open to the scientific community for screening.
