|Porch, Timothy - Tim|
Submitted to: Journal of the American Society for Horticultural Science
Publication Type: Peer reviewed journal
Publication Acceptance Date: 6/13/2008
Publication Date: 5/1/2009
Citation: Porch, T.G., Blair, M., Lariguet, P., Galeano, C., Pankhurst, C., Broughton, W.J. 2009. Generation of a mutant population for TILLING common bean genotype BAT 93. Journal of the American Society for Horticultural Science. 134:348-355. Interpretive Summary: Common bean (Phaseolus vulgaris L.) is the principal grain legume used for direct human consumption worldwide. Therefore, it is important to develop efficient approaches for genetic analysis of common bean using cutting-edge technologies. A powerful method of genetic analysis, TILLING (targeted induced local lesions in genomes), is based on the generation of mutations at specific genes of interest in order to determine the function of the gene in the plant. This study determined the appropriate concentrations of a chemical mutagen, EMS (ethyl methane sulfonate), for the generation of a mutant population in a common bean variety, BAT 93. The mutant population will subsequently be used to study the function of specific agriculturally important genes in order to improve the quality and increase the productivity of common bean.
Technical Abstract: Common bean (Phaseolus vulgaris L.) is the principal grain legume used for direct human consumption worldwide and thus is an important target for concentrated genomics research, such as reverse genetic approaches like TILLING (targeted induced local lesions in genomes). TILLING, based on the development of a large mutagenesis population, offers a unique opportunity to perform reverse genetics in common bean due to the absence of an effective transformation protocol and thus the inability to develop transposon-based or T DNA-based mutagenesis systems. Using a range of 0-60 mM EMS (ethyl methane sulfonate) mutagen concentrations, this study determined that 40mM EMS is an appropriate concentration for the generation of a mutagenesis population in genotype BAT 93 based on overall survival, development, and yield of treated seed. EMS concentrations at higher concentrations resulted in inadequate survival rates of less than 10%. Based on TILLING results from other species, a population of 5,000 lines is estimated to be sufficient for saturation of the common bean genome. Phenotypic mutation frequencies and the isolation of targeted mutations show that the population is effective for genetic analysis.