Enhancing the Efficiency of Induced Mutagenesis Through an Integrated Biotechnology
Crops Pathology and Genetics Research
2012 Annual Report
1a.Objectives (from AD-416):
The SY will serve in a technical advisor role to this IAEA-sponsored Coordinated Research Project (CRP) to share expertise in the area of rice genomics and induced mutagenesis with research contract holders from developing countries.
1b.Approach (from AD-416):
Involves participation in Research Coordination Meetings at the IAEA, Vienna Austria by presenting and discussing ongoing research in the SY's lab involving the development and analysis of rice mutants.
The agreement was established in support of objectives 1 and 2 of the in-house project, the goal being to develop improved rice (Oryza sativa L.) germplasm for use in breeding elite varieties adapted to temperate environments by identifying, characterizing, and manipulating genes that affect crop productivity. The goal of this project involves participation in Research Coordination Meetings at the International Atomic Energy Agency in Vienna, Austria by presenting and discussing ongoing research in the Scientist’s laboratory involving the development and analysis of rice mutants.
During the past year, a manuscript describing the application of reduced representation sequencing method called restriction enzyme site comparative analysis (RESCAN) to evaluate mutation density/load and thereby reduce the cost of identifying mutant populations with sufficient mutation densities to support efficient application of reverse genetics methods such as Targeting of Induced Local Lesions in Genomes (TILLING) was published and also disseminated among cooperators in this project. A second reduced representation sequencing approach (i.e. exome sequencing) is being evaluated in cooperation with researchers at University of California, Davis as a means to characterize mutants analogous to sequencing flanking regions of insertional mutations. RESCAN or exome sequencing will be employed to examine the genetically effective cell number (GECN) in rice by analyzing M2 plants derived from the same and different panicles from the same M1 plant. M2 from M1 derived by chemical and gaama-irradiation mutagenesis will be assessed to determine if the mutagen or mutagenic treatment protocol affects the GECN. M3 seeds were produced from over 1,000 independent M1 that were derived from sodium azide mutagenesis of seed of the very early-maturing rice variety Kitaake. A number of visible mutant phenotypes have been observed and additional phenotyping of M3 seeds and plants will be performed during the project period. M4 seeds are expected by the end of 2012.