Progress 10/01/05 to 09/30/06
Outputs
Progress Report 4d Progress report. 4d. This report serves to document research conducted under a reimbursable agreement between ARS and the University of Washington, Seattle, WA. Additional details of research can be found in the report for the parent CRIS 5306-21000-016-00D Rice Genetics and Germplasm Enhancement for Temperate Environments. During FY 2006, plants grown from seeds of the rice cultivar Nipponbare that were treated with the chemical ethyl methane sulfonate (EMS) at three different rates: 1%, 1.25%, and 1. 5% were harvested (i.e., M2 seeds). For the 1% EMS plants, a total of 11, 882 M1 plants yielded at least one M2 seed. For the 1.25% and 1.5% EMS treatments, seeds were harvested from 1,291 and 3,017 M1 plants, respectively. Seeds from 1,026 M1 plants from the 1.5% EMS treatment were sown and tissue samples of over 800 of these plants were sent to the University of Washington for DNA extraction and TILLING analysis. Results of TILLING analysis indicates
that this 1.5% EMS population has a mutation rate of >1/500 kb of DNA which is sufficiently high enough for efficient TILLING. M2 seeds from this population and a population derived from 1 mM sodium azide/15 mM methyl nitrosourea mutagenesis developed in 2004-2005 have been planted to form the basis of a TILLING mutant population of 5,000 lines. Tissue will be harvested and sent to our cooperator who is currently in the process of moving operations to the University of California, Davis. M3 seeds will be collected from the lines and prepared for distribution via the Genetic Stocks- Oryza collection at the Dale Bumpers National Rice Research Center. Additional mutagenesis treatments using the 1 mM sodium azide/15 mM methyl nitrosourea protocol were carried out and 4,000 to 5,000 additional M1 plants will be harvested in FY 2006 for inclusion in this project.
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Progress 07/04/04 to 08/31/06
Outputs
Progress Report Objectives (from AD-416) Develop a 10,000 individual EMS population of rice (cv. Nipponbare). Provide tissue samples to Seattle and arrange for public distribution of seeds. Characterize some tilled genes in Nipponbare by simple genetic and phenotypic analyzes. Approach (from AD-416) Seeds of the cv. Nipponbare will be subjected to treatment with various chemical mutagens known to induced point mutations and small deletions in DNA. Seeds will be planted and allowed to reproduce. Single panicles from these plants will be harvested and single seed from each of these panicles will be planted. Leaf tissue will be harvested and freeze dried (i.e. preserved) from this generation of plants, which will also be allowed to produce new seeds. Freeze dried tissue from plants producing sufficient amounts of seed will be sent to our cooperators, who will extract DNA samples and analyze them for mutations in a pre-determined set of rice genes. Seeds from plants whose tissue samples yielded detectable mutations in genes of interest will be planted and the resulting plants will be used for simple genetic and phenotypic characterization. Documents Reimbursable with University of Washington/NRI. Log 24824. Significant Activities that Support Special Target Populations The reimbursable agreement with the University of Washington that funded this research project was terminated 8/31/2006.
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Progress 10/01/04 to 09/30/05
Outputs
4d Progress report. This report serves to document research conducted under a reimbursable agreement between ARS and the University of Washington, Seattle, WA. Additional details of research can be found in the report for the parent CRIS 5306-21000-016-00D Rice Genetics and Germplasm Enhancement for Temperate Environments. During FY 2005, seeds of the rice cultivar Nipponbare were treated with the chemical ethyl methane sulfonate (EMS) at three different rates: 1%, 1.25%, and 1.5%. Approximately 20,000 plants from seeds treated with 1% EMS are growing and seed will be harvested in FY 2006. For the 1.25% and 1.5% EMS treatments, approximately 2,500 and 5,000 plants are growing and seeds will be harvested in FY 2006. Seeds obtained from plants treated with chemical mutagens in FY 2004 were planted and seedlings were transplanted into the field for tissue and seed production. These include: about 700 second generation seedlings from the 1% EMS, about 2,000 from the 0.75%
EMS, about 2,000 from the 1 mM sodium azide/15 mM methyl nitrosourea, and about 4,000 from the 1 mM sodium azide/10 mM methyl nitrosourea treatments. Tissue from about 950 of the 1 mM sodium azide/15 mM methyl nitrosourea derived plants has been sampled and sent to our cooperators at the University of Washington and remaining plants will be sampled and seed harvested at the end of FY 2005 and beginning of FY 2006.
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Progress 10/01/03 to 09/30/04
Outputs
4. What were the most significant accomplishments this past year? 4D. This report serves to document research conducted under a reimbursable agreement between ARS and the University of Washington at Seattle. Additional details of research can be found in the report for the parent CRIS 5306-21000-016-00D. During FY 2004, seeds of the cultivar Nipponbare were mutagenized using two different chemical treatments, 1) ethyl methanesulfonate at 0.25% (5, 000 seeds), 0.5% (10,000 seeds), and 0.75% (5,000 seeds) and 2) a combination of sodium azide (1 mM) and methyl nitrosourea (10 mM methyl nitrosourea - 10,000 seeds, 15 mM methyl nitrosourea - 10,000 seeds). Treated seed (M1) have been germinated and plants are growing in the greenhouse and field. M2 seed will be harvested at the end of the season and an assessment of fertility will be made. Approximately 800 M2 plants derived from ethyl methanesulfonate (0.25% and 0.5%) treated M1 seed from the previous year were planted in
the field. Leaf tissue will be harvested from the M2, lyophilized and sent to cooperators for DNA extraction and TILLING and seeds (M3) will be collected at the end of the season.
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