1a.Objectives (from AD-416):
The objective of the research is to characterize two of the genes identified as differentially expressed as candidates for genes involved in regulation of soybean oil content. The long-term goal is to facilitate targeting of regulatory genes that will enhance seed oil content in U.S. soybeans.
1b.Approach (from AD-416):
1. Determine the location of candidate gene expression and action. Prepare two constructs for in situ hybridization and evaluate 30 sections of seed for transcript localization.
2. Create transgenic soybeans for evaluation of candidate gene function. Prepare two RNAi constructs containing the two candidate genes under the control of seed-specific promoters and transform two RNAi constructs into soy to generate T1 soybean seed for evaluation.
3. Evaluate transgenic plants that over-express multidrug toxin efflux (MATE) and malate dehydrogenase (MDH) genes for yield and seed quality.
4. Annotate transcripts encoding transporters, steps in carbon and nitrogen metabolism, signal transduction components, and hormone biosynthesis specific to each of 12 tissues from both NILS lines.
5. Characterize gene sub-functionalization within selected soybean gene families.
This research addresses Objective 3 of the parent project 3640-21000-028-00D: Determine genes regulating oil and protein accumulation in soybean through whole genome transcript analysis and functionally characterize gene candidates. The genome structure of some three hundred lines of soybean fast neutron mutants was analyzed by comparative genome hybridization. Data showed that deletions ranged from quite small to extremely large. We found that a significant portion of the soybean genome could be deleted and plants remained normal in growth and development.