Location: Plant Genetics Research
Project Number: 5070-21000-043-04-N
Project Type: Non-Funded Cooperative Agreement
Start Date: Oct 1, 2017
End Date: Mar 31, 2019
1. Track carbon partitioning with isotopes through metabolism including: gluconeogenesis, glyoxylate cycle, beta oxidation, lipid turnover, and dietary fiber-carbohydrate production pathways to discern the fate of to assess the sources of carbon for non-digestible carbohydrates. 2. In concert with labeling studies, identify the enzymatic steps responsible for production of non-digestible carbohydrates through monitoring targeted RNA and key enzyme activity levels over development. 3. Development of soybean lines with altered conversion production of non-digestible carbohydrates by targeting the suppression (through RNAi) or mutation (through characterization of FN mutant lines) of genes encoding key enzymatic steps.
Isotopic labeling pulse-chase experiments will be used to study the flow of carbon during seed dessication. Transcriptomics provided by collaborators will complement the labeling experiments. We will focus on central carbon metabolism that causes changes in carbohydrate levels. Labeling in the form of both radioactive (14C) and stable isotope (13C) experiments will be used to judge metabolic activity with quantitative analyses by liquid chromatography linked to tandem mass spectrometers. The project will result in metabolic flux maps of soybean seeds during late stages of seed development and highlight pathway operation to guide metabolic engineering efforts.