Location: Soybean Genomics & Improvement Laboratory
Project Number: 8042-21220-261-000-D
Project Type: In-House Appropriated
Start Date: Mar 28, 2023
End Date: Mar 27, 2028
Objective:
Objective 1: Conduct research to develop high throughput proteomic and metabolomic methods that can be used to discover and characterize potentially useful compounds, such as functional nutritional and defense components of soybean and common bean.
Subobjective 1-1: Test the metabolomics platform and identify compounds that can contribute to improved protein quality traits in soybean and nutritional traits in dry beans.
Subobjective 1-2: Discover novel defense compounds induced in dry beans in response to bacterial and fungal pathogen infections and characterize metabolites of these compounds to discover molecular pathways with potentially high antibiotic efficacy and/or disease resistance.
Objective 2: Conduct research to identify, characterize and compare proteins and metabolites of soybean seeds containing high protein, high oleic, low linolenic, and modified carbohydrate profiles developed through chemical or bioengineered technology, and conventional hybridization.
Sub-objective 2: Proteomics and metabolomics approaches will be used to characterize seeds from genetically altered soybeans with improved protein, oil, and carbohydrate traits.
Approach:
For Objective 1, basic mass spectrometry instrument validation and software utilization will be performed to establish a metabolomics research platform. Validation will be performed on chemical standards, a yeast reference standard, a soybean seed reference, a bean seed reference, and other useful and relevant models for soybean and bean biology and pathology. Metabolite extraction methodology will be optimized per experiment. Targeted and non-targeted metabolomics will be performed to identify novel phytoalexins produced in beans during immune responses to halo blight bacterial infection and bean rust infection. Compounds will be identified on the basis of their chemical structure relationships to known standards. Candidate phytoalexins will be tested for efficacy in microbiology culture assays. Broader changes in bean metabolism associated with disease resistance will be evaluated by assembling metabolic network pathways. For Objective 2, we will derive proteomic and metabolomic profiles of mutated soybean lines that produce high amounts of seed protein, high oleic acid, low linolenic acid, and modified carbohydrates. The soybean lines were developed through chemical or bioengineered, and conventional hybridization. The seed composition will be measured initially by near-infrared spectroscopy. The detailed analysis will be performed using high-throughput proteomic and metabolomic mass spectrometry technology. We will 1) quantitatively measure changes in protein composition of seeds using tandem mass tag isobaric labels and mass spectrometry, 2) quantitatively measure changes in metabolites such as isoflavones (genistein, daidzein, and glycitein), sulfur-containing amino acids (methionine and cysteine), and sugars (glucose, sucrose, raffinose, and stachyose) using comparative mass spectrometry, and 3) integrate proteomics and metabolomics data by pathway analysis and link findings informatically to genotype.
