Objective 1: Identify novel targets for increased oil that minimize the negative relationship between oil and protein.
• (FY18, FY19) Mapping of QTL for oil and oil/protein content from developed populations.
• (FY19) Confirmation of QTL for oil and oil/protein content from developed populations.
• (FY18) Development of carbon flux map for conventional soybean- 09/2018.
• (FY18,FY19) Development of carbon flux maps for selected transgenic events aiding in the identification of potential bottlenecks limiting accumulation of oil in the seed.

Objective 2: Develop and characterize germplasm with = 20.5% seed oil content (13% moisture) while maintaining protein content (= 35%) and yield.
• (FY17-FY19) Report of yield and NIR data from field trials.
• (FY17-FY19) Development and evaluation of germplasm with increased oil.
• Release of germplasm with increased oil- 03/2019 & subsequent (data will be summarized after project conclusion).

Objective 3: Develop transgenic germplasm with essential fatty acids and enhanced value added constituents
• (FY17-FY18) Creation of novel transgenic soybean that harbor gene stacks designed to simultaneously pull carbon towards oil reserves, and increase photosynthesis (carbon capture).
• (FY18-FY19) Report on traits of the aforementioned transgenic events which will include protein/oil content, along with small scale agronomic information (plot size estimates).

Objective 4: Development of near isogenic lines of increased oil loci.
• (FY19) Development of NIL germplasm in specific genotypic and MG classes based on markers.
• (FY19) Release of NILs as a community resource (releases may continue to take place following the project period).
• Report of stability of alleles across MG regions (data collected outside the timeline of this project and will be summarized after project conclusion).

Updated November 21, 2019:
Through this project, we were able to develop breeding lines with a “balanced” composition of protein and oil. From AK, one such line is in the last stage of testing and being ramped up for release as germplasm for the greater breeding community; four lines from OH require 1-2 additional years of testing, but are very close to project objectives, including one line which meets project objectives (108% ave check yield, 35.3% protein, 20.7% oil); six released lines from MO have elevated oil, nearly meeting project objectives; and 1 line from NC will be released that nearly meets project objectives. In addition, the support from USB allowed the identification of new genetic sources and key alleles that have been used in crosses to develop future germplasm. This information has been published and presented so that other breeders are aware of these resources and they can be incorporated into programs.

This program is reducing to practice the synthesis of novel co-products in soybean seed, namely the production of very long chain omega-3 fatty acids EPA (DHA in progress), high value carotenoid astaxanthin, a form of Vit E (tocotrienol) along with oil elevated in stearic and oleic acids. These novel output traits hold great potential for applications in feed, food, industrial and cosmetic industries.

A sequential protocol of extraction of soluble sugars, starch, hemicellulose, and crystalline cellulose was tested and validated in mature soybean seeds. Mature seeds, harvested in Fall 2018 from four different states were analyzed (total 220 samples). Correlation analyses identified: i) lines with higher average oil and protein content and lower concentration of total soluble sugars, and ii) lines with consistently low oligosaccharide content across locations. During this project, a mathematical model describing biomass accumulation in Thorne embryos was built, biomass accumulation rates were determined, in vitro culture conditions for embryos development were identify, and substrate uptake rates were evaluated. Each one of these accomplishments is getting us closer to pinpoint the causes of the negative correlation between oil and protein content in soybean embryos.