Update:
This FY21 Meal Protein project is based on the progress made during our FY20 project. This is the Q1 report (Oct – Dec, 2020), almost all soybean fields planted for this project have been harvested and cooperators from all states reported a good harvest. Despite the extra-ordinary challenges due to COVID-19, we are on target on our proposed research activities. Data collection and data analyses for the 2020 soybean crop is still on-going. We hope to report the results in our March report. Our progress from FY20 project is presented below:

We report excellent progress in all areas in this project in FY20, including the release of nine high yielding cultivars/germplasm with high-meal protein and/or high genetic diversity, thus exceeding our target for FY20. Twelve papers were published on research results in refereed journals and many presentations were made in professional and stakeholder meetings.

Key Activities:
Development of U.S. adapted high-yielding germplasm with >48% meal protein, =36% seed protein and >95% check yield and test environmental stability of seed protein
Cultivar/germplasm releases: We released nine new lines during FY20, in addition to eight lines released during FY19 that meet the targeted criteria above (Table 1). Each of the lines was tested for stability of seed protein and seed yield in more than 20 environments across several states and years.
Private Company Outreach: Several of our recent releases have been used by private companies. Highpro1 (Mian), E11128T (Wang), S14-9653 (Scaboo), S09-13185 (Chen), and M10-2017102 (Lorenz) are some of the lines being used by the private seed companies. Additionally, two advanced breeding lines M13-264022 and M10-314001 developed by Lorenz at UMN was transferred to two private companies for evaluation via MTA. Seed of the high protein variety TN15-5007 is being increased by Tennessee Advanced Genetics, Inc. and will be available for sale to farmers in winter/spring 2021.
Table 1. Released lines in FY20
Line name MG Yield Protein Oil Meal Protein Exotic Pedigree breeder/state
%check 13% M % %
M11-314101 I 96 37.0 18.3 49 - Lorenz, MN
E15079T II 98 37.0 19.0 50 - Wang, MI
HM15-H006 II 95 37.5 18.2 49.8 - McHale, OH
HM16-M019 III 95 35.8 19.4 48.2 - McHale, OH
S16-16641R IV 96 38.0 18.0 50 - Chen, MO
TN14-4304 IV 100 36.1 19.2 48 - Pantalone, TN
S09-13185C V 100 37.7 18.5 50 - Chen, MO
USDA-N6005 VI 100 36.9 18.3 49 25 Mian, NC
G11-7103 VII 98 38 19 50 50 Li, GA

At least 6 additional lines in MG 0-VIII will be ready for release by during FY 2021. We also have robust pipelines of breeding materials in all maturity groups (MG0-VII) from F1 hybrids to advanced breading lines that have been tested for yield in multiple environments for at least 2-3 years.
Activity. Develop U.S. adapted lines from G. max x G. soja crosses producing =36% seed protein and >48% meal protein
High performing lines derived from wild soybean: From the USDA Regional trials, we identified 2 breeding lines, N14- 8537 (MG7) and N16-9198 (MG7), that have elevated protein and similar yields of the checks. Based on the summary result over locations from 2019, N14- 8537 showed 37.4% and 17.8% protein and oil content respectively (data represented at 13% moist). N16-9198 showed elevated protein content of 38.4% however, the oil content was around 16.8%. this line also showed higher Meal value of 50.1% which was higher or similar to the tested checks with the same maturity group. The yield value of N14-8537 and N16-9198 was 37 and 43 bu/ac respectively. Both lines have 25% pedigree derived from wild soybean PI 366122. In 2020, we also yield tested hundreds of new breeding lines derived from wild soybean in local yield trials. Several lines with 25-50% G. soja backgrounds by pedigree producing more than 90% check yield and >48% meal protein will be ready for release during FY21. These are in addition to 17 lines with 50% soja backgrounds that were released during FY19.
Activity. Map and characterize QTL alleles/genes responsible for high protein in domestic and exotic cultivated and wild soybean and/or determine their impact on seed yield
Mapping of protein genes/quantitative trait loci: several novel Quantitative Trait Loci for protein have been identified and mapping in additional 12 G. max cultivated x G. max PI populations are being completed. At least 10 new QTLs were reported by the group in FY20. Additionally, mapping in 10 G. max x G. soja (domesticated x wild) RIL populations (~2600 progeny) are also underway (Drs. Song, Taliercio, Carter and Mian). Mapping in these populations should be completed in FY21. Dr. Li has identified a fast neutron deletion on chrom 12 that increases protein by 3-4% without reducing oil or yield. A DNA tag for this mutation has been developed.

Activity. Map amino acid QTL alleles in cultivated and wild soybean
Mapping of amino acid QTL from G. soja (Scaboo, Taliercio et al.): Mapping of meaningful QTL for amino acids in max x max crosses has been challenging, due to the small genetic variation for individual amino acids. However, some successes were achieved in mapping larger effect amino acid QTL in some max x soja mapping populations.
Activity. Identify and characterize QTL/genes for environmentally stable high protein
QTL mapping of the environmentally stable high protein:
The Nguyen Lab, MU, identified and confirmed several soybean PIs with environmentally stable high protein. Among these, two PI 555396 and PI 567425B were crossed with MG I elite lines to develop recombinant inbred lines (RIL) mapping populations, which were grown in replicated field experiments in summer, 2020, at two locations, the Bradford Farm, Columbia, MU, and the Experimental Farm, MSU, in collaboration with Dr. D. Wang. Protein/oil content assays for phenotypic data and Soy6KSNP chip-based genotyping will be conducted for preliminary genetic analysis.
Identification of temperature-regulated genes:
The Nguyen Lab completed RNA-Seq experiments in controlled conditions of growth chambers and completed WGRS analysis of six soybean genotypes. Using an integrative approach, we completed bioinformatics analysis of RNA-Seq transcriptome and WGRS datasets in collaboration with the bioinformatics laboratory of the University of Missouri. We identified 21 candidate genes that are located in different genomic loci and potentially involved in the regulation of protein stability. We initiated gene expression analysis, based on RNA-Seq profiles of cv. Forrest, in conjunction with haplotype analysis using the existing WGRS datasets of the 106 set, leading to association analysis to identify SNPs/genes significantly associated with high protein phenotypes. We plan to further investigate functionality of these genes using genome editing technology.
Activity. Optimize and test molecular markers for confirmed major high protein Danbaekkong or BARC-7 allele on chromosome 20 and assess the impacts of these allele on seed yield
Two major protein QTL on chromosome 20: Significant progress has been made through fine mapping to delineate Danbaekkong and BARC-7 high protein alleles on chromosome 20, we now have data to support that DAN and BARC-7 high protein alleles are different from each other. DNA markers for both of these alleles are now available for marker assisted breeding.
Development and testing of near isogenic lines (NILs) for yield comparison and fine mapping of high protein genes: The question of the yield drag from any major high protein gene looms large due the general negative relationship of protein with seed yield in soybean. So, our group has been developing the genetic materials needed to answer this very important question for at least two major seed protein genes on chromosome 20 mapped from Danbaekkong and BARC-7 sources. We have developed and tested a MG-VII NILs pair in ‘Benning’ background and demonstrated that the Danbaekkong high protein allele does not reduce the yield in this background while increasing seed protein by >3%. We have developed NILs with the same allele in MG-III ‘Wyandot’ background which are currently being tested for yield in IA, MI, and OH. NILs for the Danbaekkong and BARC-7 alleles have also been developed in MG-IV, V, and VI and should be ready for yield testing during the FY21-23 duration.
Activity: Incorporate high (>75%) oleic acid into high yielding lines with >36% seed and >48% meal protein
Crosses were made in several maturity groups to combine high protein with high oleic in the same lines. We currently have F2-F3 seeds that are being advanced by single seed descent.

The Annual Meeting: of the project was held on February 17 and 18, 2020 in Chesterfield, MO with excellent support from USB. All Co-PIs, joined the meeting. Additionally, USB Meal Project Manager, Lisa Weaver; Director of Supply Programs, Jennifer Jones; and Vice President, Keenan McRoberts from SmithBucklin joined the meeting. Michael Kidd and Dr. Sam Rochell from University of Arkansas and Scott Raubenstine from Purdue Agribusiness made presentations via ZOOM on importance of soymeal for poultry. The meeting was a great success and significant progress on all objectives were reported by the researchers in the group.