Project Details:

Discover Sources, Genes and Develop U.S. Adapted Germplasm with Improved Protein Quantity and Quality Utilizing Diversity in Cultivated and Wild soybeans (1920-152-0103-A)

Parent Project: This is the first year of this project.
Checkoff Organization:United Soybean Board
Categories:Seed composition, Breeding & genetics, Industrial use – Meal
Organization Project Code:1920-152-0103-A
Project Year:2019
Lead Principal Investigator:Rouf Mian (USDA/ARS-Ohio State University)
Co-Principal Investigators:
Benjamin Fallen (Clemson University)
Asheesh Singh (Iowa State University)
Blair Buckley (Louisiana State University)
Dechun Wang (Michigan State University)
E. Tallercio (North Carolina State University)
Leah McHale (The Ohio State University)
Leandro Mozzoni (University of Arkansas)
Zenglu Li (University of Georgia)
Aaron Lorenz (University of Minnesota)
Pengyin Chen (University of Missouri)
Henry Nguyen (University of Missouri)
Andrew Scaboo (University of Missouri)
Vince Pantalone (University of Tennessee-Institute of Agriculture)
Yong-Qiang An (USDA-ARS)
Tommy Carter (USDA-ARS)
Qijian Song (USDA-ARS)
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Keywords: protein, amino acid, seed composition, protein composition, amino acid composition, germplasm

Contributing Organizations

Funding Institutions

Information and Results

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Project Summary

Protein diversity in un-adapted exotic sources can be used to reverse the slow decline in seed protein and solve the meal protein problem of U.S. soybean. This project will discover diverse protein sources, identify and characterize protein genes, and develop molecular markers for increased protein and essential amino acids. We have made demonstrable progress in all of these areas in our previous protein project. Our research will develop U.S. adapted high-yielding high-protein lines, genetic information and molecular markers that will be useful to both private and public soybean breeders to address the low meal protein problems in future U.S. soybean cultivars.

Project Objectives

Objective 1. Explore diverse soybean germplasm, including wild soybeans, for sources of genetic diversity responsible for seed protein content

Activities 1a: Develop U.S. adapted high-yielding germplasm with 48 - 49% meal protein and =18.5 % seed oil and test environmental stability of seed protein

Activities 1b: Identify genetically diverse cultivated and wild sources of soybean with higher (>36%) protein that can produce >48% meal protein

Objective 2: Identify, confirm and characterize genes responsible for seed protein synthesis and storage

Activities for Objective 2. 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

Objective 3: Develop breeder-friendly genetic markers for targeted protein genes to assist development of high quality germplasm with high protein levels and/or increases in select amino acids

Activities 3a. 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

Activities 3b. Map amino acid QTL alleles in cultivated and wild soybean

Activities 3c. Map QTL for seed storage proteins (7s and 11s) and determine the impact of modified storage protein on amino acid

Objective 4: Deploy biotechnology tools to influence protein or amino acid changes in soybean seed

Activities 4: Use CRISPR-Cas9 genome editing and other technologies to validate the high-protein Danbaekkong allele on chromosome 20 in at least two elite soybean lines

Project Deliverables

Quarterly reports to USB on the project in December, March, June and Sept of each year. Specific outputs and deliverables for 2019 are as follows:

Objective 1.
• Release at least five germplasm lines with =48% meal protein, =18.5% seed oil and =95% seed yield of elite checks
• Enter at least 20 lines with =48% meal protein and =18.5% seed oil in USDA Uniform Tests
• Identify and/or characterize new diverse sources of seed protein
• Finger print >12 lines (that are candidate for germplasm release) with 50K or 6K SNP chips
Objective 2.
• Map protein QTL in at least six bi-parental and two GWAS populations
• Further characterize protein QTL on chromosome 20, 10 and 5
• Develop breeder-friendly markers for high-protein alleles at more than one major protein QTL
Objective 3:
• Map new QTL for Sulphur-containing amino acids in wild soybean
• Map QTL amino acid in cultivated soybean
• Knowledge on usefulness and contribution of 7S and 11S protein sub-units on essential amino acid concentration in soy protein
Objective 4:
• Validate the Danbaekkong allele on chromosome 20 via gene editing technology

Progress of Work

Updated May 1, 2020:
The protein content of U.S. commercial soybean seed has been declining for decades and is on a path to decline to 33.7% (45.6% meal protein) by 2030 (USB VTF, 2015). Decades of commercial breeding focused on increased yield with little attention to protein has resulted in decreased protein in U.S. soybean. The lower protein of soybean puts U.S. growers at a disadvantage in the global marketplace. Although increased yield is associated with reduced seed protein, the negative correlation between the two is not absolute. Scientific evidence shows that soybean breeders have the opportunity for meaningful (2-3%) increases in seed protein without loss of yield. This opportunity is due in large part to the treasure trove of genetic diversity in seed protein that exists in the exotic soybean accessions preserved in the USDA collection. Current ‘on farm’ soybean varieties trace their ancestry to 17 of the 20,000 ancestors in the collection, revealing a very narrow genetic base.

With grant supports from United Soybean Board, our national team of 17 public sector soybean breeders and genomic scientists from 10 major soybean producing states has recently (since 2017) started a coordinated research initiative to solve these major challenges to the US soybean industry. We are achieving success through germplasm selection, discovery and utilization of genes controlling seed protein synthesis and storage, innovative breeding technologies, and new genetic and genomic tools. Our national team with advanced breeding and gene discovery pipelines is producing novel high-yielding cultivars/germplasm with improved protein.

Below are few of the highlights of our progress since 2018.

Cultivar/germplasm releases:
• Several releases were made by our group in 2018-19: M10-2017102, E11128T, S14-9653, S09-13185, R11-7999, and N10-711.
• Several more breeding lines will be released by 2020. Additionally, we have many advanced breeding lines with better yield performance potential than the current batch of releases.

Mapping of protein genes/quantitative trait loci:
• Several new Quantitative Trait Loci (QTL) for protein have been identified and mapping in additional 12 populations are progressing on schedule. Dr. Li has identified a fast neutron deletion on chrom 12 that increase protein by 3-4% without reducing oil or yield. If this data hold up this will be a great noble high protein source.
• Significant progress has been made 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.

High-protein high-meal protein lines derived from wild soybean:
• 17 lines with 50% G. soja pedigree was recently published in Crop Science Journal (in press).
• N14-8537, a MGVIII line with 25% wild soybean (PI366122) has 100% yield of check cultivars and 50% meal protein its being releases as a germplasm.
• Another MG VII line, N16-9198 with 25% wild soybean (PI366122) also has competitive yield with elite checks with 50% meal protein.

Final Project Results

Updated May 1, 2020:
[Participants: Drs. Lorenz, UMN; Wang, MSU; McHale, The OSU; Singh, ISU; Chen, Nguyen, and Scaboo, Mizzou; Mozzoni, UARK; Pantalone, UT; Li, UGA; Carter, Taliercio, Song, AN, and Mian, USDA-ARS; and Fallen, Clemson Univ.]

Soybean meal accounts for 60-70% of the total value of soybean. The meal protein contents of recent commercial soybean cultivars in the U.S. have dipped below the minimum of 47.5%, needed for the national and international soybean meal markets and are reflective of a long-term pattern of slow decline in seed protein content. This is a major concern of U.S. soybean growers, who want to reverse this trend. The negative correlations of seed protein with seed yield and oil have been major constraints to the development of soybean lines with high meal protein without reduction in yield or oil. Another major concern is that U.S. varieties trace to a very narrow genetic base with low seed protein. This project aims to develop U.S. adapted soybean which has improved meal protein content, amino acid composition, and high-yield by expanding the genetic diversity. Variety development is supported by discovering new accessions, genes, and molecular markers for increased protein and essential amino acids utilizing the genetic diversity in cultivated and wild soybean. We report remarkable progress in all areas in this project in FY19, including the release of 7 high yielding cultivars/germplasm with high-meal protein and/or high genetic diversity.

Cultivar/germplasm releases: seven high yielding germplasm/cultivars with >48% meal protein and/or high genetic diversity were released/registered in FY19. Each of the lines was tested for stability of seed protein and seed yield in more than 20 environments across several states and years.

New sources of protein diversity were identified and characterized. Several high-protein lines were developed via fast neutron and characterized. Several exotic cultivated soybean lines with stable protein contents under cold and warm environments were identified and characterized. New QTL for high-protein and sulfur containing amino acids were mapped and markers for marker assisted breeding were developed.

Two separate studies indicated that seed protein contents can be increased by 2-3% using the Danbaekkong high-protein allele on chromosome 20 without compromising seed yield.

The cultivars/germplasm released by our group can be requested from the respective scientists for breeding and experimental purposes through appropriate MTAs.

Fourteen papers were published on our research results in refereed journals and many presentations were made in professional and stakeholder meetings in FY19.

Benefit to Soybean Farmers

In time, soybean varieties with more protein will be available to growers. This soybean will produce a superior meal that will be in demand by animal producers for inclusion in poultry and swine feed.

Performance Metrics

• Private and public soybean breeders/scientists agree to evaluate the high protein germplasm released from this research and consider them as potential parents for their crosses to develop commercial high yielding, high protein soybean varieties.
• Poultry and swine production companies show interest in our germplasm to evaluate in their own feeding trials.
• High protein and amino acid QTL are discovered and DNA markers are developed for the seed composition traits identified in this project used by private and public soybean breeding programs for research and product development.

Project Years