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Molecular Breeding

, Volume 32, Issue 2, pp 291–298 | Cite as

Identification of a plant introduction soybean line with genetic lesions affecting two distinct glycinin subunits and evaluation of impacts on protein content and composition

  • Won-Seok Kim
  • Jason D. Gillman
  • Hari B. Krishnan
Article

Abstract

Unlike other oilseeds, soybean (Glycine max [L.] Merr) is also valuable due to its direct conversion into human food. One notable example is the cheese-like product tofu. The quality of tofu is improved when protein subunits derived from two glycinin genes, Gy1 and Gy4, are reduced or absent. Here we report the discovery that one exotic soybean plant introduction line, PI 605781 B, has not only a previously described loss-of-expression mutation affecting one glycinin gene (gy4), but also bears an extremely rare, potentially unique, frameshift mutation in the Glycinin1 gene (gy1-a). We analyzed glycinin gene expression via qRT-PCR with mRNA from developing seeds, which revealed that the novel allele dramatically reduced Gy1 mRNA accumulation. Similarly, both A4A5B3 and A1aB1a protein subunits were absent or at undetectable levels, as determined by two-dimensional protein fractionation. Despite the reduction in glycinin content, overall seed protein levels were unaffected. The novel gy1-a allele was found to be unique to PI 605871B in a sampling of 247 diverse germplasm lines drawn from a variety of geographic origins.

Keywords

Glycinin Soybean Tofu Seed proteins 

Notes

Acknowledgments

The authors would like to acknowledge the superb technical assistance of Sungchan Jang, Jeremy Mullis and Nathan Oehrle. This research was supported by the USDA-Agricultural Research Service.

Supplementary material

11032_2013_9870_MOESM1_ESM.pptx (154 kb)
Supplementary Figure 1 Typical genotyping “melting peak” analyses for Gy1 and Gy4 SimpleProbes. Colors: Light blue line indicates negative control, dark blue indicates homozygote mutant, red indicates homozygote wild type, green indicates heterozygote. Analysis was run on a Lightcycler 480 II (Roche). (PPTX 153 kb)
11032_2013_9870_MOESM2_ESM.pptx (351 kb)
Supplementary Figure 2 One dimensional protein gel electrophoresis of lines selected via genotyping assays. 40 micrograms of seed protein were separated on 12.5 % SDS-PAGE gels. C indicates protein from ‘Patriot’ seeds and PI indicates protein from PI 605871 B seeds, numbered samples were F2:3 seeds from F2 selections for gy4 or Gy4 homozygotes identified by SimpleProbe assay (genotype is indicated at bottom of chart). * indicates either A4 or B5 glycinin subunit, encoded by the Gy4 gene. (PPTX 351 kb)
11032_2013_9870_MOESM3_ESM.xlsx (12 kb)
Supplemental Table 1 Glycinin primers used in this work (XLSX 12 kb)
11032_2013_9870_MOESM4_ESM.xlsx (33 kb)
Supplemental Table 2 Distribution of Gy1 alleles in a diverse sampling of 247 genotypes (XLSX 32 kb)

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Copyright information

© Springer Science+Business Media Dordrecht (outside the USA) 2013

Authors and Affiliations

  • Won-Seok Kim
    • 1
  • Jason D. Gillman
    • 1
  • Hari B. Krishnan
    • 1
  1. 1.United States Department of Agriculture-Agricultural Research ServiceUniversity of Missouri-ColumbiaColumbiaUSA

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