Single-base deletion in GmCHR5 increases the genistein-to-daidzein ratio in soybean seed

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  • Sarkar Md. Abdur Rauf
    The United Graduate School of Agricultural Sciences, Kagoshima University Faculty of Agriculture, Saga University Department of Genetic Engineering and Biotechnology, Faculty of Biological Science and Technology, Jashore University of Science and Technology
  • Otsu Wakana
    Faculty of Agriculture, Saga University
  • Suzuki Akihiro
    The United Graduate School of Agricultural Sciences, Kagoshima University Faculty of Agriculture, Saga University
  • Hashimoto Fumio
    The United Graduate School of Agricultural Sciences, Kagoshima University
  • Anai Toyoaki
    The United Graduate School of Agricultural Sciences, Kagoshima University Faculty of Agriculture, Saga University
  • Watanabe Satoshi
    The United Graduate School of Agricultural Sciences, Kagoshima University Faculty of Agriculture, Saga University

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  • Single-base deletion in <i>GmCHR5</i> increases the genistein-to-daidzein ratio in soybean seed

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<p>Novel mutant alleles related to isoflavone content are useful for breeding programs to improve the disease resistance and nutritional content of soybean. However, identification of mutant alleles from high-density mutant libraries is expensive and time-consuming because soybean has a large, complicated genome. Here, we identified the gene responsible for increased genistein-to-daidzein ratio in seed of the mutant line F333ES017D9. For this purpose, we used a time- and cost-effective approach based on selective genotyping of a small number of F2 plants showing the mutant phenotype with nearest-neighboring-nucleotide substitution–high-resolution melting analysis markers, followed by alignment of short reads obtained by next-generation sequencing analysis with the identified locus. In the mutant line, GmCHR5 harbored a single-base deletion that caused a change in the substrate flow in the isoflavone biosynthetic pathway towards genistein. Mutated GmCHR5 was expressed at a lower level during seed development than wild-type GmCHR5. Ectopic overexpression of GmCHR5 increased the production of daidzein derivatives in both the wild-type and mutant plants. The present strategy will be useful for accelerating identification of mutant alleles responsible for traits of interest in agronomically important crops.</p>

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