Abstract
In soybean, the W4 gene encoding dihydroflavonol-4-reductase controls anthocyanin pigment biosynthesis in flowers. The mutant allele, w4-m, is characterized by variegated flowers and was evolved from the insertion of an endogenous transposable element, Tgm9, in intron II of the W4 gene. In the w4-m mutant line, reversion of the unstable allele from variegated to normal purple flower in revertants would indicate Tgm9’s excision accompanied by its insertion into a second locus. We identified a male-sterile, female-sterile mutant from such germinal revertant bearing purple flowers. The objectives of our investigation were to map the sterility locus, identify candidate genes for the male-fertile, female-fertile phenotype, and then determine if sterility is associated with the insertion of Tgm9 in the sterility locus. We used bulked segregant analysis to map the locus to molecular linkage group J (chromosome 16). Fine mapping enabled us to flank the locus to a 62-kb region that contains only five predicted genes. One of the genes in that region, Glyma16g07850.1, codes for a helicase. A rice homolog of this gene has been shown to control crossing over and fertility phenotype. Thus, Glyma16g07850.1 is most likely the gene regulating the male and female fertility phenotype in soybean. DNA blot analysis of the segregating individuals for Tgm9 showed perfect association between sterility and the presence of the transposon. Most likely, the sterility mutation was caused by the insertion of Tgm9. The transposable element should facilitate identification of the male- and female-fertility gene. Characterization of the fertility gene will provide vital molecular insight on the reproductive biology of soybean and other plants.
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Acknowledgments
We express our sincerest thanks to Dr. Karin Bodensteiner for critically reading the manuscript, her helpful comments, and encouraging remarks. Funding has been received from the UWSP Undergraduate Education Initiative fund, UWSP Student Research Fund, and USDA, Agricultural Research Service, and USDA-NRI.
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This is a joint contribution from the Department of Biology, University of Wisconsin-Stevens Point, and the Department of Agronomy, Iowa State University. The mention of a trademark or proprietary product does not constitute a guarantee or warranty of the product by the Department of Biology, UWSP, or Iowa State University, and the use of the name by the Department of Biology, UWSP, and Iowa State University implies no approval of the product to the exclusion of others that may also be suitable.
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Supplemental Fig. S1
Heatmap of RNA-seq analysis of the five candidate genes at different developmental stages. RFKM reads per kilobase of transcript per million mapped reads (PPTX 75 kb)
Supplemental Fig. S2
DNA gel blot analysis of plants segregating for fertility and sterility phenotypes. The 5′ end of the Tgm9 transposon was used as a probe. Plants were scored for fertility and sterility and fertile plants were progeny tested to score homozygous fertile and heterozygous plants. Fertility/sterility phenotype is shown on the top of the gel. A homozygous fertile, B homozygous sterile, H heterozygous. The arrow represents a band that is missing in all the homozygous fertile plants and is always present in homozygous sterile plants. Scoring for that band is written at the bottom of the gel. B/H band present, A band missing (PPTX 62 kb)
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Raval, J., Baumbach, J., Ollhoff, A.R. et al. A candidate male-fertility female-fertility gene tagged by the soybean endogenous transposon, Tgm9. Funct Integr Genomics 13, 67–73 (2013). https://doi.org/10.1007/s10142-012-0304-1
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DOI: https://doi.org/10.1007/s10142-012-0304-1