Department of Biology, University of Wisconsin-Stevens Point, 800 Reserve Street, Stevens Point, WI, 54481, USA.
Funct Integr Genomics. 2013 Mar;13(1):67-73. doi: 10.1007/s10142-012-0304-1. Epub 2012 Nov 27.
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.
在大豆中,W4 基因编码二氢黄酮醇 4-还原酶,控制花中花色苷的生物合成。突变等位基因 w4-m 的特征是花斑,是由内源性转座元件 Tgm9 在 W4 基因的内含子 II 中的插入而进化而来的。在 w4-m 突变系中,不稳定等位基因从花斑到正常紫色花的回复突变表明 Tgm9 的切除伴随着它插入到第二个基因座。我们从带有紫色花的这种生殖性回复突变体中鉴定出一个雄性不育、雌性不育的突变体。我们研究的目的是定位不育基因座,鉴定雄性可育、雌性可育表型的候选基因,然后确定不育是否与 Tgm9 在不育基因座中的插入有关。我们使用混池分离分析将该基因座定位到分子连锁群 J(染色体 16)。精细定位使我们能够将该基因座侧翼到一个包含仅五个预测基因的 62-kb 区域。该区域中的一个基因 Glyma16g07850.1 编码一个解旋酶。该基因的水稻同源物已被证明控制交叉和育性表型。因此,Glyma16g07850.1 很可能是调节大豆雄性和雌性育性表型的基因。对分离个体进行 Tgm9 的 DNA 印迹分析显示,不育与转座子的存在之间存在完美的关联。很可能,不育突变是由 Tgm9 的插入引起的。该转座元件应有助于鉴定雄性和雌性育性基因。育性基因的特征将为大豆和其他植物的生殖生物学提供重要的分子见解。