Wang C S, Todd J J, Vodkin L O
Department of Agronomy, University of Illinois, Urbana 61801.
Plant Physiol. 1994 Jun;105(2):739-48. doi: 10.1104/pp.105.2.739.
The seed of all wild Glycine accessions have black or brown pigments because of the homozygous recessive i allele in combination with alleles at the R and T loci. In contrast, nearly all commercial soybean (Glycine max) varieties are yellow due to the presence of a dominant allele of the I locus (either I or i) that inhibits pigmentation in the seed coats. Spontaneous mutations to the recessive i allele occur in these varieties and result in pigmented seed coats. We have isolated a clone for a soybean dihydroflavonol reductase (DFR) gene using polymerase chain reaction. We examined expression of DFR and two other genes of the flavonoid pathway during soybean seed coat development in a series of near-isogenic isolines that vary in pigmentation as specified by combinations of alleles of the I, R, and T loci. The expression of phenylalanine ammonia-lyase and DFR mRNAs was similar in all of the gene combinations at each stage of seed coat development. In contrast, chalcone synthase (CHS) mRNA was barely detectable at all stages of development in seed coats that carry the dominant I allele that results in yellow seed coats. CHS activity in yellow seed coats (I) was also 7- to 10-fold less than in the pigmented seed coats that have the homozygous recessive i allele. It appears that the dominant I allele results in reduction of CHS mRNA, leading to reduction of CHS activity as the basis for inhibition of anthocyanin and proanthocyanin synthesis in soybean seed coats. A further connection between CHS and the I locus is indicated by the occurrence of multiple restriction site polymorphisms in genomic DNA blots of the CHS gene family in near-isogenic lines containing alleles of the I locus.
由于纯合隐性i等位基因与R和T位点的等位基因组合,所有野生大豆种质的种子都有黑色或棕色色素。相比之下,几乎所有商业大豆(Glycine max)品种都是黄色的,这是因为I位点存在一个显性等位基因(I或i),该等位基因抑制种皮色素沉着。这些品种中会发生向隐性i等位基因的自发突变,导致种皮有色素沉着。我们使用聚合酶链反应分离出了一个大豆二氢黄酮醇还原酶(DFR)基因的克隆。我们在一系列近等基因系中研究了DFR以及类黄酮途径的其他两个基因在大豆种皮发育过程中的表达情况,这些近等基因系的色素沉着因I、R和T位点等位基因的组合而有所不同。在种皮发育的每个阶段,所有基因组合中苯丙氨酸解氨酶和DFR mRNA的表达都相似。相比之下,在携带导致黄色种皮的显性I等位基因的种皮发育的所有阶段,查尔酮合酶(CHS)mRNA几乎检测不到。黄色种皮(I)中的CHS活性也比具有纯合隐性i等位基因的有色素种皮低7至10倍。看来显性I等位基因导致CHS mRNA减少,进而导致CHS活性降低,这是抑制大豆种皮中花青素和原花青素合成的基础。在含有I位点等位基因的近等基因系中,CHS基因家族的基因组DNA印迹中出现多个限制性位点多态性,这表明CHS与I位点之间存在进一步的联系。