Molecular Characterization of a HMW Glutenin Subunit Allele Providing Evidence for Silencing of x-type Gene on Glu-B1

YANG Zu-Jun , LI Guang-Rong , LIU Chang , FENG Juan , ZHOU Jian-Ping , REN Zheng-Long
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引用次数: 10

Abstract

Understanding the molecular structure of high-molecular-weight glutenin subunit (HMW-GS) may provide useful evidence for the study on the improvement of quality of cultivated wheat and the evolution of Glu-1 alleles. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) shows that the subunits encoded by Glu-B1 were null, named 1Bxm, in a Triticum turgidum var. dicoccoides line PI94640. Primers based on the conserved regions in wheat HMW-GS gene promoter and coding sequences were used to amplify the genomic DNA of line PI94640. The PCR products were sequenced, and the total nucleotide sequence of 3 442 bp including upstream sequence of 1 070 bp was obtained. Compared with the reported gene sequences of Glu-1Bx alleles, the promoter region of the Glu-1Bxm showed close resemblance to 1Bx7. The Glu-1Bxm coding region differs from the other Glu-1Bx alleles for a deduced mature protein with only 212 residues, and a stop codon (TAA) at 637 bp downstream from the start codon was present, which was probably responsible for the silencing of x-type subunit genes at the Glu-B1 locus. Phylogenetic tree based on the nucleotide sequence alignment of HMW glutenin subunit genes showed that 1Bxm was the most ancient type of Glu-B1 alleles, suggesting that the evolution rates are different among Glu-1Bx genes. Further study on the contribution of the unique silenced Glu-B1 alleles to quality improvement was also discussed.

一个HMW谷蛋白亚基等位基因的分子特征为Glu-B1上x型基因的沉默提供证据
了解高分子量谷蛋白亚基(hwh - gs)的分子结构可以为研究栽培小麦品质的改良和谷蛋白1等位基因的进化提供有益的依据。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)结果表明,在一株小麦(Triticum turgidum var. dicoccoides) PI94640中,Glu-B1编码的亚基为零,命名为1Bxm。以小麦HMW-GS基因启动子保守区和编码序列为引物,扩增小麦PI94640的基因组DNA。对PCR产物进行测序,得到总核苷酸序列为3 442 bp,其中上游序列为1 070 bp。与已报道的Glu-1Bx等位基因序列比较,Glu-1Bxm的启动子区域与1Bx7非常相似。与其他Glu-1Bxm等位基因不同的是,Glu-1Bxm编码区只有212个残基,并且在起始密码子下游637 bp处存在一个停止密码子(TAA),这可能是导致Glu-B1位点x型亚基基因沉默的原因。基于HMW谷蛋白亚基基因核苷酸序列比对的系统发育树显示,1Bxm是最古老的Glu-B1等位基因类型,表明Glu-1Bx基因之间的进化速率存在差异。进一步探讨了独特的Glu-B1沉默等位基因对品质改善的贡献。
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