节节麦HWM-GS D^tx5与普通小麦HWM-GS Dx5亚基的AS-PCR鉴别  

AS-PCR for Distinguishing HMW-GS D^tx5 from Aegilops tauschii and Dx5 from Common Wheat

作　　者：曲继鹏[1] 温雯[2] 李俊[3] 郑建敏[3] 杨武云[3] 万洪深[3] 

机构地区：[1]西昌学院,西昌615013 [2]农业部科技发展中心,北京100122 [3]四川省农业科学院作物研究所农业部西南地区小麦生物学与遗传育种重点实验室,成都610066

出　　处：《分子植物育种》2017年第10期4024-4032,共9页Molecular Plant Breeding

基　　金：国家重点研发项目(2016YFD0100102);国家自然科学基金(31401383);优质专用小麦新品种培育四川省青年科技创新研究团队(2014TD0014)共同资助

摘　　要：高分子量谷蛋白(HMW-GS)是小麦的重要储藏蛋白之一,决定小麦烘烤品质,其中HMW-GS Glu-D1位点与面包烘烤品质的关系最为密切。人工合成小麦(synthetic hexaploid wheat,SHW)继承了节节麦D基因组丰富的遗传变异,是高效利用野生祖先种优良基因的重要桥梁资源;近年来随着人工合成小麦在小麦遗传育种中的不断应用,来自节节麦类群的大量高分子量谷蛋白亚基类型越来越多的涌向普通小麦。而传统SDS-PAGE无法区分节节麦HMW-GS Dtx5和普通小麦Dx5亚基,鉴于此本研究利用节节麦HMW-GS Dtx5亚基和普通小麦HMW-GS Dx5亚基DNA序列之间存在的几个SNPs差异开发出等位特异PCR(allele specific polymerase chain reaction,AS-PCR)引物,通过PCR方法来区分两种不同来源的5亚基。结果显示,所设计出的两对引物都能够扩增出清晰稳定的目标条带,并且两对引物的扩增结果一致,含HMW-GS Dx5亚基的表现为阳性带,含节节麦来源的Dtx5亚基表现为阴性;同源性分析表明Dtx5亚基与Dx2亚基氨基酸序列的同源性要高于Dtx5与Dx5之间的同源性。High-molecular-weight（HMW） glutenin, one of the important storage proteins of common wheat,confers processing quality of wheat grain. Among the loci associated with HMW-GS, the locus D1 determines the breaking quality of bread-wheat dough mostly. Synthetic hexaploid wheat（SHW） inherits the abundant genetic diversity of the D genome from Aegilops tauschii, and has gradually been used as a bridge-tool for common wheat improvement. Therefore, more and more HMW-GS subunit types of Ae. tauschii have been introgressed into the common wheat with the irresistible application of SHW in common wheat breeding. However, the traditional SDS-PAGE couldn＇t distinguish Dtx5 from Ae. tauschii and Dx5 from common wheat. In this study, we developed two pairs of AS-PCR primers, based on three SNPs in the HMW-GS D1 sequence of Ae. tauschii and common wheat. The sharp and stable fragments amplified by designed AS-PCR primers indicated their availability and convenience in distinguishing Dtx5 and Dx5. And Dx5 from common wheat obtained the amplified fragments by AS-PCR primers, while no fragment was amplified for HMW-GS Dtx5 from Ae. tauschii. And the amplified results were consistent in both pairs of AS-PCR primers. Moreover, Dtx5 is more highly homologous with Dx2 than Dx5 in the derived amino acid sequence.

关 键 词：节节麦 普通小麦 Dtx5亚基 Dx5亚基 AS-PCR 

分 类 号：S512.1[农业科学—作物学]