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 glutenin (HMW-GS) is one of the most important storage proteins in wheat and determines the baking quality of wheat. The HMW-GS Glu-D1 locus is most closely related to bread baking quality. Synthetic hexaploid wheat (SHW) inherits abundant genetic variation of D genome and is an important bridge resource for efficient use of the excellent gene of wild ancestral species. With the continuous application of synthetic wheat in wheat breeding in recent years , A large number of high-molecular-weight glutenin subunit types from Arthropod populations grow more and more into common wheat. However, traditional SDS-PAGE can not differentiate the HMW-GS Dtx5 and Dx5 subunit of common wheat. In view of this, the present study uses the relationship between HMW-GS Dtx5 subunit and HMW-GS Dx5 subunit DNA sequence Several SNPs Differentiate Allele specific polymerase chain reaction (AS-PCR) primers were developed to distinguish the two subunits of the 5 subunits by PCR. The results showed that the designed two pairs of primers were able to amplify a clear and stable target band, and the amplification results of the two pairs of primers were consistent. The HMW-GS Dx5 subunit showed a positive band with nodular sources Dtx5 subunit showed negative results. Homology analysis indicated that the homology of Dtx5 subunit with Dx2 subunit amino acid sequence was higher than that between Dtx5 and Dx5.