节节麦HWM-GS Dtx5与普通小麦HWM-GS Dx5亚基的AS-PCR鉴别.doc

节节麦HWM-GS Dtx5与普通小麦HWM-GS Dx5亚基的AS-PCR鉴别
曲继鹏温雯李俊郑建敏杨武云万洪深
西昌学院农业部科技发展中心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
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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。
关键词:
节节麦; 普通小麦; Dtx5亚基; Dx5亚基; AS-PCR;
作者简介:万洪深, wanhongshen@
基金:国家重点研发项目(2016YFD0100102)
AS-PCR for Distinguishing HMW-GS Dtx5 from Aegilops tauschii and Dx5 mon Wheat
Qu Jipeng Wen Wen Li Jun Zheng Jianmin Yang Wuyun Wan Hongshen
Xichang College; Development Center of Science and Technology, Ministry of Agriculture; Key Laboratory of Biology and ic Breeding in Wheat (Southwest) , Ministry of Agriculture, Research Institute of Crop Science, Sichuan Academy of Agricultural Sciences;
Abstract:
High-molecular-weight (HMW) glutenin, one of the important storage proteins mon 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 ic diversity of the D genome from Aegilops tauschii, and has gradually been used as a bridge-tool mon wheat improvement. Therefore, more and more HMW-GS subunit types of Ae. tauschii have been introgressed into mon wheat with the irresistible application of SHW mon wheat breeding. However, the traditional SDS-PAGE couldn't distinguish Dtx5 from Ae. tauschii and Dx5 mon wheat. In this study, we developed two pairs of AS-PCR primers, based on three SNPs in