小麦优质谷蛋白亚基分子标记多重PCR体系的建立与应用  

Establishment and Application of Multiplex-PCR for Wheat Glutenin Subunits Relative to Superior End-Use Quality

作　　者：郑寒[1,2] 陈静[1] 任妍[1,2] 余懋群[1] 付体华[2] 

机构地区：[1]中国科学院成都生物研究所,四川成都610041 [2]四川农业大学农学院,四川雅安625014

出　　处：《作物学报》2009年第10期1831-1835,共5页Acta Agronomica Sinica

基　　金：国家自然科学基金项目(30871527);国家转基因生物新品种培育重大专项(2009ZX08009-010B);中国科学院知识创新工程方向性项目(KSCX2-YW-N-052);四川省育种攻关项目资助

摘　　要：Ax1/Ax2*、Dx5和过量表达的Bx7亚基(Bx7OE)被认为是对小麦品质有正向效应的优质亚基。根据以上亚基特异性分子标记建立相应的多重PCR体系,经品种和群体检验,证明利用该体系鉴定亚基的结果稳定可靠、成本较低。利用该多重PCR技术对89份西藏小麦育成和推广品种(系)的优质亚基频率进行鉴定,结果表明,Dx5和Ax1/Ax2*亚基的频率均为12.4%,没有检测到Bx7OE,同时携带两个优质亚基的材料频率为10.1%,该麦区品质育种必须加强优质亚基的引入。针对优质亚基Ax1/Ax2*、Dx5和Bx7OE的多重PCR体系,为小麦品质育种亲本评价和通过杂交方法聚合优质亚基基因提供了一种实用可靠的标记辅助选择技术。Wheat processing qualities are largely dependent on the number and composition of high-molecular-weight glutenin subunits （HMW-GS）. It has been demonstrated that Ax1/Ax2^＊, Dx5, and overexpressed Bx7 （Bx7^OE） are normally associated with superior end-use quality, especially dough strength. To date, Bx7^OE allele has not been exploited widely in China wheat breeding program. With the optimization of reaction components and amplification condition, three pairs of PCR primers targeting for Ax 1/Ax2^＊, Bx7^OE, and Dx5 genes were used to establish a multiplex PCR for the purpose of enhanced molecular marker-assisted breeding with good reliability and low cost. The validation with wheat cultivars （lines） carrying known genes displayed that the developed multiplex PCR permitted the discrimination of these major HMW-GS in a single PCR reaction and agarose gel assay. A total of 89 wheat cultivars and advanced lines from Tibet, China were tested by the multiplex PCR-based assay. The results showed that the frequencies of Axl/Ax2^＊ and Dx5 were both 12.4%, and only 10.1% of the accessions possessed both Axl/Ax2＊ and DxS, whereas no accessions carried Bx7^OE. Introduction of HMW-GS relative to good gluten quality should enhance the im- provement of wheat quality in this region.

关 键 词：小麦品质 高分子量麦谷蛋白亚基(HMW-GS) 多重PCR 

分 类 号：S512.1[农业科学—作物学]