基于173份小麦评价糯蛋白亚基基因(Wx)的分子标记和构建其多重PCR体系  

Evaluation of Molecular Markers for Wx Genes and Development of Their Multiplex PCR Systems Based on the Compositions of Wx Proteins for 173 Wheat Cultivars and Lines

作　　者：张晶[1] 张晓科[1] 梁强[1] 任万杰[1] 张钰玉[1] 

机构地区：[1]西北农林科技大学农学院/国家小麦改良中心杨凌分中心,杨凌712100

出　　处：《农业生物技术学报》2011年第3期417-426,共10页Journal of Agricultural Biotechnology

基　　金：农业部948项目小麦分子育种实用化研究(No.2006-G2);农业部小麦产业技术体系建设专项(No.nycytx-03);西北农林科技大学唐仲英育种基金共同资助

摘　　要：糯蛋白亚基基因(Wx)组成与小麦淀粉品质特性密切相关,筛选和建立其高效的鉴定方法和体系,对小麦品质育种改良和种质资源快速评价具有重要意义。本研究基于SDS-PAGE方法检测173份小麦(Triticum aestivum L.)品种(系)糯蛋白亚基组成的结果,评价序列标签位点(sequence tagged sites,STS)标记的有效性,利用有效的STS标记构建Wx基因分子检测的多重PCR体系。SDS-PAGE方法检测结果表明,20份小麦品种(系)缺失Wx-B1蛋白亚基,1份3个Wx蛋白亚基全部缺失,依次占11.6%和0.6%。4个共显性和2个显性Wx基因STS标记检测供试小麦的结果,与SDS-PAGE结果完全一致。构建了两套分子标记检测的多重PCR体系,其中多重PCR体系Ⅰ能够同时检测Wx-A1和Wx-B1位点的4种等位变异,体系Ⅱ可同时检测Wx-B1和Wx-D1位点的4种等位变异,两套多重PCR体系检测的结果与SDS-PAGE和单一PCR的检测结果也完全一致。筛选的6个STS标记和构建的两套PCR检测体系,用于小麦Wx基因的分子标记辅助选择,将有助于提高小麦品种淀粉品质评价和选育的效率。Quality properties of starch are strongly affected by the compositions of Wx genes in wheat.It is very important to establish stable and efficient methods and systems to detect Wx genes for improvement of quality and rapid evaluation of germplasm resources in wheat breeding programs.In this study,based on the compositions of Wx proteins in 173 wheat（Triticum aestivum L.） cultivars and advanced lines using SDS-PAGE method,the validity of sequence tagged sites（STS） markers for Wx genes were evaluated,and two types of multiplex PCR systems were built up.The results using the SDS-PAGE method showed that twenty of these cultivars and advanced lines were the null Wx-B1 type（11.6%）,and one of them was waxy wheat lacked all three Wx proteins（Wx-A1,Wx-B1 and Wx-D1）（0.6%）.The results identified by four co-dominant and two dominant STS markers for Wx genes were all identical to those detected by the SDS-PAGE method.Using the six effective molecular markers above,two multiplex PCR systems were established.The multiplex PCRⅠ could simultaneity identify four alleles at Wx-B1 and Wx-D1 loci,and multiplex PCRⅡ were used to detect four alleles in Wx-B1 and Wx-D1 loci.The genotypes of all wheat cultivars and advanced lines identified by the two multiplex PCRs were coincided with those detected by the SDS-PAGE and corresponding single PCR method,respectively.The six STS markers and two multiplex PCRs can be used to increase the efficiency of evaluation and improvement of wheat starch properties in breeding programs.

关 键 词：普通小麦 糯蛋白 SDS-PAGE 分子标记 多重PCR 

分 类 号：S511[农业科学—作物学]