簇毛麦HMW-GS基因的位点特异PCR(AS-PCR)标记及其多态性  

The Allelic Specific PCR Marker and Diversity of HMW-GS Genes in Dasypyum villosum

作　　者：刘守斌[1] 梁荣奇[1] 尤明山[1] 李保云[1] 刘广田[1] 

机构地区：[1]中国农业大学农学与生物技术学院植物遗传育种系,农业生物技术国家重点实验室,北京市作物遗传改良重点实验室,农业部作物基因组与遗传改良重点实验室,北京100193

出　　处：《华北农学报》2011年第1期1-5,共5页Acta Agriculturae Boreali-Sinica

基　　金：国家自然科学基金项目(39930110);北京市自然科学基金项目(6990001)

摘　　要：簇毛麦含有丰富的HMW-GS基因变异。本研究旨在建立簇毛麦HMW-GS基因的位点特异PCR(AS-PCR)标记,并对18个居群簇毛麦HMW-GS基因遗传多样性进行系统评价,为开发和利用簇毛麦HMW-GS改良普通小麦品质奠定基础。根据先前克隆的3个簇毛麦HMW-GS基因序列(1Va、1Vb和1Vc)和已发表的普通小麦HMW-GS基因序列,经过同源性比较后设计了簇毛麦HMW-GS基因的位点特异性引物P6+P7。对中国春及其簇毛麦附加系、6个不同来源簇毛麦的扩增结果表明,该对引物能在普通小麦背景下特异扩增簇毛麦HMW-GS基因,可以鉴别簇毛麦之间的HMW-GS等位基因变异,且其长度变异主要存在于中部重复结构区。利用该标记对来自美国国家种质库的18份材料的108个单株进行了研究,结果发现,每个单株均能扩增出1条或2条片段,共检测出7种等位基因变异的类型。不同等位基因类型的频率也不一样,等位基因b存在于除W6 7288外的所有簇毛麦居群中,占检测单株总数的59.55%,其次为等位基因d,占25.19%,等位基因a和等位基因f频率最低,仅出现在1个单株中。说明由于簇毛麦为异花受粉植物,同一居群不同单株间出现了不同的等位基因。D.villosum is a rich reservoir for genic diversity of High-Molecular-Weight glutenin subunit（HMW-GS）.In this paper,one Allelic Specific PCR（AS-PCR）marker for the amplification of the genes coding HMW-GS of D.villosum was developed on the basis of the sequences of 1Va,1Vb,1Vc and common wheat HMW glutenin genes.PCR analysis on HMW glutenin genes of CS and its addition lines showed the specialty of this AS-PCR marker,while analysis on HMW-GS genes of six accessions of D.villosum showed a high degree of length polymorphism,which suggested that the size variation observed in different allelic subunits of D.villosum was mainly due to the different length of the central repetitive domain.Using this AS-PCR marker,seven alleles at Glu-V1 were found among 108 H.villosum individuals from 18 accessions.The frequencies of those alleles were different.The allele b was found in all accessions except W6 7288,and its frequency was 59.55%of 108 individuals,while that of allele d was 25.19%.But allele a as well as f was only found once.Therefore,different genotypes were found in the different individuals from one accession due to the cross-pollination.

关 键 词：簇毛麦 高分子量谷蛋白亚基(HMW-GS) 位点特异PCR(AS-PCR)标记 多态性 

分 类 号：S512.1[农业科学—作物学] S331