中国冬小麦品种Waxy蛋白分析及分子标记研究

Analysis of Waxy Proteins in Chinese Winter Wheat Cultivars Using SDS-PAGE and Molecular Markers

利用SDS-PAGE分析了国内外306份小麦品种(系)Waxy蛋白的缺失类型,筛选出缺失Wx-B1蛋白亚基的材料46份;通过对济麦19×豫麦47的120个F2单株Waxy蛋白亚基分析,发现缺失Wx-B1的比例接近1/4,符合孟德尔分离规律。利用3个STS标记和1个SSR标记对不同Waxy蛋白缺失类型进行鉴定,验证其在分子标记辅助育种中的有效性。结果表明,Wx-7A位点的STS引物在野生型中扩增出1条1172bp的特异带,而缺失Wx-A1蛋白亚基的突变材料中没有扩增出该特异带;Wx-4A位点的STS引物在野生型中扩增出1条440bp的特异带,缺失Wx-B1蛋白亚基的突变材料中没有该扩增片段;Wx-7D位点的STS引物在野生型中扩增出1条940bp的特异带,而在缺失Wx-D1蛋白亚基的突变材料中则扩增出1条360bp的特异带;Wx-7D位点的SSR引物在野生型中扩增出1条204bp的特异带,在缺失Wx-D1蛋白亚基的突变材料中没有扩增出该特异带。这4个标记可以用于分子标记辅助育种。

A total of 306 wheat cultivars and advanced lines from China, US and Australia were screened for the Waxy protein by SDS-PAGE. The results indicated that 46 of these cu｜tivars were the null Wx-B ｜ type. The proportion of individuals with null Wx-Bl in 120 F2 progenies derived from the Jimai 19×Yumai47 was 1/4 approximately, in accordance with the theoretical ratio. Three STS and 1 SSR markers were used to analyze the wheat cultivars with different types of Waxy proteins. Validation of the markers was carded out for the detection of Wx-7A, Wx-4A and Wx-TD genes. The results indicated that a 1172 bp-fragment was amplified with Wx-7A specific STS marker from the wild genotypes, while the fragment was absent for the mutant with null Wx-A1; a 440 bp-fragment was detected with Wx-4A specific STS marker from the wild genotypes, whereas, the fragment was absent for the mutant with null Wx-B1; a 940 bp-fragment was amplified with Wx-TD specific STS marker from the wild genotypes, and a 360 bp-fragment was found for the mutant such as ‘Baihuomai’ with null Wx-D1; and a 204 bp-fragment was detected with Wx-7D specific SSR marker from the wild genotypes, while the fragment was absent for the mutant with null Wx-D1. These markers are useful tools to identify wheat cultivars with mutant and normal alleles of the Waxy genes in marker-assisted selection of wheat breeding programme.