葡萄SNP标记的CAPS和TSP分析

CAPS and TSP Analyses of Grape SNP Markers

为了建立起葡萄SNP标记分析的一般方法和探索EST-SNP标记在葡萄中的应用,对前期开发的葡萄EST-SNP位点进行了筛选,设计了28对CAPS(cleaved amplified polymorphic sequences,酶切扩增多态性序列)标记引物和22对TSP(Temperature-switch PCR,温度转变PCR)标记引物,并在31份葡萄材料中进行了分析,其中20对CAPS引物和10对TSP引物电泳条带较清晰,多态性较好。基因分型统计显示:所有的SNP位点均含有2个等位基因,平均多态性信息含量(PIC)为0.336,平均Shannon’s信息指数为0.511。基因分型和葡萄材料聚类结果均显示这两种方法所获结果提供的信息量大且结果可靠。本方法简便、成本低,可作为SNP分型有效方法用于葡萄品种鉴定、基因分型和遗传多样性分析等。

The grape EST-SNP loci developed previously were firstly screened in order to establish the ordinary analyzing method and investigate the performance of grape SNP markers. Twenty-eight CAPS （cleaved amplified polymorphic sequences）primers and twenty-two TSP（temperature-switch PCR） primers were designed to analyze the screened SNP loci. Finally,twenty CAPS primers and ten TSP primers produced clear and polymorphic bands in 31 grape genotypes. The genotyping results showed that all SNP loci have two alleles. The average polymorphism information content and Shannon index are 0.336 and 0.511,respectively. The genotyping and cluster results of this study showed that CAPS and TSP method were efficient and reliable. The CAPS and TSP primers developed in this study could be used in SNP genotyping,cultivar identification and genetic diversity analysis of grape varieties in future.