大豆EST-SSR标记开发及与Genomic-SSR的比较研究  

Development of soybean EST-SSR marker and comparison with genomic-SSR marker

作　　者：常玮[1] 赵雪[1] 李侠[1] 邱波[1] 韩英鹏[1] 縢伟丽 李文滨[1] 

机构地区：[1]东北农业大学大豆研究所,国家教育部大豆生物学重点实验室,黑龙江哈尔滨150030

出　　处：《中国油料作物学报》2009年第2期149-156,共8页Chinese Journal of Oil Crop Sciences

基　　金："十一五"863目标导向项目(2006AA10ZF1Z1);863计划重点项目(2006AA100104-4);国家自然科学基金(30400285)

摘　　要：对458 220条大豆EST序列进行SSR搜索,共检测出EST-SSR序列39 989条,经拼接得到无冗余EST-SSR序列8 190条,包括357种重复基元。其中二、三核苷酸重复基元类型居多,分别占无冗余EST总数的11.13%和16%,统计得到二核苷酸重复类型12种,三核苷酸重复类型60种。以含有简单重复序列的无冗余EST序列设计200对引物,其中148对引物有清晰且单一条带扩增产物,以30份大豆品种资源进行引物筛选,获得多态性引物31对。以21份大豆不同基因型的基因组DNA为模板选取30对显示多态性的大豆EST-SSR引物和30对大豆基因组SSR引物进行扩增,带型统计结果显示:大豆EST-SSR与基因组SSR在供试基因型间多态性指数均值分别为0.55和0.44,二者揭示的多态性水平差异不大。从而说明利用生物信息学方法基于大豆EST开发SSR标记是切实可行的,大豆EST-SSR可以用于大豆遗传多样性分析,是大豆DNA分子标记体系的一个重要补充。458 220 EST sequences of soybean were used to screen SSR. 39 989 EST-SSR sequences were detected and 8 190 noni-redundant EST sequences,including 357 motifs were obtained through splicing. Among these sequences, tri-nucleotide repeat and di-nueleotide repeat were the most common types,and accounted for 11.13% and 16% of the total non-redundant EST respectively. 12 di-nucleotide repeat motifs and 60 tri-nucleotide repeat motifs were screened. 200 pairs of primers were designed based on the non - redundant SSR-EST sequence data of soybean, 148 pairs of the primers can successfully amplify,31 pairs of which present polymorphism among 30 different soybean genotypes. 21 different genotypes of soybean were used to investigate the polymorphism for comparing EST-SSR markers and genomic SSR markers. The results showed that the average polymorphism index from EST-SSR markers was 0.55 rather than 0.44 from the genomic SSR markers. These two markers demonstrated similar level of polymorphism. Our study suggested that the development of soybean EST-SSR markers based on EST data by bioinformatics methods was practical, efficient and economical. Soybean EST-SSR marker could be used for analysing genetic diversity of soybean, and this new type of molecular marker was an important component of soybean DNA molecular marker system.

关 键 词：大豆 EST—SSR 生物信息 多态性 

分 类 号：Q751[生物学—分子生物学] S565.103[农业科学—作物学]