SRAP、ISSR技术的优化及在甘蓝类植物种子鉴别中的应用  

Optimization of SRAP & ISSR Technology and Its Application in the Identification of Seeds of Brassica oleracea L.

作　　者：刘冲[1,2] 葛才林[2] 任云英[1] 陈锦秀[1] 杨晓锋[1] 薄天岳[1] 

机构地区：[1]上海市农业科学院园艺研究所,上海市设施园艺技术重点实验室,上海201106 [2]扬州大学农学院

出　　处：《生物工程学报》2006年第4期657-661,共5页Chinese Journal of Biotechnology

基　　金：上海市科委基础性研究计划项目(No.03JC14060);上海市农委科技兴农重点攻关项目(No.2004-2-5)。~~

摘　　要：将SRAP与ISSR2种分子标记技术应用于8种甘蓝类植物(BrassicaoleraceaL.)的种子鉴别中。先以甘蓝(Brassicaoleraceavar.capitata)基因组DNA为模板,通过对SRAP、ISSR反应体系中各影响因素的逐一筛选,优化了甘蓝类植物SRAP、ISSR反应体系。进而采用30个SRAP引物组合和15个ISSR引物对白甘蓝、皱叶甘蓝、红甘蓝、羽衣甘蓝、花椰菜、青花菜、抱子甘蓝、球茎甘蓝的基因组DNA进行了PCR扩增,结果表明M3-E5与M4-E5两个SRAP引物组合可以在8种甘蓝类植物之间显示较高的多态性;844和888两个ISSR引物也可在8种甘蓝类植物之间产生很好的多态,特别是844引物单独应用即可区分所有材料。In this study, the molecular marker technology of SRAP and ISSR were applied in rapid identification of seeds from eight species of Brassica oleracea L. Firstly, using the genomic DNA of cabbage as template, SRAP and ISSR reaction systems were optimized through testing every factor, respectively, that affects PCR amplification. Then, using the optimized reaction systems, 30 SRAP primer pairs and 15 ISSR primers were applied to amplify genomic DNA of cabbage, savoy, purple cabbage, borecole, cauliflower, broccoli, Brussels sprouts, and kohlrabi The results showed that high polymorphisms were exhibited among the eight species of Brassica oleracea L. by SRAP primer pairs of M3-E5 and M4-E5, as well as ISSR primers of 844 and 888, especially primer 844 which can identify all eight materials efficiently.

关 键 词：甘蓝类植物 SRAP ISSR 反应体系 鉴别 

分 类 号：S635[农业科学—蔬菜学]