白菜EST-SSR信息分析的标记建立.docVIP

白菜EST-SSR信息分析与标记的建立( 忻雅1 ，崔海瑞*1 ，卢美贞1 ，姚艳玲1，金基强1，林容杓2，崔水莲2 (1.浙江大学农业与生物技术学院原子核农业科学研究所，杭州 310029；2.忠南大学园艺系，大田305764，韩国) 摘要：对4584条白菜EST进行了搜索，共发掘474个SSR，出现的频率为10.3%，包括40种重复基元。其中二核苷酸和三核苷酸重复类型占主导地位，二者出现的频率基本相近，占总SSR的近83%；其它重复类型所占比例均不足5%。GA和GAA是二、三核苷酸中的优势重复类型，分别占二、三核苷酸重复类型的71.5%和37.5%。挑选其中符合引物设计的15条设计了EST-SSR引物。在合适的PCR反应体系下，以构建EST的白菜自交系A的DNA为模板，对设计的EST-SSR引物进行筛选，发现15对EST-SSR引物都能扩增出产物。进一步用这些可扩增的引物对29个白菜品种进行PCR扩增，发现7对引物显示多态性，占可扩增引物的46.7%。此结果表明，根据EST建立EST-SSR标记是一条简便而又有效的途径。 关键词：EST；SSR信息；标记建立；白菜 Data mining for SSRs in ESTs and EST-SSR marker development in Chinese cabbage Xin Ya1，Cui Hairui*1，Lu Meizhen1 , Yao Yanling1, Jin Jiqiang1 , Lim yongpyo 2 , Choi suryun 2 Institute of Nuclear and Agricultural Sciences，College of Agriculture and Biotechnology，Zhejiang University， Hangzhou 310029；2. Department of Horticulture, Chungnam National University， Taejeon 305-764，Korea) Abstract：4584 ESTs of Chinese cabbage were screened and 474 SSRs including 40 kinds of repeat motifs were mined out, accounting for 10.3% of ESTs. Dinucleotide and trinucleotide repeats, with similar frequency and accounting for 83% together in all SSRs, were dominant, while the frequency for other repeat type is below 5% each. GA and GAA are the most frequent motifs, accounting for 71.5% and 37.5% in dinucleotide and trinucleotide repeats respectively. 15 primer pairs for EST-SSRs were designed. Under a suitable PCR system, primers were screened against genomic DNA of inbreed line A from which the cDNA library was constructed, and all the 15 primer pairs showed the amplification. Then all the primers available were subjected to PCR for DNAs from 29 Chinese cabbage varieties and 7 primer sets showed polymorphisms, accounting for 46.7% of primers available. Results indicate it is a simple and effective approach to develop SSR markers based on ESTs. Key words: EST；SSR information；Marker development；Chinese cabbage 简单重复序列（SSR），也叫微卫星（Microsatellites），由于多态信息含量高、共显性遗传、技术简单、重复性好、特异性强等特点，被认为是植物育种中最有用的分子标记系统。表达序列标签（E