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茶叶科学 ›› 2006, Vol. 26 ›› Issue (1): 17-23.doi: 10.13305/j.cnki.jts.2006.01.003

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茶树EST-SSR的信息分析与标记建立

金基强1,崔海瑞1*,陈文岳2,卢美贞1,姚艳玲1,忻雅2,龚晓春1   

  1. 1.浙江大学原子核农业科学研究所, 浙江杭州 310029;
    2.杭州市农业科学院, 浙江杭州 310024
  • 收稿日期:2005-08-02 修回日期:2005-10-10 出版日期:2006-03-25 发布日期:2019-09-10
  • 通讯作者: *崔海瑞
  • 作者简介:金基强(1983— ),男,河南信阳人,硕士研究生,研究方向为植物诱变遗传与分子改良。

Data Mining for SSRs in ESTs and Development of EST-SSR Marker in Tea Plant (Camellia sinensis)

JIN Ji-qiang1, CUI Hai-rui1, *, CHEN Wen-yue2, LU Mei-zhen1, YAO Yan-ling1, XIN Ya2, GONG Xiao-chun1   

  1. 1.Institute of Nuclear and Agricultural Sciences, Zhejiang University, Hangzhou 310029, China;
    2. Hangzhou Academy of Agricultural Sciences, Hangzhou 310024, China
  • Received:2005-08-02 Revised:2005-10-10 Online:2006-03-25 Published:2019-09-10

摘要: 在1589条茶树EST中,共发掘出了281个EST-SSR,分布于246条EST中,出现频率是17.68%,平均长度为33.06βbp,平均分布频率是1/2.61βkb。在茶树EST-SSR中,二核苷酸重复是主要的重复类型,出现最多的重复基元类型是AG/CT重复。设计了19对SSR引物,在对引物、dNTP、MgCl2的浓度及退火温度等参数进行测试后,建立了合适的PCR反应体系。以衍生绝大多数EST-SSR的龙井43βDNA为模板,对引物进行了筛选,有16对引物显示扩增,可用率为84.2%; 进一步在10个茶树品种中进行多态性测试,显现出多态性的引物占可扩增引物的62.5%。本文研究结果证明了根据茶树EST建立SSR标记是有效、可行的。

关键词: 茶树, EST, SSR信息, 标记建立

Abstract: :Totally 281 SSRs distributed in 246 ESTs were mined out, accounting for 17.68% of 1589 ESTs updated in tea. The average length of tea plant EST-SSRs searched out is 33.06βbp and the average distance of distribution is 1/2.16βkb. The dinucleotide repeat is the dominant type with repeat motif AG/CT being the most common. 19 primer pairs for EST-SSRs were designed. After testing on the annealing temperature and the concentration of primers, dNTP and MgCl2, a suitable PCR system was established. Under the condition of reaction system developed, the primers designed were screened against genomic DNA of Longjing 43 from which most EST-SSRs were derived, and 16 primer pairs showed the amplification,accounting for 82.4% of total primers. Then the primers showing amplification were subjected to PCR for DNAs from 10 tea plant cultivars and 10 primer sets showed polymorphisms, accounting for 62.5% of primers available. Results prove that it is an effective and feasible approach to develop SSR markers based on ESTs in tea.

Key words: tea plant, ESTs, SSR information, marker development

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