生物多样性 ›› 2008, Vol. 16 ›› Issue (6): 555-561.doi: 10.3724/SP.J.1003.2008.08132

• 论文 • 上一篇    下一篇

基于棉花参比种质的SSR多态性核心引物筛选

潘兆娥, 孙君灵, 王西文, 贾银华, 周忠丽, 庞保印, 杜雄明*   

  1. (中国农业科学院棉花研究所/农业部棉花遗传改良重点开放实验室, 河南安阳 455000)
  • 出版日期:2008-11-20

Screening of SSR core primers with polymorphism on a cotton panel

Zhao-E Pan, Junling Sun, XiwenWang, Yinhua Jia, Zhongli Zhou, Baoyin Pang, Xiongming Du*   

  1. Cotton Research Institute, Chinese Academy of Agricultural Sciences; Key Laboratory of Cotton Genetic Improvement of Agricultural Ministry, Anyang, Henan 455000
  • Online:2008-11-20

棉花品种间遗传多样性的评价和遗传图谱构建都需要大量多态性的SSR标记。本研究通过构建棉花参比种质(panel), 高效快速地筛选出棉花多态性核心引物。本实验选用12份表型性状和遗传差异较大的棉花种质作为参比种质, 对来自Cotton Microsatellite Database (CMD)网站上 (http://www.cottonssr.org) 的5,914对棉花SSR引物进行了PCR扩增。结果表明: 在不同棉种间多态性的引物有4,800对, 约占有效扩增引物(5,300对)的90.6%; 能区分不同品种间差异的引物大约有500对, 占有效扩增引物的9.4%。我们推荐其中319对扩增效果好、条带清晰的引物作为棉花种质资源鉴定和分子指纹分析的核心引物, 其中277对能把陆地棉品种间的差异鉴别出来。此外, 我们还找到了在陆地棉、海岛棉及亚洲棉三大栽培棉中都有差异的13对共同SSR引物, 建议作为分子指纹分析和种质鉴定的首选引物。本研究公开了319对核心SSR引物及其多态性信息, 有利于规范引物筛选程序、提高引物筛选效率, 对棉花种质资源的遗传多样性分析、指纹图谱构建, 以及种子真伪性和纯度的鉴定等都具有现实意义。

A large number of polymorphic Simple Sequence Repeat (SSR) primers are needed to assess genetic diversity and construct genetic maps for cotton varieties. Our objective was to screen the cotton core primers with polymorphism based on a standard cotton panel. In the experiment, 12 accessions with significantly different phenotypes and genetic background were used as panel germplasm for screening 5,914 pairs of SSR primers obtained from the Cotton Microsatellite Database (http://www.cottonssr.org). A total of 4,800 pairs of SSR primers accounting for 90. 6% of the 5,300 effectively amplified primers were polymorphic among different species. Of these, about 500 pairs of primers, 9.4% of effectively amplified primers, could distinguish among cotton varieties. We suggest the use of 319 pairs of polymorphic primers with suitable amplification and clear bands as core primers for evaluating cotton germplasm and depicting fingerprints. We found 277 pairs of polymorphic primers that can identify upland cotton varieties. Moreover, we recommend 13 SSR primer pairs with polymorphism in all three species of upland cotton, sea island cotton and Asiatic cotton as the first-choice markers for molecular fingerprint and germplasm identification. Here we publish 319 core primer pairs and their polymorphic information, which will be helpful to regulate the procedure and promote the efficiency for screening SSR primers, and will play an important role in cotton genetic diversity evaluation, fingerprint construction, seed genuineness and purity identification of the cotton varieties.

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