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园艺学报 ›› 2010, Vol. 37 ›› Issue (2): 221-228.

• 蔬菜 • 上一篇    下一篇

甘蓝EST-SSR 标记的开发与应用

陈 琛;庄 木;李康宁;刘玉梅;杨丽梅;张扬勇;程 斐;孙培田;方智远   

  1. 1 中国农业科学院蔬菜花卉研究所,北京100081;2 青岛农业大学,山东青岛266109;3 内蒙古农业大学,呼和浩特010081)
  • 收稿日期:2009-12-07 修回日期:2010-01-25 出版日期:2010-02-25 发布日期:2010-02-25

Development and Utility of EST-SSR Marker in Cabbage

CHEN Chen;ZHUANG Mu;LI Kang-ning;LIU Yu-mei;YANG Li-mei;ZHANG Yang-yong;CHENG Fei;SUN Pei-tian;FANG Zhi-yuan
  

  1. (1Institute of Vegetables and Flowers,Chinese Academy of Agricultural Sciences,Beijing 100081,China;2Qingdao Agricultural University,Qingdao,Shandong 266109,China;3Inner Mongolia Agricultural University,Huhhot 010081,China)
  • Received:2009-12-07 Revised:2010-01-25 Online:2010-02-25 Published:2010-02-25

摘要: 从NCBI 数据库下载62 567 条甘蓝相关EST,处理后得到19 611 条无冗余的EST,对其进行SSR搜索,共得到1 219 条SSR,分布在1 176 条EST 上,分布密度为1/11.48 kb,包括273 种重复基元。在甘蓝EST-SSR中,二核苷酸(353个SSR)和三核苷酸(423个SSR)占主导地位,所占比例分别为28.96%和34.70%;出现最多的重复基元是AG/CT,占总数的25.59%,其次为AAG/CTT(94 个,占7.71%)。针对1 176 条含有SSR 的EST 设计合成了978 对引物。用两份甘蓝自交系对978 对引物进行初筛,897 对引物有扩增产物,
共扩增出1 026 条扩增带;128 对引物表现出多态性,共有258 条多态性带(占总带数的25.15%)。利用4对多态性引物,初步构建了中甘11、8398、中甘15、中甘21 杂交新品种及其亲本的指纹图谱。以上结果说明从甘蓝相关EST 数据库中开发的SSR 引物有较好的可用性。

关键词: 甘蓝, EST-SSR, 标记开发, 指纹图谱

Abstract: A total of 62 567 ESTs in Brassica oleracea were downloaded from NCBI for the identification and development of SSR marker. In total, 1 219 SSRs distributed in 1 176 ESTs were identified from 19 611 non-redundant ESTs, with an average of one SSR per 11.48 kb, and included 273 SSR motifs.Analysis
of SSR motifs revealed that the dinucleotides (353 SSR) and tricleotides (423 SSR) were major motifs, accounted for 28.96% and 34.70% respectively. The AG/CT was the most frequent motifs and accounted for 25.59%,followed by AAG/CTT (94 SSR,7.71%). Based on the 1 176 SSR-containing ESTs, a total of 978 primer pairs were successfully designed and assessed validation of the amplification using two cabbage inbreed lines (397 and 20-2-5). The results showed that 897 primer pairs yielded 1 026 amplification bands, of which 128 primer pairs exhibited polymorphism with 258 polymorphic bands accounted for 25.15%. SSR fingerprint was obtained for 4 cabbage F1, Zhonggan 11, 8398, Zhonggan 15 and Zhonggan 21, and theirparents,and all the above materials could be discriminated by four specific EST-SSR primer pairs. The results
indicated that the developed SSRs from ESTs of Brassica oleracea were validate and practicable.

Key words: cabbage, EST-SSR, marker development, fingerprinting