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茶树EST-SSRs分布特征及引物开发

Characterization of EST-derived Microsatellites and Development of SSR-markers in Tea(Camellia sinensis)

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【作者】 王丽鸳姜燕华段云裳成浩周健曾建明

【Author】 WANG Li-yuan1,JIANG Yan-hua1,DUAN Yun-shang2,CHENG Hao1,ZHOU Jian1,ZENG Jian-ming1(1 Tea Research Institute,Chinese Academy of Agricultural Sciences/National Center for Tea Improvement,Hangzhou 310008; 2Nanjing Agricultural University,Nanjing 210095)

【机构】 中国农业科学院茶叶研究所/国家茶树改良中心南京农业大学

【摘要】 为了在茶树中开发EST-SSR s功能性标记,利用生物信息学方法对NCB I网上公开的3288条茶树(Cam ellia sinensis)ESTs序列进行EST-SSR s特征分析。剔除冗余序列,得到非冗余序列2083条。在非冗余序列中发现含不同重复基元SSR s的EST序列有385条,共486个EST-SSR s,平均相隔2.10 kb出现1个SSR。在2~6 bp的重复基元中,二核苷酸重复基元的SSR s出现频率最高(51.97%),其次是三核苷酸(19.55%)。对所有的重复基元类型进行统计分析发现,所占比例最高的是AG/CT(47.74%),其次分别是AT/TA(4.73%)和AAG/CTT(4.73%)。利用Prim e 5软件,设计了206对EST-SSR s引物,随机选用72对引物进行SSR扩增,发现31对引物可以扩增出条带,其中29对引物具有多态性,多态性比率为93.5%。这些EST-SSR s将有助于茶树基因组学方面的研究。

【Abstract】 To develop EST-SSRs functional markers in tea(Camellia sinensis),characterization of simple sequence repeats(SSRs)were analyzed by 3288 EST(expressed sequence tag)sequences in Camellia sinensis from NCBI.A total of 2083 non-redundant EST sequences were obtained from 3288 sequences by redundant analysis and there were 486 EST-SSRs present in 385 ESTs out of the non-redundant sequences with one EST-SSR in every 2.10 kb of EST sequence in average.Among the different motifs ranging from 2bp to 6bp,dinucleotide repeats were most abundant(51.97%),followed by trinucleotide repeats(19.55%)and mononucleotide repeats(23.35%).Among all identified motif types,AT/TA had the highest frequency(4.73%),followed by AAG/CTT(4.73 %).206 pairs of SSR primers were designed by the software,primer 5.Among 72 pairs of primers selected randomly from them,31 primer pairs were successfully amplifying DNA fragments with 29 polymorphic primers.The proportion of primers that produced polymorphic fragments was 93.5%.Furthermore,the availability of microsatellite markers can be expected to enhance the power and resolution of genomic analysis in tea.

【关键词】 茶树EST-SSRs分子标记多态性
【Key words】 Camellia sinensisEST-SSRsMolecular markerPolymorphism
【基金】 国家科技基础条件平台建设计划(2005DKA21002)
  • 【文献出处】 植物遗传资源学报 ,Journal of Plant Genetic Resources , 编辑部邮箱 ,2009年04期
  • 【分类号】S571.1
  • 【被引频次】48
  • 【下载频次】269
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