基于荧光标记的紫娟茶树转录组EST-SSR标记开发

EST-SSR marker development of Zijuan tea tree transcriptome based on the fluorescent labeling

为开发紫娟茶树转录组EST-SSR标记,基于前期对紫娟茶树芽、第2叶、开面叶、成熟叶转录组高通量测序所得到的242 757条Unigene进行多态性分析与评价,再利用荧光标记PCR技术,规模化开发茶树EST-SSR标记。结果表明:从紫娟茶树转录组中搜索得到46 041条Unigene含有57 976个SSR位点,出现频率为23.88%。EST-SSR类型以一核苷酸、二核苷酸、三核苷酸重复为主,占总SSR的98.54%。设计合成138对荧光SSR引物,采用荧光标记PCR技术对4个差异较大的茶树品种进行引物筛选,其中44对引物得到高质量的EST-SSR标记位点。这些EST-SSR可用于茶树遗传多样性分析、分子育种等。

To develop the transcriptome EST-SSR markers of Zijuan tea tree,the polymorphism of 242 757 unigenes got by high-throughput sequencing based on the earlier-stage study of the bud,the second leaf,the leaves without bud and mature leaves of Zijuan tea tree was analyzed and evaluated,and then fluorescent tags PCR technology was used for scale development of tea tree EST-SSR markers. The results showed that 46 041 unigenes containing 57 976 simple sequence repeats（ SSR） loci were obtained by searching from Zijuan tea plant transcriptomes with the frequency of 23. 88%. The main repeat types of EST-SSR were mononucleotide,dinucleotide and trinucleotide accounted for 98. 54% of total SSR. 138 pairs of fluorescent marker primers were designed and synthesized,then four different tea varieties were screened by using fluorescent marker PCR technology,44 pairs of primers got high quality of EST-SSR markers loci. These EST-SSRs can be used for tea tree genetic analysis and molecular breeding.