梅EST-SSR特征及其对红叶李的可转移性研究

【目的】分析梅EST-SSR特征及其对红叶李的可转移性。【方法】从National Center for Biotechnology Information(NCBI)下载4 589条梅表达序列标签(Expressed Sequence Tag;EST),去除无效碱基后进行拼接和简单重复序列(Simple Sequence Repeat)位点查找;设计引物进行PCR扩增。【结果】获得4 392条unigene,长度为2 420.422kb;搜索到776个SSR位点,分布在650条EST上,出现频率为14.8%。随机设计了15对引物,以4个红叶李品种基因组DNA为模板,进行了PCR扩增,有9对引物可以扩增到清晰稳定的产物,6对多态性引物共检测出20个位点,平均每对引物可以扩增出3.3条多态性片段,平均多态性信息含量(PIC)为0.8。【结论】SSR位点在梅EST序列上分布频率较高,梅EST-SSR在红叶李中具有较高的多态性,可用于红叶李亲缘关系分析和遗传图谱构建等方面的研究。

On the characteristics of EST-derived SSR marker development of Prunus mume and its transferability to Prunus cerasifera

[Objective] This study aims to characterize Prunus mume EST-SSRs and analyze their transferability to Prunus cerasifera.[Methods] A total of 4,589 ESTs were downloaded from NCBI and then the no redundant sequences were screened for SSR sites.[Results]SSR sites were found in 650 ESTs with an amount of 776 and occurrence frequency of 14.8％.Fifteen pairs of primers were designed for polymerase chain reaction using four cultivars of Prunus cerasifera as templates.Nine pairs of primers were successful for expected amplicons,and 6 pairs showed polymorphism with average values of 3.3 sites per one primer pair (average PIC=0.8).[Conclusion] These data showed that SSR sites showed a high occurrence frequency in Prunus mume,and the corresponding markers could be usable for the related studies in Prunus cerasifera.