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目的初步从滇丹参转录组文库中筛选出EST序列简单重复序列(SSR)位点,为进一步筛选出多态性丰富的SSR引物奠定基础。方法使用Micro SAtellite(MISA)软件分析滇丹参高通量转录组SSR的分布频率和重复基元的类型特征,利用软件Primer3设计引物,实现SSR引物初次筛选,再使用PCR扩增技术对滇丹参SSR引物进行二次筛选。结果40对引物共有25对出现特异性扩增片段,其中20对扩增产物与预期产物相符度较高。结论利用初步筛选出可用的SSR分子标记,为下一步筛选出多态性丰富的SSR引物奠定基础。转录组SSR分子标记开发将有助于滇丹参遗传多样性的研究。
OBJECTIVE: To screen SSR loci from the cDNA library of Salvia miltiorrhiza Bge., And lay the foundation for further screening of SSR primers rich in polymorphisms. Methods Micro SAtellite (MISA) software was used to analyze the distribution frequency and type of repetitive motif of Salvia miltiorrhiza high-throughput transcriptome. Primer3 was designed by software Primer3 to achieve primary screening of SSR primers, and then PCR amplification of SSR Primer for secondary screening. Results A total of 25 pairs of primers had 25 pairs of specific amplified fragments. Among them, 20 pairs of amplified products were highly consistent with the expected products. Conclusion The available SSR markers were screened initially, which laid the foundation for the further screening of polymorphic SSR primers. Development of transcriptome SSR molecular markers will be helpful for the genetic diversity of Salvia yunnanensis.