利用电子PCR分析草菇基因组SSR标记多态性

Polymorphism analysis of genomic SSR markers of Volvariella volvacea by electronic PCR

摘要:

【目的】利用电子PCR分析草菇基因组SSR标记多态性，并通过PCR验证分析结果的可靠性。【方法】利用MISA程序定位草菇基因组SSR位点并结合primer3.0程序设计SSR分子标记引物，运用电子PCR进行SSR分子标记多态性分析，基于分析结果进行PCR验证。【结果】随机选取658对SSR引物在草菇同核体菌株V23-1和PD19中进行真实PCR检测，结果表明28.6%的SSR引物具有多态性。数据分析表明，如果SSR标记来源于电子PCR产物长度没有差异的类型，仅4.8%的SSR引物在真实PCR中表现出多态性；如果SSR标记来源于电子PCR产物长度差异大于或者等于3 bp的类型，其中至少48.3%的SSR引物在真实PCR中表现出多态性。【结论】 PCR验证结果表明利用电子PCR可以提高SSR多态性引物的筛选效率。

Abstract:

[Objective] To verify the polymorphism of SSR markers analyzed with electronic PCR in Volvariella volavacea genome by using real PCR. [Methods] SSR loci in V. volvacea genome were identified with MISA software and the primers of the SSR molecular markers were developed with Primer3.0 software. The polymorphism of SSR markers were analyzed with electronic PCR and then verified by PCR. [Results] The 658 pairs of randomly selected SSR primers were checked by real PCR in 2 homokaryon strains, V23-1 and PD19, and the results show that 28.6% of SSR primers have the polymorphism. When the tested SSR loci belong to the type that had no length difference analyzed by the electronic PCR, only 4.8% SSR primers had the polymorphism. When the tested SSR loci selected from the group in which the length difference among the electronic PCR products of different genome was more than or equal to 3 bp, the true polymorphous SSR primers reached 48.3%. [Conclusion] The real PCR confirmed that it is more efficient to select polymorphous SSR markers with electronic PCR.