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【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。
【Objective】 The objective of this study was to analyze the SSR marker polymorphism of Straw mushroom genome by electronic PCR and confirm the reliability of the results by PCR. 【Method】 The SSR locus of V. rufestris was mapped by MISA program and Primer 3.0 program was used to design SSR marker. SSR molecular marker polymorphism was analyzed by electronic PCR, and PCR was performed based on the analysis results. 【Result】 A total of 658 pairs of SSR primers were selected for real-time PCR analysis in the nucleus of V. simulans V23-1 and PD19. The results showed that 28.6% of SSR primers were polymorphic. Data analysis showed that only 4.8% of SSR primers showed polymorphism in real PCR if the SSR markers originated from the genotypes where the length of the electronic PCR products did not differ. If the SSR markers were derived from PCR products with a length difference greater than or equal to 3 bp Type, of which at least 48.3% of SSR primers showed polymorphism in real PCR. 【Conclusion】 PCR validation results show that the use of electronic PCR can improve SSR polymorphism primer screening efficiency.