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禾本科植物内生真菌研究13:禾本科植物内生真菌的分离鉴定及基因组DNA的快速提取

DOI: 10.7685/j.issn.1000-2030.2012.03.010, PP. 52-56

Keywords: 禾本科植物内生真菌,基因组DNA的提取,NRPS基因,PCR扩增,序列比对

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Abstract:

调查我国7省12个不同地区的禾本科植物内生真菌,经分离、培养后鉴定得到2种Epichlo?属和4种属内生真菌。采用改良的QS(improvedquickandsafe,IQS)法进行禾本科植物内生真菌基因组DNA的提取,并通过PCR进行验证。以生长在平板上的菌丝为起始材料,仪器要求简单、操作简便,快速安全、省时省力,整个提取过程大约40min。该方法和常规SDS法相比,时间缩短到2周,为原来的1/4。以IQS法提取的DNA为模板,检测禾本科植物内生真菌中的NRPS基因,并在NCBI上进行比对分析,发现与已有的NRPS基因高度相似。这种快速提取法所获得的基因组DNA质量高,可用于后续PCR、基因克隆及其他高通量测试,特别适合获取大量真菌菌株基因组DNA时使用。

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