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  

Grass endophytic fungi researches 13:Endophytic fungi identification and rapid extraction of genomic DNA

作　　者：王永[1] 纪燕玲[1] 王晗[1] 陈永敢[1] 王志伟[1] 

机构地区：[1]南京农业大学生命科学学院,江苏南京210095

出　　处：《南京农业大学学报》2012年第3期52-56,共5页Journal of Nanjing Agricultural University

基　　金：国家自然科学基金项目(30670008;30800156;30970081)

摘　　要：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Neotyphodium属内生真菌。采用改良的QS(improved quick and safe,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40 min。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。Endophytic fungi were surveyed in 12 different regions of 7 provinces in China,and 2 Epichlo species and 4 Neotyphodium species were identified.In this study,we used improved method for genomic DNA extraction from slow-growing fungal endophytes and validated it by PCR.The IQS（improved quick and safe）method was achieved by using plate-grown mycelia as the starting material.This new method was in virtue of simple equipment,easy operation,rapid and safe extraction,and low cost.The whole extraction process could be accomplished in 40 minutes.It took 2 weeks which was 1/4 of SDS procedures.Then DNA was used as template to detect the NRPS genes of the endophytes.The resulted sequences had a high sequence similarity with the NRPS gene.The quality of DNA samples from this method could be used for subsequent PCR,cloning,and other high-throughput testing requirements,particularly suitable for a large number of fungal strains.

关 键 词：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 

分 类 号：Q933[生物学—微生物学]