适用于rDNA ITS分析的兰属菌根真菌培养及DNA提取方法  

A Rapid and Simple Method for Culturing of Cymbidium mycorrhizal Fungi and Extracting of DNA from the Fungi for the Analysis of rDNA ITS

作　　者：庄彩云 李潞滨[1] 胡陶[1] 唐征[2] 刘振静[1] 杨凯[3] 

机构地区：[1]中国林业科学研究院林业研究所国家林业局林木培育实验室,北京100091 [2]北京农学院植物科学技术系,北京102206 [3]北京农学院生物技术系

出　　处：《北京农学院学报》2007年第3期4-6,31,共4页Journal of Beijing University of Agriculture

基　　金：948项目(植物基因测序及功能改良技术引进);北京市属市管高校人才强教计划资助项目

摘　　要：为探讨兰属植物与其共生真菌之间专一性关系,采用覆盖纤维素滤膜于PDA培养基的方法培养兰属菌根真菌,提取其DNA后用于rDNA ITS分析。在培养过程中克服PDA液体培养及固体培养获得菌丝体少的缺点,操作方便,获得大量菌丝体;采用改进的提取植物基因组DNA的SDS方法提取真菌的DNA,通过提高缓冲液的pH值、增加EDTA的浓度以提高DNA的提取效果。结果表明,此培养方法及改进的DNA提取方法均简便易行,提取的DNA产量高、质量较好;可满足菌根真菌rDNA ITS分析的要求。In order to analyze the specificity relationship between the Cymbidium and the Cymbidium mycorrhizal fungi ,a rapid method for culturing the Cymbidium mycorrhizal fungi on PDA covered with cellulose acetate membrane was reported in this paper. The extracted DNA of Cymbidium mycorrhizal fungi was used for the rDNA ITS analysis. The method obtained a great quantity hyphostroma. The improved SDS method which used in plant genomic DNA was applied in extraction of fungi DNA. The DNA was satisfied for the rDNA ITS analysis of the Cymbidium mycorrhizal fungi.

关 键 词：兰属菌根真菌 醋酸纤维素滤膜 真菌DNA 

分 类 号：Q939.11[生物学—微生物学]