松干基褐腐病菌的分子检测方法  

Molecular Detection of Inonotus weirii Gilgertson

作　　者：吴翠萍[1,2] 高振兴[3] 吴晶[4] 陈贯源[1] 杨静[2] 安榆林[2] 叶建仁[1] 

机构地区：[1]南京林业大学森林资源与环境学院,江苏南京210037 [2]江苏出入境检验检疫局,江苏南京215021 [3]泰州出入境检验检疫局,江苏泰州225300 [4]南京出入境检验检疫局,江苏南京211106

出　　处：《西南林学院学报》2010年第5期53-57,共5页Journal of Southwest Forestry College

基　　金：国家质检总局科技项目(2005IK070)资助;国家科技部项目(2009DFA31950)资助

摘　　要：松干基褐腐病菌是我国重要的植物检疫性病原菌,为建立该病原菌准确、灵敏、快速的检测方法,根据该病菌与同属内形态与寄主上最接近的4个种的ITS差异区域,设计出一对特异性引物IW1、IW2,经PCR扩增,只有松干基褐腐病菌可得到约165 bp的产物,而其余作为对照的菌株均无此产物,其检测灵敏度为100 pg基因组DNA。此外,特异性引物IW1和IW2还适合于Real-timePCR,其检测灵敏度更低,为0.1 pg基因组DNA(20μL反应体系)。应用此2种方法均能成功的从人工污染了的松干基褐腐病菌的木块中检测出病原菌。Inonotus weirii is an important plant quarantine pathogen in China.In order to set up an accurate,sensitive and quick way to detect this pathogen,a pair of specific primers,i.e.,IW1,IW2 were designed on the basis of the differences in the internal transcribed spacer（ITS） sequences between I.weirii and four species in the genus Inonotus,which were mostly close to Inonotus weirii in terms of morphology.The results of PCR showed that a 165bp product was only obtained from the sample of I.weirii.The detection sensitivity was 100 pg genomic DNA per 25μL PCR reaction volume.In addition,IW1,IW2 primers were successfully applied to real-time PCR with higher sensitivity of 0.1 pg genomic DNA per 20μL PCR reaction volume.The I.weirii pathogen could be rapidly and accurately detected from artificially blurred wood by these two methods.

关 键 词：松干基褐腐病菌 分子检测 REAL-TIME PCR 

分 类 号：S719.240.7[农业科学—林学]