菜豆萎蔫病菌的PCR检测技术  

Development of a PCR test for the detection of Curtobacterium flaccumfaciens pv.flaccumfaciens

作　　者：张晓梅[1] 林友伟[1] 吴新华[2] 郭坚华[1] 

机构地区：[1]南京农业大学农业部病虫监测与治理重点开放实验室,江苏南京210095 [2]江苏省出入境检验检疫局,江苏南京210090

出　　处：《南京农业大学学报》2004年第3期42-45,共4页Journal of Nanjing Agricultural University

基　　金：国家自然科学基金 (3 9970 481);国家 863计划项目(2 0 0 1AA2 490 2 1);江苏省出入境检验检疫局项目(KJ15 2 0 0 1)

摘　　要：根据在GenBank已登录的萎蔫短小杆菌 (Curtobacterium flaccumfaciens)的基因间重复序列 ,用DNAman 5 0软件比较同源性 ,设计了一对引物CffF1和CffR2 ,并以菜豆萎蔫病菌的基因组DNA为模板进行扩增 ,得到了一段长 2 80bp的PCR特异产物。参试的其他属如棒形细菌属 (Clavibacter)、节杆菌属 (Arthrobacter)、拉氏杆菌属 (Rathayibac ter)、红球菌属 (Rhodococcus)细菌和萎蔫短小杆菌的其他致病变种 (Cur f pv oortii、Cur f pv poinsettiae、Cur f pv betae) ,以及其他植物病原细菌的基因组DNA均无扩增产物。该检测方法特异性强、灵敏度高 ,在 10According to the repetitive sequence obtained with Curtobacterium flaccumfaciens strains in GenBank,a pair of specific polymearase chain reaction(PCR)primers CffF1 and CffR2 were designed with the software DNAman 5 0.The primers(CffF1 and CffR2)specifically amplified a 280 bp fragment when Cur flaccumfaciens pv flaccumfaciens (Cff)genomic DNA was used as template,but did not amplify any PCR product from other bacteria.They included some phenotypically related bacteria,the species of Clavibacter,Arthrobacter,Rathayibacter,Rhodococcus and pathovars of Cur.flaccumfaciens pv oortii,Cur flaccumfaciens pv poinsettiae , Cur flaccumfaciens pv betae, and other pathogenic bacteria.Moreover,this PCR protocol can detect Cff sufficiently in 100 pg template DNA.It was demonstrated to be a highly sensitive,specific and rapid PCR assay for the detection of Cff.

关 键 词：菜豆 萎蔫病菌 PCR 检测技术 萎蔫短小杆菌 基因间重复序列 基因组DNA 

分 类 号：S436.43[农业科学—农业昆虫与害虫防治] S41-30[农业科学—植物保护]