番茄细菌性叶斑病菌超分支滚环扩增快速检测技术  

Rapid detection of Pseudomonas syringae pv.tomato using hyperbranched rolling circle amplification

作　　者：王念武[1,2] 王婷[1] 沈建国[2] 胡方平[1] 

机构地区：[1]福建农林大学植物保护学院,福州350002 [2]福建出入境检验检疫局,福州350001

出　　处：《植物保护》2014年第2期90-94,122,共6页Plant Protection

基　　金：国家质检总局项目(20011K158);国家自然科学基金项目(30671463);福建省发改委项目(KY0030060)

摘　　要：根据番茄细菌性叶斑病菌(Pseudomonas syringae pv.tomato,Pst)的一段特异蛋白基因序列,按照锁式探针的设计原理设计探针和扩增引物,优化系列反应条件,建立了特异性的Pst超分支滚环扩增技术。试验结果表明:该检测技术能够从供试的10种不同的病原菌中特异性地检测出番茄细菌性叶斑病菌,DNA检测的最低浓度为500fg/μL,检测灵敏度高于常规PCR。A padlock probe was designed based on the sequence of the unique hypothetic protein gene in the complete genome of Pseudomonas syringae pv.tomato （Pst）,according to the design principle of padlock probe and primers.Detection system of hyperbranched rolling circle amplification （HRCA） was established and optimized.The results showed that Pst could be specifically detected by this method,while other reference plant pathogens could not be detected.The detection sensitivity of HRCA was about 500 fg/μL of DNA concentration,higher than that of the conventional PCR method.

关 键 词：番茄细菌性叶斑病菌 超分支滚环扩增 锁式探针 

分 类 号：S436.412[农业科学—农业昆虫与害虫防治]