瓜黑星病菌、枯萎病菌和蔓枯病菌的三重PCR检测  

Triplex PCR detection of Cladosporium cucumerinum,Fusarium oxysporum f. sp. niveum and Mycosphaerella melonis in infected plant tissues

作　　者：高永洋[1] 王楠[1] 高观朋[1] 王伟[1] 

机构地区：[1]华东理工大学生物反应器工程国家重点实验室,上海200237

出　　处：《植物病理学报》2010年第4期343-350,共8页Acta Phytopathologica Sinica

基　　金：国家自然科学基金项目(30871664);上海市重点攻关项目(08391911400);国家重点实验室专项经费(2060204)

摘　　要：通过测定黄瓜黑星病菌(Cladosporium cucumerinum)rDNA的ITS序列,比对近缘种及瓜类几种重要病原菌的ITS序列,设计出特异性引物HX-1/HX-2,经过对引物HX-1/HX-2PCR条件的优化,可以扩增出1条190bp的黄瓜黑星病菌特异性DNA条带,灵敏度达到1pg/μL。进一步将引物HX-1/HX-2和瓜类枯萎病菌、瓜类蔓枯病菌特异检测引物Fn-1/Fn-2、Mn-1/Mn-2组合,建立三重PCR体系,可一次检测出瓜类黑星病菌、瓜类枯萎病菌、瓜类蔓枯病菌3种瓜类植物重要的病原菌。建立了可以应用于田间瓜类黑星病菌PCR检测技术和瓜类主要病害三重PCR检测技术,对瓜类病害的诊断和防治具有重要的指导作用。rDNA ITS sequences of Cladosporium cucumerinum were sequenced.A pair of specific primers HX-1/HX-2 was designed from homology comparison of ITS sequences of C.cucumerinum with closely-rela-ted species and some pathogenic fungi on cucurbitaceous plants.A single special PCR band of 190 bp existed in C.cucumerinum was amplified under the optimized PCR parameters using HX-1/HX-2 primers and the detection sensitivity was 1 pg/μL of genomic DNA.Then a multiplex PCR system was developed to detect C.cucumerinum,Fusarium oxysporum f.sp.niveum and Mycosphaerella melonis simultaneously with combining primers HX-1/HX-2,Fn-1/Fn-2 and Mn-1/ Mn-2.It was the first time to establish the triplex PCR system for detecting C.cucumerinum,F.oxysporum f.sp.niveum and M.melonis in infected plant tissues simultaneously.The PCR-based method developed here could direct the diagnosis of plant disease and pathogen monitoring.

关 键 词：瓜黑星病菌 瓜枯萎病菌 瓜蔓枯病菌 分子检测 三重PCR 

分 类 号：S432.44[农业科学—植物病理学]