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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。
The specific primers HX-1 / HX-2 were designed by determining the ITS sequence of Cladosporium cucumerinum rDNA ITS sequences and comparing the ITS sequences of several important pathogenic bacteria in melon and related species. / HX-2PCR conditions can be optimized to amplify a 190bp cucumber black spot bacteria-specific DNA bands, the sensitivity of 1pg / μL. The primer HX-1 / HX-2 and Fusarium oxysporum f. Sp., Fusarium solani specific primers Fn-1 / Fn-2 and Mn-1 / Mn-2 were further combined to establish a triplex PCR system, Melons black star bacteria, Fusarium wilt bacteria, melon gourd bacteria three kinds of melons important pathogens. The establishment of a triplex PCR detection technique that can be applied to the field detection of melon black spot bacteria and the main diseases of melons has an important guiding role in the diagnosis and prevention of melon diseases.