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根据黄瓜棒孢叶斑病菌多主棒孢(Corynespora cassiicola)与棒孢属下女贞棒孢(Corynespora lieustri)、威尔士棒孢(Corynespora cambrensis)和其他常见病原真菌Actin基因序列差异,设计多主棒孢的特异性引物Caa5F/Caa5R,建立了黄瓜棒孢叶斑病菌的PCR检测方法,可对引起黄瓜棒孢叶斑病的病原菌扩增出160 bp的特异性条带,检测灵敏性为4 pg·μL-1 DNA,并可从接种后发病的黄瓜叶片总DNA中检测到特异条带。该引物的PCR检测方法灵敏性较高,可直接检测植株总DNA,无需病原菌的分离培养,适用于对黄瓜棒孢叶斑病特异快速的检测。
According to the sequence differences of Corynespora cassiicola and Corynespora lieustri, Corynespora cambrensis and other common pathogenic fungi Actin, The specific primers Caa5F / Caa5R of Cucumber were used to establish the PCR detection method of Cucumber leaf spot cucumber, which could amplify the specific band of 160 bp for the pathogen causing cucumber leaf spot cucumber disease. The detection sensitivity was 4 pg · ΜL-1 DNA, and specific bands could be detected from the total DNA of cucumber leaves after inoculation. The PCR detection method of the primer has high sensitivity and can directly detect the total DNA of the plant without isolating and culturing the pathogen and is suitable for detecting the cucumber leaf spot leaf spot disease rapidly and specifically.