The Colletotrichum nicotianae specimen was collected from main tobacco fields in ShandongrnProvince, and then was isolated, detected and identified. The results showed that the pathogen ofrnColletotrichum nicotianae was Colletotrichum destructivum 0' Gara. Based on the differences in internalrntranscribed spacer (ITS) sequence of C. nicotianae and other tobacco fungal pathogens, a pair of specificrnprimers Col/Co2 was designed. With this pair of primers, a single product of about 300 bp was cloned fromrnthe total DNA of C. destructivum. C. destructivum could be specifically detected with the primers Col/Co2rnfrom tobacco tissues infected naturally and soil samples. The sensitivity of detection was 0. 1 pg/(jJ. Itrnprovided basis for rapid and reliable detection of C. destructivum.%从山东临沂等地采集烟草炭疽病害标本,并进行分离、检测、鉴定.结果表明,山东烟草炭疽病原为毁灭炭疽菌.根据烟草炭疽菌与其它烟草病害病原菌核糖体转录间隔区(internal transcribed spacer,ITS)序列间的差异,设计了一对特异性引物Co1/Co2.利用此引物可从烟草炭疽菌总DNA中扩增约300 bp的特异性片段,其余菌株和对照PCR结果为阴性.灵敏度实验证明,可以检测到目的片段的DNA浓度为0.1 pg/μl.对土样检测结果表明,该引物能特异性地检测到Colletotrichum destructivum的存在,为快速、灵敏地检测烟草炭疽菌提供了应用依据.
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