山东省主产烟区烟草炭疽病菌的鉴定与分子检测

Identification and Molecular Detection of Colletotrichum nicotianae Pathogen from Main Tobacco Fields in Shandong Province

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 spacerI,TS)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。

The Colletotrichum nicotianae specimen was collected from main tobacco fields in Shandong Province, and then was isolated, detected and identified. The results showed that the pathogen of Colletotrichum nicotianae was Colletotrichum destructivum O＇ Gara. Based on the differences in internal transcribed spacer （ITS） sequence of C. nicotianae and other tobacco fungal pathogens, a pair of specific primers Col/Co2 was designed. With this pair of primers, a single product of about 300 bp was cloned from the total DNA of C. destructivum. C. destructivum could be specifically detected with the primers Col/Co2 from tobacco tissues infected naturally and soil samples. The sensitivity of detection was 0. 1 pg/μl. It provided basis for rapid and reliable detection of C. destructivum.