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Molecular identification and detection of tobacco anthracnose pathogen

CHAI Xin;XU Yeping;YAO Jian;YU Xiaofeng;ZONG Kai;LI Yunfei;JIANG Tong;School of Plant Protection, Anhui Agricultural University;Technical Center, Anhui Entry-Exit Inspection and Quarantine Bureau;  
The r DNA complete sequence of tobacco anthracnose pathogen was cloned and sequenced. The full length of the r DNA sequence is 2870 bp. Sequence comparison results showed that the rDNA of tobacco anthracnose pathogen shared a high sequence similarity(96.0%-96.2%) with that of Colletotrichum gloeosporioides. Furthermore, from the constructed phylogenetic tree, it was indicated that tobacco anthracnose pathogen and C. gloeosporioides clustered into a separate branch, which indicated that tobacco anthracnose pathogen had the closest relationship with C. gloeosporioides. Therefore, combined with its morphology characteristic, tobacco anthracnose pathogen isolated from tobacco plant in Guizhou could be preliminarily identified as C. gloeosporioides. A pair of specific primer was designed according to the rDNA sequence of tobacco anthracnose pathogen. Not only tobacco anthracnose pathogen could be identified from 8 different pathogens isolated from tobacco plants, but also it could be identified from 7 different anthracnose pathogens. Plants of Nicotiana tabacum were inoculated with tobacco anthracnose pathogen. Using the total DNA extracted from the diseased tissue as a template, a specific band could be amplified by PCR with the specific primer. It was suggested that the method could be applied for the identification and rapid detection of C. gloeosporioides from tobacco plants.
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