柏木枯萎病病原菌的形态特征及分子鉴定

Identification on Pathogen of Chinese Weeping Cypress Dieback Disease

采用温室接种致病性测定、形态学观察和分子鉴定方法对柏木枯萎病病原进行了研究。从云南省昆明市、石林县及玉溪市采集柏木枯萎病病样13份,分离获得来自柏木树根部、茎干及枝的18个真菌分离物。经过致病性试验证实,分离物CWC1-CWC10均为柏木枯萎病病原菌。经形态学鉴定,CWC1-CWC10均为Neofusicoccum parvum。利用rDNA internal transcribed spacer region(ITSI-5.8S-ITS2)进行PCR扩增,并将测序结果在GenBank中进行同源性比对分析显示,其rDNA ITS序列分析结果表明该菌与N.parvum的同源性为99%,分子鉴定与形态学鉴定结果一致。Neofusicoccum parvum作为柏木枯萎病的病原菌系首次报道。

The pathogens of Chinese weeping cypress（ Cupressus funebris Endl）dieback disease were identified on the basis of pathogenicity testing, as well as morphological characters and molecular identification. Thirteen diseased Chinese weeping cypress samples were observed in Kunming, Shilin, and Yuxi in Yunnan Province（southwestern China）. Eighteen isolates of possible pathogenic fungi were isolated from roots, trunks and branches of infected Chinese weeping cypress plants. Bases on the pathogenicity experiment, the results showed that CWC1- CWC10 were the pathogens of Chinese weeping cypress wilt disease. The morphological identification showed that CWC1-CWC10 was identified as Neofusicoceum parvum. By analysis of the rDNA internal transcribed spacer region （ITSI-5.8S-ITS2）. BLAST searches at GenBank showed a high identity with reference sequences（ ITS： 〉 99% ）. Representative sequences of both regions were deposited to GenBank （ITS：Accession No. FJ842960）. Morphological and molecular results confirmed this species as N. parvum, N. parvum was first reported as the pathogen of Chinese weeping cypress dieback disease.