杨晓琦, 周小军, 朱丽燕, 陈向阳, 何晓婵, 张传清. 金线莲炭疽病病原菌的分离鉴定及其对9种杀菌剂的敏感性[J]. 农药学学报, 2020, 22(6): 951-958. DOI: 10.16801/j.issn.1008-7303.2020.0127
    引用本文: 杨晓琦, 周小军, 朱丽燕, 陈向阳, 何晓婵, 张传清. 金线莲炭疽病病原菌的分离鉴定及其对9种杀菌剂的敏感性[J]. 农药学学报, 2020, 22(6): 951-958. DOI: 10.16801/j.issn.1008-7303.2020.0127
    YANG Xiaoqi, ZHOU Xiaojun, ZHU Liyan, CHEN Xiangyang, HE Xiaochan, ZHANG Chuanqing. Isolation and identification of pathogen causing anthracnose on Anoectochilus roxburghii and its sensitivity to nine fungicides[J]. Chinese Journal of Pesticide Science, 2020, 22(6): 951-958. DOI: 10.16801/j.issn.1008-7303.2020.0127
    Citation: YANG Xiaoqi, ZHOU Xiaojun, ZHU Liyan, CHEN Xiangyang, HE Xiaochan, ZHANG Chuanqing. Isolation and identification of pathogen causing anthracnose on Anoectochilus roxburghii and its sensitivity to nine fungicides[J]. Chinese Journal of Pesticide Science, 2020, 22(6): 951-958. DOI: 10.16801/j.issn.1008-7303.2020.0127

    金线莲炭疽病病原菌的分离鉴定及其对9种杀菌剂的敏感性

    Isolation and identification of pathogen causing anthracnose on Anoectochilus roxburghii and its sensitivity to nine fungicides

    • 摘要: 为明确金线莲炭疽病的病原菌,根据科赫法氏则采用组织分离法对采集的病叶进行了病原菌分离,确定其致病性后,结合形态学特征及基于肌动蛋白基因 (actin gene-ACT)、β2微管蛋白基因 (β2-tubulin gene-TUB2)、几丁质合酶基因 (chitin synthase A gene-CHS-I)、磷酸甘油醛脱氢酶基因 (glyceraldehyde-3-phosphatedehydrogenase gene-GAPDH) 和核糖体内部转录间隔区 (ITS) 联合系统发育分析对该病原菌进行了鉴定,并测定了9种杀菌剂对该病原菌的离体抑菌活性。结果表明:分离得到的炭疽菌菌株在金线莲离体叶片上表现出的症状与田间自然发病症状相似,证明所分离的菌株为引起金线莲炭疽病的致病菌。根据病原菌的菌落、分生孢子及附着胞形态等初步判断其为长直孢炭疽菌 Colletotrichum gigasporum;进一步基于ACTCHS-I、GAPDH、ITS和TUB2联合系统发育分析的分子鉴定结果与形态学鉴定结果一致,确定了引起该病害的病原菌为长直孢炭疽菌。离体抑菌活性测定结果表明,咪鲜胺、戊唑醇、肟菌酯、氯苯醚酰胺、苯醚甲环唑、丙硫菌唑、氟啶胺、甲基硫菌灵和百菌清对供试病原菌的EC50值分别为0.0145、0.240、18.2、29.9、0.114、0.402、0.0191、2.80和38.5 μg/mL,其中咪鲜胺和氟啶胺的抑菌活性最强。

       

      Abstract: In this study, the causative pathogen of anthracnose on Anoectochilus roxburghii was classified. According to Koch's law, tissue separation method was used to isolate the pathogen from the diseased leaves. The pathogenic fungus was identified based on the morphologic characteristics and multi-locus phylogenetic analysis by combing five genes, including actin ACT, β2-tubulin gene TUB2, chitin synthase A gene CHS-I, glyceraldehyde-3-phosphate dehydrogenase gene GAPDH, and ribosome internal transcription spacer ITS. And the in vitro inhibitory activities of nine common fungicides against this pathogen were determined to provide a basis for its control. In the pathogenicity tests, the isolates caused similar symptoms as the natural symptom in the fields, indicating they were the pathogenic fungi. According to the colony, conidial and appressorium morphology of pathogenic fungi, it was preliminarily judged as Colletotrichum gigasporum. The molecular identification based on the phylogenetic analysis of ACT, CHS-I, GAPDH, ITS and TUB2 was consistent with the morphological identification. Thus, the pathogen causing the anthracnose disease on A. roxburghii was identified to be C. gigasporum. The results of the in vitro fungicidal activity showed that the EC50 values of prochloraz, tebuconazole, trifloxystrobin, Y13149, difenoconazole, prothioconazole, fluazinam, thiophanate-methyl and chlorothalonil against C. gigasporum were 0.0145, 0.240, 18.2, 29.9, 0.114, 0.402, 0.0191, 2.80, and 38.5 μg/mL, respectively. Among them, prochloraz and fluazinam had the strongest fungicidal activities.

       

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