董伟清 何芳练 江 文 高美萍 张尚文 王 艳 韦绍龙
蒋慧萍 陈丽娟 闭志强 欧昆鹏 颜梅新*
|单位： （广西农业科学院 生物技术研究所 南宁 530007）|
|关键词： 荔浦芋 疫病 芋疫霉菌 鉴定 原生质体|
|出版年,卷(期):页码：2016 ,47 ( 11 )： 页码：1861-1866|
【目的】鉴定荔浦芋疫病病原及制备原生质体，为荔浦芋病原检测、致病机理研究及健康种苗生产提供科学依据。【方法】对采集自广西荔浦县芋头种植区的芋疫病标样进行分离，通过形态特征和rDNA-ITS分子生物学相结合的方法对其病原进行鉴定，同时对获得的病原菌进行原生质体制备。【结果】通过对分离的芋疫病病原进行致病性测定、形态特征观察，将引起荔浦芋疫病的病原初步鉴定为芋疫霉菌（Phytophthora colocasiae）；rDNA-ITS序列分析结果表明，菌株DNA序列与GenBank已发表的P. colocasiae不同分离物序列同源性达99%，进一步确定所测菌株为芋疫霉菌。制备获得的芋疫霉菌原生质体呈透明圆形或近圆形，大小不一。【结论】引起荔浦芋疫病的病原为芋疫霉菌，制备的芋疫霉菌原生质体可用于芋疫霉病致病机理研究。
【Objective】The present study was conducted to identify the causal pathogen of taro leaf blight disease by using morphology and molecular methods， and establish a protoplast preparation protocol， in order to provide scientific basis for pathogen detection of taro leaf blight， pathogenesis and virus-free and healthy seedling production. 【Method】The samples were collected from taro production area in Lipu county， Guangxi. The pathogen was isolated， and identified through morphological examination combined with rDNA-ITS sequence analysis. Protoplast of pathogen was prepared by enzymatic digestion.【Result】The taro leaf blight pathogen was preliminarily identified as Phytophthora colocasiae based on its morphological characteristics and pathogenicity test. The results of rDNA-ITS sequence analysis indicated that the nucleotide sequence of the isolate was 99% similar to the isolates of P. colocasiae strains available in GenBank， which confirmed further that the isolate was P. colocasiae. Furthermore， the prepared protoplasts of P. colocasiae were circular or nearly circular， small or large， transparent.【Conclusion】The causal pathogen of taro leaf blight in Lipu of Guangxi is identified as P. colocasiae and its prepared protoplast could be used for pathogenesis study of taro leaf blight disease.