猕猴桃间座壳菌Diaporthe actinidiae的绿色荧光蛋白基因标记及侵染过程Green fluorescent protein gene markers and infection of Diaporthe actinidiae
冯丹丹;邓蕾;苏琦雅;刘德江;钟彩虹;李黎;
摘要(Abstract):
果实软腐病是猕猴桃贮藏期间最严重的真菌病害,猕猴桃间座壳Diaporthe actinidiae是该病检出率最高且致病力最强的病原菌。该病菌从花前期开始侵染,至果实贮藏期才表现症状,侵染至发病周期较长,可借助荧光标记对其侵染过程进行研究。本研究采用PEG介导原生质体转化的技术,运用携带GFP及潮霉素抗性基因的双元载体pCAMBgfp对D. actinidiae菌株进行遗传转化。首先,对转化子进行潮霉素及绿色荧光筛选,随后进行菌落的生物学特征观察、荧光稳定性验证及致病力检测,最终得到3株菌丝形态、生长速度及致病力与野生型菌株一致的稳定荧光菌株。将KFDA-17转化子接种于贮藏期猕猴桃果实‘金梅’,1 d后果实出现水渍症状,随后出现软腐症状且逐步加重;发病部分菌丝体在荧光显微镜下呈现明显的绿色荧光,可清晰观察侵染进程。本研究成功构建了PEG介导的间座壳菌丝体的原生质体遗传转化体系,且KFDA-17荧光菌株可用于侵染过程研究。
关键词(KeyWords): 猕猴桃软腐病;荧光蛋白;遗传转化;转化子;致病力
基金项目(Foundation): 国家重点研发计划项目(2019YFD1000200);; 国家自然科学基金青年科学基金(31701974,31901980);; 长江猕猴桃产业技术研究项目(CJZX20210102)~~
作者(Authors): 冯丹丹;邓蕾;苏琦雅;刘德江;钟彩虹;李黎;
DOI: 10.13346/j.mycosystema.210410
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