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作 者:盛家伟 满益龙 欧阳超 刘思珍 郭盛 孙乾隆 陈岳[3] 张鑫[3] 谭新球 王运生[1] SHENG Jiawei;MAN Yilong;OUYANG Chao;LIU Sizhen;GUO Sheng;SUN Qianlong;CHEN Yue;ZHANG Xin;TAN Xinqiu;WANG Yunsheng(Plant Protection College,Hunan Agricultural University,Changsha 410128,China;Longping Branch,Graduate School of Hunan University,Changsha 410125,China;Plant Protection Institute,Hunan Academy of Agricultural Science,Changsha 410125,China;Agricultural Biotechnology Institute,Hunan Academy of Agricultural Science,Changsha 410125,China)
机构地区:[1]湖南农业大学植物保护学院,长沙410128 [2]湖南大学研究生院隆平分院,长沙410125 [3]湖南省农业科学院植物保护研究所,长沙410125 [4]湖南省农业科学院农业生物技术研究所,长沙410125
出 处:《植物病理学报》2023年第1期110-118,共9页Acta Phytopathologica Sinica
基 金:国家自然科学基金项目(31972223);国家自然科学青年基金项目(31701814);湖南农业科技创新资金项目(2020CX39、2020CX43、2021CX86)。
摘 要:由辣椒尖孢炭疽菌(Colletotrichum acutatum)侵染引起的炭疽病是辣椒生产中最具有破坏性的真菌病害,严重影响辣椒的品质和产量。本研究以辣椒尖孢炭疽病菌HHDL02为对象,采用1 mol·L^(-1)NH_(4)Cl 2%的酶裂解液裂解分生孢子萌发2 h的芽管,裂解2~3 h可以高效制备原生质体,结合PEG介导转化法成功将GFP基因导入,其转化菌株菌落形态、菌丝生长速率、分生孢子形态、孢子萌发、附着胞形成、产孢量和致病性与野生型菌株无明显差异,且后代荧光信号遗传稳定。PEG介导的原生质体转化适合辣椒尖孢炭疽菌遗传操作,有助于其致病机理的研究。Pepper anthracnose, caused by Collectotrichum acutatum infection, is the most destructive fungal disease in pepper production, which seriously threatens yield and quality of pepper. The genetic transformation with higher efficiency for the pepper pathogen C. acutatum is still not available. Our research provided optimal conditions to obtain protoplasts with higher efficiency at 2% of the enzyme in 1 mol·L^(-1)NH_(4)Cl solution, 2 h of germination for the germ tube of conidium, and 2-3 h of incubation with application of the protoplasts, the GFP gene was successfully introduced into the wild type C. acutatum strain HHDL02 through modified PEG-mediated transformation. We further evaluated some of the transformants for colony morphology, growth rate, conidium morphology, conidial germination, appressorium formation, conidial production quantity and pathogenicity, which exhibited no obvious differences as compared to the wild type strains. In addition, visualizing the fluorescence signals of their progenies of the transformants suggested that the GFP gene integrated in C. acutatum strain HHDL02 was genetically stable. These results indicated that the protoplasts produced efficiently with the optimal approach from germ tube and the subsequent transformation mediated by PEG was suitable for the genetic operation of C. acutatum, and also helpful for the study on the mechanism of pathogenesis on pepper plants.
关 键 词:辣椒尖孢炭疽菌 PEG介导转化 绿色荧光蛋白 芽管
分 类 号:S436.418.11[农业科学—农业昆虫与害虫防治]
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