转绿色荧光蛋白基因的尖孢镰刀菌生物学特性 - yzxzcom.DOC

香蕉枯萎病原菌转gfp基因及其转化子生物学特性研究 李燕丹，肖荣凤刘波*，许文耀2 （福建省农业科学院农业生物资源研究所，福建 福州 350003），并能用于香蕉体内检测。关键词：尖孢镰刀菌专化型；绿色荧光蛋白基因；A Biochemical Characterization of Fusarium oxysporum f. sp. cubense Agent of Fusarium Wilt of Banana ,with the Green Fluorescent Protein(GFP)Gene LI Yan-dan , XIAO Rong-feng, LIU Bo,XU Wen-yao (1.Institute of Agrobiological Resouces, Fujian Academy of Agricultural Sciences, Fuzhou 350003, China;2. Institute of Plant Protect ,Fujian Agriculture and Forest University,Fuzhou 350002, China) Biochemical Characterization of Fusarium oxysporum f. sp. cubense Agent of Fusarium Wilt of Banana ,with the Green Fluorescent Protein(GFP)Gene LI Yan-dan , XIAO Rong-feng, LIU Bo,XU Wen-yao (1.Institute of Agrobiological Resouces, Fujian Academy of Agricultural Sciences, Fuzhou 350003, China;2. Institute of Plant Protect ,Fujian Agriculture and Forest University,Fuzhou 350002, China) Abstract:【Objective】To transform Fusarium oxysporum f. sp. cubense strain with gfp gene by protoplast methods.The biochemical characterization and the pathogenicity of the tagged strains and the wild type srtain were compared in this paper. The gfp-tagged srtain was used to mark this fungus in order to visualize and analyze the colonization and infection processes in vivo.【Method】PEG-mediated transformation system of Fusarium oxysporum f. sp. cubense was established. The process of the infection of the gfp-tagged srtain was observed in the banana plants using confocal laser scanning microscopy.【Result】Transformation Fusarium oxysporum f. sp. cubense Foc-3076 strain with gfp gene was successful. Morphologically，the cells of the transformed strain were similar to wild type srtain as determined. It was the same for the gfp-tagged strians and the wild type strain in the colony morphology，pH value and growth rate. The presence of the green fluorescent protein DNA in the fungal cells was confirmed by PCR. Resistance to hygromycin B of the transformants has improved. Fluorescence microscopy showed expr