绿色荧光蛋白基因在小麦禾谷镰刀菌中的表达与鉴定

为了获得稳定表达绿色荧光蛋白（GFP）的我国代表性小麦禾谷镰刀菌菌株，用于小麦抗赤霉病接种鉴定与分析，以禾谷镰刀菌武昌菌株Fg175原生质体为受体，经PEG介导将序列优化的水母GFP基因转入该菌株中，分析鉴定了转基因菌株GFP基因的表达。PCR分析表明，Fg175基因组中整合了GFP基因，Western blot检测到高效表达的GFP特异蛋白；激光共聚焦显微镜及体视镜观察进一步证实，转基因菌株Fg175-GFP的分生孢子、菌丝及接种小麦麦穗后，均可发出清晰的绿色荧光。

Expression and characterization of a green fluorescence protein gene in a transgenic Fusarium graminearum from wheat

To obtain a representative Fusarium head blight (FHB) strain from wheat in China that can stably express a green fluorescence protein and be used for FHB evaluation and analysis, a Fusarium graminearum strain, Fg175, isolated from wheat spike in Wuchang, China was subjected to generation of transgenic strains. Protoplasts from this strain were prepared and used for PEG-mediated genetic transformation with a vector carrying a codon-optimized GFP sequence. The transgenic F. graminearum strains were generated and the expression of GFP in the strains was molecularly characterized, together with fluorescence image analysis. The results indicated that the GFP gene was indeed integrated into the genome of the Fg175 strain as shown by PCR identification. Western blot revealed an expected, specific protein with a strong signal in all the transgenic strains, indicating a high expression level of the GFP. Further image analyses with fluorescence microscope confirmed a clear fluorescence in conidiaspores, mycelia and infected wheat spikes of the transgenic strain Fg175-GFP.