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 analysis 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.