Functional Analysis of FolSwe1 Gene in Fusarium oxysporumn f.sp.lini
To identify the function of the FolSwe1 gene in Fusarium oxysporum f.sp.lini,the coding region of the gene and its flanking sequence were amplified and sequenced,and a gene dele-tion cassette was constructed by using Split-Maker strategy.The FolSwe1 gene deletion mutant strains were obtained by transforming wild-type protoplasts mediated by polyethylene glycol(PEG).The phenotype and pathogenicity of the deletion mutant were assayed through conventional biological observation and measurement.A fluorescent vector pZESH1(pZWH1+EGFP+FolSwe1)was constructed for subcellular localization of the FolSwe1 protein.The sequencing results showed that the coding region of FolSwe1 gene is 3219 bp,containing one intron with length of 54 bp,and the obtained flanking sequence contained upstream 1704 bp and downstream 1694 bp of the coding region.Phenotypic observation revealed that the hyphae of the deletion mutant were shorter and grew denser compared with the wild-type,and the colony of deletion mutant was signifi-cantly smaller than that of the wild type.The most remarkable change was that the deletion mutant did not produce spores anymore,indicating that the gene was closely related to the conidiogenesis of the fungus.The infection assay to the seedling of the flax indicated that the pathogenicity of the deletion mutant decreased significantly.The subcellular localization showed that the fluorescence signals mainly appeared in the cell membrane.In summary,the FolSwe1 gene is mainly responsible for the conidiogenesis,vegetative growth and pathogenicity of the fungus.
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