Fluorescent Labeling the Peroxisome and Nucleus in Fusarium oxysporum f. sp. niveum
SHI Xiaoxiao1,2, WANG Jiaoyu2*, XIAO Chenwen3, WANG Yanli2, LI Dayong4, CHAI Rongyao2, SUN Guochang2*
FON (Fusarium oxysporum f. sp. niveum) causes watermelon Fusarium wilt, a destructive disease on watermelon worldwide. Research on the development and pathogenesis of FON lays the foundation for the control of the disease. Fluorescent labeling of the organelles and cell structures using fluorescent proteins is an important strategy in the investigations on fungal development and pathogenesis. In the present work, the nuclei and peroxisomes of FON were labeled with green or red fluorescent proteins. Via AtMT (Agrobacterium tumefaciens -mediated transformation), we generated three types of FON transformants which carried nuclei labeled with mCherry, peroxisomes labeled with GFP or peroxisomes labeled with DsRED, respectively. In the strains with mCherry labeled nuclei, the bright red fluorescence in round dots were detected in hyphae and conidia, overlaying well with the fluorescence formed by DAPI staining. In the strains with GFP or DsRED labeled peroxisomes, green or red small fluorescent dots were present in hyphal and conidial cells, corresponding with the distribution of peroxisomes in fungal cells. Further, the numbers of the fluorescent dots was increased significantly on lipids. Calcofluor white staining was also performed on the three transformants. Under confocal microscopy, the blue fluorescence of Calcofluor white cooperated well with the fluorescence of green or red fluorescent proteins, producing ideal multi-fluorescent images. In addition, the fluorescent proteins could be stably expressed and well distributed during the subcultivation of the transformants. The growth and phenotypes of the transformants were unaltered compared with the wild type strain. We provided a useful tool for the study on the organelle dynamics, development and pathogenesis of FON.
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