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    Author(s): Marko Riedel; Gautier Calmin; Lassaad Belbahri; Francois Lefort; Monika Gotz; Stefan Wagner; Sabine Werres
    Date: 2009
    Source: Journal of Eukaryotic Microbiology. 56: 130-135
    Publication Series: Scientific Journal (JRNL)
    PDF: Download Publication  (1.0 MB)

    Description

    Transgenic Phytophthora ramorum strains that produce green fluorescent protein (GFP) constitutively were obtained after stable DNA integration using a polyethylene glycol and CaCl2-based transformation protocol. Green fluorescent protein production was studied in developing colonies and in different propagules of the pathogen to evaluate its use in molecular and physiological studies. About 12% of the GFP transformants produced GFP to a level detectable by a confocal laser scanning microscope. Green fluorescent protein could be visualized in structures with vital protoplasm, such as hyphal tips and germinating cysts. In infection studies with Rhododendron, one of the GFP expressing strains showed aggressiveness equal to that of the corresponding non-labelled isolate. Thus, GFP could be used as a reporter gene in P. ramorum. Limitations of the technology are discussed.

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    Citation

    Riedel, Marko; Calmin, Gautier; Belbahri, Lassaad; Lefort, Francois; Gotz, Monika; Wagner, Stefan; and Werres, Sabine. 2009. Green Fluorescent Protein (GFP) as a reporter gene for the plant pathogenic oomycete Phytophthora ramorum. Journal of Eukaryotic Microbiology. 56: 130–135

    Keywords

    Chromists, DNA transformation, stramenopiles.

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