Green Fluorescent Protein (GFP) as a Reporter Gene for the Plant Pathogenic Oomycete Phytophthora ramorum |
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Authors: | MARKO RIEDEL GAUTIER CALMIN LASSAAD BELBAHRI FRANCOIS LEFORT MONIKA GÖTZ STEFAN WAGNER SABINE WERRES |
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Institution: | Julius Kuehn-Institute (JKI), Federal Research Centre for Cultivated Plants, Institute for Plant Protection in Horticulture and Forests (GF), Messeweg 11/12, 38104 Braunschweig, Germany, and; Plants and Pathogens Group, Institute Earth Nature and Landscape, University of Applied Sciences of Western Switzerland, 150 Route de Presinge 1254 Jussy, Switzerland, and; Julius Kuehn-Institute (JKI), Federal Research Centre for Cultivated Plants, Institute for Biosafety of Genetically Modified Plants, Messeweg 11/12, 38104 Braunschweig, Germany |
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Abstract: | ABSTRACT. 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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Keywords: | Chromists DNA transformation stramenopiles |
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