Green fluorescent protein (GFP) as a reporter gene for the plant pathogenic oomycete Phytophthora ramorum.
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.
Riedel
Marko
M
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.
Calmin
Gautier
G
Belbahri
Lassaad
L
Lefort
Francois
F
Götz
Monika
M
Wagner
Stefan
S
Werres
Sabine
S
D002122:Calcium Chloride; D018929:Cell Culture Techniques; D004271:DNA, Fungal / Q000235:genetics / Q000378:metabolism; D017930:Genes, Reporter; D005821:Genetic Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000378:metabolism; D054884:Host-Pathogen Interactions; D018613:Microscopy, Confocal; D010838:Phytophthora / Q000235:genetics* / Q000254:growth & development; D016133:Polymerase Chain Reaction; D029793:Rhododendron / Q000382:microbiology*; D014170:Transformation, Genetic
D004271:DNA, Fungal; D049452:Green Fluorescent Proteins; D002122:Calcium Chloride