Transformation of the oomycete pathogen Phytophthora megasperma f. sp. glycinea occurs by DNA integration into single or multiple chromosomes.
A procedure for stable transformation was developed for Phytophthora megasperma f. sp. glycinea, an oomycete pathogen of soybean. Transformants were obtained using a bacterial hygromycin resistance gene fused to a promoter and terminator from the ham34 gene of another oomycete, Bremia lactucae. Vector DNA, alone or complexed to cationic liposomes, was introduced into protoplasts using polyethylene glycol and CaCl2. DNA and RNA hybridization, and phosphotransferase assays, confirmed the presence and expression of vector DNA in the transformants. Hybridization to electrophoretically separated chromosomes of P. m. glycinea showed that vector DNA had integrated into only one chromosome in four transformants, and into multiple chromosomes in one transformant.
Judelson
H S
HS
Department of Vegetable Crops, University of California, Davis 95616.
Coffey
M D
MD
Arredondo
F R
FR
Tyler
B M
BM
D002874:Chromosome Mapping; D004271:DNA, Fungal / Q000235:genetics*; D004352:Drug Resistance, Microbial; D015870:Gene Expression; D005822:Genetic Vectors; D010770:Phosphotransferases / Q000235:genetics; D017853:Phosphotransferases (Alcohol Group Acceptor); D010838:Phytophthora / Q000235:genetics*
D004271:DNA, Fungal; D010770:Phosphotransferases; D017853:Phosphotransferases (Alcohol Group Acceptor); C063242:hygromycin-B kinase