Transformation of the oomycete pathogen, Phytophthora infestans.

H. S. Judelson, B. M. Tyler, Richard W Michelmore

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Abstract

A stable transformation procedure has been developed for Phytophthora infestans, an oomycete fungus that causes the late blight diseases of potato and tomato. This is the first description of reliable methods for transformation in an oomycete pathogen. Drug-resistant transformants were obtained by using vectors that contained bacterial genes for resistance to hygromycin B or G418 fused to promoters and terminators from the Hsp70 and Ham34 genes of the oomycete, Bremia lactucae. Using polyethylene glycol and CaCl2, vector DNA was introduced into protoplasts as a complex with cationic liposomes or with carrier DNA only. Transformants were obtained at similar frequencies with each combination of promoter and selectable marker and were confirmed by DNA and RNA hybridization and phosphotransferase assays. Transformation occurred through the integration of single or tandemly repeated copies of the plasmids into genomic DNA, conferring mitotically stable drug-resistant phenotypes. The sizes of the marker gene mRNAs in each transformant and the results of transcript mapping studies were consistent with the function of the B. lactucae regulatory sequences in P. infestans. A hygromycin-resistant transformant was tested and found to maintain pathogenicity, indicating that the gene transfer procedure will be useful for the molecular analysis of genes relevant to disease.

Original languageEnglish (US)
Pages (from-to)602-607
Number of pages6
JournalMolecular plant-microbe interactions : MPMI
Volume4
Issue number6
StatePublished - Nov 1991

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ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Biotechnology
  • Microbiology

Cite this

Judelson, H. S., Tyler, B. M., & Michelmore, R. W. (1991). Transformation of the oomycete pathogen, Phytophthora infestans. Molecular plant-microbe interactions : MPMI, 4(6), 602-607.