A RAPD Assay for Strain Typing of the Biotrophic Grape Powdery Mildew Fungus Uncinula necator Using DNA Extracted from the Mycelium

Abstract

Délye, C., Corio-Costet, M.-F., and Laigret, F. 1995. A RAPD assay for strain typing of the biotrophic grape powdery mildew fungus Uncinula necator using DNA extracted from the mycelium. Experimental Mycology 19, 234-237. We describe, for the first time, a RAPD assay using DNA extracted from the mycelium of a powdery mildew fungus, Uncinula necator, a pathogen of grape. No contamination by plant DNA was observed, and the resulting patterns were fully repetitive. RAPD profiles were unchanged when using two different DNA polymerases or three different thermocyclers. Thirteen strains were tested for amplification, using 95 primers. Only 4% of the amplified fragments were polymorphic. Cluster analysis revealed that the strains from the same geographical origin had the higher genetic similarity, suggesting a short-range dissemination of U. necator. This RAPD assay was also successfully applied to the grape downy mildew fungus, Plasmopara viticola, indicating that it can be used for other fungi which cannot be grown on artificial media.