Analysis of total DNA by minisatellite and simple-sequence repeat primers for the use of population studies inPlasmopara halstedii

Abstract

Total DNA of Plasmopara halstedii isolates from Germany was analysed for polymorphisms potentially useful for the differentiation of field isolates with respect to epidemiological studies or pathotype characterization. The isolation of the DNA started from mitotically formed zoosporangia, which is the only cellular structure of the biotrophic pathogen accessible independently from its host. The total DNA of the pathogen was used to perform DNA fingerprints with minisatellite and simple-sequence repeat primers. Polymorphisms were found that allowed differentiation on the level of single field isolates; however, they were not correlated with either physiological races or the geographic origin of the isolates. Using such differentiating primers, single spore strains of three pathogen isolates were also analysed with respect to genetic homogeneity. Minor variation was visible in the mitotically derived offspring, but the overall appearance of these patterns was mostly uniform with those of the respective parental isolate.Key words: Plasmopara halstedii, polymerase chain reaction (PCR), simple sequences, mitochondrial DNA, Helianthus annuus.