Sequence parts of the internal transcribed spacer (ITS) region of nuclear ribosomal DNA were analysed to screen for the intraspecific variability of a non-coding genomic region in 15 Plasmopara halstedii populations of different pathotype and geographic origin. Samples revealed uniformity in a ca. 790 Bp fragment comprising of the ITS-1, 5.8S and front parts of the ITS-2. In contrast, clear differences were found in a ca. 810 Bp fragment of the ITS-2 thus allowing differentiation between populations of pathotype 100, 310 and 330 and a group of populations representing pathotypes 700, 701, 703, 710 and 730. Samples of pathotypes 700 to730 originated from Slovakia, France, and Germany, but were uniform in both ITS sequence parts, thus indicating very recent origin of these highly aggressive physiological races. The potential use of ITS sequences for pathotype differentiation and phylogenetic studies in P. halstedii is discussed.