Sequence analysis of the ITS region of rDNA was used to investigate the level of genetic diversity occurring within Hyaloperonospora parasitica, and to show the relationship between phylogenies of these fungi and their hosts (Brassicaceae). Maximum parsimony and neighbour-joining analyses were performed using sequences from 32 isolates of Hyaloperonospora and Perofascia, which infect core brassicaceous hosts. For comparison, five isolates of Peronospora were also studied. The constructed phylogenetic trees showed trichotomy, showing that Hyaloperonospora, Perofascia, and Peronospora have different evolutionary histories. Although isolates from Peronospora and Perofascia clearly formed respective clades, the Hyaloperonospora group allowed separation of the isolates into four distinct clades, which shared significantly low sequence similarities. We suggest that H. parasitica infecting brassicaceous hosts should be divided into a number of distinct species. The comparison of the phylogeny of H. parasitica and that of the Brassicaceae suggests that this fungus is closely related with tribes Arabidae and Brassiceae within this host family, illustrating the potential of downy mildews for co-evolution with their hosts.