With ∼7000 species in ∼220 genera, the Dolichopodidae is one of the most speciose families of Diptera. Though the family as such is well defined, knowledge on the internal phylogenetic relationships is generally poor and although authors of successive monographs and catalogues indifferently listed most genera in specific subfamilies, their decisions were rarely based on sound phylogenetic analyses and never on molecular data. In a first attempt to unravel the phylogeny of Dolichopodidae, a combined COI + 12S rDNA dataset (1199 characters) of 119 samples of 101 European species was used in Bayesian (BAY), neighbour joining (NJ) and weighted/unweighted maximum parsimony analyses (MP). At the subfamily level, our study supports the monophyly of Dolichopodinae, Sympycninae, and Hydrophorinae (including Machaerium Haliday, 1832). Ten (Campsicnemus Haliday, 1851, Chrysotus Meigen, 1824, Dolichopus Latreille, 1796, Gymnopternus Loew, 1857, Medetera Fischer von Waldheim, 1819, Poecilobothrus Mik, 1878, Rhaphium Meigen, 1803, Teuchophorus Loew, 1857, Sciapus Zeller, 1842, Syntormon Loew, 1857) of the 14 multispecies genera formed monophyletic assemblages in all analyses and relationships among Argyra Macquart, 1834 species were supported in most analyses. At the subgeneric level, seven of the nine stable species-groups in Dolichopus as established during previous research were supported in most analyses. The validity of the recent transfer of Hercostomus chrysozygos Wiedemann, 1817 to Poecilobothrus was clearly supported in all analyses. Within Argyra and Rhaphium, interspecific relationships reflected previously used subgeneric classifications (Lasiargyra Mik, 1878, Leucostola Loew, 1857 and Argyra s.s. in Argyra; Porphyrops Meigen, 1824, Rhaphium s.s. and Xiphandrium Loew, 1857 in Rhaphium). Further, subclades within Medetera corresponded to species-groups defined by other authors featuring a different morphology and ecology. Anepsiomyia flaviventris (Meigen, 1824) most probably does not belong to Sympycninae but its current position within Peloropeodinae could not be confirmed. © CSIRO 2007.