AbstractContarinia comprises one of the largest genera among gall forming midges of Cecidomyiidae, where identification and species relationships are uncertain. Using data on phenological development, morphometric relationships and mitochondrial DNA, the status of two isomorphic species, C. vincetoxici and C. asclepiadis, which attack the perennial herb Vincetoxicum hirundinaria, were investigated. Data show that they are two distinct species. In rearing experiments, the two gall midges were shown to have different times of adult emergence. Small differences in wing morphology were revealed that separate the two species from each other, as well as from C. loti, the type species of Contarinia. Sequence differences in the mitochondrial cytochrome b gene corroborate the specific status of C. vincetoxici and C. asclepiadis. Furthermore, a phylogenetic analysis, also including three other Contarinia species, showed that the two gall midges on V. hirundinaria are not even the most closely related species, suggesting two separate evolutionary colonizations of the host plant.