The Western Mediterranean populations of D. subobscura have a very similar chromosomal inversion polymorphism to the American colonizing populations, with the exception of the O5 inversion. This inversion, which is found in appreciable frequencies in colonizing populations and is distributed according to a significant latitudinal cline, both in North and South America, has never been reported in that Palearctic region. Therefore, an analysis of the distribution of this inversion, along a latitudinal cline, has been carried out in Europe. The O Chromosome inversion polymorphism has been studied in seven populations: Gävle and Lilla Edet (Sweden), Gesten (Denmark), Ter‐Apel (Holland) and Crézy, Aizenay and Taulé (France). The O5 inversion was only detected in three populations and in low frequencies: Gävle (3.7%), Lilla Edet (1.8 %) and Taulé (1.2 %). However, none of these three populations has all the inversions found in the American populations. The clinal distribution of some O chromosomal arrangements has also been studied according to latitude and the temporal changes of this polymorphism.