Phylogeny and higher classification of the cerambycid subfamily Lepturinae remain controversial. Here we report the results of a cytogenetic study of 18 species currently classified in Lepturini and 12 species in other tribes of Lepturinae (1 in Oxymirini, 1 in Rhamnusiini and 10 in Rhagiini) from Western Europe. The male sex chromosome formula is XY in all Rhagiini, Oxymirini and in Grammoptera ruficornis in the Lepturini (whose tribal placement may be doubtful), and X0 in all the remaining Lepturini. The rarity of the X0 formula in other Cerambycidae indicates that the Y chromosome was lost in a common ancestor of the Lepturini or a subgroup thereof, indicating its monophyletic origin. The number of chromosomes is stable in the Lepturini that lack a Y chromosome (19,X/20,XX in males and females, respectively), but varies from 20 to 24 in the remaining genera, probably the consequence of evolution by chromosome fission. Whereas all the males with 19 or 20 chromosomes have an early gametogenesis, which is achieved before the imago stage, the species with more than 20 chromosomes seem to have a delayed male gametogenesis, which is still active in the young imagoes. The species of Rhagiini with 22 chromosomes may constitute a monophyletic group.