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Background  Investigating the evolution of species-specific insect genitalia is central to understanding how morphological diversification contributes to reproductive isolation and lineage divergence. While many studies evoke some form of sexual selection to explain genitalia diversity, the basis of selection and the mechanism of heterospecific mate exclusion remains vague. I conducted reciprocal mate pair trials in the  Drosophila mojavensis  species cluster to quantify the frequency of failed insemination attempts, historically referred to as pseudocopulation, between lineages with discrete size and shape differences of the male aedeagus.    Results  In cross-taxon matings aedeagus size had a significant effect on pseudocopulation frequencies, while aedeagus shape and genetic distance did not. The direction of the size difference was an important factor for successful mating. When females were mated to a cross-taxon male with a larger aedeagus than males from her own species, the pair could not establish a successful mating interaction. Females mated to cross-taxon males with a smaller aedeagus than conspecific males were able to establish the mating interaction but had issues disengaging at the end of the interaction.    Conclusions  The results of this study support a role for aedeagus size in the male-female mating interaction, with a secondary role for aedeagus shape. In natural populations, mating failure based on aedeagus size could serve as an important reproductive isolating mechanism resulting in failed insemination attempts after both the male and female show a willingness to mate.    Electronic supplementary material  The online version of this article (doi:10.1186/s12862-014-0255-3) contains supplementary material, which is available to authorized users.

The role of aedeagus size and shape in failed mating interactions among recently diverged taxa in the Drosophila mojavensisspecies cluster