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Background  Male genitals have repeatedly evolved left-right asymmetries, and the causes of such evolution remain unclear. The  Drosophila nannoptera  group contains four species, among which three exhibit left-right asymmetries of distinct genital organs. In the most studied species,  Drosophila pachea,  males display asymmetric genital lobes and they mate right-sided on top of the female. Copulation position of the other species is unknown.    Results  To assess whether the evolution of genital asymmetry could be linked to the evolution of one-sided mating, we examined phallus morphology and copulation position in  D. pachea  and closely related species. The phallus was found to be symmetric in all investigated species except  D. pachea , which displays an asymmetric phallus with a right-sided gonopore, and  D. acanthoptera,  which harbors an asymmetrically bent phallus. In all examined species, males were found to position themselves symmetrically on top of the female, except in  D. pachea  and  D. nannoptera , where males mated right-sided, in distinctive, species-specific positions. In addition, the copulation duration was found to be increased in the  nannoptera  group species compared to closely related outgroup species.    Conclusion  Our study shows that gains, and possibly losses, of asymmetry in genital morphology and mating position have evolved repeatedly in the  nannoptera  group. Current data does not allow us to conclude whether genital asymmetry has evolved in response to changes in mating position, or vice versa.    Electronic supplementary material  The online version of this article (10.1186/s12862-019-1434-z) contains supplementary material, which is available to authorized users.

Repeated evolution of asymmetric genitalia and right-sided mating behavior in the Drosophila nannoptera species group