Left-right asymmetric genitalia have appeared multiple times independently in insects and have been associated with changes in mating positions. However, there is little experimental data on how the evolution of genital asymmetries may have affected the evolution of mating positions or vice versa. As opposed to its closely-related species, Drosophila pachea has a conspicuous asymmetry in its male genitalia external lobes, with the left lobe being 1.49 ± 0.08 (SD) times longer and thinner than the right lobe. In a laboratory stock, we found that 20% of the males possess fully symmetric lobes. To better understand how asymmetric genitalia may affect mating, we compared D. pachea copulation behaviour between these mutant males and wild-type males. We found that D. pachea wild-type males adopt a one-sided mating posture with the male always one-sided 8.55° ± 1.79° (SD) towards the female’s right side. Within 45-min recordings, all wild-type males did mate whereas 39% of symmetric mutants failed to form a stable mating complex and did not mate. In successful copulations, symmetric mutants also adopted a right-sided mating posture but the angle between male and female bodies was significantly more variable compared to wild-type males. Our results suggest that lobe size asymmetry is required for the formation of a stable mating complex and for the positioning of the male according to a precise angle on the female. However, lobe size asymmetry is not required for D. pachea right-sided mating posture
