Cuticular Hydrocarbon Recognition in the Mating Behavior of Two Pissodes Species

Two sibling weevil species, Pissodes strobi Peck and P. nemorensis Germar (Coleoptera: Curculionidae), can form reduced-fitness hybrids in the laboratory, but neither their premating isolation mechanisms nor mating behaviors are well-understood. Cuticular hydrocarbons (CHCs) have been reported as crucial chemical cues in mating recognition in many insects, including weevils, and, thus, may also mediate the mating behavior of P. strobi and P. nemorensis. We conducted a series of behavioral observations, bioassays, and chemical analyses to investigate the role of CHCs in their mating behavior. Copulation behavior of both species followed similar steps: approaching, mounting, tapping, aedeagus extrusion, and copulation. In P. strobi, hexane extraction significantly reduced the number of successful male copulations compared with freeze-killed females. Conversely, significantly fewer P. nemorensis males copulated with dead females compared with live females. No significant differences were detected among hexane-extracted, freeze-killed or recoated female carcasses to P. nemorensis. These findings suggested that female cuticular extracts contain important cues in mate recognition in P. strobi but not in P. nemorensis. We identified 21 CHCs from both species with variation in abundances between sexes and seasons. Discriminant analysis revealed incomplete overlap of CHC compositions in females of the two species in summer, when hybridization potentially occurs.