Deciphering the signature of cuticular lipids with contact sex pheromone function in a parasitic wasp

The surface of insects is covered by a complex mixture of cuticular hydrocarbons (CHCs) to prevent desiccation. In many species these lipids also have communicative functions, but often it is unknown which components are crucial for the behavioural response. Furthermore, it is often ignored that polar lipids also occur on the insects' cuticle and might interact with CHCs. In the parasitic wasp Lariophagus distinguendus, CHCs function as a contact sex pheromone eliciting wing-fanning in males. Interestingly, not only females but also newly emerged males have the pheromone, resulting regularly in homosexual courtship. However, males deactivate the pheromone within the first two days after emergence. This deactivation is accompanied by the disappearance of 3-methylheptacosane (3-MeC27) and some minor components from the CHC profile of males. Here we show that 3-MeC27 is a key component of the contact sex pheromone which, however, triggers courtship behaviour only if an olfactory background of other cuticular lipids is present. Males responded to (S)-3-MeC27 enantioselectively when applied to filter paper but on three-dimensional dummies both enantiomers were behaviourally active, suggesting that physical stimuli also play a role in sexual communication of the wasps. Finally, we report that triacylglycerides (TAGs) are also essential components of the pheromone, and present evidence that TAGs actually occur on the cuticle of L. distinguendus. Our data provide novel insights into the semiochemical function of cuticular lipids by showing that the bioactivity of CHCs may be influenced by the stereochemistry and a synergetic interaction with long time ignored TAGs.