Lipases and carboxylesterases are involved in interspecific pheromone differences between two moth species

Moth sex pheromones are a classical model for studying sexual selection. Females produce a species-specific pheromone blend that attracts males. Revealing the enzymes involved in the interspecific variation in blend composition is key for understanding the evolution of these sexual communication systems. The nature of the enzymes involved in the variation of acetate esters, which are prominent compounds in moth pheromone blends, remains unclear. We identified enzymes involved in acetate metabolism in two closely related species: Heliothis (Chloridea) subflexa and H. (C.) virescens, which differ in production of acetate esters. Through comparative transcriptomic analyses and CRISPR/Cas9 knockouts, we showed that two lipases and two esterases induce lower levels of acetate esters in female pheromones. To place our findings in an evolutionary context, we explored the molecular evolution of related lipases and esterases in Lepidoptera. Together, our results show that lipases and carboxylesterases are unexpectedly involved in tuning Lepidoptera pheromones composition.