Evolution of queen pheromone receptor tuning in four honeybee species (Hymenoptera, Apidae, Apis)

Honeybees (genus: Apis) use a plethora of pheromones for intraspecific communication. The primary compound produced by the queen’s mandibular glands, 9-ODA, is involved in mating in all Apis species. It is the ligand of the most highly expressed olfactory receptor in males of Apis mellifera: AmelOR11. Putative orthologs are found in the genomes of other Apis species: Apis dorsata, Apis florea, and Apis cerana. Modeling of OR11 proteins shows high structure conservation except for AflorOR11. Using heterologous expression in Drosophila and calcium imaging, a broad odorant screening revealed that all OR11 respond predominantly to 9-ODA, but also to secondary ligands, except AflorOR11, which remains specific to 9-ODA. Secondary ligands were confirmed by optical imaging of male antennal lobes in A. mellifera. This work supports a conserved queen sex pheromone detection channel in honeybees, albeit with an extended response spectrum possibly playing a role in reproductive isolation among species.