Functional characterization of an olfactory receptor in the Oriental fruit fly, Bactrocera dorsalis, that responds to eugenol and isoeugenol.

Most insects utilize a diverse array of olfactory cues for host finding and reproduction behaviors. Males of several Dacini fruit fly species (Tephritidae) are selectively attracted to certain phytochemicals to acquire sex pheromone precursors. Despite the importance of olfaction, only a limited number of olfactory receptors (ORs) in Dacini fruit flies have been characterized. In this study, I focused on the OR94b-2 subfamily, because a clade consisting of these homologs was distinctly localized from other OR clades in a constructed phylogenetic tree. To deorphanize the ORs, I used plant essential oils to screen ligands. Using heterologous expression in Xenopus oocytes, I analyzed the functional properties of BdorOR94b-2 and ZcucOR94b-2 from the Oriental fruit fly, Bactrocera dorsalis, and the melon fruit fly, Zeugodacus cucurbitae, respectively. I found that cinnamon leaf oil evoked responses in oocytes expressing BdorOR94b-2 and the corresponding co-receptor BdorORCO. Gas chromatography-mass spectrometry analysis indicated that eugenol is a major component of cinnamon leaf oil. In accordance with the response to cinnamon leaf oil containing eugenol, BdorOR94b-2/BdorORCO revealed a weak but significant response to eugenol. BdorOR94b-2/BdorORCO also responded to isoeugenol, but not to other related aromatic semiochemicals such as known male-specific attractants and sex pheromones. In contrast, ZcucOR94b-2/ZcucORCO did not respond to any of the phenolic compounds tested, including eugenol analogs. Therefore, BdorOR94b-2/BdorORCO is narrowly tuned to eugenol analogs, whereas ZcucOR94b-2/ZcucORCO likely binds to other compound(s).