Characterization and Functional Analysis of OcomOBP7 in Ophraella communa Lesage

SIMPLE SUMMARY: Ophraella communa Lesage is a specific biological control agent of the invasive weed Ambrosia artemisiifolia L. Understanding the molecular mechanism by which O. communa recognizes A. artemisiifolia will help improve its bio-control effect. Odorant-binding proteins (OBPs) play a vital role in insect olfactory perception. In this study, the sequence and expression characteristics of OcomOBP7 were analyzed. We obtained the pure protein of OcomOBP7 by prokaryotic expression and purification, and its binding characteristics were analyzed using a fluorescence competitive binding assay. Finally, we verified the function of OcomOBP7 in vivo using RNAi combined with an electroantennography (EAG) assay. The results showed that the binding ability of OcomOBP7 was broad-spectrum and was involved in the host plant localization of O. communa. ABSTRACT: The olfactory system plays a key role in various insect behaviors, and odorant-binding proteins participate in the first step of the olfactory process. Ophraella communa Lesage is an oligophagous phytophagous insect that is a specific biological control agent for Ambrosia artemisiifolia L. The leaf beetle must identify and locate A. artemisiifolia through olfaction; however, its odorant-binding protein (OBP) function has not yet been reported. In this study, OcomOBP7 was cloned, and its tissue expression profile and binding ability were analyzed using RT-qPCR and fluorescence binding assays, respectively. Sequence analysis demonstrated that OcomOBP7 belongs to the classical OBP family. The RT-qPCR results showed that OcomOBP7 was specifically expressed in the antennae, indicating that OcomOBP7 may be involved in chemical communication. The fluorescence binding assay showed that OcomOBP7 has an extensive binding ability to alkenes. The electroantennography experiments showed that O. communa antennal response to α-pinene and ocimene decreased significantly after interference because the two odors specifically bound to OcomOBP7. In summary, α-pinene and ocimene are odorant ligands corresponding to OcomOBP7, indicating that OcomOBP7 is involved in the chemical recognition of A. artemisiifolia. Our study lays a theoretical foundation for research into O. communa attractants, which is helpful for the better biological control of A. artemisiifolia by O. communa.