BdorOBP69a is involved in the perception of the phenylpropanoid compound methyl eugenol in oriental fruit fly (Bactrocera dorsalis) males

The oriental fruit fly ( Bactrocera dorsalis ) is a devastating fruit pest that infests more than 450 plant species . Methyl eugenol (ME) has been used as male attractant to monitor and eradicate B. dorsalis populations for 70 years, but the molecular basis of its activity remains largely unclear. Previously, BdorOBP83b and BdorOBP56f-2 as odorant binding proteins (OBPs) were identified responsible for ME perception. In this study, ME-induced expression profiles and in vitro binding assays revealed that BdorOBP69a is also produced in response to ME and binds directly to it with strong affinity (K d = 9.54 μM). BdorOBP69a −/− null mutants generated by CRISPR/Cas9 mutagenesis showed significantly lower electroantennogram and behavioral responses to ME than wild-type controls. Molecular docking analysis followed by site-directed mutagenesis showed that residues Leu89 and Phe145 are essential for the interaction between BdorOBP69a and ME. BdorOBP69a is therefore an important component involved in the perception of ME in B. dorsalis and a promising molecular target for the development of new male attractants. The molecular docking and binding assay data also provide an important reference for future OBP gene manipulation and ME chemical engineering to improve the efficiency of male attractants. • BdorOBP69a has strong direct binding affinity to male attractant ME i n vitro. • BdorOBP69a −/− null mutants showed significantly lower electroantennogram and behavioral responses to ME. • Residues Leu89 and Phe145 are essential for the binding of BdorOBP69a to ME.