Mechanistic exploration of odorant binding protein-mediated chlorpyrifos resistance in Nilaparvata lugens: Insights from insecticide sequestration and transcriptional regulation.

The effectiveness and sustainable application of insecticides are severely threatened by the rapid evolution of resistance in agricultural pests. Recent research indicates that odorant binding proteins (OBPs) may be involved in facilitating insecticide resistance, while the specific mechanisms remain poorly understood. Herein, 11 OBPs were identified from Nilaparvata lugens. Among them, OBP5 exhibited high and specific expression in the head, and showed constitutive overexpression in the chlorpyrifos-resistant strain. Knockdown of OBP5 notably restored susceptibility to chlorpyrifos in N. lugens, while overexpression of OBP5 in Escherichia coli significantly enhanced bacterial tolerance to chlorpyrifos. Fluorescence competitive binding assay confirmed the strong binding affinities of OBP5 to chlorpyrifos and its active metabolite chlorpyrifos-oxon. Molecular docking studies proposed a critical interacting amino acid (Lys147) in the binding site, which was further validated by comparative binding studies between wildtype OBP5 and the mutated protein OBP5. Furthermore, Lim1β that also presented overexpression pattern in the resistant strain, was found to regulate expression of OBP5 through a dual-luciferase reporter assay. Our findings demonstrate that the overexpression of OBP5 contributes to chlorpyrifos resistance by binding and sequestering the insecticides, shedding light on the sequestration resistance mechanism conferred by OBPs and offering potential targets for resistance management.