Biotin deficiency bridges metabolic signaling to insecticide sequestration in Nilaparvata lugens

Abstract

Insecticide resistance poses a critical challenge to global agricultural sustainability. While metabolic detoxification and target-site mutations are well-characterized resistance mechanisms, the role of micronutrient homeostasis remains understudied. This study reveals that biotin deficiency in Nilaparvata lugens drives imidacloprid resistance through a multi-tiered regulatory network coordinating chemosensory protein (CSP) dynamics. Biotin deficiency enhances CSP-mediated insecticide sequestration via high-affinity binding to CSP2, CSP4, CSP7, and CSP15, which are overexpressed in resistant strains. RNA interference and dual-luciferase assays demonstrate that the aryl hydrocarbon receptor and its nuclear translocator (AhR/ARNT) transcriptionally activate CSP2 and CSP15, with their knockdown partially restoring insecticide susceptibility. Furthermore, biotin deficiency activates reactive oxygen species (ROS)/cap ‘n’ collar C (CncC) signaling, elevating AhR/ARNT expression through transcriptional reprogramming. Yeast three-hybrid assays identify a post-translational regulatory layer, wherein biotin directly inhibits AhR–ARNT heterodimerization. Field-evolved resistant populations recapitulate this mechanism, exhibiting conserved molecular signatures including biotin deficiency, ROS/CncC pathway activation, and AhR/ARNT-CSP overexpression correlated with resistance intensity. These findings establish a unified model wherein biotin scarcity reprograms xenobiotic defense through three synergistic mechanisms: enhanced CSP–insecticide binding, transcriptional amplification via ROS/CncC-AhR/ARNT signaling, and post-translational optimization of transcriptional complexes. The operational conservation of this pathway across laboratory and ecological contexts underscores its evolutionary significance while revealing novel targets for resistance management, particularly biotin-based synergists and AhR dimerization inhibitors.