Transcriptional Signaling Cascade Orchestrates CYP6AE48 Overexpression to Drive λ-Cyhalothrin Detoxification in Spodoptera litura

Lambda-cyhalothrin (λ-cyhalothrin) is a broad-spectrum pyrethroid against lepidopteran pests, yet its efficacy is increasingly threatened by insect resistance. Although CYP6AE48 overexpression has been implicated in λ-cyhalothrin detoxification in Spodoptera litura, the regulatory mechanism remains unclear. Here, CRISPR/Cas9-generated CYP6AE48 knockout increased larval susceptibility to λ-cyhalothrin, confirming its essential role in detoxification. λ-Cyhalothrin exposure triggered the expression of several transcription factors, including CncC, MafK, EcR, USP, AhR, and ARNT . Their knockdown suppressed CYP6AE48 expression, thereby compromising larval tolerance. Dual-luciferase reporter assay demonstrated that these transcription factors regulated CYP6AE48 transcription by binding to the specific cis -acting elements. Moreover, protein kinases AKT, ERK, JNK, and RSK2 were implicated as upstream regulators, orchestrating transcription factor expression and subsequent CYP6AE48-mediated detoxification. These findings reveal a multilayered regulatory circuitry that activates CYP6AE48 expression to facilitate λ-cyhalothrin detoxification, providing novel insights into the evolutionary adaptation of detoxification mechanisms and highlighting potential molecular targets for future pest management strategies.