Biochemical, in silico , and field evaluation of emerging insecticide chemistries and their binary mixtures with orange oil against Aphis gossypii (Glover) (Hemiptera: Aphididae)

Abstract Aphis gossypii (Glover 1877) remains a major constraint to cotton production worldwide, necessitating effective, resistance-conscious management strategies. This study systematically evaluated three emerging insecticide chemistries: flupyradifurone, flonicamid, and cyantraniliprole, both individually and in binary mixtures with a commercial orange oil formulation against adult A. gossypii. The evaluation integrated laboratory toxicity bioassays, biochemical enzyme profiling, computational (in silico) docking, and multiseason field validation. Probit analysis revealed pronounced differences in insecticidal potency, with LC50 values ranging from 0.095 mg L−1 for flupyradifurone to 2.210 mg L−1 for cyantraniliprole, and corresponding toxicity indices (TI) of 100 and 11.1, respectively, confirming flupyradifurone as the most active compound. Biochemical assays demonstrated treatment-specific modulation of major detoxification enzymes. Orange oil alone significantly induced esterase, glutathione S-transferase, cytochrome P450, and acetylcholinesterase activities at LC25, whereas most insecticides and their mixtures markedly suppressed esterase activity at LC50. Molecular docking analyses corroborated these findings, showing that cyantraniliprole exhibited the highest binding affinity toward the esterase active site (−6.6949 kcal mol−1), followed by flupyradifurone and flonicamid, while d-limonene displayed the weakest affinity (−4.3736 kcal mol−1), consistent with its limited direct enzymatic interaction. Critically, two-season field trials demonstrated consistently high aphid suppression (90% population reduction) across all treatments, with no significant interannual variation, proving that laboratory-detected antagonism in mixtures did not compromise field efficacy. Collectively, these results demonstrate the robust field performance of emerging insecticide chemistries and support their strategic integration with botanical products as sustainable components of resistance-aware integrated pest management (IPM) programs targeting A. gossypii.