BackgroundFall armyworm, Spodoptera frugiperda, a formidable agricultural pest, has developed resistance to various synthetic insecticides. However, how S. frugiperda utilizes its limited energy and resources to deal with various insecticides remains largely unexplored.ResultsWe utilized transcriptome sequencing to decipher the broad‐spectrum adaptation mechanism of S. frugiperda to eight insecticides with distinct modes of action. Analysis of the Venn diagram revealed that 1014 up‐regulated genes and 778 down‐regulated genes were present in S. frugiperda treated with at least five different insecticides, compared to the control group. Exposure to various insecticides led to the significant up‐regulation of eight cytochrome P450 monooxygenases (P450s), four UDP glucosyltransferases (UGTs), two glutathione S‐transferases (GSTs), and two ATP‐binding cassette transporters (ABCs). Among them, the sfCYP340AD3 and sfCYP4G74 genes were demonstrated to respond to stress from six different insecticides in S. frugiperda, as evidenced by RNA interference and toxicity bioassays. Furthermore, homology modeling and molecular docking analyses showed that sfCYP340AD3 and sfCYP4G74 possess strong binding affinities to a variety of insecticides.ConclusionCollectively, these findings showed that S. frugiperda utilizes a battery of core detoxification genes to cope with the exposure of synthetic insecticides. This study also sheds light on the identification of efficient insecticidal targets gene and the development of resistance management strategies in S. frugiperda, thereby facilitating the sustainable control of this serious pest.This article is protected by copyright. All rights reserved.