The cereal weevil, Sitophilus zeamais is one of the most destructive pests of stored cereals worldwide. Frequent use of fumigants for managing stored-product insects has led to the development of resistance in insects. Essential oils from aromatic plants including the tea oil plant, Melaleuca alternifolia may provide environmentally friendly alternatives to currently used pest control agents. However, little is known about molecular events involved in stored-product insects in response to plant essential oil fumigation. M. alternifolia essential oil was shown to possess the fumigant toxicity against S. zeamais. The constituent, terpinen-4-ol was the most effective compound for fumigant toxicity. M. alternifolia essential oil significantly inhibited the activity of three enzymes in S. zeamais, including two detoxifying enzymes, glutathione S-transferase (GST), and carboxylesterase (CarE), as well as a nerve conduction enzyme, acetylcholinesterase (AChE). Comparative transcriptome analysis of S. zeamais through RNA-Seq identified a total of 3,562 differentially expressed genes (DEGs), of which 2,836 and 726 were up-regulated and down-regulated in response to M. alternifolia essential oil fumigation, respectively. Based on gene ontology (GO) analysis, the majority of DEGs were involved in insecticide detoxification and mitochondrial function. Furthermore, an abundance of DEGs mapped into the metabolism pathway in the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway database were associated with respiration and metabolism of xenobiotics, including cytochrome P450s, CarEs, GSTs, and ATP-binding cassette transporters (ABC transporters). Some DEGs mapped into the proteasome and phagosome pathway were found to be significantly enriched. These results led us to propose a model of insecticide action that M. alternifolia essential oil likely directly affects the hydrogen carrier to block the electron flow and interfere energy synthesis in mitochondrial respiratory chain. This is the first study to perform a comparative transcriptome analysis of S. zeamais in response to M. alternifolia essential oil fumigation. Our results provide new insights into the insecticidal mechanism of M. alternifolia essential oil fumigation against S. zeamais and eventually contribute to the management of this important agricultural pest.

Pesticide resistance has developed as a result of long‐term and extensive use of chemical pesticides. Essential oils from aromatic plants may provide a new and safe alternative to conventional insecticides. In this study, the insecticidal activities of the essential oil of Melaleuca alternifolia and their chemical constituents against Helicoverpa armigera Hubner were investigated, and the underlying mechanisms were studied. The essential oil showed distinct antifeedant (AFC₅₀ = 8.93 mg/ml) and good contact (LD₅₀ = 50.28 μg/larva) activities against H. armigera at 24 hr. Ten chemical components were identified using a gas chromatograph/mass spectrometer, and mainly included terpinen‐4‐ol (40.09%), γ‐terpinene (21.85%), α‐terpinene (11.34%), α‐terpineol (6.91%), α‐pinene (5.86%), terpinolene (3.24%) and 1,8‐cineole (1.83%). Among them, five components were determined and results showed that these constituents possessed obvious antifeedant activities. The activities of acetylcholinesterase and glutathione S‐transferase were notably inhibited by the essential oil, as compared with the control, with strong dose‐ and time‐dependent effects. The results provide a basis for their development and utilization in the control of insects in the future.