Transcriptome profiling reveals differential gene expression of detoxification enzymes in Sitophilus zeamais responding to terpinen-4-ol fumigation.

Plant essential oils with high bioactivity provide environmental friendly alternatives for synthetic pesticides. Melaleuca alternifolia essential oil and its main constituent terpinen-4-ol have high insecticidal activity to Sitophilus zeamais Motschulsky. Terpinen-4-ol may be metabolized by human and insect cytochrome P450s. However, little is known about how insects systemically respond to terpinen-4-ol. In this study, we used an RNA-seq approach to evaluate the global gene expression of S. zeamais after terpinen-4-ol fumigation. Duplicates of fumigated and control groups, for a total of four libraries, were collected for sequencing. A total of 36,117 unigenes with an average length of 1036 bp were generated in the de novo assembled transcriptome. Comparative analysis of S. zeamais libraries identified 592 differentially expressed genes (DEGs), of which 308 and 284 genes were up- and down-regulated in response to terpinen-4-ol fumigation, respectively. GO and KEGG functional analyses were performed for up and downregulated DEGs separately, showing these DEGs were enriched for terms related to catalytic activity, carbohydrate metabolism, and xenobiotics biodegradation and metabolism. DEGs encoding enzymes for detoxification were detected, including sixteen cytochrome P450s (P450s), eight glutathione S-transferase (GSTs), fourteen esterase (ESTs), ten UDP-glucuronosyltransferase (UGTs), and two ATP-binding cassette transporter (ABC transporter) genes. Real-time quantitative PCR confirmed that ten P450s, three GSTs and one EST were up-regulated dramatically after exposure to terpinen-4-ol at different concentrations and over a time course. The results provided a transcriptional overview of the changes in a stored-grain pest in response to terpinen-4-ol fumigation. The analysis revealed the expression levels of detoxification genes were altered, especially for P450s, and provided candidate genes for understanding systemic metabolic responses to terpinen-4-ol in insects.