Contact toxicity and transcriptomic analysis of terpinen‐4‐ol exposure in Tribolium castaneum

Abstract

• Activity of four detoxifying enzymes was notably affected by terpene terpinen-4-ol. • 260 differentially expressed genes were found between T4ol-treated and control. • Pathways associated with development are inhibited following T4ol exposure. • RNAi of CYP9Z6 or GSTs7 increased mortality of T. castaneum larvae after T4ol exposure. • Terpene terpinen-4-ol exhibited pronounced contact toxicity against T. castaneum. The terpene, terpinen-4-ol (T4ol), exhibits contact toxicity in Tribolium castaneum . However, the molecular mechanisms underlying this toxicity have not been elucidated. This study examined changes in the expression of four classic enzymes after exposure of T. castaneum to T4ol. Acetylcholinesterase and glutathione S-transferase activities were markedly inhibited after exposure to T4ol, while that of the detoxifying enzyme cytochrome oxidase P450 increased markedly. Carboxylesterase activity did not show significant changes. Furthermore, RNA sequencing revealed 260 differentially expressed genes (DEG) between the T4ol-treated and control samples, and qRT-PCR was used to validate the RNA-Seq data. The Gene Ontology analysis classified the DEGs into 36 functional groups, including the immune system processes, response to stimulus, and developmental processes. T4ol altered the response to stimulus and the immune system process of beetles by inducing the expression of the genes Stabilin-1, Attacin 1, and Defensin 1. Furthermore, the DEGs receptor tyrosine kinase Torso-like protein ( RTKTsl ), Frizzled 4 ( Fz4 ), Protein Wnt-5b, Ecdysone-induced protein 78C ( E78 ), Zinc finger protein GLIS1 ( ZFPGLIS1 ) were classified as participating in beetle development, and Fz4 and Protein Wnt-5b also mapped to the Wnt signaling pathway. This indicated that pathways associated with development are inhibited after exposure to T4ol. T4ol also induced CYP9Z6 / GSTs7 overexpression, and RNAi targeting these genes significantly increased larvae mortality on T4ol exposure, supporting the participation of CYP9Z6 / GSTs7 in the response to T4ol in T. castaneum . The results of this study will facilitate understanding of the toxic mechanisms of T4ol and provide a basis for controlling the pests of stored products.