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  • Title: Knockdown of NADPH-cytochrome P450 reductase and CYP6MS1 increases the susceptibility of Sitophilus zeamais to terpinen-4-ol.
    Author: Huang Y, Liao M, Yang Q, Shi S, Xiao J, Cao H.
    Journal: Pestic Biochem Physiol; 2020 Jan; 162():15-22. PubMed ID: 31836049.
    Abstract:
    Terpinen-4-ol showed highly insecticidal activity to stored-grain pest Sitophilus zeamais, and cytochrome P450s were strongly induced in response to terpinen-4-ol fumigation. Understanding of the function of P450 enzyme system in the susceptibility to terpinen-4-ol in S. zeamais will benefit the potential application of terpinen-4-ol in controlling stored-grain pests. In the present study, the synergist piperonyl butoxide increased the toxicity of terpinen-4-ol to S. zeamais, with a synergism ratio of 3.5-fold. Two isoforms of NADPH-cytochrome P450 reductase (SzCPR) were identified, with the difference at the N-terminal. SzCPR contained an N-terminal membrane anchor, FMN, FAD, and NADP binding domains. Expression levels of SzCPR were upregulated by tea tree oil (TTO) and its main constituent terpinen-4-ol under different concentrations and time periods. RNAi was generated for S. zeamais by feeding adults dsRNA and the knockdown of SzCPR increased the susceptibility of S. zeamais to terpinen-4-ol, with higher mortality of adults than control under terpinen-4-ol fumigation. Further RNAi analysis showed that P450 gene CYP6MS1 mediated the susceptibility of S. zeamais to terpinen-4-ol. These results revealed that cytochrome P450 enzyme system, especially CYP6MS1 participated in the susceptibility of S. zeamais to terpinen-4-ol. The findings provided a foundation to clarify the metabolic mechanisms of terpinen-4-ol in stored-grain pests.
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