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. 2020 Jun 25;11:589. doi: 10.3389/fgene.2020.00589

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Copyright © 2020 Gao, Zhang, Wei, Wei, Xiong, Lu, Zhang, Gao and Li.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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(A) Heat map of 758 genes differentially expressed in larvae exposed to 5% A. vulgaris essential oil (EO) and control. Rows represent a single gene and columns are comparisons between genes in the exposure treatment and the control. The left-hand column clusters samples (N = 6) based on similarity of log10 transformed gene expression. Lighter colors indicate lower levels of differential expression. (B) Changes in the gene expression profile between the control and 5% A. vulgaris EO exposure groups. (C) A comparison of the expression profiles of the selected genes as determined by RNA-sequencing and qRT-PCR. OBP, Odorant binding protein; CYP, Cytochrome P450; Ugt, UDP-glucuronosyltransferase; EST, Esterase; GST, Glutathione S-transferase; MRP, Multidrug resistance protein; BG, Beta-glucuronidase; XDH, Xanthine dehydrogenase; FMO, Flavin-containing monooxygenase. Effects of 5% A. vulgaris EO exposure on the accumulation of TcCYP4BN6 (D) and TcOBPC11 (E) in 20-day-old larvae of T. castaneum. Acetone treatment represents the negative control group, which has been ascribed an arbitrary value of 1. The 12, 24, 36, 48, 60, and 72 h are six different test time points following EO treatment. (F) RNAi-mediated gene silencing. Twenty-day-old larvae were used for dsRNA injection. RNA was extracted and quantified by qRT-PCR at fifth day. Control beetles were injected with the same amount of green fluorescent protein (GFP) dsRNA. (G) Effect of TcCYP4BN6/TcOBPC11 silencing by injection of ds-TcCYP4BN6/ds-TcOBPC11 on toxicity of EO to 20-day-old larvae. Larvae injected with ds-TcCYP4BN6/ds-TcOBPC11/ds-GFP only served as untreated controls. Following injection with ds-TcCYP4BN6/ds-TcOBPC11/ds-GFP, larvae were treated with EO for 72 h. Data shown are mean ± SE (n = 3). An asterisk above bars indicated significant differences in the mRNA expression among the control and treatments (**P < 0.01, ***P < 0.001; Student’s t-test). Different letters above bars indicate significant differences (P < 0.05) according to LSD multiple comparison tests.