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氯虫苯甲酰胺暴露对粘虫(鳞翅目:夜蛾科)的毒理学和转录组学效应及谷胱甘肽 S-转移酶的功能特征。

Toxicological and transcriptomic effects in Mythimna separata (Lepidoptera: Noctuidae) exposed to chlorantraniliprole and functional characterization of glutathione S-transferases.

机构信息

Key Laboratory of Biology and Sustainable Management of Plant Diseases and Pests of Anhui Higher Education Institutes, School of Plant Protection, Anhui Agricultural University, Hefei, China.

Department of Science and Technology, Chuzhou University, Chuzhou, China.

出版信息

Pest Manag Sci. 2022 Nov;78(11):4517-4532. doi: 10.1002/ps.7072. Epub 2022 Jul 28.

Abstract

BACKGROUND

Chlorantraniliprole (CAP) is an efficient anthranilic diamide insecticide against economically important pests such as the oriental armyworm, Mythimna separata (Lepidoptera: Noctuidae). Resistance to CAP may develop due to enhanced enzymatic detoxification. The glutathione S-transferase (GST) superfamily in M. separata has not been systematically characterized. The aim of this study was therefore to explore the effects of lethal and sublethal doses of CAP on M. separata larvae, screen differentially expressed genes (DEGs) responding to CAP exposure, identify and characterize the GST superfamily, and analyze the metabolism of CAP by recombinant GSTs.

RESULTS

The toxicity bioassay showed that CAP was active against M. separata third-instar larvae. LC50 was 17.615, 3.127, and 1.336 mg/L after 24, 48, and 72 h, respectively. Poisoned larvae showed contracted somites and disrupted midgut. Total GST activity in larvae was significantly elevated 24 h after CAP exposure. RNA-sequencing generated 43 055 unigenes with an average length of 1010 bp, and 567 up-regulated and 692 down-regulated DEGs responding to CAP treatment were screened. Thirty-five GST genes were identified from unigenes, including 31 cytosolic, three microsomal, and one unclassified. The expression profile of GST genes was analyzed using samples from different developmental stages, adult tissues, and CAP treatments. Metabolic assays indicated that CAP was depleted by recombinant MseGSTe2 and MseGSTs6.

CONCLUSIONS

This study provides insight into the toxicological and transcriptomic effects in M. separata larvae exposed to CAP. The identification and functional characterization of the GST superfamily will improve our understanding of CAP detoxification by GSTs. © 2022 Society of Chemical Industry.

摘要

背景

氯虫苯甲酰胺(CAP)是一种高效的邻苯二甲酰胺类杀虫剂,可有效防治东方粘虫(Mythimna separata)等重要经济害虫。由于增强的酶解毒作用,可能会产生对 CAP 的抗性。然而,M. separata 中的谷胱甘肽 S-转移酶(GST)超家族尚未得到系统的描述。因此,本研究旨在探讨 CAP 对 M. separata 幼虫的致死和亚致死剂量的影响,筛选响应 CAP 暴露的差异表达基因(DEGs),鉴定和表征 GST 超家族,并分析重组 GSTs 对 CAP 的代谢。

结果

毒性生物测定表明,CAP 对 M. separata 三龄幼虫具有活性。24、48 和 72 h 时 LC50 分别为 17.615、3.127 和 1.336 mg/L。中毒后的幼虫表现出体节收缩和中肠破裂。暴露于 CAP 后 24 h 时,幼虫中的总 GST 活性显著升高。RNA-seq 共产生 43055 条 unigenes,平均长度为 1010 bp,筛选出 567 个上调和 692 个下调的 DEGs 响应 CAP 处理。从 unigenes 中鉴定出 35 个 GST 基因,包括 31 个细胞质、3 个微粒体和 1 个未分类的 GST 基因。使用来自不同发育阶段、成虫组织和 CAP 处理的样本分析 GST 基因的表达谱。代谢测定表明,CAP 被重组 MseGSTe2 和 MseGSTs6 消耗。

结论

本研究提供了关于 CAP 暴露的 M. separata 幼虫毒理学和转录组学效应的见解。GST 超家族的鉴定和功能表征将提高我们对 GST 解毒 CAP 的理解。© 2022 化学工业协会。

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