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鉴定与β-氯氰菊酯解毒相关的 CYP4D2 并对其功能进行表征。

Identification and Functional Characterization of CYP4D2 Putatively Associated with β-Cypermethrin Detoxification in .

机构信息

Department of Parasitology, Zunyi Medical University, Zunyi 563000, China.

Laboratory of Evolutionary and Functional Genomics, School of Life Sciences, Chongqing University, Chongqing 401331, China.

出版信息

Genes (Basel). 2022 Dec 11;13(12):2338. doi: 10.3390/genes13122338.

Abstract

, a polyphagous pest, serves as a vector for in China. Currently, there are no effective control strategies for this vector. Agricultural pest control may cause to become a threat due to the development of pesticide resistance. Cytochrome P450s (CYP450) plays a significant role in detoxifying xenobiotics in insects. In this study, we performed RNA sequencing of exposed to β-cypermethrin for 0 and 1 h and then gene cloning of the five up-regulated CYP450 genes. Three CYP450 genes were successfully cloned, and their expression patterns in different developmental stages and in different tissues were analyzed by RT-qPCR. was observed to have the highest expression in the midgut (fold change 2.82 for , 2.62 for , and 1.77 for ). Functional analysis was carried out according to overexpression in S2 cells from the pfastbac1 vector and RNAi with siRNA. The results of the CCK8 assay indicated that the overexpression of the recombinant protein PoCYP4D2 suppressed the decrease in S2 cell viability due to β-cypermethrin. The expression levels of PoCYP4D2 decreased significantly, and the mortality rates increased from 6.25% to 15.0% at 3 h and from 15.0% to 27.5% at 6 h after -siRNA injection. These results suggest that may be an essential key gene in the metabolism of β-cypermethrin in . This study constitutes a foundation to explore further the functions of CYP450 genes in insecticide metabolism.

摘要

斜纹夜蛾,一种多食性害虫,在中国充当了 的载体。目前,针对这种载体,尚无有效的控制策略。农业害虫防治可能会导致 由于农药抗性的发展而成为威胁。细胞色素 P450s(CYP450)在昆虫解毒外来物质方面发挥着重要作用。在这项研究中,我们对暴露于β-氯菊酯 0 和 1 小时的 进行了 RNA 测序,然后对五个上调的 CYP450 基因进行了基因克隆。成功克隆了三个 CYP450 基因,并通过 RT-qPCR 分析了它们在不同发育阶段和不同组织中的表达模式。 在中肠中表达最高( 的倍数变化为 2.82, 的倍数变化为 2.62, 的倍数变化为 1.77)。根据来自 pfastbac1 载体的 S2 细胞过表达和 siRNA 的 RNAi 进行了功能分析。CCK8 测定的结果表明,重组蛋白 PoCYP4D2 的过表达抑制了β-氯菊酯对 S2 细胞活力的降低。PoCYP4D2 的表达水平显著下降,在 -siRNA 注射后 3 小时,死亡率从 6.25%增加到 15.0%,在 6 小时从 15.0%增加到 27.5%。这些结果表明, 可能是 代谢β-氯菊酯的重要关键基因。本研究为进一步探索昆虫 CYP450 基因在杀虫剂代谢中的功能奠定了基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2d6/9777750/31c6fb649457/genes-13-02338-g001.jpg

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