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烟草夜蛾()幼虫中 NADPH-细胞色素 P450 还原酶基因沉默后对辛硫磷的敏感性增加。

Tobacco Cutworm () Larvae Silenced in the NADPH-Cytochrome P450 Reductase Gene Show Increased Susceptibility to Phoxim.

机构信息

Guangdong Province Key Laboratory of Eco-Circular Agriculture, South China Agricultural University, Guangzhou 510642, China.

Key Laboratory of Agro-Environment in the Tropics, Ministry of Agriculture, South China Agricultural University, Guangzhou 510642, China.

出版信息

Int J Mol Sci. 2019 Aug 6;20(15):3839. doi: 10.3390/ijms20153839.

Abstract

Nicotinamide adenine dinucleotide phosphate (NADPH)-cytochrome P450 reductases (CPRs) function as redox partners of cytochrome P450 monooxygenases (P450s). CPRs and P450s in insects have been found to participate in insecticide resistance. However, the CPR of the moth has not been well characterized yet. Based on previously obtained transcriptome information, a full-length CPR cDNA of () was PCR-cloned. The deduced amino acid sequence contains domains and residues predicted to be essential for CPR function. Phylogenetic analysis with insect CPR amino acid sequences showed that is closely related to CPRs of Lepidoptera. Quantitative reverse transcriptase PCR (RT-qPCR) was used to determine expression levels of in different developmental stages and tissues of . expression was strongest at the sixth-instar larvae stage and fifth-instar larvae showed highest expression in the midgut. Expression of in the midgut and fat body was strongly upregulated when fifth-instar larvae were exposed to phoxim at LC (4 μg/mL) and LC (20 μg/mL) doses. RNA interference (RNAi) mediated silencing of increased larval mortality by 34.6% (LC dose) and 53.5% (LC dose). Our results provide key information on the gene and indicate that expression levels in larvae influence their susceptibility to phoxim and possibly other insecticides.

摘要

烟酰胺腺嘌呤二核苷酸磷酸(NADPH)-细胞色素 P450 还原酶(CPRs)作为细胞色素 P450 单加氧酶(P450s)的氧化还原伴侣发挥作用。昆虫中的 CPRs 和 P450s 被发现参与了杀虫剂抗性。然而,这种蛾的 CPR 尚未得到很好的描述。基于之前获得的转录组信息,通过 PCR 克隆了全长的 CPR cDNA。推导的氨基酸序列包含被预测对 CPR 功能至关重要的结构域和残基。与昆虫 CPR 氨基酸序列的系统发育分析表明,与鳞翅目昆虫的 CPRs 密切相关。定量逆转录 PCR(RT-qPCR)用于确定 在不同发育阶段和组织中的表达水平。 在 6 龄幼虫期表达最强,在 5 龄幼虫期中肠表达最高。当 5 龄幼虫暴露于 phoxim 时(LC (4 μg/mL)和 LC (20 μg/mL)剂量),中肠和脂肪体中的 表达强烈上调。RNA 干扰(RNAi)介导的 沉默使幼虫死亡率增加了 34.6%(LC 剂量)和 53.5%(LC 剂量)。我们的结果提供了 基因的关键信息,并表明幼虫中 的表达水平影响它们对 phoxim 及可能其他杀虫剂的敏感性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f511/6696589/5790b17f93b0/ijms-20-03839-g001.jpg

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