The function of two P450s, CYP9M10 and CYP6AA7, in the permethrin resistance of Culex quinquefasciatus.
The function of two P450s, CYP9M10 and CYP6AA7, in the permethrin resistance of Culex quinquefasciatus.
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DOI：
10.1038/s41598-017-00486-0
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发表时间：
2017-04-03
影响因子：
4.6
通讯作者：
Liu N
中科院分区：
文献类型：
--
作者：
Gong Y;Li T;Feng Y;Liu N
细胞色素P450单加氧酶通过使抗性昆虫代谢杀虫剂而在杀虫剂抗性中起关键作用。以往的研究表明，两个P450基因，CYP 9 M10和CYP 6AA 7，不仅上调，而且在抗性库蚊诱导。本研究利用杆状病毒介导的表达系统在昆虫草地贪夜蛾（Spodoptera frugiperda，Sf 9）细胞中分别表达了CYP 9 M10和CYP 6AA 7与细胞色素P450还原酶（cytochrome P450 reductase，CPR），并对CYP 9 M10/CPR和CYP 6AA 7/CPR的酶活性和对氯菊酯及其代谢产物3-苯氧基苯甲醇（3-phenoxybenzoic alcohol，PBOH）的代谢能力进行了表征。PBOH和PBCHO均对库蚊幼虫有毒性，可通过CYP 9 M10/CPR和CYP 6AA 7/CPR进一步代谢，最终代谢产物在此鉴定为PBCOOH，其对蚊子幼虫的毒性显著降低。基于细胞的MTT（3-[4，5-二甲基噻唑-2-基]-2，5-二苯基四唑溴化物）细胞毒性试验显示，表达CYP 9 M10/CPR或CYP 6AA 7/CPR的Sf 9细胞增加了细胞系对氯菊酯、PBOH和PBCHO的耐受性。本研究证实了CYP 9 M10和CYP 6AA 7在氯菊酯及其代谢产物PBOH和PBCHO的解毒过程中所起的重要作用。
Cytochrome P450 monooxygenases play a critical role in insecticide resistance by allowing resistant insects to metabolize insecticides. Previous studies revealed that two P450 genes, CYP9M10 and CYP6AA7, are not only up-regulated but also induced in resistant Culex mosquitoes. In this study, CYP9M10 and CYP6AA7 were separately co-expressed with cytochrome P450 reductase (CPR) in insect Spodoptera frugiperda (Sf9) cells using a baculovirus-mediated expression system and the enzymatic activity and metabolic ability of CYP9M10/CPR and CYP6AA7/CPR to permethrin and its metabolites, including 3-phenoxybenzoic alcohol (PBOH) and 3-phenoxybenzaldehyde (PBCHO), characterized. PBOH and PBCHO, both of which are toxic to Culex mosquito larvae, can be further metabolized by CYP9M10/CPR and CYP6AA7/CPR, with the ultimate metabolite identified here as PBCOOH, which is considerably less toxic to mosquito larvae. A cell-based MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) cytotoxicity assay revealed that Sf9 cells expressing CYP9M10/CPR or CYP6AA7/CPR increased the cell line’s tolerance to permethrin, PBOH, and PBCHO. This study confirms the important role played by CYP9M10 and CYP6AA7 in the detoxification of permethrin and its metabolites PBOH and PBCHO.