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鉴定调控褐飞虱细胞色素 P450 CYP6ER1 新型可变剪接异构体的 SNP 位点。

Identification of SNPs involved in regulating a novel alternative transcript of P450 CYP6ER1 in the brown planthopper.

机构信息

State Key Laboratory of Biocontrol, Sun Yat-sen University, Guangzhou, China.

Guangxi Academy of Agricultural Sciences, Nanning, Guangxi, China.

出版信息

Insect Sci. 2018 Oct;25(5):726-738. doi: 10.1111/1744-7917.12472. Epub 2017 Jul 17.

Abstract

Cytochrome P450-mediated metabolic resistance is one of the major mechanisms involved in insecticide resistance. Although the up-regulation of cytochrome P450 plays a vital role in insecticide metabolism, the molecular basis for the transcriptional regulation of cytochrome P450 remains largely unknown. The P450 gene CYP6ER1, has been reported to confer imidacloprid resistance to the brown planthopper, Nilaparvata lugens. Here, we identified a novel alternative transcript of CYP6ER1 (transcript A2) that had different expression patterns between resistant and susceptible populations, and was more stable after insecticide induction. The promoter of this transcript was sequenced and multiple single nucleotide polymorphisms (SNPs) were detected in individuals from susceptible and resistant field-collected populations. Resistant alleles of four SNPs were found to significantly enhance the promoter activity of the CYP6ER1 transcript A2. Electrophoretic mobility shift assays (EMSAs) revealed that these SNPs might regulate the binding of transcription factors to the promoter. Our findings provide novel evidence regarding the transcriptional regulation of a metabolic resistance-related gene and may be useful to understand the resistance mechanism of N. lugens in the field.

摘要

细胞色素 P450 介导的代谢抗性是昆虫抗药性的主要机制之一。虽然细胞色素 P450 的上调在杀虫剂代谢中起着至关重要的作用,但细胞色素 P450 的转录调控的分子基础在很大程度上仍然未知。P450 基因 CYP6ER1 已被报道赋予褐飞虱Nilaparvata lugens 对吡虫啉的抗性。在这里,我们鉴定了 CYP6ER1 的一种新的替代转录本(转录本 A2),它在抗性和敏感种群之间具有不同的表达模式,并且在杀虫剂诱导后更稳定。该转录本的启动子被测序,并在来自敏感和抗性田间采集种群的个体中检测到多个单核苷酸多态性(SNP)。发现四个 SNP 的抗性等位基因显著增强了 CYP6ER1 转录本 A2 的启动子活性。电泳迁移率变动分析(EMSA)表明,这些 SNP 可能调节转录因子与启动子的结合。我们的发现为代谢抗性相关基因的转录调控提供了新的证据,并可能有助于理解田间褐飞虱的抗性机制。

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