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基因表达分析和酶活性测定揭示了苹果蠹蛾羧酸酯酶基因CpCE - 1在杀虫剂解毒中的潜在作用。

Gene expression analysis and enzyme assay reveal a potential role of the carboxylesterase gene CpCE-1 from Cydia pomonella in detoxification of insecticides.

作者信息

Yang Xue-Qing

机构信息

Key Laboratory of Economical and Applied Entomology of Liaoning Province, College of Plant Protection, Shenyang Agricultural University, Shenyang, 110866, Liaoning, China.

出版信息

Pestic Biochem Physiol. 2016 May;129:56-62. doi: 10.1016/j.pestbp.2015.10.018. Epub 2015 Oct 23.

DOI:10.1016/j.pestbp.2015.10.018
PMID:27017882
Abstract

Carboxylesterases (CarEs) are responsible for metabolism of xenobiotics including insecticides in insects. Understanding the expression patterns of a such detoxifying gene and effect of insecticides on its enzyme activity are important to clarify the function of this gene relevant to insecticides-detoxifying process, but little information is available in the codling moth Cydia pomonella (L.). In this study, we investigated the expression profiles of CarE gene CpCE-1 at different developmental stages and in different tissues of C. pomonella, as well as the larvae exposed to chlorpyrifos-ethyl and lambda-cyhalothrin by using absolute real-time quantitative PCR (absolute RT-qPCR). Results indicated that CpCE-1 expression was significantly altered during C. pomonella development stages, and this expression differed between sexes, with a higher transcript in females than males. Meanwhile, CpCE-1 is overexpressed in cuticle, midgut and head than silk gland, fat body and Malpighian tubules. Exposure of third instar larvae to a non-lethal dosage of chlorpyrifos-ethyl and lambda-cyhalothrin resulted in induction of CpCE-1 transcript. The total carboxylesterase enzyme activity was inhibited by chlorpyrifos-ethyl in vivo; in contrast, the activity of Escherichia coli produced recombinant CpCE-1 was significantly inhibited by both lambda-cyhalothrin and chlorpyrifos-ethyl in vitro. These results suggested that CpCE-1 in C. pomonella is potentially involved in the development and in detoxification of chlorpyrifos-ethyl and lambda-cyhalothrin.

摘要

羧酸酯酶(CarEs)负责昆虫体内包括杀虫剂在内的外源化合物的代谢。了解这种解毒基因的表达模式以及杀虫剂对其酶活性的影响,对于阐明该基因在杀虫剂解毒过程中的功能很重要,但在苹果蠹蛾Cydia pomonella(L.)中相关信息较少。在本研究中,我们利用绝对实时定量PCR(绝对RT-qPCR)研究了苹果蠹蛾CpCE-1基因在不同发育阶段和不同组织中的表达谱,以及暴露于乙基毒死蜱和高效氯氟氰菊酯的幼虫的表达谱。结果表明,CpCE-1的表达在苹果蠹蛾发育阶段有显著变化,且这种表达在性别间存在差异,雌性转录本高于雄性。同时,CpCE-1在表皮、中肠和头部中的表达高于丝腺、脂肪体和马氏管。用非致死剂量的乙基毒死蜱和高效氯氟氰菊酯处理三龄幼虫会导致CpCE-1转录本的诱导。体内乙基毒死蜱抑制了总羧酸酯酶活性;相反,体外高效氯氟氰菊酯和乙基毒死蜱均显著抑制了大肠杆菌产生的重组CpCE-1的活性。这些结果表明,苹果蠹蛾中的CpCE-1可能参与了发育过程以及对乙基毒死蜱和高效氯氟氰菊酯的解毒。

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