Chin J Schisto Control ›› 2018, Vol. 30 ›› Issue (2): 149-154.

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Study on expression characteristics of cytochrome P450 genes in Anopheles sinensis

WEI Mo| TANG Jian-xia| LI Ju-lin| ZHANG Mei-hua| CHEN Jing| XU Sui| CAO Yuan-yuan| YANG Meng-meng| ZHU Guo-ding*| ZHOU Hua-yun| CAO Jun   

  1. Key Laboratory of National Health and Family Planning Commission on Parasitic Disease Control and Prevention| Jiangsu Provincial Key Laboratory on Parasite and Vector Control Technology| Jiangsu Institute of Parasitic Diseases| Wuxi 214064| China
  • Online:2018-05-11 Published:2018-05-14
  • Contact: ZHU Guo?ding

中华按蚊细胞色素P450基因表达特征分析

卫沫|唐建霞|李菊林|张梅花|陈静|徐岁|曹园园|杨蒙蒙|朱国鼎*|周华云|曹俊   

  1. 国家卫生和计划生育委员会寄生虫病预防与控制技术重点实验室、江苏省寄生虫与媒介控制技术重点实验室、江苏省血吸虫病防治研究所(无锡214064)
  • 通讯作者: 朱国鼎
  • 作者简介:卫沫|男|硕士研究生。研究方向:分子寄生虫学
  • 基金资助:
    国家自然科学基金(81401694、81772230);江苏省自然科学基金(BK20141106、BK20150001);江苏省重点研发计划(BE2016631);江苏省“科教强卫工程”项目(ZDXKA2016016)

Abstract: Objective To explore the expression characteristics of cytochrome P450 (CYP) candidate genes (CYP6M3, CYP6Y1, CYP6P5, CYP4H14, CYP4G17, CYP12F16) in Anopheles sinensis. Methods The samples were collected from different developmental stages (eggs, larvae, pupae and adult mosquitoes (females and males)), and different tissues (salivary glands, malpighian tubes, midguts, ovaries, and fat bodies) of An. sinensis and the female adult mosquitoes exposed by different insecticide doses (0, 1.25, 3.75, 6.25, 12.5 μg/bottle) and time points (0, 5, 15, 30, 60 minutes), then the total RNA was extracted. Quantitative reverse transcription PCR (qPCR) assay was used to analyze the relative expressions of six CYP genes in An. sinensis at different developmental stages, tissues and different insecticide exposure doses and time. Results The expressions of CYP6M3 and CYP6Y1 in the male adult mosquitoes were the highest, the expression of CYP6M3 gene in the males was 35.1 times higher than that in the females, the expression of CYP6Y1 in the males was 61.4 times higher than that in the females; the expression level of CYP4H14 in the larvae was the lowest, the expression of CYP4H14 in the females was 22.5 times higher than that in the fourth instar larvae. The expressions of candidate CYP genes in different tissues of An. sinensis were significantly different, the expression of CYP6M3 in the malpighian tubule was 38.9 times higher than that in the ovary, the expression of CYP6Y1 in the fatbody was 9.1 times higher than that in the ovary, the expression of CYP4G17 was 4.6 times higher than that in the ovary, and the expression of CYP12F16 was 4.4 times higher thanthat in the ovary. The exposure to different insecticide doses and time showed some induction effects in the expressions of candidate CYP genes, which affected the expressions of candidate CYP genes in An. sinensis. Conclusion The expression of candidate CYP genes varies significantly in different developmental stages and various tissues of An. sinensis, and exposure to deltamethrin at various doses and time points affects CYP genes expression in An. sinensis.

Key words: Anopheles sinensis; Deltamethrin; Cytochrome P450 gene; Expression characteristic

摘要: 目的 探讨6种溴氰菊酯抗性候选细胞色素P450(CYP)基因(CYP6M3、CYP6Y1、CYP6P5、CYP4H14、CYP4G17、CYP12F16)在中华按蚊体内的表达特征。方法 收集中华按蚊不同发育时期(卵、幼虫、蛹、雌性成蚊和雄性成蚊)和组织(唾液腺、马氏管、中肠、卵巢以及脂肪体)样本,以及雌性成蚊在暴露于不同溴氰菊酯剂量(0、1.25、3.75、6.25、12.5 μg/瓶)和时间(0,5、15、30、60 min)后的样本.提取总RNA,利用反转录实时定量PCR(qPCR)技术分析CYP6M3、CYP6Y1、CYP6P5、CYP4H14、CYP4G17、CYP12F16基因在中华按蚊不同发育时期、组织以及不同溴氰菊酯接触剂量和时间下的相对表达量。结果 CYP6M3与CYP6Y1基因在雄性中华按蚊成蚊体内的表达量最高,CYP6M3基因在雄性成蚊体内的表达量是雌性成蚊的35.1倍,CYP6Y1基因在雄性成蚊体内的表达量是雌性成蚊的61.4倍;CYP4H14基因在幼虫期表达量最低,且在雌性成蚊体内表达量是四龄幼虫体内表达量的22.5倍。候选CYP基因在中华按蚊不同组织内的表达量差异具有统计学意义,CYP6M3基因在马氏管内的表达量是在卵巢中的38.9倍,CYP6Y1基因在脂肪体内的表达量是在卵巢中的9.1倍,CYP6P5基因在中肠内的表达量是在卵巢中的30.3倍,CYP4G17基因在脂肪体内的表达量是在卵巢中的4.6倍,CYP12F16基因在马氏管内的表达量是在卵巢中的4.4倍。接触不同溴氰菊酯剂量和时间对候选CYP基因的表达水平表现出一定的诱导效应,影响候选CYP基因在中华按蚊体内的表达。结论 候选CYP基因在不同发育期中华按蚊体内和不同组织中差异表达,暴露于不同溴氰菊酯剂量和时间影响CYP基因在中华按蚊体内表达。

关键词: 中华按蚊;溴氰菊酯;细胞色素P450基因;表达特征

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