表达LdCYP6B53果蝇品系建立及对杀虫剂的敏感性

doi:10.11924/j.issn.1000-6850.casb16030222

中国农学通报 ›› 2016, Vol. 32 ›› Issue (19): 97-101.doi: 10.11924/j.issn.1000-6850.casb16030222

表达LdCYP6B53果蝇品系建立及对杀虫剂的敏感性

薛绪亭,孙丽丽,刘鹏,王志英,曹传旺

东北林业大学林学院,东北林业大学林学院,东北林业大学林学院,东北林业大学林学院,东北林业大学林学院

收稿日期:2016-03-31 修回日期:2016-06-06 接受日期:2016-05-20 出版日期:2016-07-15 发布日期:2016-07-15
通讯作者: 曹传旺
基金资助:
国家自然科学基金面上项目“舞毒蛾Methuselah like 受体生理功能及其G蛋白偶联信号转导机制研究”(31570642)；东北林业大学本科生创新项目“舞毒蛾Methuselah-like受体调控P450 基因表达研究”(201510225076)。

Construction of Transgenic Drosophila Expressing LdCYP6B53 and Its Susceptibility to Insecticides

Received:2016-03-31 Revised:2016-06-06 Accepted:2016-05-20 Online:2016-07-15 Published:2016-07-15

摘要/Abstract

摘要： 旨在探索舞毒蛾CYP6B53 基因对百树菊酯、溴氰菊酯、辛硫磷、氯虫苯甲酰胺4 种杀虫剂的敏感性。通过转基因技术将LdCYP6B53 基因转入果蝇体内，分别建立attP40＞CYP6B53 和VK5＞CYP6B53 转基因果蝇品系，并测定杀虫剂对转基因果蝇毒力。限制性内切酶消化、RT-PCR和核酸序列测定表明转基因果蝇品系中稳定表达LdCYP6B53 基因。与对照attP40、VK5 果蝇品系相比，转基因果蝇品系attP40＞CYP6B53 和VK5＞CYP6B53 对百树菊酯的敏感性显著降低，LC50分别为对照组的1.83和1.61 倍；而对溴氰菊酯敏感性显著增加，LC50分别为对照组的0.52 和0.21 倍；对辛硫磷和氯虫苯甲酰胺的敏感性与对照组差异不显著。本研究利用转基因技术成功构建了转舞毒蛾P450 基因CYP6B53 的果蝇品系attP40＞CYP6B53 和VK5＞CYP6B53，可用于杀虫剂筛选，为LdCYP6B53 功能研究提供了手段。

关键词: 夏玉米, 夏玉米, 涝渍指标, 作物需水量, 涝渍强度指数

Abstract: The objective was to investigate the susceptibility of CYP6B53 to four insecticides including cyfluthrin, deltamethrin, phoxim and chlorantraniliprole. The attP40＞CYP6B53 and VK5＞CYP6B53 transgenic Drosophila were constructed by transforming LdCYP6B53 into Drosophila. The toxicity of insecticides to transgenic Drosophila was measured using bioassay. The CYP6B53 gene was confirmed to stably express into Drosophila by restriction endonuclease digestion, RT-PCR and sequencing. Compared with attP40 and VK5 control Drosophila, the attP40＞CYP6B53 and VK5＞CYP6B53 transgenic Drosophila significantly decreased susceptibility to cyfluthrin with 1.83- fold and 1.61- fold of the control Drosophila for LC50, respectively. However, the attP40＞CYP6B53 and VK5＞CYP6B53 transgenic Drosophila significantly increased susceptibility to deltamethrin with 0.52- fold and 0.21- fold of the control Drosophila for LC50, respectively. No significant difference of susceptibility to phoxim and chlorantraniliprole was detected between attP40＞CYP6B53 and VK5＞CYP6B53 transgenic Drosophila and the control Drosophila. The transgenic Drosophila lines attP40＞CYP6B53 and VK5＞CYP6B53 were successfully constructed by using transgenic technology, and these transgenic Drosophila lines could be used for screening insecticides and provide a method for studying LdCYP6B53 functions.

薛绪亭,孙丽丽,刘鹏,王志英,曹传旺. 表达LdCYP6B53果蝇品系建立及对杀虫剂的敏感性[J]. 中国农学通报, 2016, 32(19): 97-101.

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表达LdCYP6B53果蝇品系建立及对杀虫剂的敏感性

Construction of Transgenic Drosophila Expressing LdCYP6B53 and Its Susceptibility to Insecticides

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