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

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att P40>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att P40、VK5果蝇品系相比,转基因果蝇品系att P40>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att P40>CYP6B53和VK5>CYP6B53,可用于杀虫剂筛选,为LdCYP6B53功能研究提供了手段。

The objective was to investigate the susceptibility of CYP6B53 to four insecticides including cyfluthrin, deltamethrin, phoxim and chlorantraniliprole. The att P40CYP6B53 and VK5CYP6B53 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 att P40 and VK5 control Drosophila, the att P40CYP6B53 and VK5CYP6B53 transgenic Drosophila significantly decreased susceptibility to cyfluthrin with 1.83- fold and 1.61- fold of the control Drosophila for LC_（50）,respectively. However, the att P40CYP6B53 and VK5CYP6B53 transgenic Drosophila significantly increased susceptibility to deltamethrin with 0.52- fold and 0.21- fold of the control Drosophila for LC_（50）,respectively. No significant difference of susceptibility to phoxim and chlorantraniliprole was detected between attP40CYP6B53 and VK5CYP6B53 transgenic Drosophila and the control Drosophila. The transgenic Drosophila lines att P40CYP6B53 and VK5CYP6B53 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.