小峰熊蜂头部乙酰胆碱酯酶测定条件的优化及其对六种常用杀虫剂的敏感性  

Optimization of conditions for assaying activity of acetylcholinesterase in Bombus hypocrita(Hymenoptera:Apidae) and its sensitivity to six common insecticides

作　　者：廖秀丽[1] 罗术东[1] 伍翔[2] 吴杰[1] 

机构地区：[1]中国农业科学院蜜蜂研究所,农业部授粉昆虫生物学重点开放实验室,北京100093 [2]中化化工科学技术研究总院,北京100083

出　　处：《昆虫学报》2011年第12期1361-1367,共7页Acta Entomologica Sinica

基　　金：国家蜜蜂产业技术体系建设专项经费(CARS-45);农业农村资源等监测统计经费(1251017220);国家“948”项目(2011-S1);公益性行业(农业)科研专项(201203080)

摘　　要：小峰熊蜂Bombus hypocrita是我国优势熊蜂种群之一,因其易于饲养、群势较强且授粉性能优良而成为我国设施农业常用优良授粉蜂种,但常受到以乙酰胆碱酯酶(AChE)为靶标酶的有机磷和氨基甲酸酯类杀虫剂的危害。为合理规避这两类杀虫剂对熊蜂的危害,同时也为完善熊蜂授粉配套技术和保护野生熊蜂资源提供理论基础,本研究利用正交试验对小峰熊蜂头部乙酰胆碱酯酶活性的测定条件进行了优化,并明确了6种常用有机磷和氨基甲酸酯类杀虫剂对乙酰胆碱酯酶活性的影响。结果表明:各测定因素对小峰熊蜂乙酰胆碱酯酶活性测定影响的大小顺序依次为:酶浓度>pH>温度>底物浓度>反应时间;小峰熊蜂头部乙酰胆碱酯酶活性的最适反应条件为:酶浓度0.25g蛋白质/L,底物浓度0.8mmol/L,pH值7.5,温度40℃,反应时间5min。毒死蜱、三唑磷、丙溴磷、异丙威、仲丁威和残杀威6种杀虫剂对小峰熊蜂头部乙酰胆碱酯酶离体抑制作用均呈现明显的剂量-效应关系,其抑制中浓度IC50分别为0.39,1.79,0.42,0.04,0.43和0.63mmol/L。这6种杀虫剂对小峰熊蜂AChE抑制作用的强弱依次为:异丙威>毒死蜱>三唑磷>仲丁威>残杀威>丙溴磷,即小峰熊蜂对异丙威最敏感,而对丙溴磷的敏感性最弱。Bombus hypocrita（Hymenoptera：Apidae） is one of the dominant bumblebees in China,and is widely used as one of the most crucial pollinators in greenhouse due to easy mass-rearing,strong population and effective pollinating performance.However,it is often threatened by organophosphate and carbamate insecticides which are widely used in China,as these insecticides can inhibit the acetylcholinesterase（AChE） activity in insects.In order to avoid harm to bumblebees by these insecticides and improve the pollination technology and conservation of bumblebees,we optimized the reaction conditions to assay the AChE activity from the head of B.hypocrita,and evaluated the effect of six common organophosphate and carbamate insecticides on the AChE activity.The results showed that the optimal conditions are as follows：enzyme concentration 0.25 g protein/L,substrate concentration 0.8 mmol/L,pH 7.5,reaction temperature 40℃,and reaction time 5 min.The inhibition of six insecticides to AChE from B.hypocrita showed a dosage effect.The medium inhibition concentration（IC50） values for chlorpyrifos,profenofos,triazophos,isoprocarb,bassa and propoxur are 0.39,1.79,0.42,0.04,0.43 and 0.63 mmol/L,respectively.The inhibitory effects of six insecticides to AChE from high to low are in the following order：isoprocarb〉 chlorpyrifos〉 triazophos〉 bassa〉 propoxur〉 profenofos,and B.hypocrita is more sensitive to isoprocarb than to the other five insecticides.

关 键 词：小峰熊蜂 乙酰胆碱酯酶 正交试验 最佳测定条件 杀虫剂敏感性 抑制中浓度 

分 类 号：Q965.9[生物学—昆虫学]