星豹蛛乙酰胆碱酯酶(AChE)活性测定及药剂对其活性的抑制作用  

Determination of the Activity of Acetylcholinesterase in Pardosa astrigera and the Inhibition Effect of Pesticide on Its Activity

作　　者：马敏[1] 张宾[2] 葛衍珍[1] 李生才[1] 

机构地区：[1]山西农业大学农学院,山西太谷030801 [2]山西农业大学文理学院,山西太谷030801

出　　处：《安徽农业科学》2011年第6期3362-3364,共3页Journal of Anhui Agricultural Sciences

基　　金：山西省留学基金(2009042);山西省青年科技研究基金(2008021041)

摘　　要：[目的]系统研究星豹蛛(Pardosa astrigera)乙酰胆碱酯酶(AChE)的性质,确定其抗药性的发生和发展及其抗性水平,建立快速、准确的AChE活性检测方法。[方法]采用正交试验设计确定星豹蛛不同部位AChE活性测定的最优条件组合,并在此基础上进一步研究豹蛛AChE的体躯分布及其对4种常用药剂的敏感度。[结果]星豹蛛头胸部、腹部和附肢AChE活性测定的最优条件组合是:酶浓度分别为12、18、29 g/L;底物浓度分别为0.6、1.0、1.0 mmol/L;反应体系pH值均为7.0;反应温度分别为30、35、35℃;反应时间均为5min。AChE主要分布在星豹蛛的头胸部,提取液中含有Triton X-100时,AChE的比活力要比无Triton X-100时大。灭多威、辛硫磷、高效氯氰菊酯、毒死蜱对星豹蛛头胸部中AChE的抑制中浓度(IC50)分别为7.76×10-5、1.76×10-4、4.12×10-4、4.94×10-4mol/L。[结论]星豹蛛体内AChE是膜结合的;4种药剂对星豹蛛头胸部中AChE的抑制具有很好的剂量效应,表明星豹蛛头胸部AChE可以作为环境中有机磷、氨基甲酸酯和拟除虫菊酯类农药污染的生物化学标记。[Objective] The paper was to systemically study the characteristics of acetyl cholinesterase（AChE） in Pardosa astrigera,and confirm the occurrence and development of its pesticide resistance and the resistance level,thereby establishing a quick and accurate detection method for enzyme activity.[Method] The optimal conditions for assaying the activity of AChE in different parts of P.strigera were determined by orthogonal experiment.The distribution conditions of acetyl cholinesterase（AChE） in different tissues of P.astrigera were further studied,and the sensitivities of the enzyme to four common pesticides were also determined.[Result] The optimal condition for assaying the activity of AChE in cephalothorax,abdomen and the appendage of P.astrigera was as follows,enzyme concentrations： 12,18 and 29 g/L;substrate concentrations： 0.6,1.0 and 1 mmol/L;pH value,7.0;reaction temperatures：30,35,35 ℃;reaction time,5 min.AChE was mainly distributed in the cephalothorax of P.astrigera.The specific activity of AChE in the extract solution with Triton X-100 was higher than that without Triton X-100.The median inhibitory concentrations（IC50） of Methomyl,Phoxim,Beta cypermethrin,chlorpyrifos against AChE in the cephalothorax of P.astrigera were 7.76×10-5,1.76×10-4,4.12×10-4and 4.94×10-4mol/L,respectively.[Conclusion] AChEs in P.astrigera were membrane-bound.The inhibition of four pesticides on AChE in the cephalothorax of P.astrigera had good dose-effect,this indicated that AChE in the cephalothorax of P.astrigera could be used as the biochemical marker to monitor the contamination of organic phosphorus,carbamate and pyrethroid pesticides in the environment.

关 键 词：星豹蛛 乙酰胆碱酯酶 生物化学标记 药剂敏感度 抑制中浓度 

分 类 号：Q965.9[生物学—昆虫学]