›› 2008, Vol. 51 ›› Issue (7): 714-71.

• 研究论文 • 上一篇    下一篇



  • 出版日期:2008-07-20 发布日期:2008-07-20
  • 通讯作者: 高希武

Establishment of optimization of assaying activity of acetylcholinesterase from Odontotermes formosanus (Isoptera: Termitidae) and their sensitivity to insecticides

TANG Fang   

  • Online:2008-07-20 Published:2008-07-20

摘要: 用正交试验方法研究了酶浓度、底物浓度、反应体系pH值、反应温度、反应时间5个因素对黑翅土白蚁Odontotermes formosanus (Shiraki)乙酰胆碱酯酶(AChE)活性测定的影响。通过对正交试验结果进行极差和方差分析,明确了测定黑翅土白蚁AChE活性的最适反应条件是酶浓度为12.5 g/L,底物浓度为8 mmol/LpH8.0,反应温度40℃,反应时间5 min。此外,研究了6种药剂对黑翅土白蚁体内AChE活性的影响。结果表明:灭多威、辛硫磷、三唑磷、丙溴磷、马拉硫磷和氧化乐果6种药剂对黑翅土白蚁AChE抑制中浓度(IC50)分别为3.52×10-41.86×10-35.13×10-39.55×10-48.81×10-3,和1.39×10-2 mol/L。在3.3×10-75×10-3 mol/L的浓度范围内,上述6种药剂对黑翅土白蚁体内AChE活性的抑制作用都具有明显的剂量效应关系。

关键词: 黑翅土白蚁, 乙酰胆碱酯酶, 最适反应条件, 正交试验, 抑制中浓度, 药剂敏感度

Abstract: The optimal conditions for assaying the activity of acetylcholinesterase (AChE) from Odontotermes formosanus (Shiraki) were determined by orthogonal experiment. The concentrations of acetylcholinesterase and substrate, pH and temperature of the reaction system, and reaction time were optimized. L25 (56) orthogonal matrix was adopted, without considering interaction. Through analyzing the data from orthogonal experiment with range analysis and variance analysis, we found that the optimal conditions were as enzyme concentration 12.5 g/L, substrate concentration 8 mmol/L, pH 8.0, reaction temperature 40, and reaction time 5 min. Furthermore, the inhibition of 6 insecticides to AChE from O. formosanus was determined.  The IC50 values of methomyl, phoxim, triazophos, profenofos, malathion and omethoate were 3.52×10-4, 1.86×10-3, 5.13×10-3, 9.55×10-4, 8.81×10-3 and 1.39×10-2 mol/L, respectively. The inhibition of AChE activity by six insecticides increased with the increasing of insecticide concentration from 3.3×10-7 to 5×10-3 mol/L.

Key words: Odontotermes formosanus, acetylcholinesterase, optimal condition, orthogonal experiment, IC50 values, sensitivity to insecticide