Bu Chun-Ya, Feng Xiao-Jiao, Wang Xiao-Qin, Cao Yang, Wang You-Nian, Chen Qing, Gao Pin, Peng Bo, Li Jin-Ling, Han Jing-Yu, Shi Guang-Lu
College of Biology Science and Engineering, Beijing University of Agriculture, Beijing 102206 China. Key Laboratory of Urban Agriculture (North) Ministry of Agriculture P. R. China, Beijing University of Agriculture Beijing, 102206 China.
Plant Science and Technology College, Beijing University of Agriculture, Beijing 102206 China.
J Econ Entomol. 2015 Apr;108(2):769-79. doi: 10.1093/jee/tou046. Epub 2015 Feb 8.
The carmine spider mite, Tetranychus cinnabarinus (Boisduval), is a major agriculture pest. It can be found worldwide, has an extensive host plant range, and has shown resistance to pesticides. Organophosphate and carbamate insecticides account for more than one-third of all insecticide sales. Insecticide resistance and the toxicity of organophosphate and carbamate insecticides to mammals have become a growing concern. Acetylcholinesterase (AChE) is the major targeted enzyme of organophosphate and carbamate insecticides. In this study, we fully cloned, sequenced and characterized the ace1 gene of T. cinnabarinus, and identified the differences between T. cinnabarinus AChE1, Tetranychus urticae Koch AChE1, and human AChE1. Resistance-associated target-site mutations were displayed by comparing the AChE amino acid sequences and their AChE three-dimensional (3D) structures of the insecticide-susceptible strains of T. cinnabarinus and T. urticae to that of a T. urticae-resistant strain. We identified variation in the active-site gorge and the sites interacting with gorge residues by comparing AChE1 3D structures of T. cinnabarinus, T. urticae, and humans, though their 3D structures were similar. Furthermore, the expression profile of T. cinnabarinus AChE, at the different developmental stages, was determined by quantitative real-time polymerase chain reaction; the transcript levels of AChE were higher in the larvae stage than in other stages. The changes in AChE expression between different developmental stages may be related to their growth habits and metabolism characteristics. This study may offer new insights into the problems of insecticide resistance and insecticide toxicity of nontarget species.
朱砂叶螨（Tetranychus cinnabarinus (Boisduval)）是一种主要的农业害虫。它在全球范围内都有分布，寄主植物范围广泛，并且已表现出对杀虫剂的抗性。有机磷和氨基甲酸酯类杀虫剂占所有杀虫剂销售额的三分之一以上。杀虫剂抗性以及有机磷和氨基甲酸酯类杀虫剂对哺乳动物的毒性已成为日益关注的问题。乙酰胆碱酯酶（AChE）是有机磷和氨基甲酸酯类杀虫剂的主要作用靶点酶。在本研究中，我们对朱砂叶螨的ace1基因进行了全长克隆、测序和特征分析，并确定了朱砂叶螨AChE1、二斑叶螨（Tetranychus urticae Koch）AChE1和人类AChE1之间的差异。通过比较朱砂叶螨和二斑叶螨杀虫剂敏感品系与二斑叶螨抗性品系的AChE氨基酸序列及其AChE三维（3D）结构，展示了与抗性相关的靶点突变。通过比较朱砂叶螨、二斑叶螨和人类的AChE1 3D结构，我们确定了活性位点峡谷以及与峡谷残基相互作用位点的差异，尽管它们的3D结构相似。此外，通过定量实时聚合酶链反应确定了朱砂叶螨AChE在不同发育阶段的表达谱；AChE的转录水平在幼虫阶段高于其他阶段。不同发育阶段AChE表达的变化可能与其生长习性和代谢特征有关。本研究可能为杀虫剂抗性和非靶标物种的杀虫剂毒性问题提供新的见解。
