Mohamed Magda A, M E Mahdy El-Sayed, Ghazy Abd-El-Hady M, Ibrahim Nihal M, El-Mezayen Hatem A, Ghanem Manal M E
Molecular Biology Department, National Research Centre, 33-El-El Bohouth Street, Dokki, Giza P.O. 12622, Egypt.
Biochemistry Department, Hellwan University, Cairo, Egypt.
Pestic Biochem Physiol. 2017 Jan;135:27-34. doi: 10.1016/j.pestbp.2016.07.002. Epub 2016 Jul 19.
Acetylcholinesterases (AChEs) from the infective juveniles (IJs) of entomopathogenic nematode (EPN) have been investigated with respect to their susceptibility to insecticides and immunological characteristics, aiming at nominating the most compatible insecticide(s) to be used in conjunction with the most insecticide-tolerant EPN strain before incorporation in integrated pest management (IPM) programs. The inhibition kinetics of two purified AChE isoenzymes, AChEAII and AChEBI isolated from Heterorhabditid bacteriophora EM2 strain, by different insecticides revealed that the insensitivity to inhibition by such insecticides could be arranged in a descending order as; methomyl>carbofuran>acetamiprid>oxamyl>malathion. Except for malathion, the insecticides competitively inhibited AChEs with Ki values ranging from 0.1 to 15mM and IC values from 1.25 to 23mM. The two AChE isoforms are several folds less sensitive to inhibition by methomyl and carbofuran compared to those previously reported for other insect species. AChEBI was used as an immunogen to raise anti-AChEBI antisera in rabbits. The prepared antisera cross-reacted with AChEs of five different heterorhabditid nematode strains implying the presence of common epitopes shared along all the examined strains. Such studies could aid in the rational selection of the compatible insecticide(s) and the prepared polyclonal anti-AChE antisera would be a valuable immunodiagnostic tool for evaluating the most insecticide-tolerant EPN strain(s) in IPM programs.
针对昆虫病原线虫(EPN)感染性幼虫(IJ)的乙酰胆碱酯酶(AChE)对杀虫剂的敏感性及其免疫学特性进行了研究,目的是在将其纳入综合虫害管理(IPM)计划之前,确定与最耐杀虫剂的EPN菌株联合使用的最兼容杀虫剂。从嗜菌异小杆线虫EM2菌株中分离出的两种纯化的AChE同工酶AChEAII和AChEBI,被不同杀虫剂抑制的动力学研究表明,对这些杀虫剂抑制作用的不敏感性可按降序排列为:灭多威>克百威>啶虫脒>杀线威>马拉硫磷。除马拉硫磷外,这些杀虫剂竞争性抑制AChE,Ki值范围为0.1至15mM,IC值范围为1.25至23mM。与先前报道的其他昆虫物种相比,这两种AChE同工型对灭多威和克百威抑制作用的敏感性要低几倍。使用AChEBI作为免疫原在兔中制备抗AChEBI抗血清。制备的抗血清与五种不同异小杆线虫菌株的AChE发生交叉反应,这意味着在所有检测菌株中存在共同的表位。此类研究有助于合理选择兼容的杀虫剂,并且制备的多克隆抗AChE抗血清将成为评估IPM计划中最耐杀虫剂的EPN菌株的有价值的免疫诊断工具。