二斑叶螨田间种群对阿维菌素的抗性及抗性相关基因表达分析  

Abamectin resistance and expression of resistance-related genes in field populations of Tetranychus urticae( Acari: Tetranychidae) in China

作　　者：刘贻聪 王玲[1] 张友军[1] 谢文[1] 吴青君[1] 王少丽[1] LIU Yi-Cong WANG Ling ZHANG You-Jun XIE Wen WU Qing-Jun WANG Shao-Li(Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, China)

机构地区：[1]中国农业科学院蔬菜花卉研究所,北京100081

出　　处：《昆虫学报》2016年第11期1199-1205,共7页Acta Entomologica Sinica

基　　金：公益性行业(农业)科研专项(201203038);国家现代西甜瓜产业技术体系(CARS-26-10)项目;农业部园艺作物生物学与种质创制重点实验室项目

摘　　要：【目的】二斑叶螨Tetranychus urticae Koch是为害多种农作物的世界性重大害螨。本研究旨在明确二斑叶螨不同田间种群对阿维菌素的抗药性、抗性相关基因的突变频率及其表达量变化。【方法】采用药管浸叶法测定了我国二斑叶螨8个地理种群对阿维菌素的抗药性并检测其抗性基因突变频率,结合荧光定量PCR技术分析了高抗种群中抗性相关基因表达量变化。【结果】测试的二斑叶螨8个田间种群均对阿维菌素具有抗药性。北京密云、山东潍坊、海南三亚和湖南长沙种群均对阿维菌素产生了极高水平的抗性,抗性倍数分别为1 526.75,481.00,315.25和160.75倍,而北京通县、北京海淀、山西运城和山东泰安种群对阿维菌素的抗性倍数达54.38-136.38倍,处于高抗性水平。二斑叶螨对阿维菌素抗性相关的谷氨酸氯离子通道基因Glu Cl的突变频率在各个田间种群中存在差异。北京密云种群中Glu Cl的突变频率最高（91.7%）,其次是山东潍坊（66.7%）和海南三亚（63.3%）种群;山西运城种群最低（13.3%）,且点突变频率与抗性倍数之间呈显著的正相关（P〈0.05）。与相对敏感种群相比,高抗性二斑叶螨种群中Glu Cl和γ-氨基丁酸受体基因GABAR表达量显著下降。【结论】二斑叶螨田间种群普遍对阿维菌素产生了高水平抗性,抗性相关基因Glu Cl的点突变及其表达量的降低可能与田间抗药性产生相关;田间防治二斑叶螨应避免使用阿维菌素。【Aim】The two-spotted spider mite,Tetranychus urticae Koch is an important pest on many agricultural crops worldwide. This study aims to understand the current abamectin resistance and the mutation frequency and expression change of resistance-related genes in different field populations of T.urticae in China. 【Methods】The resistance of eight field populations of T. urticae to abamectin was determined by vial-leaf dipping（ VLD） method,and the gene mutation frequency related to abamectin resistance was detected and the expression levels of genes likely related to abamectin resistance in highly resistant populations were determined by quantitative RT-PCR technique. 【Results】All the eight field populations of T. urticae tested developed resistance to abamectin. Extremely high resistance to abamectin was found in the populations from Miyun,Beijing（ MY-BJ）,Weifang,Shandong（ WF-SD）,Sanya,Hainan（ SY-HN）,and Changsha,Hunan（ CS-HN）,with the resistance ratios of 1 526. 75-,481. 00-,315. 25-,and 160. 75-fold,respectively. In the populations from Tongxian,Beijing（ TX-BJ）,Haidian,Beijing（ HD-BJ）,Yuncheng,Shanxi（ YC-SX）,and Tai ＇an,Shandong（ TA-SD）,high resistance was observed with the resistance ratios ranging from 54. 38-to 136. 38-fold. The mutation frequency of the glutamate-gated chloride ion channel gene Glu Cl related to abamectin resistance differed in the eight field populations. The highest and lowest mutation frequencies were found in MY-BJ（ 91. 7%） and YC-SX（ 13. 3%） populations,respectively,while moderate mutation frequencies were found in WF-SD and SY-HN populations. There was a significantly positive correlation between resistance level and the mutation frequency of Glu Cl（ P 0. 05）. The expression levels of Glu Cl and γ-aminobutyric acid receptor gene（ GABAR） in the resistant populations decreased significantly as compared with those in the relatively susceptible strain. 【Conclusion】Abamectin resistance exists widely in field populations of T

关 键 词：二斑叶螨 阿维菌素 抗药性 抗性相关基因 基因表达量 

分 类 号：Q965.9[生物学—昆虫学]