为了明确茶假眼小绿叶蝉（Empoasca vitis）对联苯菊酯代谢抗性相关的解毒酶,本研究采用室内生物测定和生化分析方法,比较了假眼小绿叶蝉抗联苯菊酯品系和敏感品系的3种解毒酶活性差异,并分析了3种增效剂对联苯菊酯的增效作用。解毒酶活性测定表明,茶假眼小绿叶蝉抗联苯菊酯品系羧酸酯酶（CarE）、细胞色素P450氧化酶O-脱甲基（PNOD）、谷胱甘肽-S-转移酶（GST）比活力都高于敏感品系,3种解毒酶活性分别为敏感品系的2.09倍、4.34倍和1.59倍。增效剂磷酸三苯酯（TPP）、增效醚（PBO）和顺丁烯二酸二乙酯（DEM）在茶假眼小绿叶蝉敏感品系中的增效比分别为1.04、1.09、1.00,而在联苯菊酯抗性品系中的增效比分别为1.80、7.97、1.03。上述结果证明,羧酸酯酶和细胞色素P450氧化酶活性的增强在茶假眼小绿叶蝉对联苯菊酯的抗性产生中起到了重要的作用。
Tea green leafhopper, Empoasca vitis, is the most serious pest in tea gardens. The detoxifying enzyme activities and synergism in the bifenthrin-resistant and susceptible leafhopper were measured via biochemical methods and bioassays to clarify their potential relationship to the resistance of tea leafhopper to bifenthrin. The detoxifying enzyme activity tests showed that the activities of carboxylesterase (CarE), Cytochrome P450-mediated O-demethylation activity toward p-nitroanisole (PNOD) and glutathione-S-transferase (GST) in the resistant strain were all higher than those in susceptible strain. The activities of the three detoxifying enzymes in bifenthrin-resistant strain were 2.09, 4.34 and 1.59 folds those of the susceptible strain respectively. Synergism tests showed that the synergist ratios of triphenyl phosphate (TPP), piperonyl buoxide (PBO) and diethyl maleate (DEM) were 1.04, 1.09, and 1.00 folds respectively in the bifenthrin-susceptible strain and 1.80, 7.97 and 1.03 foldsrespectively in the bifenthrin-resistant strain. These results indicate that CarE and PNOD play important roles in bifenthrin resistance of tea leafhoppers.
[1] 边磊, 孙晓玲, 陈宗懋. 假眼小绿叶蝉的日飞行活动性及成虫飞行能力的研究[J]. 茶叶科学, 2014, 34(3): 248-252.
[2] Pu X Y, Feng M G, Shi C H.Impact of three application methods on the field efficacy of a Beauveria bassiana-based mycoinsecticide against the false-eye leafhopper, Empoasca vitis (Homoptera: Cicadellidae) in the tea canopy[J]. Crop Protection, 2005, 24(2): 167-175.
[3] 张聪, 刘慧刚, 章晓凤. 联苯菊酯对靶标生物及非靶标生物毒性的对映体差异[J]. 浙江工业大学学报, 2009, 37(4): 366-371.
[4] 王念武, 徐金汉, 陈峥, 等. 不同茶园假眼小绿叶蝉抗药性比较[J]. 福建农林大学学报: 自然科学版, 2004, 33(2): 169-173.
[5] 庄家祥, 傅建炜, 苏庆泉, 等. 福建省茶小绿叶蝉抗药性的地区差异[J]. 茶叶科学, 2009, 29(2): 154-158.
[6] 邢剑飞, 刘艳, 颜冬云. 昆虫对拟除虫菊酯农药的抗性研究进展[J]. 环境科学与技术, 2010, 33(10): 68-74.
[7] 沈晋良, 吴益东. 棉铃虫抗药性及其治理[M]. 北京: 中国农业出版社, 1995: 259-280.
[8] Zhu YC, Snodgrass GL.Cytochrome P450 CYP6X1 cDNAs and mRNA expression levels in three strains of the tarnished plant bug Lygus lineolaris (Heteroptera: Miridae) having different susceptibilities to pyrethroid insecticide[J]. Insect Molecular Biology, 2003, 12(1): 39-49.
[9] Li Xianchun, Schuler MA, Berenbaum MR.Molecular mechanisms of metabolic resistance to synthetic and natural xenobiotics[J]. Annual Review of Entomology, 2007, 52: 231-253.
[10] 李春生. 增效剂对3种农药的增效作用及对甜菜夜蛾解毒酶活性的影响[D]. 重庆: 西南大学, 2006: 1-2.
[11] 农业部农药检定所. NY/T 1154.14—2008. 农药室内生物测定试验准则杀虫剂第14部分: 浸叶法 [S]. 北京: 中国农业出版社, 2011.
[12] Bradford MM.A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding[J]. Analytical Biochemistry, 1976, 7(1/2): 248-254.
[13] Rose R L, Barbhaiya L, Roe R M, et al. Cytochrome P450-associated insecticide resistance and the development of biochemical diagnostic assays in Heliothis virescens [J]. Pesticide Biochemistry and Physiology, 1995, 51(3): 178-191.
[14] Bautista MA, Miyata T, Miura K, et al.RNA interference-mediated knockdown of a cytochrome P450, CYP6BG1, from the diamondback moth, Plutella xylostella, reduces larval resistance to permethrin[J]. Insect Biochemistry Molecular Biology, 2009, 39(1): 38-46.
[15] 何玉仙, 黄建, 杨秀娟, 等. 烟粉虱对拟除虫菊酯杀虫剂的抗性机理[J]. 昆虫学报, 2007, 50(3): 241-247.
[16] Puinean AM, Foster SP, Denholm I, et al.Amplification of a cytochrome P450 gene is associated with resistance to neonicotinoid insecticides in the aphid Myzus persicae[J]. PLoS Genetics, 2010, 6(6): 1-11.
[17] Small GJ, Hemingway J.Molecular characterization of the amplified carboxylesterase gene associated with organophosphorus insecticide resistance in the brown planthopper Nilaparvata lugens[J]. Insect Molecular Biology, 2000, 9(6): 647-653.
[18] Cao CW, Zhang J, Gao XW, et al.Overexpression of carboxylesterase gene associated with organophosphorous insecticide resistance in cotton aphids, Aphis gossypii (Glover)[J]. Pesticide Biochemistry Physiology, 2008, 90(3): 175-180.
[19] Pan Y, Guo H, Gao XW.Comparative study on carboxylesterase activity, cDNA sequence, and gene expression between malathion susceptible and resistant strains of the cotton aphid, Aphis gossypii[J]. Comparative Biochemistry Physiology Part B Biochemistry Molecular Biology, 2010, 152(3): 266-270.
[20] Rumpf S, Hetzel F, Frampton C.Lacewings (Neuroptera: Hemerobiidae and Chrysopidae) and integrated pest management: enzyme activity as biomarker of sublethal insecticide exposure[J]. Journal of Economic Entomology, 1997, 90(1): 102-108.
[21] Fukami JI.Metabolism of several insecticides by glutathion S-transferase[J]. Pharmacology & therapeutics, 1980, 10(3): 473-514.
浙公网安备 33019902000101号 
