阿维菌素与杀铃脲对舞毒蛾幼虫的联合作用机制
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摘要
【目的】通过舞毒蛾幼虫解毒酶活力、保护酶活力测定,研究阿维菌素和杀铃脲对舞毒蛾幼虫的联合作用机制,为上述2种农药的混用提供理论依据。【方法】人工饲料中添加不等量的9 000μg·mL~(-1)阿维菌素丙酮溶液和25 000μg·mL~(-1)杀铃脲丙酮溶液,分别配制成含有阿维菌素、杀铃脲的单剂和复合剂混毒饲料。饲料中药剂含量:低浓度处理组单剂阿维菌素0.50μg·g~(-1),单剂杀铃脲8.30μg·g~(-1),复合剂阿维菌素0.50μg·g~(-1)、杀铃脲8.30μg·g~(-1);高浓度处理组单剂阿维菌素1.10μg·g~(-1),单剂杀铃脲16.60μg·g~(-1),复合剂阿维菌素1.10μg·g~(-1)、杀铃脲16.60μg·g~(-1)。采用含毒饲料处理3龄舞毒蛾幼虫,24 h后统计幼虫死亡率,测定存活幼虫解毒酶CarE和GST活力以及保护酶SOD、POD、CAT和PPO活力。【结果】对照组、阿维菌素和杀铃脲单剂处理时舞毒蛾幼虫死亡率为0,低浓度复合剂处理的舞毒蛾幼虫死亡率为1.15%,与对照组、单剂处理组无显著差异;高浓度复合剂处理的舞毒蛾幼虫死亡率为4.11%,与对照组、单剂处理组有显著差异。阿维菌素和杀铃脲单剂处理、复合剂处理的幼虫CarE和GST活力均高于对照组;阿维菌素和杀铃脲单剂处理时,幼虫CarE和GST活力显著高于对照组,复合剂处理与对照组无显著差异。阿维菌素和杀铃脲单剂处理及复合剂处理时,幼虫SOD、CAT和PPO活力均高于对照组,而POD活力均低于对照组;复合剂处理SOD、POD和CAT活力均高于单剂处理,而复合剂处理PPO活力则均低于单剂处理。【结论】阿维菌素和杀铃脲联合作用主要是通过抑制舞毒蛾幼虫解毒酶CarE和GST活力,再辅以抑制保护酶PPO活力,从而提高毒杀效果。
【Objective】 The combined use of abamectin and triflumuron has synergistic effect on the control of gypsy moths, but the synergy mechanism of abamectin and triflumuron is still unknown. In order to clarify the synergy mechanism of abamectin and triflumuron, 3 rd-instar Lymantria dispar larvae were treated with abamectin and triflumuron alone or their combination, and then the activities of detoxification enzymes and protective enzymes were determined, which would provide theoretical basis for the mixture use of the two pesticides. 【Method】 Abamectin acetone solution of 9 000 μg·mL~(-1) or triflumuron acetone solution of 25 000 μg·mL~(-1), and the two combination were added to the artificial diet for preparation of poisonous feed for L. dispar larvae. In a low concentration group, the content of drugs was 0.50 μg·g~(-1) abamectin or 8.30 μg·g~(-1) triflumuron in single pesticide treatment group, and 0.50 μg·g~(-1) abamectin plus 8.30 μg·g~(-1) triflumuron in mixed pesticide group. In a high concentration group, the content of drugs was 1.10 μg·g~(-1) abamectin or 16.60 μg·g~(-1) triflumuron in single pesticide treatment group, and 1.10 μg·g~(-1) abamectin plus 16.60 μg·g~(-1) triflumuron in mixed pesticide group. The third instar L. dispar larvae were fed with the poisonous artificial diet for 24 h. Survivors were counted following activity determinations of detoxification enzyme and protective enzyme. 【Result】 Mortality analysis showed that all larvae of the control group, single abamectin treatment, and single triflumuron treatment survived from this experiment. The mortality of mixed pesticide treatment group was 1.15% at a low concentration, and 4.11% at a high concentration. There was no significant difference between low concentration group and control, but there was significant difference between the high concentration group and control. Detoxification enzyme activities analysis showed that the activities of CarE and GST of larvae treated with abamectin or triflumuron were significantly higher than those of the control group, while there were no significant differences in the enzymes activities between the mixed pesticide treatment and the control group. Protective enzyme activities assay showed that activities of SOD, CAT and PPO of larvae treated with abamectin, triflumuron, and the two combination were higher than those of control group, however POD activities of treated larvae were lower than that of the control. Meanwhile, activities of SOD, POD and CAT of larvae treated with the mixed pesticides were higher than that treated with either single pesticide, while the case for PPO activity was opposite. 【Conclusion】 It is concluded that the combined effect of abamectin and triflumuron on Lymantria dispar larvae is mainly through inhibiting the activity of the detoxifying enzymes CarE and GST, and supplemented by inhibiting the activities of protective enzymes PPO, so as to improve the toxicity effect.
【Objective】 The combined use of abamectin and triflumuron has synergistic effect on the control of gypsy moths, but the synergy mechanism of abamectin and triflumuron is still unknown. In order to clarify the synergy mechanism of abamectin and triflumuron, 3 rd-instar Lymantria dispar larvae were treated with abamectin and triflumuron alone or their combination, and then the activities of detoxification enzymes and protective enzymes were determined, which would provide theoretical basis for the mixture use of the two pesticides. 【Method】 Abamectin acetone solution of 9 000 μg·mL~(-1) or triflumuron acetone solution of 25 000 μg·mL~(-1), and the two combination were added to the artificial diet for preparation of poisonous feed for L. dispar larvae. In a low concentration group, the content of drugs was 0.50 μg·g~(-1) abamectin or 8.30 μg·g~(-1) triflumuron in single pesticide treatment group, and 0.50 μg·g~(-1) abamectin plus 8.30 μg·g~(-1) triflumuron in mixed pesticide group. In a high concentration group, the content of drugs was 1.10 μg·g~(-1) abamectin or 16.60 μg·g~(-1) triflumuron in single pesticide treatment group, and 1.10 μg·g~(-1) abamectin plus 16.60 μg·g~(-1) triflumuron in mixed pesticide group. The third instar L. dispar larvae were fed with the poisonous artificial diet for 24 h. Survivors were counted following activity determinations of detoxification enzyme and protective enzyme. 【Result】 Mortality analysis showed that all larvae of the control group, single abamectin treatment, and single triflumuron treatment survived from this experiment. The mortality of mixed pesticide treatment group was 1.15% at a low concentration, and 4.11% at a high concentration. There was no significant difference between low concentration group and control, but there was significant difference between the high concentration group and control. Detoxification enzyme activities analysis showed that the activities of CarE and GST of larvae treated with abamectin or triflumuron were significantly higher than those of the control group, while there were no significant differences in the enzymes activities between the mixed pesticide treatment and the control group. Protective enzyme activities assay showed that activities of SOD, CAT and PPO of larvae treated with abamectin, triflumuron, and the two combination were higher than those of control group, however POD activities of treated larvae were lower than that of the control. Meanwhile, activities of SOD, POD and CAT of larvae treated with the mixed pesticides were higher than that treated with either single pesticide, while the case for PPO activity was opposite. 【Conclusion】 It is concluded that the combined effect of abamectin and triflumuron on Lymantria dispar larvae is mainly through inhibiting the activity of the detoxifying enzymes CarE and GST, and supplemented by inhibiting the activities of protective enzymes PPO, so as to improve the toxicity effect.
引文
崔博, 王春鑫, 吕妍,等.2017. 两种杀虫剂复配的固体纳米分散体的增效研究.中国生物防治学报,33(6):760-766.(Cui B, Wang C X, Lü Y, et al.2017. Synergistic effect of mixing solid nanodispersions of two insecticides.Chinese Journal of Biological Control,33(6):760-766.[in Chinese])
沈登荣, 何超, 赵远艳, 等.2017. 多杀菌素与4种杀虫剂复配对西花蓟马的联合毒力. 江苏农业科学, 45(23):91-93.(Shen D R, He C, Zhao Y Y, et al. 2017. The combined toxicity of spinosad and four insecticides to western flower thrips. Jiangsu Agricultural Sciences, 45(23):91-93. [in Chinese])
杨超.2016. 防治粘虫(Mythimna separata)和豆蚜(Aphis cracivora)的杀虫剂增效复配研究.上海: 华东理工大学硕士学位论文.(Yang C. 2016. Study on synergistic combination of insecticides for controlling Mythimna separata and Aphis cracivora larvae.Shanghai: MS thesis of East China University of Science and Technology. [in Chinese])
庾琴,王振, 封云涛, 等.2013. 不同杀虫剂对苹果黄蚜的毒力及复配研究. 植物保护, 39(3): 178-181.(Yu Q, Wang Z, Feng Y T, et al. 2013. Toxicity test of different pesticides and their mixed formulation against Aphis citricola. Plant Protection, 39(3): 178-181. [in Chinese])
张国财.2013. 森林有害生物防治. 哈尔滨: 黑龙江科学技术出版社.(Zhang G C. 2013. Forest Pest Management. Harbin: Heilongjiang Science and Technology Press. [in Chinese])
Bai S H,Ogbourne S.2016. Eco-toxicological effects of the avermectin family with a focus on abamectin and ivermectin. Chemosphere, 154: 204-214.
Barrett K, Grandy N, Harrison E. 1994.Guidance document on regulatory testing procedures for pesticides and non-target arthropods: from the ESCORT workshop (European standard characteristics of beneficials regulatory testing).IAC Wageningen, The Netherlands, 28-30.
Belinato T A, Martins A J, Lima J B P, et al. 2013. Effect of triflumuron, a chitin synthesis inhibitor, on Aedes aegypti, Aedes albopictus and Culex quinquefasciatus under laboratory conditions. Parasites & Vectors, 6(1): 83.
Bradford M M.1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72 (1): 248-254.
Chernaki-Leffer A M, Soso-Gomez D R, Almeida L M, et al.2011. Susceptibility of Alphitobius diaperinus (Panzer) (Coleoptera, Tenebrionidae) to cypermethrin, dichlorvos and triflumuron in southern Brazil. Revista Brasileira De Entomologia, 55 (1): 125-128.
Henriques B S, Genta F A, Mello C B, et al.2016. Triflumuron effects on the physiology and reproduction Ofrhodnius prolixus adult females. BioMed Research International, (2): 1-11.
Jacups S P, Paton C J, Ritchie S A.2014. Residual and pre-treatment application of starycide insect growth regulator (triflumuron) to control Aedes aegypti in containers. Pest Management Science, 70 (4): 572-575.
Jin T, Lin Y, Jin Q, et al.2014. Sublethal effect of avermectin and acetamiprid on the mortality of different life stages of Brontispa longissima (Gestro) (Coleoptera: Hispidae) and its larvae parasitoid Asecodes hispinarum Bou■ek (Hymenoptera: Eulophidae). Crop Protection, 58: 55-60.
Kotze A C, Ruffell A, Lamb J, et al.2018. Response of drug-susceptible and -resistant Haemonchus contortus larvae to monepantel and abamectin alone or in combination in vitro. Vet Parasitol, 249: 57-62.
Mermans C, DermauwW, Geibel S, et al.2017. A G326E substitution in the glutamate-gated chloride channel 3 (GluCl3) of the two-spotted spider mite Tetranychus urticae abolishes the agonistic activity of macrocyclic lactones. Pest Management Science, 73 (12): 2413-2418.
Merzendorfer H.2006. Insect chitin synthases: a review. Journal of Comparative Physiology B, 176 (1): 1-15.
Merzendorfer H.2013. Chitin synthesis inhibitors: old molecules and new developments. Insect Science, 20 (2): 121-138.
Ozdemir N, Kahraman T.2016. Rapid confirmatory analysis of avermectin residues in milk by liquid chromatography tandem mass spectrometry. Journal of Food & Drug Analysis, 24 (1): 90-94.
Sagheer M,Yasir M, Mansoor H, et al.2012. Impact of triflumuron on reproduction and development of red flour beetle, Tribolium castaneum (herbst) (Coleoptera: Tenebrionidae). Pakistan Journal of Agricultural Sciences, 49 (2): 173-178.
Siddique S, Syed Q,Saleem Y, et al.2015. Toxicity of avermectin B1b to earthworm and cockroaches. Journal of Animal & Plant Sciences, 25 (3): 844-850.
Sun R, Liu C, Zhang H, et al.2015. Benzoylurea chitin synthesis inhibitors. Journal of Agricultural and Food Chemistry, 63 (31): 6847-6865.
Waghorn T S, McKay C H, Heath A C.2013. The in vitro response of field strains of sheep blowflies Lucilia sericata and L. cuprina (Calliphoridae) in New Zealand to dicyclanil and triflumuron. New Zealand Veterinary Journal, 61 (5): 274-280.
Wann K T.2010. The cellular actions of the avermectins. Phytotherapy Research, 1 (4): 143-150.
Wanna R, Xu Z H, Yu H X.2012. Bioefficacy of a mixed biocide BtA against Helicoverpa armigera (Lepidoptera: Noctuidae) and its contact toxicity to pupae and adults of parasitoid Microplitis mediator (Hymenoptera: Braconidae). Acta Entomologica Sinica, 55(8): 941-949.
Wei P, Che W, Wang J, et al.2018. RNA interference of glutamate-gated chloride channel decreases abamectin susceptibility in Bemisia tabaci. Pesticide Biochemistry and Physiology, 145: 1-7.
Xu Z, Shi L, Peng J, et al.2016. Analysis of the relationship between P-glycoprotein and abamectin resistance in Tetranychus cinnabarinus (Boisduval). Pesticide Biochemistry and Physiology, 129: 75-82.
Zandonadi C H S, Alves T C, Silva S M. 2017. Tank mixture of pesticides for Spodoptera frugiperda control in maize with triflumuron. Bioscience Journal, 33 (1): 31-40.
Zhao Y, Sun Q, Hu K, et al.2016. Isolation, characterization, and tissue-specific expression of GABA A receptor α1 subunit gene of Carassius auratus gibelio after avermectin treatment. Fish Physiology and Biochemistry, 42 (1): 83-92.
沈登荣, 何超, 赵远艳, 等.2017. 多杀菌素与4种杀虫剂复配对西花蓟马的联合毒力. 江苏农业科学, 45(23):91-93.(Shen D R, He C, Zhao Y Y, et al. 2017. The combined toxicity of spinosad and four insecticides to western flower thrips. Jiangsu Agricultural Sciences, 45(23):91-93. [in Chinese])
杨超.2016. 防治粘虫(Mythimna separata)和豆蚜(Aphis cracivora)的杀虫剂增效复配研究.上海: 华东理工大学硕士学位论文.(Yang C. 2016. Study on synergistic combination of insecticides for controlling Mythimna separata and Aphis cracivora larvae.Shanghai: MS thesis of East China University of Science and Technology. [in Chinese])
庾琴,王振, 封云涛, 等.2013. 不同杀虫剂对苹果黄蚜的毒力及复配研究. 植物保护, 39(3): 178-181.(Yu Q, Wang Z, Feng Y T, et al. 2013. Toxicity test of different pesticides and their mixed formulation against Aphis citricola. Plant Protection, 39(3): 178-181. [in Chinese])
张国财.2013. 森林有害生物防治. 哈尔滨: 黑龙江科学技术出版社.(Zhang G C. 2013. Forest Pest Management. Harbin: Heilongjiang Science and Technology Press. [in Chinese])
Bai S H,Ogbourne S.2016. Eco-toxicological effects of the avermectin family with a focus on abamectin and ivermectin. Chemosphere, 154: 204-214.
Barrett K, Grandy N, Harrison E. 1994.Guidance document on regulatory testing procedures for pesticides and non-target arthropods: from the ESCORT workshop (European standard characteristics of beneficials regulatory testing).IAC Wageningen, The Netherlands, 28-30.
Belinato T A, Martins A J, Lima J B P, et al. 2013. Effect of triflumuron, a chitin synthesis inhibitor, on Aedes aegypti, Aedes albopictus and Culex quinquefasciatus under laboratory conditions. Parasites & Vectors, 6(1): 83.
Bradford M M.1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72 (1): 248-254.
Chernaki-Leffer A M, Soso-Gomez D R, Almeida L M, et al.2011. Susceptibility of Alphitobius diaperinus (Panzer) (Coleoptera, Tenebrionidae) to cypermethrin, dichlorvos and triflumuron in southern Brazil. Revista Brasileira De Entomologia, 55 (1): 125-128.
Henriques B S, Genta F A, Mello C B, et al.2016. Triflumuron effects on the physiology and reproduction Ofrhodnius prolixus adult females. BioMed Research International, (2): 1-11.
Jacups S P, Paton C J, Ritchie S A.2014. Residual and pre-treatment application of starycide insect growth regulator (triflumuron) to control Aedes aegypti in containers. Pest Management Science, 70 (4): 572-575.
Jin T, Lin Y, Jin Q, et al.2014. Sublethal effect of avermectin and acetamiprid on the mortality of different life stages of Brontispa longissima (Gestro) (Coleoptera: Hispidae) and its larvae parasitoid Asecodes hispinarum Bou■ek (Hymenoptera: Eulophidae). Crop Protection, 58: 55-60.
Kotze A C, Ruffell A, Lamb J, et al.2018. Response of drug-susceptible and -resistant Haemonchus contortus larvae to monepantel and abamectin alone or in combination in vitro. Vet Parasitol, 249: 57-62.
Mermans C, DermauwW, Geibel S, et al.2017. A G326E substitution in the glutamate-gated chloride channel 3 (GluCl3) of the two-spotted spider mite Tetranychus urticae abolishes the agonistic activity of macrocyclic lactones. Pest Management Science, 73 (12): 2413-2418.
Merzendorfer H.2006. Insect chitin synthases: a review. Journal of Comparative Physiology B, 176 (1): 1-15.
Merzendorfer H.2013. Chitin synthesis inhibitors: old molecules and new developments. Insect Science, 20 (2): 121-138.
Ozdemir N, Kahraman T.2016. Rapid confirmatory analysis of avermectin residues in milk by liquid chromatography tandem mass spectrometry. Journal of Food & Drug Analysis, 24 (1): 90-94.
Sagheer M,Yasir M, Mansoor H, et al.2012. Impact of triflumuron on reproduction and development of red flour beetle, Tribolium castaneum (herbst) (Coleoptera: Tenebrionidae). Pakistan Journal of Agricultural Sciences, 49 (2): 173-178.
Siddique S, Syed Q,Saleem Y, et al.2015. Toxicity of avermectin B1b to earthworm and cockroaches. Journal of Animal & Plant Sciences, 25 (3): 844-850.
Sun R, Liu C, Zhang H, et al.2015. Benzoylurea chitin synthesis inhibitors. Journal of Agricultural and Food Chemistry, 63 (31): 6847-6865.
Waghorn T S, McKay C H, Heath A C.2013. The in vitro response of field strains of sheep blowflies Lucilia sericata and L. cuprina (Calliphoridae) in New Zealand to dicyclanil and triflumuron. New Zealand Veterinary Journal, 61 (5): 274-280.
Wann K T.2010. The cellular actions of the avermectins. Phytotherapy Research, 1 (4): 143-150.
Wanna R, Xu Z H, Yu H X.2012. Bioefficacy of a mixed biocide BtA against Helicoverpa armigera (Lepidoptera: Noctuidae) and its contact toxicity to pupae and adults of parasitoid Microplitis mediator (Hymenoptera: Braconidae). Acta Entomologica Sinica, 55(8): 941-949.
Wei P, Che W, Wang J, et al.2018. RNA interference of glutamate-gated chloride channel decreases abamectin susceptibility in Bemisia tabaci. Pesticide Biochemistry and Physiology, 145: 1-7.
Xu Z, Shi L, Peng J, et al.2016. Analysis of the relationship between P-glycoprotein and abamectin resistance in Tetranychus cinnabarinus (Boisduval). Pesticide Biochemistry and Physiology, 129: 75-82.
Zandonadi C H S, Alves T C, Silva S M. 2017. Tank mixture of pesticides for Spodoptera frugiperda control in maize with triflumuron. Bioscience Journal, 33 (1): 31-40.
Zhao Y, Sun Q, Hu K, et al.2016. Isolation, characterization, and tissue-specific expression of GABA A receptor α1 subunit gene of Carassius auratus gibelio after avermectin treatment. Fish Physiology and Biochemistry, 42 (1): 83-92.