异迟眼蕈蚊对溴氰菊酯的抗性风险及解毒酶活力分析
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摘要
为了探讨异迟眼蕈蚊对溴氰菊酯的抗性风险和抗性生化机理,在室内采用溴氰菊酯对异迟眼蕈蚊进行抗性选育,对其抗性现实遗传力和解毒酶活力进行分析。结果表明:采用个体选育法和群体选育法分别对敏感品系和抗性品系进行选育,抗性选育24代后,抗性倍数达到11.36倍,现实遗传力h~2为0.077 1。当致死率为80%～90%时,预计异迟眼蕈蚊对溴氰菊酯的抗性增长10倍需要11～14代。增效剂对溴氰菊酯的增效结果表明,抗性品系中增效醚和磷酸三苯酯对溴氰菊酯的增效作用显著(P<0.05)。解毒酶活性分析表明,随着抗性选育代数的增加,多功能氧化酶O-脱甲基和羧酸酯酶的活性显著增加(P<0.05)。说明异迟眼蕈蚊对溴氰菊酯的抗性风险较小,但在食用菌生产中也要尽量降低溴氰菊酯的使用浓度;多功能氧化酶和羧酸酯酶活性的增强是异迟眼蕈蚊对溴氰菊酯产生代谢抗性的主要原因。
In order to explore the resistance risk and biochemical mechanism of deltamethrin in Bradysia difformis,selection of B.difformis resistance to deltamethrin was performed in the laboratory,and realized heritability and detoxification enzyme activity of deltamethrin in B.difformis were analyzed. The results showed that the susceptible strain and resistant strain were selected by individual selection and group selection respectively,After 24 generations of resistant selection,a resistant strain with a resistance ratio of 11.36-fold was obtained,and its realized heritability(h~2) was 0.077 1.If the lethallty rate was 80%-90%,B.difformis was expected to obtain 10-fold resistance to deltamethrin after resistant selection of 11 to 14 generations. The synergistic effect of synergist to deltamethrin showed that piperonyl butoxide and triphenyl phosphate had significant synergistic effect on deltamethrin in resistant strain(P<0.05). The activity analysis of detoxifying enzyme showed that with the increase of selective generation,the activity of O-demethylation of multi-functional oxidase and carboxylate enzyme was significantly increased(P<0.05). The results indicated that B.difformis may has low risk resistance to deltamethrin,it was necessary to reduce the concentration of deltamethrin in edible fungus production,and the enhanced activity of multifunctional oxidase and carboxylate enzyme may be the main cause of metabolic resistance to deltamethrin in B.difformis.
In order to explore the resistance risk and biochemical mechanism of deltamethrin in Bradysia difformis,selection of B.difformis resistance to deltamethrin was performed in the laboratory,and realized heritability and detoxification enzyme activity of deltamethrin in B.difformis were analyzed. The results showed that the susceptible strain and resistant strain were selected by individual selection and group selection respectively,After 24 generations of resistant selection,a resistant strain with a resistance ratio of 11.36-fold was obtained,and its realized heritability(h~2) was 0.077 1.If the lethallty rate was 80%-90%,B.difformis was expected to obtain 10-fold resistance to deltamethrin after resistant selection of 11 to 14 generations. The synergistic effect of synergist to deltamethrin showed that piperonyl butoxide and triphenyl phosphate had significant synergistic effect on deltamethrin in resistant strain(P<0.05). The activity analysis of detoxifying enzyme showed that with the increase of selective generation,the activity of O-demethylation of multi-functional oxidase and carboxylate enzyme was significantly increased(P<0.05). The results indicated that B.difformis may has low risk resistance to deltamethrin,it was necessary to reduce the concentration of deltamethrin in edible fungus production,and the enhanced activity of multifunctional oxidase and carboxylate enzyme may be the main cause of metabolic resistance to deltamethrin in B.difformis.
引文
[1] MENZEL F,SMITH J E,Colauto N B.Bradysia difformis Frey and Bradysia ocellaris (Comstock):two additional neotropical species of black fungus gnats (Diptera:Sciaridae) of economic importance:a redescription and review[J].Annals of the Entomological Society of America,2003,96(4):448-457.
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[4] 曲绍轩,李辉平,宋金俤,等.云南楚雄和丽江地区野生食用菌害虫抽查与鉴定[J].食用菌学报,2013,20(4):61-64.QU SH X,LI H P,SONG J D,et al.Pest of wild edible fungi in Chuxiong district,Kunming and Lijiang[J].Acta Edulis Fungi,2013,20(4):61-64.
[5] 樊中臣,唐俊,操海群,等.平菇及其培养料中5种拟除虫菊酯类农药的残留消解动态[J].食品科学,2013,34(11):41-44.FAN ZH CH,TANG J,CAO H Q,et al.Degradation kinetics of 5 pyrethroid pesticide residues in pleurotus ostreatus and growing medium[J].Food Science,2013,34(11):41-44.
[6] BREWER K K,KEIL C B.Permethrin resistance in Lycoriella mali (Diptera:Sciaridae)[J].Journal of Economic Entomology,1989,82(1):17-21.
[7] SHIRVANI-FARSANI N,ZAMANI A A,ABBASI S,et al.Toxicity of three insecticides and tobacco extract against the fungus gnat,Lycoriella auripila and the economic injury level of the gnat on button mushroom[J].Journal of Pest Science,2013,86(3):591-597.
[8] 丁倩,王意婷,刘俊杰,等.我国韭菜主产区韭菜迟眼蕈蚊田间种群的抗药性监测[J].应用昆虫学报,2016,53(6):1242-1249.DING Q,WANG Y T,LIU J J,et al.Monitoring the resistance of field populations of Bradysia odoriphaga Yang et Zhang in the main leek producing areas in China[J].Chinese Journal of Applied Entomology,2016,53(6):1242-1249.
[9] 陈澄宇,王翠翠,郭静静,等.韭菜迟眼蕈蚊对新烟碱类杀虫剂的抗药性及抗性机制初探[J].应用昆虫学报,2016,53(6):1250-1254.CHEN CH Y,WANG C C,GUO J J,et al.Neonicotinoid insecticide resistance in Bradysia odoriphaga Yang et Zhang[J].Chinese Journal of Applied Entomology,2016,53(6):1250-1254.
[10] 兰珊珊,林涛,林昕,等.食品安全指数法评估西南地区食用菌中农药残留风险[J].江苏农业学报,2014,30(1):199-204.LAN SH SH,LIN T,LIN X,et al.Risk assessment of pesticide residues in edible mushrooms in Southwest China based on food safety indexes[J].Jiangsu Journal of Agricultural Sciences,2014,30(1):199-204.
[11] 江雪飞,刘丹丹,樊晓琳,等.食用菌4个主产区常见农药的残留量分析[J].中国食用菌,2012,31(3):53-54.JIANG X F,LIU D D,FAN X L,et al.Analysis of common pesticides residues in four main areas of edible fungi[J].Edible Fungi of China,2012,31(3):53-54.
[12] 陈澄宇,史雪岩,髙希武.昆虫对拟除虫菊酯类杀虫剂的代谢抗性机制研究进展[J].农药学学报,2016,18(5):545-555.CHEN CH Y,SHI X Y,GAO X W.Mechanism of insect metabolic resistance to pyrethroid insecticides[J].Chinese Journal of Pesticide Science,2016,18(5):545-555.
[13] 薛明,袁林,徐曼琳.韭菜迟眼蕈蚊成虫对挥发性物质的嗅觉反应及不同杀虫剂的毒力比较[J].农药学学报,2002,4(2):50-56.XUE M,YUAN L,XU M L.The olfactory response of adults to volatiles and compare of toxicity of different insecticides to the adults and larvae of Bradysia odoriphaga[J].Chinese Journal of Pesticide Science,2002,4(2):50-56.
[14] 郭天凤,马野萍,丁荣荣,等.棉蚜对吡虫啉的抗性选育和现实遗传力分析[J].昆虫学报,2014,57(3):330-334.GUO T F,MA Y P,DING R R,et al.Selection and realized heritability analysis of resistance to imidacloprid in cotton aphid (Aphis gossypii)[J].Acta Entomologica Sinica,2014,57(3):330-334.
[15] TABASHNIK B E.Resistance risk assessment:realized heritability of resistance to Bacillus thuringiensis in diamondback moth (Lepidoptera:Plutellidae),tobacco budworm (Lepidoptera:Noctuidae),and colorado potato beetle (Coleoptera:Chrysomelidae)[J].Journal of Economic Entomology,1992,85:1551-1559.
[16] 余慧灵,向兴,袁贵鑫.溴氰虫酰胺亚致死剂量对甜菜夜蛾生长发育及体内解毒酶活性的影响[J].昆虫学报,2015,58 (6):634-641.YU H L,XIANG X,YUAN G X.et al.Effects of sublethal doses of cyantraniliprole on the growth and development and the activities of detoxifying enzymes in Spodoptera exigua (Lepidoptera:Noctuidae)[J].Acta Entomologica Sinica,2015,58 (6):634-641.
[17] BRADFORD M M.A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye blinding[J].Analytical Biochemistry,1976,72(1):248-254.
[18] 刘洪霞,冷培恩,徐劲秋,等.白纹伊蚊对溴氰菊酯的抗性选育及抗性风险评估[J].中华卫生杀虫药械,2015,21(2):125-127.LIU H X,LENG P E,XU J Q,et al.Resistance selection of Aedes albopictus to deltamethrin and risk assessment[J].Chinese Journal of Hygienic Insecticides and Equipments,2015,21(2):125-127.
[19] 魏绪强,李宗麟,潘京海,等.北京市东城区家蝇对溴氰菊酯抗性风险评估与预报[J].中国媒介生物学及控制杂志,2013,24(5):402-405.WEI X Q,LI Z L,PAN J H,et al.Risk assessment and prediction of deltamethrin resistance in Musca domestica in Dongcheng district of Beijing,China[J].Chinese Journal of Vector Biology and Control,2013,24(5):402-405.
[20] 徐鹿.灰飞虱对毒死蜱和溴氰菊酯的抗性及其机理的研究[D].南京:南京农业大学,2013.XU L.Studies on chlorpyrifos and deltamethrin resistance in small brown planthopper,Laodelphax striatellus (Fallen) and possible Mechanisms[D].Nanjing:Nanjing Agricultural University,2013.
[21] BARTLETT G R,KEIL C B O.Identification and characterization of a permethrin resistance mechanism in populations of the fungus gnat Lycoriella mali (Fitch) (Diptera:Sciaridae)[J].Pesticide Biochemistry and Physiology,1997,58(3):173-181.
[22] EI-LATIF A O A,SUBRAHMANYAM B.Pyrethroid resistance and esterase activity in three strains of the cotton bollworm,Helicoverpa armigera (Hubner)[J].Pesticide Biochemistry and Physiology,2010,96(3):155-159.
[23] YANG Y,WU Y,CHEN S,et al.The involvement of microsomal oxidases in pyrethroid resistance in Helicoverpa armigera from Asia[J].Insect Biochemistry and Molecular Biology,2004,34(8):763-773.
[2] HURLEY B P,SLIPPERS B,WINGFIELD B D,et al.Genetic diversity of Bradysia difformis (Sciaridae:Diptera) populations reflects movement of an invasive insect between forestry nurseries[J].Biological Invasions,2010,12(4):729-733.
[3] 沈登荣,张宏瑞,李正跃,等.云南食用菌眼蕈蚊分类及优势种分析[J].昆虫学报,2009,52(8):934-940.SHEN D R,ZHANG H R,LI ZH Y,et al.Taxonomy and dominance analysis of sciarid fly species (Diptera:Sciaridae) on edible fungi inYunnan[J].Acta Entomologica Sinica,2009,52(8):934-940.
[4] 曲绍轩,李辉平,宋金俤,等.云南楚雄和丽江地区野生食用菌害虫抽查与鉴定[J].食用菌学报,2013,20(4):61-64.QU SH X,LI H P,SONG J D,et al.Pest of wild edible fungi in Chuxiong district,Kunming and Lijiang[J].Acta Edulis Fungi,2013,20(4):61-64.
[5] 樊中臣,唐俊,操海群,等.平菇及其培养料中5种拟除虫菊酯类农药的残留消解动态[J].食品科学,2013,34(11):41-44.FAN ZH CH,TANG J,CAO H Q,et al.Degradation kinetics of 5 pyrethroid pesticide residues in pleurotus ostreatus and growing medium[J].Food Science,2013,34(11):41-44.
[6] BREWER K K,KEIL C B.Permethrin resistance in Lycoriella mali (Diptera:Sciaridae)[J].Journal of Economic Entomology,1989,82(1):17-21.
[7] SHIRVANI-FARSANI N,ZAMANI A A,ABBASI S,et al.Toxicity of three insecticides and tobacco extract against the fungus gnat,Lycoriella auripila and the economic injury level of the gnat on button mushroom[J].Journal of Pest Science,2013,86(3):591-597.
[8] 丁倩,王意婷,刘俊杰,等.我国韭菜主产区韭菜迟眼蕈蚊田间种群的抗药性监测[J].应用昆虫学报,2016,53(6):1242-1249.DING Q,WANG Y T,LIU J J,et al.Monitoring the resistance of field populations of Bradysia odoriphaga Yang et Zhang in the main leek producing areas in China[J].Chinese Journal of Applied Entomology,2016,53(6):1242-1249.
[9] 陈澄宇,王翠翠,郭静静,等.韭菜迟眼蕈蚊对新烟碱类杀虫剂的抗药性及抗性机制初探[J].应用昆虫学报,2016,53(6):1250-1254.CHEN CH Y,WANG C C,GUO J J,et al.Neonicotinoid insecticide resistance in Bradysia odoriphaga Yang et Zhang[J].Chinese Journal of Applied Entomology,2016,53(6):1250-1254.
[10] 兰珊珊,林涛,林昕,等.食品安全指数法评估西南地区食用菌中农药残留风险[J].江苏农业学报,2014,30(1):199-204.LAN SH SH,LIN T,LIN X,et al.Risk assessment of pesticide residues in edible mushrooms in Southwest China based on food safety indexes[J].Jiangsu Journal of Agricultural Sciences,2014,30(1):199-204.
[11] 江雪飞,刘丹丹,樊晓琳,等.食用菌4个主产区常见农药的残留量分析[J].中国食用菌,2012,31(3):53-54.JIANG X F,LIU D D,FAN X L,et al.Analysis of common pesticides residues in four main areas of edible fungi[J].Edible Fungi of China,2012,31(3):53-54.
[12] 陈澄宇,史雪岩,髙希武.昆虫对拟除虫菊酯类杀虫剂的代谢抗性机制研究进展[J].农药学学报,2016,18(5):545-555.CHEN CH Y,SHI X Y,GAO X W.Mechanism of insect metabolic resistance to pyrethroid insecticides[J].Chinese Journal of Pesticide Science,2016,18(5):545-555.
[13] 薛明,袁林,徐曼琳.韭菜迟眼蕈蚊成虫对挥发性物质的嗅觉反应及不同杀虫剂的毒力比较[J].农药学学报,2002,4(2):50-56.XUE M,YUAN L,XU M L.The olfactory response of adults to volatiles and compare of toxicity of different insecticides to the adults and larvae of Bradysia odoriphaga[J].Chinese Journal of Pesticide Science,2002,4(2):50-56.
[14] 郭天凤,马野萍,丁荣荣,等.棉蚜对吡虫啉的抗性选育和现实遗传力分析[J].昆虫学报,2014,57(3):330-334.GUO T F,MA Y P,DING R R,et al.Selection and realized heritability analysis of resistance to imidacloprid in cotton aphid (Aphis gossypii)[J].Acta Entomologica Sinica,2014,57(3):330-334.
[15] TABASHNIK B E.Resistance risk assessment:realized heritability of resistance to Bacillus thuringiensis in diamondback moth (Lepidoptera:Plutellidae),tobacco budworm (Lepidoptera:Noctuidae),and colorado potato beetle (Coleoptera:Chrysomelidae)[J].Journal of Economic Entomology,1992,85:1551-1559.
[16] 余慧灵,向兴,袁贵鑫.溴氰虫酰胺亚致死剂量对甜菜夜蛾生长发育及体内解毒酶活性的影响[J].昆虫学报,2015,58 (6):634-641.YU H L,XIANG X,YUAN G X.et al.Effects of sublethal doses of cyantraniliprole on the growth and development and the activities of detoxifying enzymes in Spodoptera exigua (Lepidoptera:Noctuidae)[J].Acta Entomologica Sinica,2015,58 (6):634-641.
[17] BRADFORD M M.A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye blinding[J].Analytical Biochemistry,1976,72(1):248-254.
[18] 刘洪霞,冷培恩,徐劲秋,等.白纹伊蚊对溴氰菊酯的抗性选育及抗性风险评估[J].中华卫生杀虫药械,2015,21(2):125-127.LIU H X,LENG P E,XU J Q,et al.Resistance selection of Aedes albopictus to deltamethrin and risk assessment[J].Chinese Journal of Hygienic Insecticides and Equipments,2015,21(2):125-127.
[19] 魏绪强,李宗麟,潘京海,等.北京市东城区家蝇对溴氰菊酯抗性风险评估与预报[J].中国媒介生物学及控制杂志,2013,24(5):402-405.WEI X Q,LI Z L,PAN J H,et al.Risk assessment and prediction of deltamethrin resistance in Musca domestica in Dongcheng district of Beijing,China[J].Chinese Journal of Vector Biology and Control,2013,24(5):402-405.
[20] 徐鹿.灰飞虱对毒死蜱和溴氰菊酯的抗性及其机理的研究[D].南京:南京农业大学,2013.XU L.Studies on chlorpyrifos and deltamethrin resistance in small brown planthopper,Laodelphax striatellus (Fallen) and possible Mechanisms[D].Nanjing:Nanjing Agricultural University,2013.
[21] BARTLETT G R,KEIL C B O.Identification and characterization of a permethrin resistance mechanism in populations of the fungus gnat Lycoriella mali (Fitch) (Diptera:Sciaridae)[J].Pesticide Biochemistry and Physiology,1997,58(3):173-181.
[22] EI-LATIF A O A,SUBRAHMANYAM B.Pyrethroid resistance and esterase activity in three strains of the cotton bollworm,Helicoverpa armigera (Hubner)[J].Pesticide Biochemistry and Physiology,2010,96(3):155-159.
[23] YANG Y,WU Y,CHEN S,et al.The involvement of microsomal oxidases in pyrethroid resistance in Helicoverpa armigera from Asia[J].Insect Biochemistry and Molecular Biology,2004,34(8):763-773.