棉蚜吡虫啉、啶虫脒不同品系解毒酶活性测定和增效剂作用的研究

通过棉蚜Aphis gossypii吡虫啉和啶虫脒抗性、敏感品系解毒酶活性测定和增效剂试验,明确与抗药性产生密切相关的解毒酶。采用室内生物测定和生化分析方法,研究棉蚜吡虫啉和啶虫脒品系解毒酶活性变化和增效剂的增效作用。解毒酶活性测定表明,棉蚜吡虫啉和啶虫脒抗性品系羧酸酯酶、谷胱甘肽-S-转移酶、细胞色素P450s O-脱乙基比活力都高于敏感品系,其中抗感品系中羧酸酯酶、细胞色素P450-O-脱乙基比活力差异都达到了显著水平(P0.05)。吡虫啉抗性品系三种解毒酶活性分别为敏感品系的3.26倍、1.08倍、1.60倍和1.58倍;啶虫脒抗性品系三种解毒酶活性分别为敏感品系的2.91倍、1.04倍、1.69倍和1.46倍。增效剂磷酸三苯酯(TPP)、增效醚(PBO)、顺丁烯二酸二乙酯(DEM),在吡虫啉敏感品系中的增效比分别为1.12、1.09、0.97,在吡虫啉抗性品系中的增效比分别为2.02、1.75、1.05;在啶虫脒敏感棉蚜品系中的增效比为1.02、1.03、1.02,在啶虫脒抗性棉蚜品系中的增效比为1.77、1.61、1.04。增效剂试验和解毒酶试验表明,酯酶和多功能氧化酶在棉蚜对吡虫啉、啶虫脒的抗性产生中起到了重要的作用。     

棉铃虫抗溴氰菊酯品系选育、生化机理及遗传方式研究

用溴氰菊酯选育抗性水平中等的河北冀县棉铃虫,经5代4次筛选获得一抗性品系(Del—R),抗性倍数由筛选前F_1代的52.8倍上升到筛选后F_5代的585.9倍(与1983年东台敏感品系相比)。利用此抗性品系进行增效剂(PBo、TPP、DEF)的活体增效作用测定和离体的酶活力研究,无论是对田间品系还是实验室选育的抗性品系,PBo对多功能氧化酶(MFO)的抑制作用是明显的,增效比从3.0—16.5不等,DEF、TPP的增效比分别为2.7和1.0。Del—R品系的多功能氧化酶环氧化活力(AE)是敏感品系的1.5倍,而Del—R品系的酯酶活力是敏感品系的1.01倍,因此多功能氧化酶活力增强是棉铃虫对溴氰菊酯产生抗性的原因之一,酯酶和羧酸酯酶在棉铃虫对溴氰菊酯的抗性中无多大作用。采用剂量对数-死亡机率值(LD—P线)分析法,将棉铃虫对溴氰菊酯的抗性品系和室内培育的敏感品系进行正交、反交和回交试验,得到的抗性遗传研究结果表明,棉铃虫对溴氰菊酯的抗性为常染色体多基因不完全显性遗传。     

哒螨灵对抗吡虫啉棉蚜种群的负交互抗性及对其生物学特性的影响

为明确哒螨灵对抗吡虫啉棉蚜Aphis gossypii种群的防治效果,采用浸渍法、微量点滴法、生化测定法和系统观察法,研究了哒螨灵对抗吡虫啉棉蚜种群(RF27)、敏感棉蚜种群(SS)和夏津种群(XJ)的负交互抗性及对其生物学特性的影响。结果表明:哒螨灵对RF27、XJ和SS的LD50分别为1.065×10-5、6.616×10-5和4.862×10-5μg/头;抗吡虫啉棉蚜种群与哒螨灵、吡蚜酮具有明显的负交互抗性,与啶虫脒、烯啶虫胺等具有交互抗性;CarE、GST和AChE比活力的提高是引起棉蚜对吡虫啉产生抗性的主要生化原因;哒螨灵对棉蚜的AChE和GST均有显著抑制作用;哒螨灵处理的抗吡虫啉棉蚜种群表现出明显的生殖缺陷,相对适合度较小,为0.67。     

棉铃虫抗辛硫磷品系的代谢抗性机理

通过重复回交和药剂选择,将棉铃虫Phoxim-R抗性品系对辛硫磷的抗性导入到BK77敏感品系中,得到棉铃虫BK77-R抗性品系,BK77-R和BK77为一对近等基因系。BK77-R抗性品系对辛硫磷的抗性达155倍,对溴氰菊酯有高水平交互抗性(抗性倍数248倍),对灭多威和硫丹有中等水平交互抗性, 分别为31倍和11倍,对丙溴磷有低水平交互抗性(4倍)。在BK77-R抗性品系中,脱叶磷(DEF,酯酶抑制剂)对辛硫磷、灭多威和硫丹具有增效作用,增效倍数分别为7倍、2倍 和1.9倍;增效醚(PBO,氧化酶抑制剂)对溴氰菊酯、灭多威和辛硫磷的增效倍数分别为21倍、2.2倍和1.7倍。与BK77敏感品系相比,BK77-R抗性品系的酯酶和多功能氧化酶活性均显著提高,而谷胱甘肽S-转移酶活性没有明显变化。上述结果表明,酯酶解毒代谢在棉铃虫BK77-R品系对辛硫磷的抗性中起重要作用,酯酶和多功能氧化酶解毒作用增强是该抗性品系对不同类型药剂产生交互抗性的重要原因。     

土耳其斯坦叶螨对杀螨剂的抗性选育及解毒酶活力变化

为探索土耳其斯坦叶螨的抗药性及其生化机理,在室内对敏感系土耳其斯坦叶螨分别用螺螨酯、甲氰菊酯和阿维菌素逐代处理,选育出抗性种群。结果表明,选育至15代,土耳其斯坦叶螨对螺螨酯、甲氰菊酯和阿维菌素的抗性指数分别达到268.63、37.98和112.68倍。分别测定敏感品系(SS)、抗螺螨酯(RS)、抗甲氰菊酯(RF)、抗阿维菌素(RA)品系的解毒酶活性显示,3种不同抗性品系相对SS品系的羧酸酯酶(CarE)、谷胱甘肽S-转移酶(GSTs)和多功能氧化酶(MFO)的比活力均有不同程度的提高,差异均达到显著水平(P0.05)。其中,RF品系的MFO比活力上升最快,是SS品系的12.7倍;RA品系的MFO比活力次之,是SS品系的5.76倍;RS品系的3种解毒酶比活力均增长较慢,其中CarE比活力上升最慢,是SS品系的1.31倍。由此表明,CarE、GSTs、MFO的活性增大可促进土耳其斯坦叶螨对3种杀虫剂的抗性形成;螺螨酯的抗性增强可能与CarE关系甚微;MFO活性的增加可能与甲氰菊酯抗性升高密切相关;GSTs、MFO的活性升高可能是土耳其斯坦叶螨对阿维菌素产生抗性的主要原因。     

棉蚜对吡虫啉等杀虫剂抗药性品系的室内选育及抗药性风险评价

通过室内杀虫剂处理水培棉苗选育抗性棉蚜,研究棉蚜在药剂诱导下抗性的变化,比较吡虫啉、吡虫啉 增效磷、氧乐果和氰戊菊酯对棉蚜抗性诱导的差异,评价棉蚜对吡虫啉抗药性的风险.结果表明,用吡虫啉等药剂分别选育棉蚜至18代都能够诱导抗性上升,棉蚜对吡虫啉抗性达到10.08倍,对吡虫啉 增效磷的抗性达9.70倍;而用氧乐果和氰戊菊酯分别选育棉蚜至12代时,其抗性分别达28.48倍和35158倍.各抗性品系停止选育12代,抗性下降.     

棉铃虫对久效磷抗性的毒理学机制

就棉铃虫田间种群对久效磷抗性的毒理学机制进行了研究。久效磷对敏感品系和高抗品系的乙酰胆碱酯酶抑制率存在显著差异,在同等剂量浓度下,对敏感品系的抑制率为７３．４８％,对高抗品系的抑制率仅为５９．６７％,说明棉铃虫对久效磷产生高水平抗性与乙酰胆碱酯酶不敏感性有关。在不同田间抗性品系中,ＰＢＯ和ＴＰＰ对久效磷均具有明显的增效作用,增效比分别为５．６～２３．４倍和２．９～４．６倍;而相应地ＢＰＯ和ＴＰＰ对     

棉铃虫Bt抗感种群间数种解毒酶和中肠蛋白酶活性的比较

比较Bt抗性种群和敏感种群棉铃虫解毒酶和中肠蛋白酶的变化，测定两种群的α-乙酸萘酯酶、乙酰胆碱酯酶、谷胱甘肽-S-转移酶的酶活力，比较α-乙酸萘酯酶和乙酰胆碱酯酶的Km和Vmax值，并采用特异性酶抑制剂进行鉴定，结果发现，抗性种群棉铃虫的乙酰胆碱酯酶活力、Km和Vmax值都 显著升高，甲基对氧磷对抗性种群棉铃虫乙酰胆碱酯酶的抑制程度比敏感种群明显霸产加，抗生种群5和敏感种群总蛋白酶活力差异不显著，弱碱性类胰蛋白酶和类凝乳蛋白酶活力差异也不显著，但抗性种群3龄幼虫的强碱性类胰蛋白酶活图形显著高于敏感种群。     

山东省不同地区棉蚜对新烟碱类杀虫剂的抗药性检测及酶抑制剂的增效作用研究

为明确山东省棉蚜对新烟碱类杀虫剂的抗性水平,采用毛细管微量点滴法测定了泰安、聊城和东营3个田间种群及1个敏感种群对吡虫啉、烯啶虫胺、啶虫脒、噻虫嗪、噻虫啉、噻虫胺6种新烟碱类杀虫剂的敏感性,同时测定了磷酸三苯酯(TPP)、顺丁烯二酸二乙酯(DEM)和增效醚(PBO)3种酶抑制剂的增效作用。结果表明:泰安棉蚜种群对烯啶虫胺的抗性倍数为16.95,处于中等抗性水平,对吡虫啉和啶虫脒的抗性倍数分别为5.69和9.57,已产生低水平抗性,对噻虫胺、噻虫嗪和噻虫啉的抗性倍数均小于3.0,仍较敏感;聊城棉蚜种群对吡虫啉、啶虫脒和噻虫嗪的抗性倍数分别为28.51、25.88和18.16,属中等抗性水平,对噻虫啉和噻虫胺的抗性倍数分别为6.01和6.37,已产生低水平抗性,对烯啶虫胺仍处于敏感阶段;东营棉蚜种群对吡虫啉、啶虫脒和噻虫胺的抗性倍数分别为37.95、21.52和12.95,已产生中等水平抗性,对噻虫啉、烯啶虫胺和噻虫嗪的抗性倍数分别为7.07、6.38和4.75,处于低水平抗性阶段。多功能氧化酶抑制剂PBO和羧酸酯酶抑制剂TPP对6种供试新烟碱类杀虫剂的增效作用明显,谷胱甘肽-S-转移酶抑制剂DEM对这6种药剂也具有一定的增效作用。研究表明,山东省泰安等3地区棉蚜种群对6种新烟碱类杀虫剂均产生了不同程度的抗药性,多功能氧化酶和羧酸酯酶可能在棉蚜对该类杀虫剂的抗性中起主要作用,谷胱甘肽-S-转移酶可能也具有一定的作用。     

不同地区灰飞虱对吡虫啉的抗性

为了明确全国不同稻区灰飞虱对吡虫啉的抗药性水平及其抗性机制,2012年采用点滴法测定了16个灰飞虱田间种群对吡虫啉的抗性倍数和3种增效剂在10个抗性种群中对吡虫啉的增效作用。结果表明,安徽安庆、合肥、江苏南京、盐城和无锡5个地区种群达到高抗水平(抗性倍数为74.1~91.2);浙江杭州、台州、湖州3个种群达到中等水平抗性(抗性倍数为35.8~47.7),福建福州和广东广州达到低水平抗性(抗性倍数为18.4~21.3),其他地区种群处于敏感性下降阶段。增效剂增效结果表明,氧化胡椒基丁醚(PBO)和磷酸三苯酯(TPP)在5个高抗种群和2个低抗种群中对吡虫啉均具有增效作用,表明P450单加氧酶和羧酸酯酶是抗性产生的主要代谢机制,而3个中抗种群中,仅PBO对吡虫啉表现出显著的增效作用,说明这3个种群仅有P450单加氧酶是其抗性上升的主要代谢机制。顺丁烯二酸二乙酯(DEM)在测定的10个种群中对吡虫啉均没有增效作用,说明谷胱甘肽S-转移酶没有参与其抗性上升。建议在中等水平至高水平抗性地区暂停使用吡虫啉防治灰飞虱。     

Selection for imidacloprid resistance in Nilaparvata lugens: cross-resistance patterns and possible mechanisms

A field population of brown planthoppers (Nilaparvata lugens St?l) was collected and selected for imidacloprid resistance in the laboratory. The resistance increased by 11.35 times in 25 generations and the resistance ratio reached 72.83 compared with a laboratory susceptible strain. The selected resistant strain showed obvious cross-resistance to all the acetylcholine receptor targeting insecticides tested (monosultap 1.44-fold, acetamiprid 1.61-fold, imidacloprid homologues JS599 2.46-fold and JS598 3.17-fold), but not to others. Further study demonstrated that TPP and DEM had no synergism on imidacloprid. However, PBO displayed significant synergism in some different strains, and the synergism increased with resistance (S strain 1.20, field population 1.43 and R strain 2.93). PBO synergism to cross-resistant insecticides was also found in the resistant strain (monosultap 1.25, acetamiprid 1.39, JS598 1.94 and JS599 2.02). We concluded that esterase and glutathione S-transferase play little role in imidacloprid detoxification. The increase of the P450-monooxygenases detoxification is an important mechanism for imidacloprid resistance and target resistance may also exist in this species.     

苹果绵蚜田间种群的抗性监测

为了掌握果园重要害虫苹果绵蚜Eriosoma lanigerum的抗药性水平及动态变化,于2012–2015年间利用浸叶法监测了中国新疆察布查尔、山东济南、陕西乾县、河北昌黎和江苏丰县苹果绵蚜对常用药剂吡虫啉、毒死蜱和高效氯氟氰菊酯的敏感性,同时测定了不同苹果绵蚜种群的酯酶和谷胱甘肽S-转移酶活力。以2012年察布查尔苹果绵蚜为相对敏感品系,结果发现:所有种群在不同年度均对吡虫啉产生了低到高水平抗性（最大抗性倍数 >2000倍）;对于高效氯氟氰菊酯,昌黎、乾县及察布查尔2014种群均为敏感,济南、丰县和察布查尔2015种群均产生了中到高水平抗性;对于毒死蜱,乾县种群保持敏感,察布查尔、昌黎、丰县和济南种群产生了低到高水平抗性。酶活力测定结果显示:谷胱甘肽S-转移酶活力与苹果绵蚜对吡虫啉和高效氯氟氰菊酯的抗性水平呈正相关性,而酯酶活力与苹果绵蚜抗药性水平无明显相关性。本研究表明,新疆察布查尔、山东济南、陕西乾县、河北昌黎和江苏丰县的苹果绵蚜对吡虫啉、毒死蜱和高效氯氟氰菊酯的抗性水平均呈逐年提高趋势,其中对吡虫啉的抗性程度最严重,防治中应注意合理轮换用药。     

4种棉蚜抗药性种群的生命力及繁殖力

用4种杀虫剂以浸渍法分别累代处理敏感棉蚜,以选育抗药性。每选育4代进行1次毒力测定及生命力和繁殖力的观察。选育至12代,棉蚜对溴氰菊酯的抗性倍数达4031.4倍,对久效磷、灭多威及硫丹的抗性倍数分别仅为6.3、6.1及4.0倍。棉蚜对溴氰菊酯抗性水平较低时,其生命力及繁殖力略低于敏感品系,随着抗性水平的提高,其生命力和繁殖力也随之增强,RF_(8)接近敏感品系的水平,RF_(12)及RF_(16)则明显超过。灭多威及硫丹抗性种群随抗性水平的提高,其生命力及繁殖力表现不同程度的降低。久效磷抗性种群,各抗性水平下其生命力和繁殖力与敏感品系相比无显著差异。     

Triazophos resistance mechanisms in the rice stem borer (Chilo suppressalis Walker)

A field population of the rice stem borer (Chilo suppressalis Walker) with 203.3-fold resistance to triazophos was collected. After 8-generation of continuous selection with triazophos in laboratory, resistance increased to 787.2-fold, and at the same time, the resistance to isocarbophos and methamidophos was also enhanced by 1.9- and 1.4-fold, respectively, implying some cross-resistance between triazophos and these two organophosphate insecticides. Resistance to abamectin was slightly enhanced by triazophos selection, and fipronil and methomyl decreased. Synergism experiments in vivo with TPP, PBO, and DEM were performed to gain a potential indication of roles of detoxicating enzymes in triazophos resistance. The synergism results revealed that TPP (SR, 1.92) and PBO (SR 1.63) had significant synergistic effects on triazophos in resistant rice borers. While DEM (SR 0.83) showed no effects. Assays of enzyme activity in vitro demonstrated that the resistant strain had higher activity of esterase and microsomal O-demethylase than the susceptible strain (1.20- and 1.30-fold, respectively). For glutathione S-transferase activity, no difference was found between the resistant and the susceptible strain when DCNB was used as substrate. However, 1.28-fold higher activity was observed in the resistant strain when CDNB was used. These results showed that esterase and microsomal-O-demethylase play some roles in the resistance. Some iso-enzyme of glutathione S-transferase may involve in the resistance to other insecticides, for this resistant strain was selected from a field population with multiple resistance background. Acetylcholinesterase as the triazophos target was also compared. The results revealed significant differences between the resistant and susceptible strain. The V_max and K_m of the enzyme in resistant strain was only 32 and 65% that in the susceptible strain, respectively. Inhibition tests in vitro showed that I₅₀ of triazophos on AChE of the resistant strain was 2.52-fold higher. Therefore, insensitive AChE may also involved in triazophos resistance mechanism of rice stem borer.     

Cross-resistance ofAphis gossypii to selected insecticides on cotton and cucumber

Cross-resistance of a fenvalerate-resistant (R-fenvalerate) strain and an imidacloprid-resistant (R-imidacloprid) strain of the cotton aphidAphis gossypii Glover to methomyl, endosulfan and omethoate was determined on cotton and cucumber in the laboratory. The R-fenvalerate strain had been selected for 16 consecutive generations on cotton, and the R-imidacloprid strain for 12. Compared with the susceptible (S) strains, the resistance in the 17th generation of the R-fenvalerate strain to fenvalerate was 29,035-fold on cotton and 700-fold on cucumber, whereas the resistance in the 13th generation of the R-imidacioprid strain to imidacloprid was 4.7-fold on cotton and 3.6-fold on cucumber. The R-fenvalerate strain did not show any cross-resistance to imidacloprid, methomyl or omethoate on cotton and cucumber, but became slightly cross-resistant to endosulfan (RR [resistance ratio] = 5.8 and 7.4 on cotton and cucumber, respectively). On cotton the R-imidacloprid strain became highly cross-resistant to fenvalerate (RR = 108.9) and slightly cross-resistant to methomyl, endosulfan and omethoate (RR = 6.3, 3.0 and 8.5, respectively). In contrast, on cucumber the R-imidacloprid strain became highly cross-resistant to fenvalerate (RR = 33.5) and slightly cross-resistant to methomyl and omethoate (RR = 7.0 and 3.3, respectively), but did not show any cross-resistance to endosulfan. It appears thatA. gossypii of both resistant strains was more resistant to the tested insecticides on cotton than on cucumber, except for imidacloprid —where the resistance on cotton was not significantly different from that on cucumber.     

Toxicological and biochemical characterization of coumaphos resistance in the San Roman strain of Boophilus microplus (Acari: Ixodidae)

The San Roman strain of the southern cattle tick, Boophilus microplus, collected from Mexico was previously reported to have a high level of resistance to the organophosphate acaricide coumaphos. An oxidative detoxification mechanism was suspected to contribute to coumaphos resistance in this tick strain, as coumaphos bioassay with piperonyl butoxide (PBO) on larvae of this resistant strain resulted in enhanced coumaphos toxicity, while coumaphos assays with PBO resulted in reduced toxicity of coumaphos in a susceptible reference strain. In this study, we further analyzed the mechanism of oxidative metabolic detoxification with synergist bioassays of coroxon, the toxic metabolite of coumaphos, and the mechanism of target-site insensitivity with acetylcholinesterase (AChE) inhibition kinetics assays. Bioassays of coroxon with PBO resulted in synergism of coroxon toxicity in both the San Roman and the susceptible reference strains. The synergism ratio of PBO on coroxon in the resistant strain was 4.5 times that of the susceptible strain. The results suggested that the cytP450-based metabolic detoxification existed in both resistant and susceptible strains, but its activity was significantly enhanced in the resistant strain. Comparisons of AChE activity and inhibition kinetics by coroxon in both susceptible and resistant strains revealed that the resistant San Roman strain had an insensitive AChE, with a reduced phosphorylation rate, resulting in a reduced bimolecular reaction constant. These data indicate a mechanism of coumaphos resistance in the San Roman strain that involves both insensitive AChE and enhanced cytP450-based metabolic detoxification.     

Synergism of insecticides provides evidence of metabolic mechanisms of resistance in the obliquebanded leafroller Choristoneura rosaceana (Lepidoptera: Tortricidae)

The interactions between six insecticides (indoxacarb, cypermethrin, chlorpyrifos, azinphosmethyl, tebufenozide and chlorfenapyr) and three potential synergists, (piperonyl butoxide (PBO), S,S,S-tributyl phosphorotrithioate (DEF) and diethyl maleate (DEM)) were studied by dietary exposure in a multi-resistant and a susceptible strain of the obliquebanded leafroller, Choristoneura rosaceana (Harris). The synergists did not produce appreciable synergism with most of the insecticides in the susceptible strain. Except for tebufenozide, PBO synergized all the insecticides to varying degrees in the resistant strain. A very high level of synergism by PBO was found with indoxacarb, which reduced the resistance level from 705- to 20-fold when PBO was administered alone and to around 10-fold when used in combination with DEF. DEF also synergized indoxacarb, cypermethrin, chlorpyrifos, azinphosmethyl and tebufenozide in the resistant strain. DEM produced synergism of indoxacarb, chlorpyrifos, azinphos-methyl and chlorfenapyr in the resistant strain. DEM was highly synergistic to cypermethrin, and to some extent to tebufenozide in both the susceptible and resistant strains equally, implying that detoxification by glutathione S-transferases was not a mechanism of resistance for these insecticides. The high level of synergism seen with DEM in the case of cypermethrin may be due to an increase in oxidative stress resulting from the removal of the antioxidant, glutathione. These studies indicate that enhanced detoxification, often mediated by cytochrome P-450 monooxygenases, but with probable esterase and glutathione S-transferase contributions in some cases, is the major mechanism imparting resistance to different insecticides in C. rosaceana.     

Variation of acephate susceptibility and correlation with esterase and glutathione S-transferase activities in field populations of the tarnished plant bug, Lygus lineolaris

The tarnished plant bug (TPB) has increasingly become an economically important pest of cotton. Heavy dependence on insecticides, particularly organophosphates and pyrethroids, for TPB control facilitated resistance development to multiple classes of insecticides. To better understand resistance and explore ways to monitor resistance in field populations, this study examined acephate susceptibility and the activities of two major detoxification enzymes in nine field populations collected in the Delta region of Mississippi and Arkansas in 2010. Two Arkansas populations from Reed and Backgate had 3.5- and 4.3-fold resistance to acephate, as compared to a susceptible laboratory strain. Extensive planting of cotton and heavy chemical sprays is a major driving force for resistance development to acephate in Mid-south cotton growing areas. Reduced susceptibility to acephate was highly correlated with elevated esterase activities. The acephate-resistant populations from Backgate, Lula, and Reed consistently had higher (up to 5.3-fold) esterase activities than susceptible populations. Regression analysis of LC₅₀s with kinetic esterase activities revealed a significant polynomial quadratic relationship with R² up to 0.89. Glutathione S-transferase (GST) also had elevated activity in most populations, but the variations of GST activities were not significantly correlated with changes of acephate susceptibility. Finally, examination of esterase and GST inhibitors indicated that suppression rates (up to 70%) by two esterase inhibitors in 2010 were slightly lower than those detected in 2006, and ethacrynic acid (EA) inhibited GST effectively in both years. Two other GST inhibitors (sulfobromophthalein and diethyl maleate) displayed significantly lower suppression rates in 2010 than those detected in 2006, suggesting a potential genetic shift in pest populations and a necessity of continued monitoring for insecticide resistance with both bioassay and biochemical approaches. Results indicated that using major detoxification enzyme activities for resistance monitoring may provide insight into acephate resistance in field populations of TPB.     

Pyrethroid synergists suppress esterase-mediated resistance in Indian strains of the cotton bollworm, Helicoverpa armigera (Hübner)

The role of esterase in pyrethroid resistance was studied in the final larval instar of different strains of the cotton bollworm, Helicoverpa armigera. The resistant strains viz., Nagpur strain and the Delhi strain were found to have elevated midgut esterase activity in comparison to the susceptible strain. Nagpur strain and Delhi strain have 2.24 and 1.73-fold higher esterase activity, respectively, than that of the susceptible strain. The Native PAGE displayed important differences in the midgut esterase isozyme pattern between the susceptible and the pyrethroid-resistant strains. Out of the 10 esterase isozyme observed, susceptible strain lacked three bands, E2, E6 and E10 that were found in the resistant strains. The potency of the synergists piperonyl butoxide (PBO) and dihydrodillapiole (DDA) as esterase inhibitor were also studied both in vitro and in vivo. The in vitro results clearly show that both PBO and DDA inhibited esterase activity in the two resistant strains, while there was almost no esterase inhibition in the homogenate of the susceptible strain. The in vivo inhibition studies (topical application of PBO and DDA followed by biochemical analysis) illustrated that PBO- and DDA-esterase binding is rather slow and non permanent process. Esterase inhibition did not occur immediately after the synergist treatment but at 4 and 8 h post treatment in case of PBO and DDA, respectively. Native PAGE revealed that the in vivo esterase inhibition caused by both PBO and DDA was due to the binding of the synergist with the E6 isozyme which was not present in the susceptible strain.