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四种蚜虫AChE的活性及其对抑制剂的敏感度 总被引:3,自引:0,他引:3
<正> 乙酰胆碱酯酶AChE是有机磷和氨基甲酸酯杀虫剂的靶标酶。测定单个昆虫AChE的活性及其对抑制剂的敏感性,对于种群AChE变构的抗性遗传学的研究以及抗性昆虫个体的生物化学检测都具有重要意义。作者参照 相似文献
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滤纸法测定有机磷杀虫剂对棉蚜酯酶的抑制频率(英) 总被引:5,自引:0,他引:5
滤纸法(FPT)测试单个棉蚜酯酶活性可以检测棉蚜对有机磷杀虫剂的抗性。为准确测试不同杀虫剂的抗性水平,本文改进了FPT的方法,测定了甲基对硫磷、久效磷和氧乐果对棉蚜个体α-NA酯酶的抑制作用,比较了BCA(敏感)和GCA(抗性)种群的抑制频率。结果指出二种不同浓度的杀虫剂对BCA种群的抑制频率明显高于GCA种群。特别在测定久效磷对经久效磷处理后棉蚜F_2代个体的抑制率,仅有一个棉蚜F_1代的抑制率达19%,其他均无抑制作用,而对未经处理的棉蚜F_1代个体的抑制率均可达70%-90%,但在α-NA酯酶活性上差异不明显。测定同地区三种有机磷杀虫剂对已喷过杀虫剂的棉蚜种群,其个体酯酶抑制频率明显低于未喷杀虫剂的棉蚜种群。这些均表明用FPT测定有机磷杀虫剂对棉蚜个体α-NA酯酶的抑制频率可作为抗性测报技术。 相似文献
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小菜蛾抗性个体不敏感乙酰胆碱酯酶的鉴定 总被引:14,自引:0,他引:14
乙酰胆碱酯酶(AChE)敏感性降低是小菜蛾对有机磷和氨基甲酸酯产生抗性的重要机制之一,已得到广泛的承认和报道。一种用硝酸纤维素膜的斑点法鉴定个体小菜蛾的抗性AChE不敏感性的应用技术,提供了早期侦测和随后监测田间种群抗性的可能性。此法操作简便灵敏。小菜蛾抗性品系(GBR)和田间种群成虫头部AChE活力,在残杀威抑制时,抑制率分别为50.97%和43.96%,有对氧磷存在时,分别为63.78%和35.87%,较敏感品系的AChE为不敏感。 相似文献
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为了探讨烟粉虱Bemisia tabaci不同种群个体乙酰胆碱酯酶敏感性差异及其与抗药性的关系, 我们选用室内饲养的烟粉虱SUD S敏感品系和6个田间抗性种群, 采用酶标板酶动力学法测定了各品系 (种群)乙酰胆碱酯酶对抑制剂的敏感性反应以及抑制剂存在时各抗性种群个体乙酰胆碱酯酶残余活性频率分布。结果表明: 在抑制剂浓度为300 μmol/L时, 敏感品系乙酰胆碱酯酶的活性基本上被完全抑制, 可以明显地区分敏感品系与田间抗性种群。在抑制剂浓度为2 000 μmol/L时, 各抗性种群个体乙酰胆碱酯酶残余活性频率分布差异明显, 其中ZZ-R种群和FZ-R种群的乙酰胆碱酯酶残余活性频率分布相似, 大部分个体的乙酰胆碱酯酶残余活性分布在1.00~1.80 mOD/min之间; SM-R种群和ND-R种群的乙酰胆碱酯酶残余活性频率分布也相似, 大部分个体的乙酰胆碱酯酶残余活性分布在0.40~1.00 mOD/min之间; LY-R和NP-R种群大部分个体的乙酰胆碱酯酶残余活性分别分布在1.00~1.60 mOD/min和0.80~1.20 mOD/min之间。各抗性种群乙酰胆碱酯酶高残余活性 (大于1.00 mOD/min)个体频率与对敌敌畏的抗性水平之间具有明显相关性, 相关系数为0.86 (P<0.05)。考虑到乙酰胆碱酯酶对抑制剂作用不敏感是一些昆虫对有机磷和氨基甲酸酯类杀虫剂抗性的重要机制之一, 建议可以将乙酰胆碱酯酶对敌敌畏的敏感性作为烟粉虱抗药性生化检测的一个参考指标。 相似文献
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菜蛾绒茧蜂和小菜蛾对杀虫剂的敏感性及酶学特性的比较研究 总被引:9,自引:2,他引:7
分别采用药膜法和浸叶法测定了菜蛾绒茧蜂Apanteles plutellae和小菜蛾Plutella xylostella对杀虫剂的敏感度。结果显示: 有机磷、氨基甲酸酯、拟除虫菊酯类杀虫剂、阿维菌素和锐劲特对菜蛾绒茧蜂高毒,而抑太保和Bt为低毒,然而,短时间(1 h)接触常规防治剂量的锐劲特、氰戊菊酯、氯氰菊酯和乙酰甲胺磷对菜蛾绒茧蜂低毒。增效剂胡椒基丁醚(PB)、磷酸三苯酯(TPP)和马来酸二乙酯(DEM)对菜蛾绒茧蜂的甲胺磷、克百威、氰戊菊酯、氯氰菊酯、阿维菌素和锐劲特敏感性增效显著,但对抑太保无增效作用。PB的增效作用显著高于TPP 和DEM。PB和TPP对菜蛾绒茧蜂羧酸酯酶(CarE),以及DEM对谷胱甘肽S转移酶(GST)具显著的活体抑制作用,但PB,TPP和 DEM对菜蛾绒茧蜂乙酰胆碱酯酶(AChE)无抑制作用。菜蛾绒茧蜂AChE的米氏常数(Km)、最大反应速度(Vmax)、CarE和GST活性分别为小菜蛾的0.22、2.08、4.60和0.45倍,甲胺磷、敌敌畏和克百威对菜蛾绒茧蜂AChE的双分子速度常数(Ki)分别为对小菜蛾的14.7、10.5 和26.0倍。酶与抑制剂反应温度增高将导致酶抑制率增高,尤其对菜蛾绒茧蜂AChE的抑制作用更为显著。上述结果表明,菜蛾绒茧蜂对有机磷和氨基甲酸酯类杀虫剂的高敏感性与其显著高的AChE敏感性有关,氧化代谢的解毒作用对菜蛾绒茧蜂耐药性的影响大于水解作用。此外,对小菜蛾和菜蛾绒茧蜂杀虫剂敏感性差异的毒理学原因进行了讨论。 相似文献
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小峰熊蜂头部乙酰胆碱酯酶测定条件的优化及其对六种常用杀虫剂的敏感性 总被引:1,自引:0,他引:1
小峰熊蜂Bombus hypocrita是我国优势熊蜂种群之一, 因其易于饲养、 群势较强且授粉性能优良而成为我国设施农业常用优良授粉蜂种, 但常受到以乙酰胆碱酯酶(AChE)为靶标酶的有机磷和氨基甲酸酯类杀虫剂的危害。 为合理规避这两类杀虫剂对熊蜂的危害, 同时也为完善熊蜂授粉配套技术和保护野生熊蜂资源提供理论基础, 本研究利用正交试验对小峰熊蜂头部乙酰胆碱酯酶活性的测定条件进行了优化, 并明确了6种常用有机磷和氨基甲酸酯类杀虫剂对乙酰胆碱酯酶活性的影响。结果表明: 各测定因素对小峰熊蜂乙酰胆碱酯酶活性测定影响的大小顺序依次为: 酶浓度>pH>温度>底物浓度>反应时间; 小峰熊蜂头部乙酰胆碱酯酶活性的最适反应条件为: 酶浓度0.25 g 蛋白质/L, 底物浓度0.8 mmol/L, pH值7.5, 温度40℃, 反应时间5 min。毒死蜱、 三唑磷、 丙溴磷、 异丙威、 仲丁威和残杀威6种杀虫剂对小峰熊蜂头部乙酰胆碱酯酶离体抑制作用均呈现明显的剂量-效应关系, 其抑制中浓度IC50分别为0.39, 1.79, 0.42, 0.04, 0.43和0.63 mmol/L。这6种杀虫剂对小峰熊蜂AChE抑制作用的强弱依次为: 异丙威>毒死蜱>三唑磷>仲丁威>残杀威>丙溴磷, 即小峰熊蜂对异丙威最敏感, 而对丙溴磷的敏感性最弱。 相似文献
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乙酰胆碱酯酶(Acetylcholinesterase,ACh E;EC3.1.1.7)作为有机磷和氨基甲酸酯类杀虫剂的作用靶标,一直是昆虫毒理学领域研究的热点。本研究采用微量滴度酶标板法研究了褐色橘蚜Toxoptera citricida不同发育阶段ACh E的生化特性及个体活性频率,离体条件下分析了3种药剂对ACh E的影响。结果表明,无翅成蚜对底物碘化硫代乙酰胆碱亲和力显著高于若蚜;就个体活性分布频率而言,25%-30%的无翅成蚜分布在9.5-10.5 nmol/(min·insect)区间,而50%-55%的若蚜分布在2-3 nmol/(min·insect)区间;离体条件下,毒扁豆碱、残杀威、异丙威等3种杀虫剂对褐色橘蚜ACh E有显著的抑制作用,与成蚜相比,若蚜ACh E对3种药剂更敏感,而毒扁豆碱对褐色橘蚜无翅成蚜与若蚜ACh E的抑制作用最好。本研究表明褐色橘蚜成蚜与若蚜生化特性及对药剂的敏感性存在显著差异。 相似文献
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为探索瓜实蝇Zeugodacus cucurbitae不同寄主种群对杀虫剂的敏感性,本研究在室内通过药膜法测定了瓜实蝇节瓜、黄瓜、丝瓜和苦瓜种群4日龄成虫对4种杀虫剂的敏感性,随后将瓜实蝇不同寄主种群致死中浓度LC50进行比较,计算抗性倍数。结果表明,4个寄主种群对甲维盐的敏感性为节瓜 > 苦瓜 > 丝瓜 > 黄瓜,其中节瓜、苦瓜种群对甲维盐最敏感,其次是丝瓜、黄瓜,LC50值分别为0.250、0.391、0.809和1.035 mg/L。4个寄主种群对灭多威的敏感性表现为节瓜 > 黄瓜 > 苦瓜 > 丝瓜,LC50值分别为0.302、0.318、0.652和0.804 mg/L。对多杀菌素的敏感性大小依次为黄瓜、节瓜、丝瓜、苦瓜,LC50值分别为1.157、1.198、1.232和2.029 mg/L。对啶虫脒的敏感性为苦瓜 > 节瓜 > 黄瓜 > 丝瓜,LC50值分别为17.946、20.166、20.190和21.986 mg/L,其中对甲维盐的敏感性差异幅度最大,为4.140倍,其次是灭多威,为2.654倍,对啶虫脒的敏感性差异幅度最小,为1.225倍。表明寄主植物可引起瓜实蝇对杀虫剂的敏感性变化。 相似文献
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The Asian citrus psyllid, Diaphorina citri Kuwayama, is a worldwide pest of citrus, which vectors the putative causal pathogen of huanglongbing. Current management practices warrant continuous monitoring of field populations for insecticide resistance. Baseline activities of acetylcholinesterase (AChE), general esterase, and glutathione S-transferase as well as sensitivity of AChE to selected organophosphate and carbamate insecticides were established for a susceptible laboratory strain (Lab) and compared with several field populations of D. citri from Florida. The specific activity of AChE in various D. citri populations ranged from 0.77 to 1.29 microM min(-1) mg of protein(-1); the Lab strain was characterized by the highest activity. Although reduced AChE sensitivity was observed in the Lab strain compared with field populations, overlap of 95% confidence intervals of I50 values (concentration required for 50% AChE activity inhibition) suggests no significant difference in AChE sensitivity among all populations tested for a given insecticide. There was no significant evidence of target site insensitivity in field populations that were exposed to the selected organophosphate and carbamate insecticides tested. The specific activity of general esterase and glutathione S-transferase was lowest in the Lab strain and was generally comparable to that of the field populations evaluated. The current data provide a mode-of-action specific baseline for future monitoring of resistance to organophosphate and carbamate insecticides in populations of D. citri. 相似文献
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USING FILTER PAPER TEST FOR DETECTING INHIBITORY FREQUENCY OF ORGANOPHOSPHATE TO ESTERASES IN COTTON APHIDS, APHIS GOSSYPII (GLOVER) 总被引:1,自引:0,他引:1
Abstract The individual esterase activity which is measured by filter paper test (FPT) method may determine the resistance of cotton aphids Aphis gossypii (Glover) against organophosphorus (OP) insecticides. For testing accurately resistant level caused by different insecticides, we applied FPT method for measuring inhibitory action of methyl-parathion, monocrotophos and omethoate to α-NA esterase of individual cotton aphids, and compared the inhibitory frequencies of these three insecticides to susceptible population (BCA) and resistant population (GCA). Results showed that their inhibitory frequencies of the susceptible population were evidently higher than that of the resistant population. The inhibitory rate of α-NA esterase in F1 generation individual cotton aphids by monocrotophos was low when the cotton aphid population had been treated in advance with monocrotophos, but it got to 75%-90% when the cotton aphid population had been untreated in advance with monocrotophos. Besides, the differences in esterase activity were not obvious between them. In same region when cotton aphids were treated with insecticides the inhibitory frequency of esterases in individual by the insecticides was lower than counterparts in individual cotton aphids which were not treated with insecticides. All these demonstrated that inhibitory frequency of α-NA esterase in individual cotton aphids by OP insecticides could be used as a technique of forecasting pest resistance. 相似文献
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Fuentes-Contreras E Figueroa CC Reyes M Briones LM Niemeyer HM 《Bulletin of entomological research》2004,94(1):11-18
The tobacco-feeding race of Myzus persicae (Sulzer), formerly known as M. nicotianae Blackman, was introduced into Chile during the last decade. In order to evaluate the genetic diversity and insecticide resistance status of Chilean tobacco aphid populations, a field survey was conducted in 35 tobacco fields covering a 300 km latitudinal survey. The populations sampled were characterized using microsatellite markers and morphometric multivariate analysis. Insecticide resistance levels were assessed through a microplate esterase assay and the mutation status of the kdr gene. All samples collected corresponded to the same anholocyclic aphid genotype, and showed morphological variation within the range expected for the tobacco-feeding race of M. persicae. Esterase activity showed the level and variability expected for an R1 clone lacking mutations in the sodium channels (susceptible kdr), thus corresponding to a type slightly resistant to organophosphate and carbamate, and susceptible to pyrethroid insecticides. 相似文献
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Abstract The cotton bollworms, Helicoverpa armigera Hübner, collected from Handan of Hebei Province, have evolved high resistance to pyrethroid, organophosphate and carbamate insecticides, The sensitivity of acetylcholinesterase (AChEs) to paraoxon and methomyl varied with the development stages of the cotton bollworm. After the treatments with LD5 and LD50 of parathion and methomyl to the cotton bollworms, the affinity of AChE to acetylthiocholine (ATCh) and acetyl-β-bmethyl-thio choline (MeTCh) increased significantly except the treatment of parathion using LD50 dosage while the sensitivity of AChEs to paraoxon significantly decreased. The sensitivity of AChEs to methomyl strongly increased in the treatment of parathion using LD50 dosages while strongly decreased in other treatments. The affinity of carboxylesterase to β-naphthyl acetate (β-NA) was higher in groups of treatment with insecticides than in group of control. The glutathione-s-transferase (GST) activity significantly decreased in the induced groups using LD5 dosages, while increased in the selection groups using LD50 dosages. The effects of parathion and methomyl on the phosphatases of cotton bollworm were related to the dosages of application and the time after treatment and the effect on the alkaline phosphatase was stronger than on acid phosphatase. 相似文献
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Jahangir Khajehali Thomas Van Leeuwen Luc Tirry 《Experimental & applied acarology》2009,49(3):185-192
Bifenazate, a new and frequently used carbazate, is a pro-acaricide which needs to be activated by carboxylesterases. We evaluated
the possible antagonism of organophosphate and carbamate insecticides on bifenazate toxicity in Tetranychus urticae applied in mixtures. Two organophosphate resistant strains were used (WI and MR-VL) and several organophosphate (chlorpyrifos,
azinphosmethyl and phosmet) and carbamate (carbaryl and methomyl) insecticides were evaluated. Mixing chlorpyrifos with bifenazate
decreased bifenazate toxicity in both tested strains. However, in the strain with a higher esterase activity, antagonism decreased
after 2 days. Of all other tested chemicals, only methomyl displayed an antagonistic effect 1 day after treatment. These findings
indicate that mixing organophosphate and carbamate insecticides with bifenazate may inhibit bifenazate efficacy under field
conditions, especially when resistant strains are present. 相似文献
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Mutations in acetylcholinesterase genes of Rhopalosiphum padi resistant to organophosphate and carbamate insecticides. 总被引:1,自引:0,他引:1
Apple grain aphid, Rhopalosiphum padi (Linnaeus), is an important wheat pest. In China, it has been reported that R. padi has developed high resistance to carbamate and organophosphate insecticides. Previous work cloned from this aphid 2 different genes encoding acetylcholinesterase (AChE), which is the target enzyme for carbamate and organophosphate insecticides, and its insensitive alteration has been proven to be an important mechanism for insecticide resistance in other insects. In this study, both resistant and susceptible strains of R, padi were developed, and their AChEs were compared to determine whether resistance resulted from this mechanism and whether these 2 genes both play a role in resistance. Bioassays showed that the resistant strain used was highly or moderately resistant to pirimicarb, omethoate, and monocrotophos (resistance ratio, 263.8, 53.8, and 17.5, respectively), and showed little resistance to deltamethrin or thiodicarb (resistance ratio, 5.2 and 3.4, respectively). Correspondingly, biochemistry analysis found that AChE from resistant aphids was very insensitive to the first 3 insecticides (I50 increased 43.0-, 15.2-, and 8.8-fold, respectively), but not to thiodicarb (I50 increased 1.1-fold). Enzyme kinetics tests showed that resistant and susceptible strains had different AChEs. Sequence analysis of the 2 AChE genes cloned from resistant and susceptible aphids revealed that 2 mutations in Ace2 and 1 in Ace1 were consistently associated with resistance. Mutation F368(290)L in Ace2 localized at the same position as a previously proven resistance mutation site in other insects. The other 2 mutations, S329(228)P in Ace1 and V435(356)A in Ace2, were also found to affect the enzyme structure. These findings indicate that resistance in this aphid is mainly the result of insensistive AChE alteration, that the 3 mutations found might contribute to resistance, and that the AChEs encoded by both genes could serve as targets of insecticides. 相似文献