Oxydemeton-methyl resistance, mechanisms, and associated fitness cost in green peach aphids (Hemiptera: Aphididae)

Susceptibility to oxydemeton-methyl and imidacloprid, and the inhibitory effects of oxydemeton-methyl and some organophosphate compounds on acetylcholinesterase (AChE) and carboxylesterase activity were studied in two populations (Karaj and Rasht) of green peach aphids, Myzus persicae (Sulzer). Results show that the Karaj population was resistant to oxydemeton-methyl but susceptible to imidacloprid. The esterase activity of the resistant and susceptible populations suggests that one of the resistance mechanisms to oxydemeton-methyl was esterase-based. The inhibition assay shows that the AChE of the Karaj population is less sensitive to oxydemeton-methyl and paraoxon derivatives. Regarding the paraoxon derivatives, the smaller paraoxon side chain is more potent against the modified AChE than against the AChE from the susceptible strain. Fertility life table parameters of green peach aphid populations resistant and susceptible to oxydemeton-methyl also were studied under laboratory conditions. The standard errors of the population growth parameters were calculated using the Jackknife method. Results showed that susceptible strain exhibits a significantly higher r(m) than the resistant strain, probably because the resistant strain had a higher generation time than the susceptible strain. These results suggested that the resistant Karaj strain may be less fit than the susceptible strain.     

Characterization of general esterases from methyl parathion-resistant and -susceptible populations of western corn rootworm (Coleoptera: Chrysomelidae)

A consistent correlation between elevated esterase activity and methyl parathion resistance among Nebraska western corn rootworm, Diabrotica virgifera virgifera LeConte, populations has previously been documented. Characterization of general esterase activity using naphtholic esters as model substrates indicated that differences between resistant and susceptible strains could be maximized by optimizing assay conditions. The optimal conditions identified here were similar to those reported for other insect species. The majority of general esterase activity was found in the cytosolic fractions of resistant populations, whereas the activity was more evenly distributed between cytosolic and mitochondrial/nuclear fractions in the susceptible population. General esterase activity was predominately located in the adult thorax and abdomen. Although there were significant differences in general esterase activities between resistant and susceptible populations, the differences exhibited in single beetle activity assays did not provide sufficient discrimination to identify resistant individuals. In contrast, single larva activity assays provided greater discrimination and could be considered as an alternative to traditional bioassay techniques.     

Enhanced esterase gene expression and activity in a malathion-resistant strain of the tarnished plant bug, Lygus lineolaris

Extensive use of insecticides on cotton in the mid-South has prompted resistance development in the tarnished plant bug, Lygus lineolaris (Palisot de Beauvois). A field population of tarnished plant bugs in Mississippi with 11-fold higher resistance to malathion was used to examine how gene regulation conferred resistance to this organophosphate insecticide. In laboratory bioassays, synergism by the esterase inhibitors S,S,S,-tributylphosphorotrithioate (DEF) and triphenylphosphate (TPP) effectively abolished resistance and increased malathion toxicity by more than 80%. Esterase activities were compared in vitro between malathion susceptible and resistant (selected) strains. More than 6-, 3- and 10-fold higher activities were obtained with the resistant strain using alpha-naphthyl acetate, beta-naphthyl acetate, and p-nitrophenyl acetate, respectively. Up to 95% and 89% of the esterase activity in the susceptible and resistant strains, respectively, was inhibited by 1 mM DEF. Inhibition of esterase activity up to 75% and 85% in the susceptible and resistant strains, respectively, was obtained with 0.03 mM TPP. Esterase activities in field populations increased by up to 5.4-fold during the fall season. The increase was synchronized with movement of the insect into cotton where exposure to pesticides occurred. Esterase cDNA was cloned and sequenced from both malathion susceptible and resistant strains. The 1818-nucleotide cDNA contained a 1710-bp open reading frame coding a 570 amino acid protein which was similar to many insect esterases conferring organophosphate resistance. No amino acid substitution was observed between susceptible and resistant strains, indicating that esterase gene mutation was not involved in resistance development in the resistant strain in Mississippi. Further examination of esterase gene expression levels using quantitative RT-PCR revealed that the resistant strain had a 5.1-fold higher level of esterase mRNA than the susceptible strain. The results of this study indicated that up-regulation of the esterase gene appeared to be related to the development of resistance in the tarnished plant bug.     

Enzymatic diagnosis of resistance to deltamethrin in diapausing larvae of the codling moth,Cydia pomonella (L.)

The resistance to deltamethrin was evaluated in diapausing larvae of 14 field populations of the codling moth (Cydia pomonella L.) From the main French orchard areas using biological assays. Resistance to deltamethrin was compared to mixed-function-oxidase (mfo) activity measured at the individual level through ethoxycoumarin-O-deethylase (ECOD) activity using a fluorescence microplate reader. The larvae were collected in corrugated paper band traps in the autumn of 1995. Analysis was previously performed on two laboratory strains, one susceptible and one resistant to deltamethrin, in order to characterize the changes in resistance during diapause development. Resistance to deltamethrin as well as the ECOD activity were stable during the diapause, and ECOD activity was always significantly lower in the susceptible strain than in the resistant one. The ECOD activity was significantly correlated to the frequency of resistant moths in the field populations. This strongly suggested the involvement of the mfo system in the resistance to deltamethrin in these populations. The intra-strain variabilities in ECOD activity of both laboratory and field resistant insects indicated that other resistance mechanisms might also be involved. Further investigations on these mechanisms are needed in order to develop complete diagnostic methods and to define suitable control strategies against each resistant population. Arch. Insect Biochem. Physiol. 39:55–64, 1998. © 1998 Wiley-Liss, Inc.     

Resistance of Spanish codling moth (Cydia pomonella) populations to insecticides and activity of detoxifying enzymatic systems

Resistance of codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae), to insecticides has become a major problem in many apple and pear production areas. Our aim was to determine the level of insecticide resistance in Spanish field populations. Seven field populations collected from apple, Malus domestica Borkhausen (Rosaceae), orchards, and three laboratory susceptible strains of codling moth were studied. Damage at harvest in all the conventional orchards from which codling moth populations were collected was higher than the economic threshold. The efficacy of eight insecticides, with five modes of action, was evaluated by topical application of the diagnostic concentrations on post‐diapausing larvae. The enzymatic activity of mixed‐function oxidases (MFOs), glutathione transferases (GSTs), and esterases (ESTs) was evaluated for each population. The susceptibility to insecticides and the biochemical activity of the three laboratory strains and one organic orchard population were not significantly different. Field populations were less susceptible to the tested insecticides than the susceptible strains, especially for azinphos‐methyl, diflubenzuron, fenoxycarb, and phosalone. The efficacy of all insecticides was significantly dependent on the activity of MFOs. Only the toxicity of the three insecticides most used in Spain when the populations were collected (azinphos‐methyl, fenoxycarb, and phosalone) was also dependent on the activity of ESTs and GSTs activity. We conclude that the control failures were because of the existence of populations resistant to the main insecticides used.     

Resistance to a nuclear polyhedrosis virus in the light-brown apple moth Epiphyas postvittana (Lepidoptera: Tortricidae)

Measurements were made of the relative susceptibility to a nuclear polyhedrosis virus of three populations of light-brown apple moth Epiphyas postvittana: a resistant laboratory strain (CAN), a susceptible laboratory strain (BAR), and a field population. CAN was found to be 50 times more resistant than BAR and 160 times more resistant than the field line. Experiments on hybrid crosses of resistant and susceptible strains showed that resistance is genetically determined. This serves as a warning of the possible selection of virus-resistant strains of insect pests, where viral insecticides are being used in the field.     

Influence of phosphine resistance genes on flight propensity and resource location in Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae): the landscape for selection

Phosphine resistance in Tribolium castaneum Herbst (Coleoptera: Tenebrionidae) has evolved through changes to enzymes involved in basic metabolic pathways. These changes impose metabolic stress and could affect energy‐demanding behaviours. We therefore tested whether phosphine resistance alleles impact the movement of these insects in their quest for new resources. We measured walking and flight parameters of four T. castaneum genotypes: (1) a field‐derived population, (2) a laboratory cultured, phosphine‐susceptible reference strain, (3) a laboratory cultured, phosphine‐resistant reference strain, and (4) a resistant introgressed strain that is almost identical genetically to the susceptible population. The temporal pattern of flight was identical across all populations, but resistant beetles took flight significantly less, walked more slowly, and located resources less successfully than did susceptible beetles. Also, the field‐derived beetles (proved not to be carrying resistance genes) walked significantly faster and more directly towards food resources, and had a higher propensity for flight when compared to the susceptible laboratory beetles. These negative effects suggest survival of beetles with the resistance alleles will be compromised should they leave phosphine application sites. The field for selection therefore extends beyond the site at which phosphine fumigant imposed its effect, and other mutations are also likely to be affected in this way.     

Biochemical studies on malathion resistance,inheritance and association of carboxylesterase activity in brown planthopper,Nilaparvata lugens complex in Peninsular Malaysia

Abstract Two sympatric populations of brown planthopper (BPH), one from rice and the other from Leersia hexandra were collected from each of five locations in Malaysia. All the tested malathion-resistant individuals of the rice BPH population and F₁ generation (cross between malathion-resistant [usually caught on rice] and malathion-susceptible [usually caught on Leersia]) showed high esterase activity, while all malathion-susceptible individuals on L. hexandra showed low esterase activity. In the F₂ generation, all the individuals tested against malathion were approximately 75% resistant and 25% susceptible and the inheritance pattern of esterase activity (high and low esterase activity) segregated in the same manner to a 3: 1 ratio. This confirms that resistance to malathion is mono-factorial and inheritance pattern of esterase activity is also linked to malathion resistance. Carboxylesterase or total esterase activity in BPH is inherited in a simple Mendelian fashion that is encoded by a single dominant gene. For the total esterase assay, average esterase activity levels in the rice-infesting population ranged from 17.64 to 19.37 nmoles 1-napthol/mg protein while that in the Leersia-infesting population ranged from 5.29 to 6.11 nmoles 1-napthol/mg protein. In terms of esterase activity, the two sympatric Nilaparvata lugens populations separated into two distinct groups. Results based on the tube color intensity test showed 96% and 98% resistant and susceptible individuals were present in the rice- and Leersia-infesting populations, respectively. In a filter paper test, the rice-infesting population had 94% with high esterase activity while the Leersia-infesting population had 96% with low esterase activity.     

新疆棉铃虫对溴氰菊酯和硫丹 抗性的生化机理研究

通过生物测定和生化分析研究了新疆棉铃虫Helicoverpa armigera (Hübner)敏感种群和室内筛选获得的抗性种群对硫丹和溴氰菊酯的反应及其α-乙酸萘酯酶和乙酰胆碱酯酶的动态活性反应。结果表明,筛选后新疆棉铃虫对硫丹和溴氰菊酯产生的抗性倍数分别为13倍和66倍。两个抗性种群的α-乙酸萘酯酶和乙酰胆碱酯酶比活力均高于敏感种群。相应杀虫剂预处理后,α-乙酸萘酯酶酶活力受到抑制。抗性种群的α-乙酸萘酯酶对底物的亲和力高于敏感种群,但Vmax低于敏感种群。抗性种群的乙酰胆碱酯酶对底物的亲和力显著低于敏感种群,Vmax比敏感种群高。聚丙烯酰胺凝胶电泳显示,两个抗性种群都有一条特异性酶带,其迁移率相近,且均可被甲基对氧磷抑制。因此推测,α-乙酸萘酯酶参与了新疆棉铃虫对硫丹和溴氰菊酯的抗性,具有代谢和阻断作用;乙酰胆碱酯酶对抗性的产生也起到了重要作用。     

Assessment of the potential for and development of organophosphorus resistance in field populations of Myzus persicae

A survey of esterases in field populations of the peach-potato aphid, Myzus persicae was made during the spring of 1975. Assay was by electrophoresis of single aphid homogenates, and the known association between the activity of an esterase and resistance to organophosphorus insecticide (OP) was used to infer resistance in field populations. The resistant variant replaced the susceptible in populations which had been treated with OP and another variant with threefold (approximately) more esterase activity appeared to be replacing the resistant variant in populations which have been treated twice with OP. The significance of this for control of M. persicae is discussed. Differences in resistance between aphids in different parts of the same field, and the widespread association of these esterase variants in favoured combinations with two electrophoretic variants at another locus have also been investigated.     

东亚飞蝗山西两地理种群酯酶特性的比较研究

对东亚飞蝗山西临猗和永济2个地理种群的酯酶特性进行了比较研究.非变性聚丙烯酰胺凝胶电泳图谱显示:以a-乙酸萘酯为底物染色,2个东亚飞蝗种群谱带差别不明显.但是,酯酶动力学研究结果表明:以a-乙酸萘酯和α-丁酸萘酯为底物时,永济种群的酯酶活性分别是临猗种群的1.81倍和1.20倍.永济种群酯酶活性的增高可能与非变性聚丙烯酰胺凝胶电泳图谱显示出较临猗种群多出的酶带有关.体外酯酶抑制动力学研究表明:永济和临猗2种群所含酯酶大都为B型酯酶,其含量分别为84.94%和91.47%.永济种群对对氧磷的耐受性要高于临猗种群,我们推测可能与2种群马拉硫磷使用背景不同有关.     

不同种群棉铃虫三龄幼虫酯酶活性频率分布与抗药性的关系

选用室内饲养的棉铃虫Helicoverpa armigera偃师和湖北2个敏感品系、对辛硫磷高抗的PCP20品系、对氰戊菊酯高抗的YG45品系及1999或2000年采自山东阳谷、河北邯郸和河南安阳的田间高抗种群,江苏徐州、湖北武汉的田间中等抗性水平种群和新疆沙湾的田间敏感种群,采用酶标板酶动力学法测定了各品系（种群）的3龄幼虫个体酯酶活性频率分布和平均酯酶活性。结果表明,偃师敏感品系、湖北敏感品系和新疆沙湾田间敏感种群的酯酶活性个体频率分布相似,三个品系（种群）的平均酯酶活性相近,分别为991、1 138、1 055 mOD·min^-1·larva^-1。室内选育的PCP20抗性品系、YG45抗性品系及山东阳谷、河北邯郸 、河南安阳田间高抗种群的高酯酶活性（活性在1 800 mOD·min^-1·larva^-1以上）个体频率明显高于三个敏感品系（种群）,平均酯酶活性在1 510～2 482 mOD·min^-1·larva^-1之间。江苏徐州、湖北武汉的田间中抗水平种群高酯酶活性个体频率及平均酯酶活性都介于敏感和高抗品系（种群）之间,平均酯酶活性为1 258～1.404 mOD·min^-1·larva^-1。棉铃虫各品系（种群）平均酯酶活性与对拟除虫菊酯类杀虫剂抗性个体频率的相关性要比对有机磷类的高,相关系数分别为0.82和0.42。分析各品系（种群）高酯酶活性个体频率与棉铃虫对拟除虫菊酯类、有机磷类杀虫剂抗性个体频率的相关性,得到相似的结果。考虑到酯酶并不是棉铃虫对拟除虫菊酯抗性的主要机理,建议酯酶活性可作为棉铃虫抗药性生化检测的一个参考指标。本文还讨论了酯酶与棉铃虫对拟除虫菊酯类杀虫剂及有机磷类杀虫剂抗性的关系。     

Methods for field detection of resistance to temephos in simuliids. Larval esterase level and topical application of the insecticide to adults.

Two practical field methods for indirect detection of simuliid populations resistant to temephos are proposed. The first is based on high esterase activity in resistant larvae and involves adaptations of a filter paper test in which faintly stained spots indicate susceptible populations and strongly stained ones reveal populations resistant to temephos. The second is based on the resistance to the larvicide when adults are topically exposed, and involves the use of diagnostic doses obtained by the comparison between the LD50 for susceptible and resistant populations. The relevance of such methods is discussed in order to help resistance detection in Simulium pertinax Kollar control programmes.     

白纹伊蚊溴氰菊酯抗性和敏感品系羧酸酯酶性质比较

本文对白纹伊蚊Aedes albopictus溴氰菊酯抗性品系和敏感品系羧酸酯酶的生物化学性质进行了比较。白纹伊蚊抗性品系和敏感品系羧酸酯酶随底物浓度(α-乙酸萘酯或β-乙酸萘酯)的变化比活力变化趋势一致,但抗性品系对这2种底物的比活力均高于敏感品系,抗性品系羧酸酯酶的米氏常数和最大反应速度与敏感品系有显著差异。胆碱酯酶抑制剂测定结果表明,抗性品系羧酸酯酶对敌敌畏和磷酸三苯酯的敏感性高于敏感品系,对残杀威的敏感性低于敏感品系。2个品系羧酸酯酶对脱叶磷的敏感性差异不大。说明羧酸酯酶可能与白纹伊蚊对溴氰菊酯抗性有关。     

新疆和云南番茄潜叶蛾种群对六种杀虫剂的敏感性及其与解毒酶活性的关系

【目的】番茄潜叶蛾Tuta absoluta 是新入侵我国的对番茄具有毁灭性危害的入侵害虫,目前入侵我国的番茄潜叶蛾种群对杀虫剂的抗性尚无报道。本研究旨在明确新疆和云南番茄潜叶蛾田间种群对6种常用杀虫剂的敏感性及其与解毒酶活性的关系。【方法】采用浸叶法测定6种常用杀虫剂对番茄潜叶蛾新疆和云南种群2龄幼虫的室内毒力。通过对2龄幼虫的生物测定确定3种增效剂［CYP450抑制剂胡椒基丁醚(PBO)、酯酶抑制剂磷酸三苯酯(TPP)和GST抑制剂丁烯二酸二乙酯(DEM)］对氯虫苯甲酰胺的增效作用。采用酶活性分析测定室内敏感种群和田间抗性种群(新疆种群) 2龄幼虫体内解毒酶［细胞色素P450酶(CYP450)、谷胱甘肽S-转移酶(GST)和羧酸酯酶(CarE)］活性,以确定杀虫剂抗性与解毒酶活性的关系。【结果】番茄潜叶蛾云南种群对6种杀虫剂的敏感性由高到低依次为甲维盐、溴虫腈、多杀菌素、茚虫威、氯虫苯甲酰胺和高效氯氰菊酯。新疆种群对6种杀虫剂的敏感性由高到低依次为甲维盐、溴虫腈、氯虫苯甲酰胺、多杀菌素、茚虫威和高效氯氰菊酯。与室内敏感种群相比,云南和新疆种群对氯虫苯甲酰胺的抗性水平最高,抗性倍数分别为212.7和169.3倍。生物测定结果表明,3种增效剂PBO, TPP和DEM均对氯虫苯甲酰胺无明显增效作用。酶活性测定结果表明,番茄潜叶蛾室内敏感种群和田间抗性种群之间2龄幼虫中CYP450, GST和CarE活性无显著差异。【结论】番茄潜叶蛾新疆和云南种群对测试的6种杀虫剂产生不同程度的抗性,对氯虫苯甲酰胺的抗性最高,番茄潜叶蛾对杀虫剂的抗性与解毒酶活性无关。本研究的结果对番茄潜叶蛾的田间防治和杀虫剂抗性治理具有指导意义。     

USING FILTER PAPER TEST FOR DETECTING INHIBITORY FREQUENCY OF ORGANOPHOSPHATE TO ESTERASES IN COTTON APHIDS, APHIS GOSSYPII (GLOVER)

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.     

东亚飞蝗山西两地理种群酯酶特性的比较研究

对东亚飞蝗山西临猗和永济2个地理种群的酯酶特性进行了比较研究。非变性聚丙烯酰胺凝胶电 泳图谱显示：以α-乙酸萘酯为底物染色,2个东亚飞蝗种群谱带差别不明显。但是,酯酶动力 学研究结果表明：以α-乙酸萘酯和α-丁酸萘酯为底物时,永济种群的酯酶活性分别是临猗 种群的1.81倍和1.20倍。永济种群酯酶活性的增高可能与非变性聚丙烯酰胺凝胶电泳图谱显 示出较临猗种群多出的酶带有关。体外酯酶抑制动力学研究表明：永济和临猗2种群所含酯酶大 都为B型酯酶,其含量分别为84.94%和91.47%。永济种群对对氧磷的耐受性要高于临猗种群 ,我们推测可能与2种群马拉硫磷使用背景不同有关。     

Purification and characterization of a carboxylesterase involved in malathion-specific resistance from Tribolium castaneum (Coleoptera: Tenebrionidae)

Specific resistance to malathion in a strain of Tribolium castaneum is due to a 44-fold increase in malathion carboxylesterase (MCE) activity relative to a susceptible strain, whereas non-specific esterase levels are slightly lower. Unlike the overproduced esterase of some mosquito and aphid species, MCE in Tribolium castaneum accounts for only a small fraction (0.033-0.045%) of the total extractable protein respectively in resistant and susceptible strains. The enzyme was purified to apparent homogeneity from these two strains and has a similar molecular weight of 62,000. However, preparative isoelectricfocusing indicated that resistant insects possess one MCE with pI of 7.3, while susceptible insects possess a MCE with a pI of 6.6. Purified MCE from both populations had different K(m) and V(m) values for hydrolysis of malathion as well as for alpha-naphthyl acetate. The kinetic analysis suggests that MCE of resistant insects hydrolyses malathion faster than the purified carboxylesterase from susceptible beetles and that this enzyme has greater affinity for malathion than for naphthyl esters. Malathion-specific resistance is due to the presence of a qualitatively different esterase in the resistant strain.     

Stability of spinosad resistance in Frankliniella occidentalis (Pergande) under laboratory conditions

The stability of spinosad resistance in western flower thrips (WFT), Frankliniella occidentalis (Pergande), populations with differing initial frequencies of resistance was studied in laboratory conditions. The stability of resistance was assessed in bimonthly residual bioassays in five populations with initial frequencies of 100, 75, 50, 25 and 0% of resistant individuals. There were no consistent changes in susceptibility of the susceptible strain after eight months without insecticide pressure. In the resistant strain, very highly resistant to spinosad (RF50>23,000-fold), resistance was maintained up to eight months without further exposure to spinosad. In the absence of any immigration of susceptible genes into the population, resistance was stable. In the case of the population with different initial frequency of resistant thrips, spinosad resistance declined significantly two months later in the absence of selection pressure. With successive generations, these strains did not change significantly in sensitivity. Spinosad resistance in F. occidentalis declined significantly in the absence of selection pressure and the presence of susceptible WFT. These results suggest that spinosad resistance probably is unstable under field conditions, primarily due to the immigration of susceptible WFT. Factors influencing stability or reversion of spinosad resistance are discussed.