Increased activity and reduced sensitivity of acetylcholinesterase associated with malathion resistance in a field population of the oriental migratory locust, Locusta migratoria manilensis (Meyen)

The susceptibility to malathion, and the activity and sensitivity of acetylcholinesterase (AChE, EC 1.1.1.7) were compared between two populations of the oriental migratory locust, Locusta migratoria manilensis (Meyen) collected from Wudi County of Shandong Province in East China and Huangliu County of Hainan Province in South China. Huangliu population showed 8.5-fold resistance to malathion compared with Wudi population. AChE from Huangliu population showed 4.8-fold higher activity than that from Wudi population toward the model substrate acetylthiocholine (ATC). Kinetic studies indicated that AChE from Huangliu population had 2.6-fold lower affinity, but 5.0-fold higher catalytic activity toward ATC than AChE from Wudi population. Significantly increased activity of AChE in Huangliu population was also confirmed by non-denaturing polyacrylamide gel electrophoresis. Inhibition kinetics revealed that AChE from Huangliu population was 9.8-, 2.4-, 8.0- and 7.7-fold less sensitive to inhibition by paraoxon, malaoxon, chlopyrifos oxon, demeton-S-methyl, respectively, than that from Wudi population. Our studies revealed that a mild resistance to malathion in Huangliu population was associated with reduced sensitivity and increased catalytic activity of AChE. Our results suggest that alterations of AChE may play an important role conferring or contribute to malathion resistance in Huangliu population of the locust.     

Comparative studies of acetylcholinesterases purified from two field populations of the oriental migratory locust (Locusta migratoria manilensis): implications of insecticide resistance

Acetylcholinesterase (AChE) was purified by affinity chromatography from two populations of the oriental migratory locust, Locusta migratoria manilensis (Meyen), collected from Huanghua and Pingshan Counties, Hebei Province of China. The purification factors and yields were 1661-fold and 19.3%, respectively, for the Huanghua population, and 3897-fold and 39.6% for the Pingshan population. Both the purification factor and yield were significantly lower in the Huanghua population than in the Pingshan population. AChE activity was almost completely inhibited by 10⁻⁶ M eserine and BW284C51, but ?5.8% of AChE activity was inhibited by ethopropazine at the same concentration, suggesting that purified AChE from either population was a typical insect AChE. However, AChE purified from the Huanghua population was 62-, 2.0-, and 1.6-fold less sensitive to inhibition by the three organophosphate compounds, chlorpyrifos oxon, demeton-S-methyl, and paraoxon, respectively, than that from the Pingshan population. Significantly lower purification factor and low yield associated with reduced sensitivity of AChE to inhibition by the organophosphates indicated that AChE purified from the Huanghua population was biochemically and pharmacologically different from that of the Pingshan population. Reduced sensitivity of AChE appeared to contribute to organophosphate resistance in the locust from Huanghua County, where insecticides have commonly been used to manage outbreaks of the locust.     

调控东亚飞蝗体温的主要环境因子分析

为明确东亚飞蝗Locusta migratoria manilensis体温在常年发生区与环境因子之间的关系,于2015—2016年采用直接抓取和网捕方法在天津市北大港蝗区捕捉东亚飞蝗的秋蝗和夏蝗,测量其瞬时体温以及周围4类8个环境指标,应用因子分析法评价东亚飞蝗体温与环境因子的关系。结果表明,东亚飞蝗白天体温比夜间高,平均体温高于26.5℃;且体温在白天出现2次高峰,分别是上午11点左右的49.0℃和下午14点左右的47.2℃。无论是夏蝗还是秋蝗,其体温均随着龄期增加呈现上升趋势,2～5龄蝗蝻、成虫的平均体温依次为36.9、37.0、37.6、37.2、38.0℃。因子分析结果表明,植被冠层温度是影响飞蝗体温的关键因子,飞蝗所处地面温度对其体温影响最大;风速和植株高度是影响飞蝗体温的重要分因子;植株高度与体温成负相关。表明植被冠层温度与东亚飞蝗的体温变化最为密切。     

深色和浅色两种表型东亚飞蝗对蝗虫微孢子的敏感性比较

为验证群居型飞蝗深浅2种表型对蝗虫微孢子Paranosema locustae的敏感性差异,本研究以东亚飞蝗Locusta migratoria manilensis为研究对象,测定并比较了深色型和浅色型2种表型的东亚飞蝗经蝗虫微孢子侵染后其发育历期、死亡率、免疫血淋巴细胞数量以及免疫基因表达量等变化情况。结果显示,浅色型东亚飞蝗的发育历期因蝗虫微孢子侵染而延长近1.0 d左右,比深色型东亚飞蝗多延长0.5 d;浅色型东亚飞蝗发死亡率明显高于深色型的;浅色型东亚飞蝗的免疫血淋巴细胞数量低于深色型的;蝗虫微孢子侵染后,深色型东亚飞蝗免疫基因toll基因和attacin基因的表达量上升,而浅色型东亚飞蝗的toll基因表达量无明显变化,attacin基因的表达量甚至出现下调。而且,蝗虫微孢子在浅色型飞蝗体内增殖更多。表明深色型东亚飞蝗在应对蝗虫微孢子侵染时较浅色型更有优势,浅色型东亚飞蝗较深色型对蝗虫微孢子更敏感。     

Identification and characterisation of ten glutathione S-transferase genes from oriental migratory locust, Locusta migratoria manilensis (Meyen)

BACKGROUND: Synthetic pyrethroids are the primary insecticides that are widely used for controlling Locusta migratoria manilensis (Meyen), a major pest in eastern and southern Asia and the Pacific region. In this paper, ten cDNAs encoding glutathione S‐transferases (GSTs) were sequenced and characterised in L. migratoria manilensis. The effects of deltamethrin on the ten GST gene expressions were studied. RESULTS: Phylogenetic analysis revealed nine GSTs in three different classes, including seven in sigma, one in delta and one in theta. The remaining GST (LmGSTu1) was unclassified. RT‐PCR analysis showed that most GST genes were expressed in all tissues examined, including the foregut, midgut, gastric caecum, hindgut, Malpighian tubules, fat bodies, muscles, spermaries and ovaries, except that LmGSTs2, LmGSTs4, LmGSTs7 and LmGSTu1 were expressed in several tissues. LmGSTu1 appeared to be the only gene whose expressions could not be detected in eggs. Real‐time quantitative PCR showed that deltamethrin at 0.08 and/or 0.12 µg mL^?1 increased almost all ten GST gene expressions in third‐instar nymph locusts. However, deltamethrin at 0.16 and/or 0.2 µg mL^?1 decreased the expressions of LmGSTd1, LmGSTs1, LmGSTs5 and LmGSTs6. CONCLUSION: The increases in GST gene expressions after deltamethrin exposure in L. migratoria manilensis might result in its elevating tolerance to other insecticides and xenobiotics. Copyright © 2011 Society of Chemical Industry     

Genomics-based approaches to screening carboxylesterase-like genes potentially involved in malathion resistance in oriental migratory locust (Locusta migratoria manilensis)

BACKGROUND: Previous studies have indicated that increased caboxylesterase (CarE) activity is a major mechanism of malathion resistance in field populations of the oriental migratory locust, Locusta migratoria manilensis (Meyen), in China. The aim of the present study was to screen CarE‐like genes from a large locust expressed sequence tag (EST) database and to assess their potential roles in malathion resistance. RESULTS: Twenty‐five ESTs derived from different CarE‐like genes in the locust EST database were identified, and 12 candidate genes with significantly increased expressions, ranging from 2.6‐ to 11.6‐fold in a field‐derived resistant (FR) colony of the locust, were found. These candidate genes were constitutively expressed in all nymph and adult stages, and most of them were predominantly expressed in the gastric caeca and the midgut. Among the 12 genes, two representative genes (LmCarE9 and LmCarE25) were chosen for RNAi followed by malathion bioassay. The nymph mortalities increased from 34.3 to 65.2 and 54.2% respectively after LmCarE9 and LmcarE25 were silenced. These results indicated significant roles of these CarE‐like genes in conferring malathion resistance in the locust. CONCLUSION: Multiple CarE‐like genes were involved in malathion resistance in the locust. As validated by RNAi followed by malathion bioassay, LmCarE9 and LmcarE25 played a significant role in conferring malathion resistance. Copyright © 2010 Society of Chemical Industry     

几种昆虫几丁质抑制剂对东亚飞蝗的毒力测定和应用评价

在室内筛选了 5种昆虫几丁质抑制剂和 1种常规杀蝗药剂对东亚飞蝗的毒效。结果表明 ,5种昆虫生长调节剂中 ,以毒饵中药剂有效含量计 ,卡死克毒效最高 ,对 4龄蝻的LC₅₀为26.5(16.5～38.0)mg/kg ,LT₅₀为4.946d ,抑太保次之,LC₅₀为284.4(133.8～449.2)mg/kg,LT₅₀为5.227d ,其余 3种毒效较低 ;但从性能价格评估 ,则以卡死克和灭幼脲较优 ,虽然致死时间较常规杀蝗剂长 ,但由于其高选择性 ,对非靶标生物影响很小 ,对环境比较安全 ,在生产上应用价值较高。常规杀蝗药剂高效氯氰菊酯对天敌和非靶标生物影响较大     

Effects of gibberellic acid on ovarian biochemical composition and ecdysteroid amounts in the migratory locust Locusta migratoria (Orthoptera,Acrididae)

Migratory locusts represent the most important crop pests in Africa and Asia. Because environmental damages are associated with the use of synthetic insecticides in locust and grasshopper control, new environmentally acceptable approaches are becoming increasingly important. The effects of gibberellic acid (GA₃), a plant growth regulator, was investigated by topical application and ingestion on the reproductive physiology and ecdysteroids production of the migratory locust Locusta migratoria at concentrations of 125, 625, 3125, 4125, 5125 and 6125 μg/mL. The treatment applied to adult females during the pre-ovipositional phase provokes a significant adverse influence on their reproductive potential. Indeed, GA₃ significantly reduced both fecundity and fertility. Gibberellic acid also caused disturbances in the incorporation of the hemolymph metabolites (proteins, carbohydrates and lipids) in the oocyte resulting in a significant reduction in their concentrations in the ovaries. Ecdysteroid production was measured by an enzymo-immunoassay. GA₃, administered by ingestion to the adult females, significantly reduced the ecdysteroid titers in ovaries and freshly laid eggs.     

Esterase-mediated malathion resistance in the human head louse, Pediculus capitis (Anoplura: Pediculidae)

Resistance in a dual malathion- and permethrin-resistant head louse strain (BR-HL) was studied. BR-HL was 3.6- and 3.7-fold more resistant to malathion and permethrin, respectively, compared to insecticide-susceptible EC-HL. S,S,S-Tributylphosphorotrithioate synergized malathion toxicity by 2.1-fold but not permethrin toxicity in BR-HL. Piperonyl butoxide did not synergize malathion or permethrin toxicity. Malathion carboxylesterase (MCE) activity was 13.3-fold and general esterase activity was 3.9-fold higher in BR-HL versus EC-HL. There were no significant differences in phosphotriesterase, glutathione S-transferase, and acetylcholinesterase activities between strains. There was no differential sensitivity in acetylcholinesterase inhibition by malaoxon. Esterases from BR-HL had higher affinities and hydrolysis efficiencies versus EC-HL using various naphthyl-substituted esters. Protein content of BR-HL females and males was 1.6- and 1.3-fold higher, respectively, versus EC-HL adults. Electrophoresis revealed two esterases with increased intensity and a unique esterase associated with BR-HL. Thus, increased MCE activity and over-expressed esterases appear to be involved in malathion resistance in the head louse.     

Variability in resistance-related enzyme activities in field populations of the tarnished plant bug, Lygus lineolaris

Widespread use of Bt crops for control of lepidopterous pests has reduced insecticide use and provided the tarnished plant bug the opportunity to become a serious pest on mid-South cotton. Organophosphate insecticides have predominantly been used against plant bugs in recent years due to the reduced efficacy of other insecticides. In this study, a biochemical approach was developed to survey enzymatic levels associated with organophosphate resistance levels in field populations of the tarnished plant bug. Forty-three populations were collected from the delta areas of Arkansas, Louisiana, and Mississippi. Three esterase substrates and one substrate each of glutathione S-transferase (GST) and acetylcholinesterase (AChE) were used to determine corresponding detoxification enzyme activities in different populations. Compared to a laboratory susceptible colony, increases up to 5.29-fold for esterase, 1.96-fold for GST, and 1.97-fold for AChE activities were detected in the field populations. In addition to the survey of enzyme activities among the populations, we also examined the susceptibility of major detoxification enzymes to several inhibitors which could be used in formulations to synergize insecticide toxicity against the target pests. As much as 52-76% of esterase, 72-98% of GST, and 93% of AChE activities were inhibited in vitro. Revealing variable esterase and GST activities among field populations may lead to a better understanding of resistance mechanisms in the tarnished plant bug. This study also reports effective suppression of detoxification enzymes which may be useful in future insecticide resistance management program for the tarnished plant bug and other Heteropteran pests on Bt crops.     

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.     

山东省绿盲蝽田间种群对六种杀虫剂的敏感性监测

为了解山东地区绿盲蝽对常用杀虫剂的敏感性变化情况,于2010—2012年采用玻璃管药膜法监测山东聊城、菏泽、滨州、德州地区绿盲蝽田间种群对马拉硫磷、毒死蜱、灭多威、丁硫克百威、联苯菊酯和氟虫腈等6种杀虫剂的敏感性。结果显示,相对于2009年的监测数据,2010—2012 年各地绿盲蝽种群对不同杀虫剂表现出不同程度的敏感性变化,但相对毒力比值均小于10倍。其中对毒死蜱、联苯菊酯的敏感性均未降低,菏泽种群表现为敏感性增强,相对毒力比值小于1。2011—2012年德州种群对丁硫克百威、灭多威和氟虫腈的敏感性降低,相对毒力比值大于3倍,其它种群敏感性变化不大,相对毒力比值均小于3倍。3年间菏泽种群对马拉硫磷的敏感性变化不大,相对毒力比值小于3倍,其它种群敏感性均有所降低。因此,毒死蜱、马拉硫磷、联苯菊酯、丁硫克百威等仍是山东棉区防治绿盲蝽的有效药剂。     

Relationship among expression, amplification, and methylation of FE4 esterase genes in Italian populations of Myzus persicae (Sulzer) (Homoptera: Aphididae)

The wide use of insecticides containing an esteric group selected resistant Myzus persicae populations characterised by the overproduction of one of two closely related carboxylesterases (E4 and FE4). In this paper, we present data collected from Italian population indicating that all the 22 populations analysed possess amplified FE4 gene only. The estimation of FE4 copy number, carried out by densitometric scanning of dot and Southern blots, puts in evidence that the different populations possess a gene copy number ranging from 6 to 104. Statistical analysis shows the existence of a high positive correlation between gene copy number and total esterase activity. In aphid strain with low FE4 copy number, these genes are almost totally methylated. On the contrary, aphid strains with high FE4 gene number evidenced highly variable methylation levels and absence of correlation between the number of genes and their methylation state. The same result has been observed when comparing FE4 methylation levels and esterase activity.     

Differential susceptibilities to pyrethroids in field populations of Chilo suppressalis (Lepidoptera: Pyralidae)

To assess the feasibility of pyrethroids for rice insect control, we examined susceptibilities of six field populations of rice stem borer Chilo suppressalis (Walker) to 10 pyrethroids using the topical application method in laboratory in 2004 and 2005. Our results showed that the seven pyrethroids with high fish-toxicity (i.e., β-cyfluthrin, λ-cyhalothrin, β-cypermethrin, deltamethrin, S-fenvalerate, α-cypermethrin, and fenpropathrin) were more effective against C. suppressalis than the three compounds with low fish-toxicity (i.e., cycloprothrin, etofenprox, and silafluofen). The results also showed that all 10 of the pyrethroids were much more effective than methamidophos and monosultap for C. suppressalis control. In addition, we found that susceptibilities of some field populations of C. suppressalis to some high fish-toxicity pyrethroids were significantly reduced, and our results indicated that a Ruian (RA) field population showed a year-to-year variation in susceptibility to most tested pyrethroids between 2004 and 2005. Our data indicated that the tolerance levels increased dramatically in RA population, especially to β-cyfluthrin and deltamethrin. This study provided the first assessment of resistance to pyrethroids in field populations of C. suppressalis. In addition, a close correlation between resistance ratios to the 10 compounds and differences of the structures of these compounds was established in the RA05 population, which was resistant to most of the pyrethroids tested while it was still very susceptible to fenvalerate with no cross resistance. Finally, the feasibility and precaution were discussed in selecting pyrethroids as alternatives to replace high toxicity organophosphates for C. suppressalis control and insecticide resistance management.     

Partial purification and characterization of a methyl-parathion resistance-associated general esterase in Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae)

Increased hydrolytic metabolism of organophosphate insecticides has been associated with resistance among Nebraska western corn rootworm populations. In this study, resistance-associated esterases were partially purified by differential centrifugation, ion exchange, and hydroxyapatite column chromatography, with a final purification factor of 100-fold and recovery of approximately 10%. Kinetic analysis of the partially purified enzyme indicated that the K_m of the group II esterases was identical for the two populations, although V_max was consistently threefold higher in the resistant population. A putative esterase, DvvII, was further purified to homogeneity by preparative polyacrylamide gel electrophoresis. DvvII is a monomer with a molecular weight of approximately 66 kDa, although three distinct isoforms with similar pIs were evident based on isoelectric focusing gel electrophoresis. Immunoassays with the Myzus persicae E4 antiserum indicated that group II esterases from D. v. virgifera were cross-reactive and expressed at much higher titers in the resistant population relative to the susceptible counterpart. These results suggest that the resistance is likely associated with overproduction of an esterase isozyme in resistant D. v. virgifera populations.     

Pyrethroid resistance and esterase activity in three strains of the cotton bollworm, Helicoverpa armigera (Hübner)

Microplate assay technique for estimation of esterase activity in a single insect was used in combination with dose mortality bioassays to detect pyrethroid resistance in three strains of Helicoverpa armigera and to study the frequency of pyrethroid resistant individuals within the population of the same strain at two larval stages, third and fifth instar. The third and fifth instar larvae of the field strains i.e., Nagpur strain and Delhi strain that displayed high degree of resistance towards deltamethrin, had higher esterase activity compared to a susceptible laboratory strain. The frequency distribution of individuals with elevated esterase activity above 1.00 absorbance unit was correlated with the resistance level of the strains. The frequency of resistant individuals in the third instar larvae of Nagpur strain and Delhi strain were 29% and 23%, respectively compared to 4% in the susceptible strain. The resistant individuals in the resistant strains have markedly increased in the fifth instar larvae with a frequency distribution of 63% and 90% in Delhi strain and Nagpur strain, respectively, while only 14% of individuals was found to have elevated esterase activity in the susceptible strain. The results demonstrated the role of esterase in pyrethroid resistance in H. armigera. Microplate assay proved to be a rapid and reliable biochemical technique for monitoring of pyrethroid resistance in H. armigera.     

暗黑赤眼蜂对梨小食心虫卵的寄生作用

为检验暗黑赤眼蜂Trichogramma pintoi Voegele对梨小食心虫Grapholita molesta(Busck)卵的寄生效果,在室内条件下,以松毛虫赤眼蜂Trichogramma dendrolimi Matsumura为对照,比较暗黑赤眼蜂对其自然寄主棉铃虫Helicoverpa armigera(Hübner)和非自然寄主梨小食心虫的寄生效果。暗黑赤眼蜂与松毛虫赤眼蜂对梨小食心虫卵的寄生率和在被寄生卵中的羽化率差异均不显著。暗黑赤眼蜂对梨小食心虫卵的寄生率显著高于对棉铃虫卵的寄生率;在15、45、60和75粒卵/管条件下,暗黑赤眼蜂寄生梨小食心虫和棉铃虫卵后羽化率差异不显著,但在30和90粒卵/管条件下,羽化率差异显著。拟合分析显示,暗黑赤眼蜂和松毛虫赤眼蜂对梨小食心虫卵的寄生符合Holling-Ⅱ型功能反应圆盘方程,说明暗黑赤眼蜂是一种防治梨小食心虫的潜在寄生蜂。     

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.     

Monitoring of beet armyworm Spodoptera exigua (Lepidoptera: Noctuidae) resistance to chlorantraniliprole in China

Beet armyworm, Spodoptera exigua is a major insect pest of vegetables in China, and has been reported to develop resistance to many broad-spectrum insecticides. Recently registered chlorantraniliprole provides a novel option for control of this pest resistant to other conventional insecticides. The susceptibilities of field collected populations were measured by diet incorporation assay with neonate, obvious variation of susceptibility was observed among the 18 field populations with LC₅₀ values varying from 0.039 to 0.240 mg/liter. Moderate resistant level was discovered in 8 of 18 field populations, other 8 populations had become low level tolerance to chlorantraniliprole, and only one population in all the field colonies remained susceptible. Biochemical assays were performed to determine the potential mechanisms involved in tolerance variation. Field populations displayed varied detoxification enzyme activities, but the regression analysis between chlorantraniliprole toxicities and enzyme activities demonstrated each field population might have specific biochemical mechanisms for tolerance. Artificial selection in laboratory with chlorantraniliprole was carried out, 23 generations of continuous selections resulted in 11.8-fold increase in resistance to chlorantraniliprole, and 3.0-fold and 3.7-fold increases in mixed function oxidase and esterase, respectively. Compared with the susceptible strain kept in laboratory the selection strain had developed 128.6-fold resistance to this insecticide. Synergism assays showed the detoxification enzymes might not involved in the resistance observed in field collected populations and the selected strain.     

Molecular and biochemical mechanisms of organophosphate resistance in laboratory-selected lines of the oriental fruit fly (Bactrocera dorsalis)

Genetic and biochemical factors leading to resistance to various organophosphate (OP) based insecticides were studied in lines selected for OP-resistance in the oriental fruit fly Bactrocera dorsalis. Lines were separately selected for resistance to naled, trichlorfon, fenitrothion, fenthion, formothion, and malathion. Overall, these lines showed increased resistance ratios ranging from 13.7- to 814-fold relative to a susceptible (S) line. Also, in these newly selected lines the same three point mutations in the ace gene, previously identified in resistance studies and designated as I214V, G488S and Q643R, were found. As expected, the enzyme from the resistant lines showed lower overall activity and reduced sensitivity to inhibition by fenitrothion, methyl-paraoxon and paraoxon compared to the wild type acetylcholinesterase (AChE) enzyme. The apparent V_max values for esterase from the resistant lines were 1.2-3.69 times higher than that of the S line. Although only the naled-, trichlorfon- and fenthion-r lines showed lower esterase affinities (based on apparent K_m values) compared with the S line, all of the V_max/K_m ratios were higher in the resistant lines compared to that of the S line. The OP-resistant lines also displayed an overall similar pattern of isozyme expression, except for one additional band found only in the naled-r line and one band that was absent in the trichlorfon-, malathion-, and fenthion-r lines. Our results also show that overall, multiple examples of high OP resistance in selected lines of B. dorsalis exhibiting the same genetic alterations in the ace gene seen previously resulted in different effects on esterase enzyme activity in relation to various OP compounds.