Mechanisms of organophosphate resistance in a field population of oriental migratory locust, Locusta migratoria manilensis (Meyen)

The susceptibilities to three organophosphate (OP) insecticides (malathion, chlorpyrifos, and phoxim), responses to three metabolic synergists [triphenyl phosphate (TPP), piperonyl butoxide (PBO), and diethyl maleate (DEM)], activities of major detoxification enzymes [general esterases (ESTs), glutathione S-transferases (GSTs), and cytochrome P450 monooxygenases (P450s)], and sensitivity of the target enzyme acetylcholinesterase (AChE) were compared between a laboratory-susceptible strain (LS) and a field-resistant population (FR) of the oriental migratory locust, Locusta migratoria manilensis (Meyen). The FR was significantly resistant to malathion (57.5-fold), but marginally resistant to chlorpyrifos (5.4) and phoxim (2.9). The malathion resistance of the FR was significantly diminished by TPP (synergism ratio: 16.2) and DEM (3.3), but was unchanged by PBO. In contrast, none of these synergists significantly affected the toxicity of malathion in the LS. Biochemical studies indicated that EST and GST activities in the FR were 2.1- to 3.2-fold and 1.2- to 2.0-fold, respectively, higher than those in the LS, but there was no significant difference in P450 activity between the LS and FR. Furthermore, AChE from the FR showed 4.0-fold higher activity but was 3.2-, 2.2-, and 1.1-fold less sensitive to inhibition by malaoxon, chlorpyrifos-oxon, and phoxim, respectively, than that from the LS. All these results clearly indicated that the observed malathion resistance in the FR was conferred by multiple mechanisms, including increased detoxification by ESTs and GSTs, and increased activity and reduced sensitivity of AChE to OP inhibition.     

Toxicological and biochemical characterizations of malathion sensitivity in two field populations of Oxya chinensis （Orthoptera： Acridoidea）

We evaluate comparative toxicity of malathion in the two populations of the grasshopper Oxya chinensis, collected from Daixian and Fanshi of Shanxi province, China. General esterases and acetylcholinesterase （ACHE） from the two populations were characterized and compared. LD50 of the Daixian population （7.58 μg/g body weight） was 2.02-fold higher than that of the Fanshi population （3.75μg/g body weight）. General esterase-specific activities in the Daixian population were 1.91,130 and 1.85-fold higher than those in the Fanshi population, when α-NA, α-NB and β-NA were used as a substrate, respectively. Kinetic studies of general esterase showed that Vmax values of general esterases hydrolyzing α-NA,α-NB and β-NA in the Daixian population were 2.15-, 1.12-, and 1.47-fold, respectively, higher than those in the Fanshi population. The AChE activity of the Fanshi population was 1.54-fold higher than that of the Daixian population. Kinetic analysis of AChE showed that significant differences were presented between the two populations in the Km values; and the Vmax value in the Fanshi population was higher than that in the Daixian population. Inhibition studies of AChE indicated that AChE from the Daixian population was 2.56-, 2.80-, and 2.29-fold less sensitive to inhibition by paraoxon, chlorpyrifos-oxon, and demeton-S-methyl, respectively, than that from the Fanshi population. These biochemical characterizations of general esterases and AChE were consistent with malathion bioassay in the two populations. It is inferred that the reduced sensitivity of altered AChE and increased general esterase activities play an important role in the differences of insusceptibility of Oxya chinensis to malathion between the two populations.     

Pharmacodynamics of malathion and carbaryl in susceptible and multiresistant German cockroaches (Dictyoptera: Blattellidae)

Studies with malathion and carbaryl were done to compare toxicity; absorption, metabolism, internal accumulation, and excretion; and in vivo inhibition of acetylcholinesterase (AChE) after topical applications to adult male susceptible (S, Orlando normal) or multiresistant (R, HRDC) German cockroaches, Blattella germanica (L.). Compared with the S strain, R cockroaches were highly resistant to malathion (about 33-fold) and only moderately resistant or tolerant to carbaryl (about 5-fold). Tests with topically applied 14C-labeled malathion and carbaryl indicated that both compounds penetrated rapidly and radioactive products were readily excreted. Rates of absorption or excretion in S and R strains did not differ significantly. Both insecticides were extensively metabolized; each yielded the same array and similar concentrations of metabolites in insects from either strain. In contrast, metabolic detoxification of malathion and carbaryl was significantly greater in R cockroaches when the insects were treated by injection. Strains did not differ significantly in the in vitro inhibition of brain AChE by either malaoxon or carbaryl. However, dramatic differences were observed between strains in the in vivo inhibition of AChE during a 6-h test period after topical treatment with malathion, and moderate but significant differences occurred between strains in the in vivo inhibition of AChE by carbaryl. These data suggest that the strong resistance to malathion and moderate resistance or tolerance to carbaryl in R cockroaches is probably a result of enhanced capability for metabolic detoxification.     

The genetic basis of malathion resistance in housefly (Musca domestica L.) strains from Turkey

Organophosphate insecticide (parathion/diazinon) resistance in housefly (Musca domestica L.) is associated with the change in carboxylesterase activity. The product of MdalphaE7 gene is probably playing a role in detoxification of xenebiotic esters. In our research, we have isolated, cloned and sequenced the MdalphaE7 gene from 5 different Turkish housefly strains. High doses of malathion (600 microg/fly) were applied in a laboratory environment for one year to Ceyhan1, Ceyhan2, Adana and Ankara strains while no insecticide treatment was performed in the laboratory to Kirazli strain. Trp251 --> Ser substitution was found in the product of MdalphaE7 gene in all malathion resistant and Kirazli stocks. In addition, we checked the malathion carboxylesterase (MCE), percent remaining activities in acetylcholinesterase (AChE), glutathion-S-transferase (GST), and general esterase activities in all 5 strains used in this study. In comparing with universal standard sensitive control WHO, a high level of MCE and GST activities were observed while lower level of general esterase activities was detected in the tested strains. In addition, a higher percent remaining activities in AChE than WHO susceptible strain were observed in all malathion resistant strains.     

Purification and biochemical characterization of glutathione S-transferases from four field populations of Bactrocera dorsalis (Hendel) (Diptera: Tephritidae)

Glutathione S-transferases (GSTs) are a group of detoxification enzymes that catalyze the nucleophilic addition of glutathione to a wide variety of endogenous and xenobiotic compounds. In this study, GSTs were purified from four field populations of Bactrocera dorsalis with different insecticide susceptibilities by glutathione-agarose affinity chromatography. The populations were collected from Dongguan (DG) and Guangzhou (GZ) of the Guangdong Province, Haikou of the Hainan province (HN), and Kunming of the Yunnan province (YN), China. Differences in GST characteristics among the four populations were studied using purified enzyme samples through comparative SDS-PAGE, kinetic, and inhibition experiments. The specific activities of the purified enzymes were similar, but the purification yield of the GZ population (31.54%) was the lowest. SDS-PAGE analysis showed only one band at approximately 23 kDa for these four populations. Kinetic analyses showed that the affinities of the purified GSTs from the GZ and YN populations for 1-chloro-2.4-dinitrobenzene (CDNB) were much higher than those of GSTs from the other two populations, whereas the HN population had the highest catalytic capability in terms of V(max) value. The optimum temperature for CDNB conjugation was 37 °C and the optimum pH was 7.5 in all four populations. Inhibition kinetics showed that ethacrynic acid, diethyl maleate, tetraethylthiuram disulfide, curcumin, bromosulfalein, and β-cypermethrin had excellent inhibitory effects on GSTs in the four populations of B. dorsalis, but the low inhibitory effects of malathion and avermectin did not differ between populations. These results suggest that GSTs may have a role in detoxification of β-cypermethrin in B. dorsalis.     

Contributions of three-site mutations in acetylcholinesterase and cytochrome P450 to genetic variation in susceptibility to organophosphate insecticides within a natural population of Drosophila melanogaster

In this study, we attempted to elucidate the two resistance factors conferring resistance to organophosphates within the Katsunuma population of Drosophila melanogaster (Meigen), one of which has been mapped on the second chromosome and the other on the third chromosome. With regard to the second chromosome factor, we tested susceptibility to malathion of 54 recombinant inbred lines with recombination between ltd and vg. Analyses of variance (ANOVAs) showed highly significant variation in susceptibility to malathion between recombinant lines. In addition, susceptibility of the second-chromosome resistant line to malathion was increased with additional application of piperonyl butoxide, suggesting a member of the Cyp gene family located between ltd and vg. With regard to the third-chromosome factor, we conducted inhibition assays of acetylcholinesterase (AChE) with respect to fenitroxon and carbaryl, to evaluate the contribution of mutated AChE to organophosphate resistance within the Katsunuma population. I ₅₀ values of resistant lines, isolated from this population, were about 15 times higher for fenitroxon, and about two times higher for carbaryl, than those of susceptible lines, suggesting the contribution of mutated AChE to organophosphate resistance within the Katsunuma population. We further investigated the genetic variation in the acetylcholinesterase (Ace) gene within the newly collected Katsunuma population, by using the allele-specific polymerase chain reaction (PCR) approach, and revealed that within this population there were high frequencies of resistant-type mutations at three sites in the Ace gene, which play critical roles in altering sensitivity of AChE to organophosphate and carbamate insecticides.     

Malathion and pyrethroid resistance in Culex quinquefasciatus from Cuba: efficacy of pirimiphos-methyl in the presence of at least three resistance mechanisms

Use of malathion for mosquito control in Cuba for 7 years up to 1986 has selected for elevated non-specific esterase and altered acetylcholinesterase (AChE) resistance mechanisms in populations of the pest mosquito Culex quinquefasciatus Say. These mechanisms are still present in relatively high frequencies in the Havana area, despite the replacement of malathion by pyrethroid insecticides for the last 3 years in the mosquito control programme. Samples of Culex quinquefasciatus populations from within a 100 km radius of Havana had high levels of resistance to malathion and lower levels of resistance to propoxur, but there was little or no cross-resistance to the organophosphorus insecticide pirimiphos-methyl. Selection with malathion for twenty-two consecutive generations in the laboratory increased the level of malathion resistance to 1208-fold and propoxur level to 1002-fold, but the maximum level of pirimiphos-methyl resistance was only 11-fold. Pirimiphos-methyl is still operationally effective, despite the resistance mechanisms segregating, so this insecticide if used for control is unlikely to select either of the known resistance factors directly in the field population. Since 1986, pyrethroids have been used extensively, and low levels of pyrethroid resistance were detected in two of five field population samples tested. Malathion selection did not increase the level of pyrethroid resistance, which indicates that one or more distinct pyrethroid resistance factors are now being selected in the field populations of Culex quinquefasciatus.     

我国棉花主产区棉蚜对吡虫啉的抗性监测及抗性机理

【目的】由于生长周期短、繁殖率高,棉蚜Aphis gossypii容易对杀虫剂产生抗药性。本研究旨在明确我国棉花主产区棉蚜对吡虫啉的抗性水平及抗性机理。【方法】采用浸叶法测定了北京海淀,河北廊坊和邯郸,山东德州,河南许昌,以及新疆奎屯和阿克苏地区棉蚜对吡虫啉的抗性水平;测定了不同种群棉蚜3种解毒酶（多功能氧化酶、羧酸酯酶、谷胱甘肽S-转移酶）及乙酰胆碱酯酶的活性;并对靶标基因烟碱型乙酰胆碱受体（nAChR）β1亚基基因进行了突变检测。【结果】北京海淀、河南许昌和河北邯郸的棉蚜对吡虫啉敏感;河北廊坊、新疆阿克苏、山东德州及新疆奎屯地区的棉蚜对吡虫啉的抗性倍数(resistance ratio, RR)分别为22.6, 26.3,53.5和61.1倍,为中等水平抗性。酶活力对比研究发现,阿克苏和奎屯地区的棉蚜多功能氧化酶的比活力分别是敏感种群(北京种群)的1.7和1.8倍,羧酸酯酶的比活力分别是敏感种群的1.6和1.7倍,谷胱甘肽S-转移酶的比活力均是敏感种群的1.5倍,但是乙酰胆碱酯酶比活力在棉蚜种群间差异不显著。靶标基因突变检测表明,河北廊坊、新疆阿克苏、山东德州及新疆奎屯棉蚜种群nAChR β1亚基均存在与吡虫啉抗性相关的精氨酸到苏氨酸（R81T）突变。【结论】结果提示,多功能氧化酶、羧酸酯酶和谷胱甘肽S-转移酶活力升高以及nAChR β1亚基R81T突变与棉蚜对吡虫啉的抗性形成相关。     

Molecular screening of insecticides with sigma glutathione S-transferases (GST) in cotton aphid Aphis gossypii using docking

Glutathione S-transferases (GSTs) are one of the major families of detoxifying enzymes that detoxifies different chemical compounds including insecticides in different insect species. Among the GST subclasses, sigma GSTs are found to be the most abundant and conserved among different insect orders. These GSTs are found to play an important role in lipid peroxidation as well as detoxification. Cotton aphid, Aphis gossypii is the most damaging sucking pest with a wide range of hosts and vector of more than 50 plant viruses. Resistance to insecticides in A. gossypii is reported in India and in other countries. Glutathione S transferases (GSTs), an oxidative enzyme is understood to have a role in insecticide resistance and plant resistance breakdown. In relation to this, we have focused on the sigma 1 (GenBank Accession No: {"type":"entrez-nucleotide","attrs":{"text":"JN989964.1","term_id":"392584103","term_text":"JN989964.1"}}JN989964.1) and sigma 2 (GenBank Accession No: {"type":"entrez-nucleotide","attrs":{"text":"JN989965.1","term_id":"392584105","term_text":"JN989965.1"}}JN989965.1) GSTs of A. gossypii and their interaction with plant natural compounds and insecticides. Molecular screening of different insecticides (Chlorphinamidine, Mevinphos, Nitenpyrum, Piperonyl butoxide, Tetrachlorovinphos, Pyrethrins, Resmetrin, Pirimicarb and Dinotefuran) and known plant derived natural compounds (Catechin, Gossypol, Myrcene, Kaempferol, P-coumaric acid, Quercetin, Tannins, α-mangostin, Capsaicin, Cinnamic acid, Citronellal, Curcumin, Dicumarol, Ellagic acid, Eugenol, Geriniol, Isoeugenol, Juglone, Menadione, Methyl jasmonate, Morin, Myricetin, Myristicin, Piperine, Plumbagin, Tangitinin C, Thymol, Vanillin, Alpha pipene, α-terpineol Apigenin and β-Caryophyllene) with sigma 1 and sigma 2 GST protein models was completed using Maestro 9.3 (Schrodinger, USA). This exercise showed the binding of piperonyl butoxide with sigma 1 GST and tannin with sigma 2 GST for further consideration.     

山东省五个棉花产区绿盲蝽对药剂的敏感性检测（英文）

利用闪烁管药膜法测定了2009年山东省德州、滨州、梁山、曲阜和聊城5个棉花产区绿盲蝽Lygus lucorum Meyer-Dür对硫丹、马拉硫磷、毒死蜱、灭多威、丁硫克百威、吡虫啉、联苯菊酯和氟虫腈8种杀虫剂的敏感性,筛选出适合各地区的高效防治药剂。结果表明：5个地区绿盲蝽种群对马拉硫磷、毒死蜱、丁硫克百威、联苯菊酯和吡虫啉处于敏感性阶段。不同种群之间对灭多威、硫丹和氟虫腈的敏感性差异较大,其中聊城种群为最敏感种群,滨州种群对灭多威、 硫丹和氟虫腈的LC₅₀值分别为聊城种群的5.12,6.04和39.80倍;曲阜种群对灭多威、硫丹和氟虫腈的LC₅₀值分别为聊城种群的22.12,5.48和22.80倍。两种群对此3种药剂的敏感性下降,而其余种群仍处于较敏感阶段。8种药剂对绿盲蝽成虫的毒力按大小依次排序为：氟虫腈＞灭多威、联苯菊酯、硫丹＞马拉硫磷、毒死蜱＞丁硫克百威＞吡虫啉。2009年7-10月间德州夏津绿盲蝽种群对8种杀虫剂的敏感性变化极微。     

Mechanisms of resistance to malathion in the medfly Ceratitis capitata

Target site insensitivity and metabolic resistance mediated by esterases have been previously suggested to be involved in resistance to malathion in a field-derived strain (W) of Ceratitis capitata. In the present study, we have obtained the coding sequence for acetylcholinesterase (AChE) gene (Ccace) of C. capitata. An allele of Ccace carrying only a point mutation Gly328Ala (Torpedo numbering) adjacent to the glutamate of the catalytic triad was found in individuals of the W strain. Adult flies homozygotes for this mutant allele showed reduced AChE activity and less sensitivity to inhibition by malaoxon, showing that target site insensitivity is one of the factors of malathion resistance. In addition, all individuals from the resistant W strain showed reduced aliesterase activity, which has been associated with specific malathion resistance in higher Diptera. However, the alphaE7 gene (CcalphaE7), sequenced in susceptible and resistant individuals, did not carry any of the mutations associated with organophosphorus insecticide resistance in other Diptera. Another esterase mechanism, perhaps a carboxylesterase selective for malathion, in addition to mutant AChE, thus contributes to malathion resistance in C. capitata.     

Purification and partial characterization of glutathione S-transferase from insecticide-resistant field populations of Liposcelis paeta Pearman (Psocoptera: Liposcelididae)

Enzymes that possess glutathione S-transferase (GST) activity were purified to homogeneity by glutathione-agarose affinity chromatography from three field populations of Liposcelis paeta (Pearman). These populations were collected from Nanyang city of Henan Province (NY), Wuzhou (WZ) and Hezhou (HZ) cities of Guangxi Province, China, and had different susceptibilities to dichlorvos [LC₅₀s of the NY (281.48 mg/m²), the WZ (285.07 mg/m²), and the HZ (243.52 mg/m²), respectively]. The specific activities of purified enzymes from these three populations increased 32.24-, 99.81-, and 42.52-fold, respectively. Kinetic analyses showed that the catalytic activity of purified GST from NY population towards GSH was much higher than the others, while WZ population reached the highest in V. SDS–polyacrylamide electrophoresis revealed that the purified GST had two subunits with a molecular mass of 23.31 and 20.43 kDa for NY, 53.14 and 20.13 kDa for WZ, and 50.79 and 19.42 kDa for HZ, respectively. The in vitro inhibition studies of GSTs indicated that three kinds of insecticides (chlorpyrifos, carbosulfan, and cypermethrin) and five metallic ions (Zn²⁺, Ba²⁺, Ca²⁺, Hg²⁺, Mn²⁺, and Mg²⁺) all possessed inhibitory effects on purified GST, and ethacrynic acid (EA, a specific inhibitor of GST) expressed inhibitory effects. In the bioassay, three populations of L. paeta had different susceptibilities to different insecticides, even after they were reared on diets consisting of 25% EA. The GST activities of L. paeta from different areas also showed different temperature and pH stabilities. The differences in GST among the three populations may be attributed partially to the differences in control practices for psocids between Henan and Guangxi Provinces. © 2009 Wiley Periodicals, Inc.     

Exploration of the susceptibility of AChE from the poultry red mite Dermanyssus gallinae (Acari: Mesostigmata) to organophosphates in field isolates from France

The red fowl mite Dermanyssus gallinae (De Geer, 1778) is a hematophagous mite species, which is very commonly found in layer facilities in Europe. The economic and animal health impact of this parasite is quite important. In laying hen houses, organophosphates are almost the only legally usable chemicals. Detecting a target resistance can be useful in order to limit the emergence of resistant populations. The acetylcholinesterase (AChE) activity and the enzyme sensitivity to paraoxon was investigated in 39 field samples and compared to a susceptible reference strain (SSK). Insensitivity factor values (expressed as IC₅₀ ratio) obtained from field isolates compared to SSK revealed some polymorphism but not exceeding a 6-fold difference. The kinetic characteristics of AChE from some field samples showed some difference in K _M values for acetylthiocholine and inhibition kinetics performed with diethyl paraoxon exhibited a 5.5-fold difference in the bimolecular rate constant in one field isolate. Taken together, these data suggested that differences in AChE susceptibility to organophosphates may exist in D. gallinae but no resistant population was found.     

β-细辛醚对谷蠹成虫体内四种酶活性的影响

采用石菖蒲Acorus gramineus根茎提取物β-细辛醚对谷蠹Rhizopertha dominica成虫进行拌粮处理,测定β-细辛醚对谷蠹体内乙酰胆碱酯酶、谷胱甘肽S-转移酶、羧酸酯酶和酯酶同工酶的时间效应和剂量效应。结果表明：β-细辛醚对试虫体内4种酶的酶活性均表现出较强的时间效应。LC₅₀（94.49 mg/kg）剂量的β-细辛醚处理后,谷蠹成虫体内乙酰胆碱酯酶和谷胱甘肽S-转移酶活性随处理时间的延长整体表现为抑制作用,对羧酸酯酶和酯酶同工酶则表现诱导增加作用。低剂量（67.5 mg/kg）β-细辛醚对乙酰胆碱酯酶具有显著的诱导作用,但是随着处理剂量的升高,对乙酰胆碱酯酶的活力多数表现为抑制作用。低剂量（≤100.0 mg/kg）β-细辛醚对谷胱甘肽S 转移酶具有诱导作用,而高剂量（≥133.3 mg/kg）β-细辛醚对谷胱甘肽S-转移酶具有抑制作用。β-细辛醚对羧酸酯酶的活性多数表现为诱导作用,提高β-细辛醚的处理剂量可提高羧酸酯酶的活力。不同剂量的β-细辛醚处理对谷蠹酯酶同工酶均具有显著的诱导作用,但诱导效果与处理剂量关系并不显著。     

河北转Bt基因棉田棉铃虫对杀虫剂的抗性及相关酶活性的变化

为明确河北省推广种植植转Bt基因抗虫棉（简称Bt棉）后, 棉铃虫Helicoverpa armigera (Hübner)对常用杀虫剂的抗药性水平及其生化机理, 2011-2012年采用点滴法对保定南郊、 沧州南皮、 邢台巨鹿3个地区的田间种群以及敏感种群进行了室内毒力测定, 并采用生化分析法对4个种群相关的羧酸酯酶（carboxylesterase, CarE）、 谷胱甘肽S 转移酶(glutathione S-transferases, GSTs)和乙酰胆碱酯酶(acetylcholinesterase, AChE)的活性进行了研究。结果表明: 3个田间种群对高效氯氰菊酯和氰戊菊酯处于中至高抗水平, 抗性倍数为20.02~73.70倍; 对灭多威处于低至中抗水平, 抗性倍数为6.27~11.84倍; 对高效氯氟氰菊酯(抗性倍数: 1.07~4.20倍), 辛硫磷、 毒死蜱和马拉硫磷(抗性倍数: 1.00~2.69倍), 以及氯虫苯甲酰胺（抗性倍数: 2.00~3.67倍）均处于敏感水平。3个田间种群的CarE, GSTs和AChE活性分别是敏感种群的1.06~1.23, 1.20~1.63和1.15~1.23倍, 这可能与其对高效氯氰菊酯、 氰戊菊酯和灭多威产生的抗性有关。     

The Genetic Basis of Malathion Resistance in Housefly (Musca domestica L.) Strains From Turkey

Organophosphate insecticide (parathion/diazinon) resistance in housefly (Musca domestica L.) is associated with the change in carboxylesterase activity. The product of MdE7 gene is probably playing a role in detoxification of xenobiotic esters. In our research, we have isolated, cloned and sequenced the MdE7 gene from five different Turkish housefly strains. High doses of malathion (600 g/fly) were applied in a laboratory environment for one year to Ceyhan1, Ceyhan2, Adana, and Ankara strains while no insecticide treatment was performed in the laboratory to Kirazli strain. Trp²⁵¹ Ser substitution was found in the product of MdE7 gene in all malathion-resistant and Kirazli stocks. In addition, we checked the malathion carboxylesterase (MCE), percent remaining activities in acetylcholinesterase (AChE), glutathion-S-transferase (GST), and general esterase activities in all five strains used in this study. In comparing with universal standard sensitive control WHO, a high level of MCE and GST activities were observed while lower level of general esterase activities was detected in the tested strains. In addition, a higher percent remaining activities in AChE than WHO susceptible strain were observed in all malathion-resistant strains.     

黄曲条跳甲对毒死蜱敏感性差异的生化机制

采用生物测定方法测定黄曲条跳甲Phyllotreta striolata(F.)2个室内试验种群(蔊菜试验种群、上海青试验种群)和1个田间自然种群对毒死蜱的LC50值。结果表明,蔊菜试验种群对毒死蜱的LC50值最低,为30.3459mg.L-1;田间自然种群对毒死蜱的LC50值最高,为77.8448mg.L-1,与蔊菜试验种群相比的敏感性指数为0.39。对不同种群黄曲条跳甲乙酰胆碱酯酶(AChE)、羧酸酯酶(CarE)、谷胱甘肽-S-转移酶(GSTs)的活性测定结果表明,黄曲条跳甲田间自然种群AChE活性最低,与菜试验种群、上海青试验种群相比,差异极显著(P>0.01);田间自然种群GSTs活性最高,与蔊菜试验种群、上海青试验种群相比,差异极显著(P>0.01);黄曲条跳甲蔊菜试验种群CarE活性最低,田间自然种群CarE活性最高,二者差异极显著(P>0.01);说明黄曲条跳甲对毒死蜱的敏感性下降可能与AChE活性的降低,与CarE、GSTs的活性提高有一定的关系。     

Resistance to malathion in field populations of Ceratitis capitata

The Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), is considered one of the most economically damaging pests of citrus orchards in Spain. Insecticide treatments for the control of this pest are mainly based on aerial and ground treatments with malathion bait sprays. However, the frequency of insecticide treatments has been increased in some areas of the Comunidad Valenciana in the last years, because of problems with the control of C. capitata. We have found that field populations from citrus and other fruit crops from different geographical areas in Spain showed lower susceptibility to malathion (6- to 201-fold) compared with laboratory populations. More importantly, differences in susceptibility could be related to the frequency of the field treatments. A resistant strain (W), derived from a field population, and a susceptible laboratory strain (C) were maintained in the laboratory. The W strain showed cross-resistance to the organophosphate fenthion (10-fold) but not to spinosad. Enzymatic assays showed that acethylcholinesterase activity was less inhibited in vivo by malathion and in vitro by malaoxon (active form of malathion) in adult flies from the W-resistant strain. Experiments to evaluate the effects of synergists revealed that the esterase inhibitor S,S,S-tributyl phosphorotrithioate (DEF) partially suppressed malathion resistance. Thus, target site insensitivity and metabolic resistance mediated by esterases might be involved in the loss of susceptibility to malathion in C. capitata. Nonetheless, additional biochemical and molecular studies will be required to confirm this hypothesis.     

Detoxification of electrophilic compounds by glutathione S-transferase catalysis: determinants of individual response to chemical carcinogens and chemotherapeutic drugs?

The glutathione S-transferases (GSTs) catalyze the GSH-dependent detoxification of reactive electrophiles such as genotoxic chemical carcinogens and cytotoxic chemotherapeutic agents. Allelic polymorphism in the GSTs has been used to investigate the hypothesis that GSTs are involved in susceptibility to human cancers. Such studies have resulted in low penetrance, high prevalence associations between cancer risk and GST polymorphisms. By examination of interindividual variation of GST expression it becomes clear that GST genotype alone is not an accurate predictor of GST expression. GST expression is tissue specific and interindividual variation of expression is at least 7-fold in normal tissues. Thus, populations of the same genotype are actually heterogeneous as regards expression. Similarly, polymorphisms are not effective in all tissues and GST induction is not independent of genotype. Mechanistic models for chemical aspects of colorectal cancer and chemotherapy for breast cancer demonstrate some of the ways by which such interactions can be studied and the potential for future studies.     

B型烟粉虱田间种群对毒死蜱和敌敌畏抗性的生化机制

通过增效剂生物测定和生化分析,探讨了采自福建省的B型烟粉虱Bemisia tabaci 6个田间种群对毒死蜱和敌敌畏抗性的生化机制。结果表明：与敏感品系SUD-S相比,6个田间种群对毒死蜱和敌敌畏分别具有54.53～78.43倍和6.23～11.25倍的抗性。TPP、PBO和DEM对毒死蜱的增效比分别为3.61～24.94倍、1.14～1.76倍和1.04倍,对敌敌畏的增效比分别为1.67～2.64倍、1.33～1.65倍和1.09倍,表明羧酸酯酶的解毒代谢在烟粉虱对毒死蜱的抗性中起着重要作用。烟粉虱抗性种群乙酰胆碱酯酶的K_m值是敏感品系的1.83～4.0倍,V _max值是敏感品系的0.34～0.62倍; 敏感品系乙酰胆碱酯酶的活性在底物浓度大于1.0 mmol/L时受抑制,抗性种群乙酰胆碱酯酶的活性在底物浓度大于16 mmol/L时受抑制;抗性种群乙酰胆碱酯酶对敌敌畏和毒死蜱的敏感度分别比敏感品系低119.92～161.33倍和10.11～14.24倍,表明烟粉虱田间抗性种群乙酰胆碱酯酶可能已发生了变构,由变构引起的乙酰胆碱酯酶不敏感是烟粉虱田间种群对毒死蜱和敌敌畏产生抗性的重要原因。结果提示,乙酰胆碱酯酶不敏感性和羧酸酯酶的解毒代谢在烟粉虱对毒死蜱的抗性中均起着重要作用,而乙酰胆碱酯酶不敏感性在对敌敌畏的抗性中起重要的作用,多功能氧化酶和谷胱甘肽S转移酶在烟粉虱对毒死蜱和敌敌畏抗性中所起的作用不大。