亚致死浓度多杀菌素对西花蓟马解毒酶系活力的影响

采用亚致死浓度(LC25)多杀菌素对西花蓟马Frankliniella occidentalis相对敏感(SS)种群进行连续选育,获得亚致死(Sub)种群。处理36代后,Sub种群对多杀菌素的敏感性下降到SS种群的5.2倍。用SS和Sub种群各自的LC10和LC25浓度多杀菌素分别处理两种群的2龄若虫,1、6 、12、24和48 h后测定羧酸酯酶(CarE)、谷胱甘肽S-转移酶(GSTs) 和多功能氧化酶(MFOs)的比活力。结果表明,Sub种群对照组CarE和GSTs比活力在除第48 h外的其他时间段都高于SS种群对照组,且6 h时两者CarE比活力差异显著,Sub种群是SS种群的1.37倍;Sub种群对照组MFOs比活力在各时间段都高于SS种群对照组,在1 和6 h时差异显著,前者分别是后者的1.62和1.36倍。再经各自的LC10和LC25浓度多杀菌素处理后,在各时间段Sub种群的CarE比活力均高于SS种群;LC25浓度处理后,Sub种群的GSTs和MFOs比活力虽在短时间内低于SS种群,但随处理时间的延长其比活力均高于SS种群。说明SS种群经亚致死浓度多杀菌素选育36代后,其体内CarE、GSTs和MFOs比活力有上升趋势;继续用亚致死浓度多杀菌素处理,则Sub种群体内解毒酶活力的动态调节能力要强于SS种群。     

亚致死浓度氯虫苯甲酰胺对小菜蛾药剂敏感度和解毒酶活性的影响

采用叶片药膜法,使用亚致死浓度(LC10、LC25)的氯虫苯甲酰胺对小菜蛾Plutella xylostella(L.)3龄幼虫连续处理5代后,试虫对氯虫苯甲酰胺的敏感度分别比敏感品系下降了57.3% 和67.7%,同时对多杀菌素的敏感度也分别下降了60.2% 和51.5%,但对毒死蜱和高效氯氰菊酯的敏感度变化不明显。采用该浓度的氯虫苯甲酰胺分别处理小菜蛾3龄幼虫24、48和72 h,可诱导其羧酸酯酶(CarE)比活力上升,但对细胞色素P450 O-脱乙基酶(ECOD)、谷胱甘肽S-转移酶(GSTs)和芳基酰胺酶(AA)有明显的抑制作用;连续处理5代后,小菜蛾CarE和ECOD的比活力显著高于对照组,分别为对照组的1.16、1.40倍和1.65、1.56倍,但GSTs和AA的比活力则分别比对照下降了11.0%、27.5%和43.6%、52.5%。结果表明,小菜蛾对氯虫苯甲酰胺产生抗性的风险较高;羧酸酯酶和多功能氧化酶可能与小菜蛾对氯虫苯甲酰胺的敏感度下降有关。     

Resistance selection and mechanisms of oriental tobacco budworm (Helicoverpa assulta Guenee) to indoxacarb

The oriental tobacco budworm, Helicoverpa assulta, is one of the most destructive pests for numerous commercial crops, and these organisms are responsible for enormous economic losses in Chinese agriculture. Insect larvae often feed within host plant fruits, providing protection from many currently used insecticides and making field control of H. assulta very difficult. Owing to its novel mode of action, high insecticidal activity, and low mammalian toxicity, the nonsystemic insecticide indoxacarb has been considered a promising alternative for the control of lepidopterous pests of agricultural significance. Indoxacarb evidences an elevated insecticidal activity against H. assulta. After 13 generations of selection with indoxacarb and bifenthrin insecticides under laboratory conditions, the LC₅₀ of these compounds for H. assulta increased by 4.19-fold and 10.67-fold, respectively. The synergists diethyl maleate (DEM) and triphenyl phosphate (TPP) increased indoxacarb toxicity by 2.76-fold and 4.10-fold in resistant strains and, comparatively, 1.58-fold and 1.75-fold in susceptible strains, suggesting that carboxylesterase (CarE) and glutathione-S-transferases (GSTs) may be involved in the development of indoxacarb resistance in H. assulta. Activity and kinetic parameters observed in detoxification enzymes further demonstrated that the enhanced activity of CarE and GSTs may be critical in development of indoxacarb resistance in H. assulta. The data provides a foundation for further study of the indoxacarb resistance mechanism observed in H. assulta and the rational use of indoxacarb as a rotation insecticide with other insecticide classes for the control of H. assulta.     

多杀菌素亚致死浓度对小菜蛾解毒酶系活力的影响

采用多杀菌素亚致死浓度,以浸叶法分别处理小菜蛾Plutella xylostella (L.)敏感种群(SS)和亚致死选育种群 的3龄幼虫,分别测定饲喂处理6、12、24、48和72 h后小菜蛾体内羧酸酯酶(CarE)、谷胱甘肽S-转移酶(GST) 和多功能氧化酶(MFOs)的活性,分析了酶活性的变化动态。结果表明,SS种群小菜蛾CarE的活性在不同时间段波动较大,经多杀菌素处理后,开始时段比活力增加,随着处理时间的延长,比活力逐渐被抑制,Sub-SS种群的GarE活力高于SS种群;多杀菌素对GST具有明显的诱导作用,亚致死浓度处理后GSTs比活力呈上升趋势,且具有一定的时间效应;对细胞色素P450酶系的O-脱甲基酶活性具有明显的抑制作用,多杀菌素亚致死浓度连续处理5代后,该酶活性更低。     

Selection and characterization of spinosad resistance in Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae)

The cross-resistance and biochemical mechanism of the beet armyworm, Spodoptera exigua (Hübner), to spinosad was studied in the laboratory. S. exigua population were collected from Shanghai suburb. After five generations of selection, the resistance of S. exigua to spinosad increased 345.4 times compared with the susceptible strain. There was no cross-resistance between spinosad and fenvalerate, phoxim, methomyl, abamectin, and cyfluthrin. When the inhibitors, PBO, TPP, DEF, and DEM were used as synergist in the susceptible strain and resistant strain, the synergistic ratio was 0.7-, 0.5-, 1.0-, and 0.6- fold for the susceptible strain, and 9.8-, 1.5-, 2.6-, and 1.5-fold for the resistant strain, respectively. The results revealed that PBO had significant synergistic effect on the resistant strain. The activity in vitro of microsomal-O-demethylase and glutathione S-transferase in the resistant strain was 5.2- and 1.0-fold of the susceptible strain, respectively. The results implied that microsomal-O-demethylase might be important in conferring spinosad resistance in the S. exigua population.     

Fipronil resistance mechanisms in the rice stem borer, Chilo suppressalis Walker

Fipronil resistance mechanisms were studied between the laboratory susceptible strain and the selective field population of rice stem borer, Chilo suppressalis Walker in the laboratory. The borer population was collected from Wenzhou county, Zhejiang province. After five generations of selection, fipronil resistance ratio was 45.3-fold compared to the susceptible strain. Synergism experiments showed that the synergistic ratios of PBO, TPP and DEF on fipronil in susceptible and resistant strains of C. suppressalis were 7.55-, 1.93- and 2.91-fold, respectively, and DEM showed no obvious synergistic action on fipronil. Activities of carboxylesterase and microsomal-O-demethylase in the resistant strain were 1.89- and 1.36-fold higher that in susceptible strain, and no significant difference of glutathione-S-transferase activity was found between the resistant and susceptible strains. The K_m and V_max experiments also demonstrated that fipronil resistance of C. suppressalis was closely relative to the enhanced activities of carboxylesterase and microsomal-O-demethylase. Moreover, cross-resistance between fipronil and other conventional insecticides and the multiple resistant properties of the original Wenzhou’s population were also discussed.     

Fipronil resistance mechanisms in the rice stem borer,Chilo suppressalis Walker

Fipronil resistance mechanisms were studied between the laboratory susceptible strain and the selective field population of rice stem borer, Chilo suppressalis Walker in the laboratory. The borer population was collected from Wenzhou county, Zhejiang province. After five generations of selection, fipronil resistance ratio was 45.3-fold compared to the susceptible strain. Synergism experiments showed that the synergistic ratios of PBO, TPP and DEF on fipronil in susceptible and resistant strains of C. suppressalis were 7.55-, 1.93- and 2.91-fold, respectively, and DEM showed no obvious synergistic action on fipronil. Activities of carboxylesterase and microsomal-O-demethylase in the resistant strain were 1.89- and 1.36-fold higher that in susceptible strain, and no significant difference of glutathione-S-transferase activity was found between the resistant and susceptible strains. The K_m and V_max experiments also demonstrated that fipronil resistance of C. suppressalis was closely relative to the enhanced activities of carboxylesterase and microsomal-O-demethylase. Moreover, cross-resistance between fipronil and other conventional insecticides and the multiple resistant properties of the original Wenzhou’s population were also discussed.     

Beta-cypermethrin resistance associated with high carboxylesterase activities in a strain of house fly, Musca domestica (Diptera: Muscidae)

A housefly strain, originally collected in 1998 from a dump in Beijing, was selected with beta-cypermethrin to generate a resistant strain (CRR) in order to characterize the resistance and identify the possible mechanisms involved in the pyrethroid resistance. The resistance was increased from 2.56- to 4419.07-fold in the CRR strain after 25 consecutive generations of selection compared to a laboratory susceptible strain (CSS). The CRR strain also developed different levels of cross-resistance to various insecticides within and outside the pyrethroid group such as abamectin. Synergists, piperonyl butoxide (PBO) and S,S,S-tributyl phosphorotrithioate (DEF), increased beta-cypermethrin toxicity 21.88- and 364.29-fold in the CRR strain as compared to 15.33- and 2.35-fold in the CSS strain, respectively. Results of biochemical assays revealed that carboxylesterase activities and maximal velocities to five naphthyl-substituted substrates in the CRR strain were significantly higher than that in the CSS strain, however, there was no significant difference in glutathione S-transferase activity and the level of total cytochrome P450 between the CRR and CSS strains. Therefore, our studies suggested that carboxylesterase play an important role in beta-cypermethrin resistance in the CRR strain.     

Assessment of resistance risk in Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae) to chlorantraniliprole

BACKGROUND: Beet armyworm, Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae), is a major pest of numerous cultivated crops. Chlorantraniliprole, the first commercialised ryanodine receptor insecticide from the anthranilic diamide class, has exceptional insecticidal activity on a range of lepidopteran pests. The aim of this study was to assess the resistance of S. exigua to chlorantraniliprole in the laboratory. RESULTS: A field‐collected population of S. exigua was selected after repeated exposure to chlorantraniliprole to determine the risk of resistance evolution. After 22 generations of selection, there was a 12.0‐fold increase in LC₅₀. The realised heritability (h²) of resistance was estimated as 0.1082 by using threshold trait analysis. The projected rate of resistance evolution indicated that, if h² = 0.1082 and 70% of the population was killed at each generation, then a tenfold increase in LC₅₀ would be expected in 21.7 generations for chlorantraniliprole. CONCLUSION: These results show that the risk of resistance development to chlorantraniliprole exists in S. exigua after continuous application. Copyright © 2011 Society of Chemical Industry     

Resistance selection and biochemical mechanism of resistance to two Acaricides in Tetranychus cinnabarinus (Boiduval)

A Tetranychus cinnabarinus strain was collected from Chongqing, China. After 42 generations of selection with abamectin and 20 generations of selection with fenpropathrin in the laboratory, this T. cinnabarinus strain developed 8.7- and 28.7-fold resistance, respectively. Resistance to abamectin in AbR (abamectin resistant strain) and to fenpropathrin in FeR (fenpropathrin resistant strain) was partially suppressed by piperonyl butoxide (PBO), diethyl maleate (DEM) and triphenyl phosphate (TPP), inhibitors of mixed function oxidase (MFO), glutathione S-transferases (GST), and hydrolases, respectively, suggesting that these three enzyme families are important in conferring abamectin and fenpropathrin resistance in T. cinnabarinus. The major resistant mechanism to abamectin was the increasing activities of carboxylesterases (CarE), glutathione-S-transferase (GST) and mixed function oxidase (MFO), and the activity in resistant strain developed 2.7-, 3.4- and 1.4-fold contrasted to that in susceptible strain, respectively. The activity of glutathione-S-transferase (GST) in the FeR strain developed 2.8-fold when compared with the susceptible strain, which meant the resistance to fenpropathrin was related with the activity increase of glutathione-S-transferase (GST) in T. cinnabarinus. The result of the kinetic mensuration of carboxylesterases (CarE) showed that the structure of CarE in the AbR has been changed.     

Selection for imidacloprid resistance in silverleaf whiteflies from the Imperial Valley and development of a hydroponic bioassay for resistance monitoring

A field-collected population of the silverleaf whitefly, Bemisia argentifolii, was selected with the nicotinyl compound, imidacloprid, over 32 generations to determine if resistance would develop when maintained under continuous selection pressure in a greenhouse. Resistance was slow to increase at first with low to moderate levels of resistance (RR from 6- to 17-fold) in the first 15 generations of selection. Further selection steadily led to higher levels of resistance, with the greatest resistance ratio at 82-fold, the gradual rise suggesting the involvement of a polygenic system. At the end of the selection, slopes of probit regressions were substantially steeper than earlier, indicating increased homogeneity of imidacloprid resistance in this strain. A hydroponic bioassay featuring systemic uptake of imidacloprid through roots was developed to monitor the changes in resistance to imidacloprid in the selected whitefly strain and in seven field-collected strains from Imperial Valley, California. Six out of seven field-collected strains exhibited low LC₅₀ values (0·002 to 0·512 mg ml^-1) compared to the selected resistant strain, with one exception where the LC₅₀ was 0·926 mg ml^-1 (RR=15·0). Variation in responses to imidacloprid in the field strains suggest that this technique is sufficiently sensitive to detect differences in susceptibilities of whitefly populations. The imidacloprid-resistant strain showed no cross-resistance to endosulfan, chlorpyrifos or methomyl (RR ranging from 0·4- to 1·5-fold). A low level of cross-resistance was observed to bifenthrin in the IM-R strain at 7-fold. The success of selection for resistance to imidacloprid has serious implications for whitefly control programs that rely heavily on imidacloprid. ©1997 SCI     

Influence of three diets on susceptibility of selected insecticides and activities of detoxification esterases of Helicoverpa assulta (Lepidoptera: Noctuidae)

The oriental tobacco worm, Helicoverpa assulta Guenée, is one of the most destructive pests of tobacco and peppers in China. We determined the susceptibility of H. assulta reared on an artificial diet, chili pepper and tobacco to four insecticides (fenvalerate, phoxim, methomyl, indoxacarb) under laboratory conditions associated with the activities of acetylcholinesterase (AChE), carboxylesterase (CarE) and glutathione S-transferase (GST) in its larvae. H. assulta larvae that were fed with chili pepper were more susceptible to fenvalerate, indoxacarb, and phoxim than those that were fed with tobacco and the artificial diet, but not to methomyl. The larvae that were fed with chili pepper were 3.65-, 2.49-, 1.92- and 2.44-fold more susceptible to fenvalerate, phoxim, methomyl, and indoxacarb than those fed with tobacco, respectively. The AChE activities of H. assulta larvae that were fed with chili pepper and tobacco were 2.12 and 1.07 μmol mg⁻¹ 15 min⁻¹, respectively, almost 2-fold difference. The CarE activity of H. assulta larvae that were fed with chili pepper, tobacco and the artificial diet was 4.12, 7.40 and 7.12 μmol mg⁻¹ 30 min⁻¹, respectively. Similarly, the GST activities of H. assulta larvae that were fed with chili pepper, tobacco and the artificial diet was 52.02, 79.37 and 80.02 μmol mg⁻¹ min⁻¹, respectively. H. assulta larvae that were fed with chili pepper were more resistance to the tested insecticides. The low activities of AChE and the high activities of CarE and GST lead to H. assulta become more susceptible to the tested insecticides.     

甜菜夜蛾敏感品系的获得及其对12种杀虫剂的敏感基线

以高效氯氰菊酯为筛选药剂,通过室内单对汰选获得了甜菜夜蛾敏感品系,并采用浸叶法和点滴法测定该敏感品系对甲氨基阿维菌素苯甲酸盐、氯虫苯甲酰胺等12种常用药剂的敏感基线。浸叶法测得该品系对供试药剂的敏感性由高到低依次为:甲氨基阿维菌素苯甲酸盐(LC50为0.0340 mg/L)、茚虫威、氟啶脲、甲氧虫酰肼、氯虫苯甲酰胺、氟虫双酰胺、高效氯氰菊酯、虫酰肼、多杀菌素、毒死蜱、虫螨腈、灭多威(124.0482mg/L)。点滴法测得12种杀虫剂LD50由小到大依次为:高效氯氰菊酯、茚虫威、甲氧虫酰肼、甲氨基阿维菌素苯甲酸盐、虫酰肼、毒死蜱、多杀菌素、氯虫苯甲酰胺、虫螨腈、氟虫双酰胺、灭多威、氟啶脲。结果表明,该敏感基线可用于甜菜夜蛾的抗药性监测。     

Toxicity,persistence and efficacy of indoxacarb and two other insecticides on Plutella xylostella (Lepidoptera: Plutellidae) immatures in cabbage

Toxicities of indoxacarb on eggs and 5-day-old larvae of diamondback moth, Plutella xylostella L., on cabbage and those of field-aged leaf residues on 5-day-old larvae were determined in the laboratory. The persistence and efficacies of indoxacarb and two other newer insecticides (spinosad and emamectin benzoate) to P. xylostella were tested under field conditions. Results from laboratory bioassays indicate that indoxacarb was highly toxic to P. xylostella larvae through food ingestion, with LC₅₀ and LC₉₀ values of 24.1 and 90.1 mg AI l ^{- 1}, respectively. However, indoxacarb had no significant effects on eggs and larvae through direct contact compared with water control. The toxicity of field-aged leaf residues of indoxacarb (0-, 3-, 5-, 7-, 10-, 14-, 17- and 21-day-old residues) declined slowly and gradually under the field conditions in South Texas. Almost all larvae died on day 5 after feeding on the leaves with 0 - 14-day residue, and the mortalities were as high as 94 and 78% for the 14- and 17-day-old leaf residues. With one application, indoxacarb suppressed P. xylostella larvae below the economic threshold for 14 - 21 days. Two field trials showed that indoxacarb at 0.05 - 0.07 kg AI ha ^{- 1} was effective against P. xylostella, providing marketable cabbage with three applications per season. In addition, indoxacarb was as effective as spinosad, and significantly more effective than emamectin benzoate.     

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.     

甜菜夜蛾对虫酰肼的抗药性研究初报

在室内用虫酰肼对甜菜夜蛾幼虫进行抗药性选育,经过12代汰选,甜菜夜蛾对虫酰肼产生了5.47倍的抗性,抗性发展缓慢。该种群对甲氧虫酰肼有一定交互抗性,对有机磷、氨基甲酸酯、拟除虫菊酯类等多种杀虫剂都不存在交互抗性,抗性比值在0.83~1.63倍之间。     

低剂量乙基多杀菌素对小菜蛾解毒酶的影响

为探讨低剂量乙基多杀菌素对小菜蛾Plutella xylostella（L.）解毒酶的影响,采用叶片浸渍法,测定了乙基多杀菌素和多杀菌素对小菜蛾敏感种群的毒力,并比较了低剂量（LC25和LC50）处理6、12、24、48和72 h时小菜蛾体内羧酸酯酶（CarE）、谷胱甘肽S-转移酶（GST）和多功能氧化酶系（MFOs）活性的变化动态。结果表明:乙基多杀菌素对小菜蛾的杀虫活性优于多杀菌素,处理48 h后其LC25和LC50浓度分别为0.018和0.048 mg/L,经此低剂量浓度处理后,小菜蛾CarE活性波动较大,6~24 h,处理组CarE活性高于对照组,且均呈先升后降趋势,24~72 h,处理组CarE活性均低于对照组,并且具有一定的时间效应;对GST具有明显的诱导作用,GST活性均高于对照组;对MFOs具有明显的抑制作用,除在48 h时相差不大外,其他时间MFOs活性均显著低于对照组。结果表明,GST可能参与了乙基多杀菌素在小菜蛾体内的代谢。     

Pyrethroid insecticide resistance in Rhipicephalus bursa (Acari, Ixodidae)

Rhipicephalus bursa is one of the most common hard tick species in Iran. It is a vector of animal and human diseases. Tick control strategies in Iran rely heavily on pyrethroid insecticide use. Hence, susceptibility status of multiple field collected populations to λ-cyhalothrin and cypermethrin was investigated using the FAO recommended larval packet test. Resistance ratios at the LC₉₉ of field populations of R. bursa compared to a susceptible strain ranged from 0.95 to 4.78 for λ-cyhalothrin and 1.43-8.6 for cypermethrin. They were 2- to 6.9-fold higher than the maximum dose recommended by the acaricide formulating companies. Biochemical assays on different field collected populations showed significant elevation in esterase and glutathione S-transferase activities and monooxygenase contents compared with the susceptible strain. Therefore, close monitoring and resistance management strategies should be employed to delay the operational loss of pyrethroids for tick control in Iran.     

Action of the ecdysteroid agonist tebufenozide in susceptible and artificially selected beet armyworm

Toxicity assays with tebufenozide, the first commercial non-steroidal ecdysteroid agonist, against a laboratory strain of the beet armyworm, Spodoptera exigua (Hübner), demonstrated the promise of this new compound for the control of this important pest. Experiments to select insects artificially from the laboratory strain by continuous exposure of larval instars to corresponding LC₂₅ doses of tebufenozide for over 12 generations (G₀→G₁₂: 14–15 months), revealed no loss in susceptibility to the insecticide for up to five generations. Moreover, retention and fate of ¹⁴C-labelled tebufenozide were investigated using G₆ larvae from the selection experiments and the results compared with those for the susceptible (G₀) larvae. In addition, piperonyl butoxide, an inhibitor of monooxygenases, when ingested by larvae along with tebufenozide, increased the susceptibility of intoxicated larvae to this ecdysteroid agonist, indicating its oxidative metabolism in Spodoptera larvae. © 1998 Society of Chemical Industry.