Selection for resistance, reversion towards susceptibility and synergism of chlorantraniliprole and spinetoram in obliquebanded leafroller, Choristoneura rosaceana (Lepidoptera: Tortricidae)

BACKGROUND: Obliquebanded leafroller, Choristoneura rosaceana (Harris), is an economic pest of pome fruits throughout North America. Repeated applications of broad‐spectrum insecticides have resulted in the development of resistance, cross‐resistance and multiple resistance in this pest. Studies were conducted to determine the possibility of resistance evolution, stability of resistance and the effect of metabolic synergists on toxicity of new reduced‐risk insecticides chlorantraniliprole and spinetoram in C. rosaceana. RESULTS: Larvae of C. rosaceana were selected for resistance to chlorantraniliprole and spinetoram in the laboratory. Significant levels of resistance to each insecticide were observed after 12 generations of selection. In the absence of selection pressure, susceptibility of a subset of larvae from both chlorantraniliprole‐ and spinetoram‐selected populations reverted to preselection levels after five and six generations respectively, indicating that resistance to both chlorantraniliprole and spinetoram was unstable in C. rosaceana. In synergist bioassays performed after 12 generations of selection, S,S,S‐tributylphosphoro trithioate (DEF) and piperonyl butoxide (PBO) synergized the toxicity of chlorantraniliprole and spinetoram respectively, suggesting the involvement of esterases in chlorantraniliprole resistance and the involvement of mixed‐function oxidases in spinetoram resistance. CONCLUSION: These findings suggest that chlorantraniliprole and spinetoram could be incorporated into C. rosaceana resistance management programs by using rotational strategies. Copyright © 2011 Society of Chemical Industry     

Baseline toxicity and stage specificity of recently developed reduced‐risk insecticides chlorantraniliprole and spinetoram to obliquebanded leafroller,Choristoneura rosaceana (Harris) (Lepidoptera: Tortricidae)

BACKGROUND: Studies were designed to assess baseline toxicity of the two recently developed reduced‐risk insecticides chlorantraniliprole and spinetoram to obliquebanded leafroller (OBLR), Choristoneura rosaceana (Harris), and to determine stage‐specific effects of these products on OBLR, so that these new chemicals could be judiciously incorporated into IPM programs by targeting only the most susceptible stages of OBLR. RESULTS: Both chlorantraniliprole and spinetoram were highly effective against neonate OBLR larvae at much lower doses than conventional standard azinphosmethyl. Most of the mortality caused by spinetoram occurred during the 4 day exposure period, while significant delayed mortality was observed as a result of chlorantraniliprole treatment during the 7 day recovery period, indicating that chlorantraniliprole is a slower‐acting compound compared with spinetoram. Unlike broad‐spectrum predecessors, the toxicity of chlorantraniliprole significantly increased with larval age. Similarly, the toxicity of spinetoram increased as larvae grew from neonates to third instars. CONCLUSION: Chlorantraniliprole and spinetoram are highly effective against OBLR larvae, regardless of larval age. Incorporation of these reduced‐risk chemistries into IPM programs for OBLR would lead to effective management of this pest in tree fruit systems. Copyright © 2011 Society of Chemical Industry     

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.     

Toxicity of chlorantraniliprole to Cry1Ac-susceptible and resistant strains of Helicoverpa armigera

Transgenic Bt cotton expressing Cry1Ac is important in controlling various agricultural pests, including Helicoverpa armigera. Especially for transgenic crops that are cultivated in large expanses, avoiding resistance development is a key for ensuring sustainability of Bt technologies. Integrated pest management, in which transgenic crops are strategically combined with rational pesticide use, may help to prevent H. armigera resistance acquisition in Bt cotton. In this study, we evaluated the toxicity of a novel insecticide (chlorantraniliprole) on Cry1Ac-susceptible and resistant individuals of H. armigera. More specifically, we assessed the effect of chlorantraniliprole on the activity of two enzymes and conducted laboratory bioassays to determine its toxicity on H. armigera larvae. Chlorantraniliprole increased esterase and glutathione-S-transferase activities in Cry1Ac susceptible and resistant populations of H. armigera. Cry1Ac resistant populations XJ-F (Cry1Ac resistance ratio 21.8-fold), XJ-10.0 (95.8-fold) and BTR (3536.5-fold) did not show cross-resistance to chlorantraniliprole, with LC₅₀ values of 0.0733 (μg/mL) in XJ-F, 0.0545 (μg/ml) in XJ-10.0 and 0.0731 (μg/mL) in BTR, which were close to that in the susceptible strain 96S (0.0954 μg/mL). Our work shows that chlorantraniliprole could be considered to be integrated in Bt cotton management schemes to delay the H. armigera resistance development.     

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。     

Metabolic mechanisms involved in the resistance of field populations of Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) to spinosad

Tuta absoluta (Meyrick) resistance to insecticides has become a significant problem in many tomato production areas in South America. New insecticides are now available for the management of this pest (i.e. spinosad), however there is scarce information about their efficacy on field populations.With the aim of determining the susceptibility of T. absoluta to spinosad we evaluated the response of second instar larvae, from five field populations (Azapa 1, Azapa 2, Lluta, Colín and Valdivia) and a laboratory reference strain (S), to a diagnostic concentration of the insecticide. We also determined the activity of the detoxifying enzymes mixed-function oxidases (MFO), glutathione-S-transferases (GST) and esterases (EST) in the same larval stage. Larval mortality in field populations was significantly lower in Azapa 1 (50.0%), Azapa 2 (44.9%), Lluta (39.9%) and Colín (53.5%) when compared to the laboratory strain (91.7%). MFO activities in field populations were between 1.8 and 4.6 times higher than those observed in the S strain, while for EST, the ratio varied from 1.7 to 14.7. The lowest ratios were observed for the GST (0.5-2.7), however, significant differences were detected for the three enzyme systems. We conclude that the evaluated mechanisms would be involved in spinosad resistance of populations of T. absoluta, presenting an increased MFO activity in all populations.     

草地贪夜蛾对甲氨基阿维菌素苯甲酸盐和虱螨脲的抗性风险评估

甲氨基阿维菌素苯甲酸盐(甲维盐)和虱螨脲是目前生产上防治草地贪夜蛾的主要杀虫剂,为评估其抗性风险,以福建省草地贪夜蛾田间种群为研究对象,在实验室抗性汰选品系选育基础上,采用数量遗传学域性状分析法并结合交互抗性测定,进行草地贪夜蛾对上述两种杀虫剂的抗性风险评估。结果表明：非连续汰选11代和10代后,草地贪夜蛾对甲维盐(F₁₁)和虱螨脲(F₁₀)的抗性倍数分别达30.57倍和11.35倍;抗性现实遗传力分别为0.403和0.555,且前半段筛选的抗性遗传力都远大于后半段;药剂在室内对草地贪夜蛾致死率为50%～90%时,对甲维盐和虱螨脲抗性倍数上升10倍需要汰选8～15代和6～12代。交互抗性测定显示,甲维盐汰选品系对氯虫苯甲酰胺、虱螨脲、虫螨腈无交互抗性,对茚虫威和乙基多杀菌素存在一定交互抗性。虱螨脲汰选品系对氯虫苯甲酰胺、甲维盐、茚虫威、虫螨腈和乙基多杀菌素均无交互抗性。结果表明：草地贪夜蛾对甲维盐和虱螨脲存在快速产生抗性的风险,但可通过与无交互抗性药剂轮用来延缓抗性发展。     

Combined detoxification mechanisms and target mutation fail to confer a high level of resistance to organophosphates in Cydia pomonella (L.) (Lepidoptera: Tortricidae)

Despite the frequent and widespread applications of organophosphates against Cydia pomonella this species has developed low levels of resistance to this chemical group. Investigations concerning the mechanisms involved in resistance are scarce, and usually consider only one of the potential mechanisms. With the aim of a better understanding the resistance mechanisms and their possible interaction, four of these mechanisms were investigated simultaneously in one sensitive (Sv) and two resistant strains (Raz and Rdfb) of this insect. Resistant strains displayed an increased mixed function oxidase activity, whereas carboxylesterase activity varied upon the substrate used. The three strains had similar β-naphtyl acetate activity, and the hydrolysis of α-naphthyl acetate and p-nitrophenyl valerate was higher in the Sv strain. The p-nitrophenyl acetate activity was highest in the resistant strains and was strongly inhibited by azinphos and DEF. The Raz strain has a modified acetylcholinesterase (AChE), which resulted in a 0.7-, 3.2- and 21.2-fold decrease in the susceptibility to chlorpyriphos-ethyl-oxon, azinphos-methyl-oxon, and paraoxon-methyl, respectively. These combined resistance mechanisms only conferred to Raz a 0.6-, 7.9- and 3.1-fold resistance to the related insecticides. Organophosphates resistance in C. pomonella results from a combination of mechanisms including modified affinities to carboxylesterase substrates, and increased metabolisation of the insecticide. The apparent antagonism between increased functionalisation and reduced sensitivity of the AChE target is discussed.     

Assessment of insecticide resistance in eggs and neonate larvae of Cydia pomonella (Lepidoptera: Tortricidae)

Spanish Cydia pomonella (L.) field populations have developed resistance to several insecticide groups. Diagnostic concentrations were established as the LC₉₀ calculated on a susceptible strain (S_Spain) for five and seven insecticides and tested on eggs and neonate larvae field populations, respectively. The three most relevant enzymatic detoxification systems (mixed-function oxidases (MFO), glutathione S-tranferases (GST) and esterases (EST)) were studied for neonate larvae.In eggs, 96% of the field populations showed a significantly lower efficacy when compared with the susceptible strain (S_Spain) and the most effective insecticides were fenoxycarb and thiacloprid. In neonate larvae, a significant loss of susceptibility to the insecticides was detected. Flufenoxuron, azinphos-methyl and phosmet showed the lowest efficacy, while lambda-cyhalothrin, alpha-cypermethrin and chlorpyrifos-ethyl showed the highest. Biochemical assays showed that the most important enzymatic system involved in insecticide detoxification was MFO, with highest enzymatic activity ratios (5.1-16.6 for neonates from nine field populations). An enhanced GST and EST activities was detected in one field population, with enzymatic activity ratios of threefold and fivefold for GST and EST, respectively, when compared with the susceptible strain. The insecticide bioassays showed that the LC₉₀ used were effective as diagnostic concentrations. Measures of MFO activity alongside bioassays with insecticide diagnostic concentrations could be used as tools for monitoring insecticide resistance in neonate larvae of C. pomonella.     

Biochemical studies on sheep body louse Bovicola ovis (Schrank) (Phthiraptera: Trichodectidae): comparison of pyrethroid and organophosphate resistant and susceptible strains

Strains of sheep louse Bovicola ovis (Schrank) with various levels of resistance to pyrethroid and one strain with high degree of resistance to organophosphate (OP) insecticides were used to investigate the biochemical mechanisms of insecticide resistance, i.e., enhanced levels of general esterases, specific acetylcholinesterases (AChE), glutathione S-transferase (GST), and mixed function oxidases. Native gel electrophoresis combined with quantitative enzyme assays showed analogous expression profiles of several esterase isozymes in all the strains tested. The determination of the sensitivity of each esterase isozyme to five inhibitors (acetylthiocholine iodide, butyrylthiocholine iodide, paraoxon eserine sulfate, and pCMB) led to the identification of nine esterases in the B. ovis strain. Gel electrophoresis results are supported by enzyme assay studies where, except for the OP resistant strain, no differences in esterase activities were detected in all the pyrethroid resistant and susceptible strains assayed. Statistical analyses demonstrated that some strains have elevated GST activities compared to the susceptible reference strain.     

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.     

溴氰虫酰胺和氯虫苯甲酰胺对三种鳞翅目害虫的毒力作用比较

为比较溴氰虫酰胺和氯虫苯甲酰胺对甜菜夜蛾、玉米螟和小地老虎的作用差异,采用浸卵法、浸叶法、饲喂毒营养液法等测定了溴氰虫酰胺和氯虫苯甲酰胺对3种害虫的卵、3龄幼虫和成虫的毒力,以及对其3龄幼虫的拒食活性。结果表明,溴氰虫酰胺和氯虫苯甲酰胺均无杀卵作用,但能降低初孵幼虫存活率。溴氰虫酰胺对甜菜夜蛾、玉米螟和小地老虎3龄幼虫的LC₅₀值分别为0.11、0.05和0.13 mg/L,相对毒力分别是氯虫苯甲酰胺的2.8、2.7和5.1倍。溴氰虫酰胺对3种害虫成虫的LC₅₀值分别为0.11、0.09和0.22 mg/L,相对毒力分别是氯虫苯甲酰胺的3.9、3.8和2.7倍。溴氰虫酰胺在亚致死浓度0.10、0.05和0.14 mg/L的剂量下分别对甜菜夜蛾、玉米螟和小地老虎3龄幼虫表现出较强的拒食作用,处理48 h后达到最高,拒食率分别为85.58%、81.91%和86.11%。药剂处理试虫虫体皱缩,体节缩短,体重增加率明显低于对照处理。表明溴氰虫酰胺对3种鳞翅目害虫的毒力作用比氯虫苯甲酰胺好,可作为氯虫苯甲酰胺的替代药品。     

松毛虫赤眼蜂对三种农田常用杀虫剂的敏感性

为明确常用杀虫剂对松毛虫赤眼蜂Trichogramma dendrolimi的安全性,室内评估了高效氯氰菊酯、阿维菌素和氯虫苯甲酰胺3种药剂对其成蜂的急性毒力以及田间推荐使用剂量对赤眼蜂各发育阶段的间接影响,并模拟自然情况,测定了不同残留期的3种药剂对赤眼蜂存活和寄生能力的影响.结果表明:以LC₅₀进行评估,对松毛虫赤眼蜂毒力最高的为阿维菌素,其次是高效氯氰菊酯,氯虫苯甲酰胺毒力最小;在田间推荐使用剂量下,阿维菌素、高效氯氰菊酯和氯虫苯甲酰胺的安全系数分别为0.017、0.066和69.175;在推荐使用剂量下,氯虫苯甲酰胺处理含有不同发育阶段赤眼蜂的米蛾Corcyra cephalonica卵后,平均羽化率最高为90.43%,其次是高效氯氰菊酯为56.46%,阿维菌素仅为9.84%;接触0~7 d残留期阿维菌素的玉米叶片24 h,松毛虫赤眼蜂全部死亡,无米蛾卵被寄生,但接触3~7 d残留期氯虫苯甲酰胺的玉米叶片,赤眼蜂的死亡率和寄生米蛾卵数量与清水对照无明显差异.表明在实施释放赤眼蜂的害虫综合治理方案中,应尽量避免使用阿维菌素,而氯虫苯甲酰胺对赤眼蜂风险性极低,具有很好的相容性,可以推广使用.     

Various mechanisms responsible for permethrin metabolic resistance in seven field-collected strains of the German cockroach from Iran, Blattella germanica (L.) (Dictyoptera: Blattellidae)

Insecticide resistance in the German cockroach can be mediated by a number of mechanisms, the most common being enhanced enzymatic metabolism. Seven field-collected strains of German cockroach, Blattella germanica (L.) with various levels of resistance to pyrethroids, five out of which were also cross-resistant to DDT were used in this study. The investigation of possible mechanisms responsible for permethrin resistance was carried out using the synergists PBO, DEF and DMC and biochemical assays, including general esterases, glutathione S-transeferases and monooxygenases assays, using an automated microtitre plate reader. PBO and DEF, the inhibitors of cytochrome p450 monooxygenases and general esterases, respectively, affected permethrin resistance to varying degrees depending on the strain. DDT resistance in five strains were not completely eliminated by the synergist DMC, an inhibitor of glutathione S-transferase enzymes, suggesting that a further non-metabolic resistance mechanism such as kdr-type may be present. This suggestion was further supported by GST assay data, where a little elevation in GST activity was detected in only two strains. The synergist data supported by biochemical assays implicated that cytochrome p450 monooxygenases or hydrolases are involved in permethrin resistance in some strains. However, these results implicated both enhanced oxidative and hydrolytic metabolism of permethrin as resistance mechanism in the other strains. The results of synergist and biochemical studies implicated that all the field-collected permethrin resistant strains have developed diverse mechanisms of resistance, although these strains have been collected from the same geographic area. The change in resistance ratios of some strains by using PBO or DEF is discussed. It is of interest to note that because resistance to permethrin was not completely eliminated by DEF and PBO, it is likely that one or more additional mechanisms are involved in permethrin resistance in every strain studied.     

超高效液相色谱法测定土壤中多杀菌素和乙基多杀菌素的残留

建立了土壤中多杀菌素(spinosad)A和D以及乙基多杀菌素(spinetoram)J和L残留量的超高效液相色谱(UPLC)检测方法。土壤样品经乙腈-5%氯化钠溶液-1 mol/L氢氧化钾溶液提取,固相萃取柱净化,UPLC检测,外标法定量。结果表明:土壤中多杀菌素和乙基多杀菌素的定量限分别为0.1和0.05 mg/kg;最小检出量分别为8.0×10-7和2.8×10-7g;在0.1~2 mg/kg添加水平下,多杀菌素在土壤中的平均回收率为89%~96%,相对标准偏差(RSDs)为2.1%~4.9%;在0.05~0.5 mg/kg添加水平下,乙基多杀菌素在土壤中的平均回收率为86%~93%,RSDs为1.2%~8.1%。该方法提取效果好,具有良好的灵敏度、回收率和重复性。     

Elevated α-esterase levels associated with permethrin tolerance in Aedes aegypti (L.) from Baja California, Mexico

A discriminating dose for permethrin for Aedes aegypti from Baja California, Mexico was determined, and was used to select individuals. Mosquitoes were collected from four different municipalities located in the north and south end of the Baja California Peninsula. Individuals were chosen for further study based on their similar response to the insecticide. We exposed 10 groups of 90 Ae. aegypti females to the discriminating dose (172 μg/ml) and after producing 50% mortality, individuals were divided into two categories: killed and survivors. Each of these groups was dissected to separate the head, thorax, and abdomen. Biochemical tests were performed on the head and thorax to determine resistance-related enzyme activities including: α- and β-esterases, glutathione S-transferase, acetyl cholinesterase, insensitive acetyl cholinesterase, and mixed-function oxidases. The results were compared with those for the susceptible New Orleans strain of Ae. aegypti. All the populations studied showed the consistent presence of α-esterases, with elevated levels in permethrin-selected populations. Although β-esterases and GST levels were present in high proportions, they did not reveal a clear pattern in relation to resistance.     

Reproductive and developmental defects in a malathion-resistant,laboratory-selected population of Drosophila melanogaster

We have examined the chromosomal basis for reproductive and developmental defects that are associated with malathion resistance in a laboratory-selected population of Drosophila melanogaster. Strains homozygous for second or third chromosomes from this population were more resistant to malathion and had greater mixed-function oxidase activity, decreased fertility, and lower egg production when compared with first chromosome or susceptible strains. Some of the strains carrying resistant third chromosomes were developmentally delayed and required a significantly longer time to pupate. Delayed pupation was not associated with increased in vitro degradation of ecdysone by larvae having increased mixed-function oxidase activity, nor could it be reversed by feeding larvae ecdysone. Differences in mixed function oxidase activity among strains homozygous for second or third chromosomes were strongly correlated with malathion resistance but not with fitness. Although both second and third chromosome strains had high mixed-function oxidase activity, only fly extracts from the third chromosome strains oxidatively degraded [³H]juvenile hormone in vitro to a significant extent. A deficit of vitellogenic oocytes and increased egg laying by females in response to topically applied juvenile hormone-I supported the hypothesis that juvenile hormone titer was lower than normal in these strains. The results indicate that different polygenic systems control malathion resistance and associated fitness defects in this selected population of D. melanogaster. Although these systems are partly independent, they overlap due to pleiotropic effects of third chromosomal genes controlling mixed-function oxidase activity on female reproduction.     

Cloning of the acetylcholinesterase 1 gene and identification of point mutations putatively associated with carbofuran resistance in Nilaparvata lugens

Molecular mechanisms of carbofuran resistance in the brown planthopper, Nilaparvata lugens Stål, were investigated. A carbofuran-resistant strain (CAS) showed approximately 45.5- and 15.1-fold resistance compared with a susceptible strain (SUS) and a non-selected field strain (FM), respectively. Activities of the esterase and mixed-function oxidase were approximately 2.8- and 1.6-fold higher, respectively, in the CAS strain than in the SUS strain, suggesting that these enzymes play a minor role in carbofuran resistance. Interestingly, the insensitivity of acetylcholinesterase (AChE) to carbofuran was approximately 5.5- and 3.7-fold higher in the CAS strain compared to the SUS and FM strains, respectively, indicating that AChE insensitivity is associated with carbofuran resistance. Western blot analysis identified two kinds of AChEs, of which the type-1 AChE (encoded from Nlace1, which is paralogous to the Drosophila AChE gene) was determined to be the major catalytic AChE in N. lugens. The open reading frame of Nlace1 is composed of 1989 bp (approximately 74 kD) and revealed 52.5% and 24.3% amino acid sequence identities to those of Nephotettix cincticeps and Drosophila melanogaster, respectively. Screening of point mutations identified four amino acid substitutions (G119A, F/Y330S, F331H and H332L) in the CAS strain that likely contribute to AChE insensitivity. The frequencies of these mutations were well correlated with resistance levels, confirming that they are associated with reduced sensitivity to carbofuran in N. lugens. These point mutations can be useful as genetic markers for monitoring resistance levels in field populations of N. lugens.     

Triazophos resistance mechanisms in the rice stem borer (Chilo suppressalis Walker)

A field population of the rice stem borer (Chilo suppressalis Walker) with 203.3-fold resistance to triazophos was collected. After 8-generation of continuous selection with triazophos in laboratory, resistance increased to 787.2-fold, and at the same time, the resistance to isocarbophos and methamidophos was also enhanced by 1.9- and 1.4-fold, respectively, implying some cross-resistance between triazophos and these two organophosphate insecticides. Resistance to abamectin was slightly enhanced by triazophos selection, and fipronil and methomyl decreased. Synergism experiments in vivo with TPP, PBO, and DEM were performed to gain a potential indication of roles of detoxicating enzymes in triazophos resistance. The synergism results revealed that TPP (SR, 1.92) and PBO (SR 1.63) had significant synergistic effects on triazophos in resistant rice borers. While DEM (SR 0.83) showed no effects. Assays of enzyme activity in vitro demonstrated that the resistant strain had higher activity of esterase and microsomal O-demethylase than the susceptible strain (1.20- and 1.30-fold, respectively). For glutathione S-transferase activity, no difference was found between the resistant and the susceptible strain when DCNB was used as substrate. However, 1.28-fold higher activity was observed in the resistant strain when CDNB was used. These results showed that esterase and microsomal-O-demethylase play some roles in the resistance. Some iso-enzyme of glutathione S-transferase may involve in the resistance to other insecticides, for this resistant strain was selected from a field population with multiple resistance background. Acetylcholinesterase as the triazophos target was also compared. The results revealed significant differences between the resistant and susceptible strain. The V_max and K_m of the enzyme in resistant strain was only 32 and 65% that in the susceptible strain, respectively. Inhibition tests in vitro showed that I₅₀ of triazophos on AChE of the resistant strain was 2.52-fold higher. Therefore, insensitive AChE may also involved in triazophos resistance mechanism of rice stem borer.     

Lethal and sublethal effects of chlorantraniliprole on three species of Rhagoletis fruit flies (Diptera: Tephritidae)

BACKGROUND: Chlorantraniliprole formulated as a 350 g kg^?1 WG (Altacor 35WG) for management of apple maggot Rhagoletis pomonella (Walsh), blueberry maggot R. mendax Curran and cherry fruit fly R. cingulata (Loew) (Diptera: Tephritidae) was evaluated in laboratory assays and field trials. RESULTS: A tarsal contact toxicity bioassay showed that a surface residue of 500 mg L^?1 of chlorantraniliprole caused significantly higher mortality of male and female flies of all species compared with a control. Male apple maggot and blueberry maggot mortality was significantly higher than that for females, but there was similar mortality of male and female cherry fruit flies. An ingestion toxicity bioassay showed that 500 mg L^?1 of chlorantraniliprole in diet caused significantly higher mortality of male and female flies of all species than the control, but there were no significant differences among the sexes. Delayed egglaying by females that had ingested chlorantraniliprole was found, but there were no significant sublethal effects on either the number of eggs laid or the egg hatch. Field trials with apple maggot and cherry fruit fly showed that protection of fruit by chlorantraniliprole was comparable with that of standard broad‐spectrum insecticides. CONCLUSIONS: The present data indicate that chlorantraniliprole has suppressant activity against Rhagoletis fruit flies, preventing fruit infestation primarily through direct lethal effects. Copyright © 2008 Society of Chemical Industry