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.     

中国疫区内苹果蠹蛾微卫星位点的扩增稳定性及遗传多样性

为揭示欧洲苹果蠹蛾微卫星位点在中国苹果蠹蛾群体遗传学研究中的有效性,以12对微卫星引物对采自中国主要疫区新疆、甘肃、黑龙江的8个苹果蠹蛾地理种群的120头个体在各位点的遗传多样性及扩增稳定性进行研究。所选取的12个微卫星位点中,有8个能够稳定扩增,各位点在大多数种群中均显示偏离哈迪-温伯格平衡,各位点上的平均等位基因数量为3.750～12.500,平均观察杂合度Ho为0.025～0.783,平均期望杂合度He为0.284～0.892。位点Cp2.P和Cp4.56分别具有较低的观察杂合度(0.109、0.025)和较高的近交系数Fis(0.806、0.954),说明这2个位点上的杂合子非常缺乏,其余6个位点均具有较高的杂合度水平和等位基因数量,适用于中国疫区内苹果蠹蛾微卫星分子标记研究。     

Insecticide resistance status of the codling moth Cydia pomonella (Lepidoptera: Tortricidae) from Greece

The codling moth Cydia pomonella L. is controlled mostly with chemical insecticides in Greece and control failures have been reported. However, there are no insecticide resistance studies in the country as yet. We examined the insecticide resistance status of 33 and 38 populations of fifth-instar non-diapausing and diapausing larvae, respectively by applying bioassays, biochemical and DNA diagnostics. Diagnostic concentrations of azinphos-methyl, phosalone, deltamethrin, thiacloprid, fenoxycarb, tebufenozide, methoxyfenozide and diflubenzuron were used in bioassays. Almost all populations showed reduced susceptibility to at least one insecticide and approximately half of them to all insecticides examined compared to a laboratory susceptible strain used as reference. However, only one out of six populations tested showed reduced susceptibility in ovicidal tests with fenoxycarb. Cross-resistances were observed among most insecticides, except from the pairs fenoxycarb–phosalone and thiacloprid–phosalone, in non-diapausing larvae. The more obvious biochemical marker associated with the reduced susceptibility observed in both larval instars was elevated cytochrome P₄₅₀ polysubstrate monooxygenases activity, followed by elevated glutathione-S-transferase activity and reduced carboxylesterases activity. Neither sodium channel nor AChE known resistance mutations were found in any of the approximately 1000 individuals of each larval instar screened with diagnostic PCR. Actions for Integrated Resistance Management and application of alternative control methods are discussed.     

不同温度下苹果蠹蛾实验种群生命表

在15、20、25、28、31、34℃的恒温条件下,测定了苹果蠹蛾Cydiapomonella（L．）各发育历期、存活率及产卵量,组建了苹果蠹蛾实验种群生命表,以探讨温度对苹果蠹蛾生长发育和繁殖的影响。在15～31℃范围内,苹果蠹蛾各阶段发育历期随温度的升高而缩短,世代平均发育历期在15、20、25、28、31℃时分别为112．0、46．8、33．3、29．6、26．2天;在34℃时,苹果蠹蛾发育受阻,世代平均发育历期延长为31．4天。成虫寿命亦随温度的升高而缩短;单雌平均产卵量在25℃时最高,为92．7粒;在15和34℃条件下,没有有效卵产生。生命表分析显示,种群趋势指数（I）在20～31℃时均大于1,25℃时苹果蠹蛾种群内禀增长率（rm）和种群趋势指数最高,分别为0．0972和39．70。研究表明,25℃左右是苹果蠹蛾生长发育和繁殖的适宜温度,温度过高或偏低均不利于苹果蠹蛾的生长繁殖。     

光周期和温度对苹果蠹蛾滞育诱导的影响

为探讨光周期和温度对苹果蠹蛾Cydia pomonella滞育诱导的影响,通过人工诱导的方法对其滞育的光周期、温度诱导条件及滞育的敏感光照虫期进行了研究。结果表明:20℃时,光照时间短于12 h,苹果蠹蛾的滞育率可达到100%;25℃时,光周期为L 8∶D 16时滞育率最高,为98.3%;30℃时,光照时间少于10 h,苹果蠹蛾的滞育率可达85%以上。在20、25和30℃条件下,幼虫滞育的临界日长分别为14.21、13.68和13.73 h。苹果蠹蛾对光照反应有2个最敏感时期,即幼虫初孵化阶段和预蛹期阶段,卵期对光照反应不敏感。表明苹果蠹蛾是一种典型的短日照滞育型种类,光周期、温度及其交互作用均对其滞育诱导具有重要影响,其中光周期占主导作用,温度随着光周期起作用,低温对滞育诱导有很好的促进作用。     

苹果蠹蛾对高效氯氟氰菊酯的抗性现状、机制及治理策略

苹果蠹蛾Cydia pomonella属鳞翅目卷蛾科,是全球仁果类水果种植地区最重要的果树害虫之一,也是我国重大农业入侵物种,对全球水果生产造成严重威胁。当前主要采用高效氯氟氯氰菊酯等杀虫剂对苹果蠹蛾进行防治,杀虫剂的频繁使用导致其对高效氯氟氰菊酯产生了抗性。针对苹果蠹蛾对高效氯氟氰菊酯的抗性问题,该文综述了全球苹果蠹蛾种群对高效氯氟氰菊酯的抗性现状,指出由编码细胞色素P450（cytochrome P450,P450）、羧酸酯酶（carboxylesterase,CarE）和谷胱甘肽S-转移酶（glutathione S-transferase,GST）的解毒酶基因过表达导致代谢能力增强是其最普遍的抗性机制,提出合理的抗性治理策略,并对苹果蠹蛾的绿色防控进行展望。     

苹果蠹蛾不同防治方法的控害效应比较

为了及时地在苹果蠹蛾Cydia pomonella(L.)入侵地实施有效的防控策略,通过评价活雌蛾诱捕器诱捕雄蛾数量、在树干上化蛹或越冬的幼虫数量以及对苹果果实的为害程度,比较性信息素诱杀技术、迷向技术和化学防治技术对苹果蠹蛾的控害效应。结果显示,采用迷向法控制苹果蠹蛾的防治效果与化学防治相当,其中在第1代幼虫为害盛末期,蛀果率可控制在1%以下,在第2代幼虫为害盛末期,可控制在2%左右;而诱捕区与对照区(无处理)的蛀果率相当,几乎无控制效应。     

中国及欧洲各国的苹果蠹蛾种群遗传多样性分析

为研究不同国家和地区苹果蠹蛾Cydia pomonella种群的遗传多样性,对采自我国新疆维吾尔自治区和黑龙江省,以及德国、法国和西班牙等欧洲国家的11个种群共60个苹果蠹蛾个体进行基因组重测序,有效测序样本数为56个,共获得8 455 843个群体单碱基突变（single nucleotide polymorphism,SNP）位点。利用SNP数据进行了苹果蠹蛾群体遗传进化树的构建、主成分分析以及遗传结构分析。结果显示,入侵我国黑龙江省的苹果蠹蛾与德国BD种群更为接近,我国新疆维吾尔自治区与黑龙江省苹果蠹蛾种群有一定的分化,前者种群遗传多样性更低。表明我国东北地区和西北地区的苹果蠹蛾种群具有不同的遗传多样性特征,入侵我国的苹果蠹蛾种群可能来源于欧洲。     

苹果蠹蛾半人工饲料与饲养温度的优化

为实现苹果蠹蛾Cydia pomonella(L.)的人工大规模饲养,对国外学者已经报道的半人工饲料配方进行改良,比较了取食5种饲料对苹果蠹蛾生长发育和繁殖的影响,并组建苹果蠹蛾在20、24、28、32℃条件下取食最优配方时的实验种群生命表。结果表明,来自我国西北疫区的苹果蠹蛾,在以豆粕、蔗糖、面粉、小麦胚芽为主要成分并添加酵母粉配制成的配方B饲喂后,与传统饲料配方相比显著影响苹果蠹蛾的发育历期和种群参数,提高存活率至50.24%、缩短幼虫发育历期至20.18 d、增加蛹重至26.54 mg,并提高单雌产卵量至85.30粒。生命表结果显示,24℃时种群趋势指数最高为16.42、单雌产卵量最大为62.44粒,28℃内禀增长率最高为0.08、雌性比例最大为0.55。表明我国西北疫区的苹果蠹蛾在24~28℃条件下,取食优化的半人工饲料利于其种群数量增加。     

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.     

Toxicity of azinphos-methyl to various development stages of the codling moth Cydia pomonella (Lepidoptera: Tortricidae)

The response to azinphos-methyl of different life-stages of the codling moth Cydia pomonella (L.) was studied. A similarity in response to azinphos-methyl (LC(50), LC(90)) was observed in neonate larvae obtained from the first and second generations of field populations. Mortality rates for neonate larvae of a field population cross-bred with a laboratory colony were lower (by a factor of 1.2-2.2) in comparison with field neonate larvae. The mortality rate of larvae from a laboratory colony exposed to artificial diet containing azinphos-methyl gradually decreased at older instars. The mortality rates of fifth-instar larvae were remarkably low when exposed to artificial diet mixed with azinphos-methyl or when topically treated with the insecticide. One- to three-day-old females were more sensitive than males of the same age, whereas the opposite was observed in 14-to 17-day-old adults. Mortality rates of 14- to 17-day-old adults were higher than those under 10 days old. No significant difference in sensitivity to the organophosphorus compound was noticed between the sexes of 7- to 10-day-old adults. Neonate larvae of the codling moth can serve as a target life-stage for various groups of pesticides, and the importance of using such a stage as a standardized methodology for monitoring resistance in the codling moth is discussed.     

枣镰翅小卷蛾成虫的寄主趋向和产卵选择

采用固相微萃取(SPME)和GC-MS分析研究木枣和酸枣挥发物成分,并测试枣镰翅小卷蛾触角电位(EAG)反应、寄主趋向和产卵选择,以探明枣镰翅小卷蛾的寄主选择机制。结果显示,在萌芽期,木枣和酸枣嫩叶的挥发成分均为罗勒烯、2-甲基-2-菠烯、α-法呢烯和邻苯二甲酸二丁酯4种,但相对含量稍有不同。枣镰翅小卷蛾成虫对木枣和酸枣2种寄主都有强烈的EAG反应,且同一寄主上雌蛾的EAG反应值极显著地高于雄蛾,其EAG值是雄蛾的3.3倍。枣镰翅小卷蛾对木枣表现强烈的趋向反应,而对酸枣的趋向不明显,且雌虫的趋向反应显著高于雄虫。木枣上的产卵量显著高于酸枣,且木枣枣吊上的单雌产卵量为307.9粒,极显著地高于酸枣枣吊上的产卵量(182.9粒)。研究表明,枣镰翅小卷蛾雌蛾在寄主选择中起主导作用,木枣是其嗜好寄主。     

苹果蠹蛾卵在梨园中的分布特性和空间格局

布于叶背面.苹果蠹蛾对梨树空间层次的产卵选择在第1代和第2代成虫产卵高峰期均表现为:中层>上层>下层.在第1代卵空间格局的基础上,采用Iwao方法确定了苹果蠹蛾卵量调查的最适抽样数和序贯抽样表.     

Reduced egg viability in codling moth Cydia pomonella (L.) (Lepidoptera: Tortricidae) following adult exposure to novaluron

BACKGROUND: The codling moth, Cydia pomonella (L.), is one of the principal pests of pome fruits in the world. The effects of novaluron, a benzoylurea chitin synthesis inhibitor insecticide registered for use on apples in the USA, on fecundity and egg viability in codling moth were studied under laboratory conditions. Three different exposure methods were investigated: ingestion, contact and topical spray. Additionally, the duration of novaluron sublethal effects was measured subsequent to the three modes of exposure. RESULTS: The fecundity of codling moth adults was not significantly affected by novaluron with any of the exposure methods. However, novaluron did cause significant reductions in the proportion of egg hatch with all three exposure methods. The duration of sublethal effects was 9 days or more for all modes of exposure, but with the topical spray these effects began to diminish after 6 days. CONCLUSION: Novaluron does not affect fecundity in codling moth, but has significant sublethal activity by reducing egg viability subsequent to adult exposure. The topical, contact and ingestion exposures all induce sublethal effects after exposure, and these persist to various degrees throughout codling moth oviposition. A more complete understanding of novaluron's lethal and sublethal activities will help IPM practitioners optimize its use for management of the codling moth. Copyright © 2008 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.     

Insecticide resistance in the tropical bedbug Cimex hemipterus

Insecticide resistance in the bedbug Cimex hemipterus was investigated using 4211 bedbugs collected from three districts of Sri Lanka. Insecticide bioassays were carried out with discriminating dosages of deltamethrin, permethrin, DDT, malathion, and propoxur. Activity levels of insecticide metabolizing enzymes and the insecticide target site acetylcholinesterase were monitored using biochemical assays. Percentage survivals after DDT, malathion, and propoxur exposure were 41-88%, 18-64%, and 11-41%, respectively. For deltamethrin and permethrin, KT₅₀/KT₉₀ (time to knock-down 50%/90% of the population) values were 0.5-24/1.0-58 and 1.3-10/2.5-47 h, respectively. Both elevated esterase and malathion carboxylesterase mechanisms were present in bedbug populations. Monooxygenase levels were heterogeneous. Organophosphate and carbamate target site acetylcholinesterase, was insensitive in 29-44% of the populations. High DDT resistance was probably due to glutathione S-transferases. Malathion carboxylesterases are mainly responsible for high malathion resistance. High tolerance to both DDT and pyrethroids suggests the presence of ‘kdr’ type resistance mechanism in one population.     

Disruption of mating in codling moth (Lepidoptera: Tortricidae) by chlorantranilipole, an anthranilic diamide insecticide

The influence of the anthranilic diamide insecticide chlorantranilipole (DPX-E2Y45; Rynaxypyr against the adult stage of codling moth, Cydia pomonella L., was examined. Insecticide residues in plastic cups sprayed with 56 mg AI L(-1) (equivalent to the recommended field rate) had a minimal effect on adult survival or fecundity, but significantly fewer female moths were mated in treated than in untreated cups. Similar results were found in field studies with adults placed in screened cages on treated apple foliage for up to 3 weeks. Mating was disrupted when either sex was topically dosed with technical material (0.01-0.1 microg). Males were more sensitive to the disruptive effects of chlorantranilipole than females. A 2 h exposure significantly reduced male activity (walking and wing fanning), but not female calling. A 25 h exposure significantly reduced the activity of both sexes. The proportion of males landing near a sex pheromone source in a flight tunnel was significantly reduced following exposures of 2-4 h and completely eliminated following a 17 h exposure. Male moths partially recovered from either topical or residual exposure to chlorantranilipole, but the extent of their recovery was dose and time dependent. Male response to sex pheromone recovered within 2 h following a 4 h exposure, but following a 17 h exposure only a partial recovery occurred after 48 h. Mating by moth pairs exposed to chlorantranilipole residues for 24 h did not significantly increase during recovery periods of 24-96 h, except in cups treated with 10% of the standard concentration. The proportion of field-collected female codling moths caught in pear ester-baited traps that were mated was significantly lower in apple orchards treated season-long with chlorantranilipole compared with females caught in either untreated orchards or those treated with sex pheromones to cause mating disruption.     

Control of the codling moth (cydia pomonella) in apple and pear orchards in Israel by mating disruption

Control of the codling moth,Cydia pomonella (L.) (Lepidoptera: Tortricidae), by pheromone-mediated mating disruption, was studied in Israel using two types of dispensers: Shin-Etsu twist-tie (normal and long-life) and TNO matrix formulation (square wafers). Factors affecting mating disruption: initial pest population density, timing of the pheromone application, early season insecticide treatments and monitoring of the pest, were evaluated. Efficacy of mating disruption was assessed by comparing catch of males in pheromone traps located in pheromone-treated and chemically sprayed control plots. Fruit infestation during the season and at picking time was the major criterion to assess mating disruption. The preferred time for pheromone application was before the first flight of the moth. When pheromones were applied at a later stage, one or two early insecticide sprays had to be used to reduce the initial level of the codling moth population. In all tests, trap catch in pheromone-treated plots was drastically reduced as compared with respective control plots. In pheromone-treated plots where total seasonal trap catch (after pheromone application) was less than 12 males/trap, fruit infestation was less than 1.5%; zero trap catch resulted in 0.1-0.3% infestation. At high population levels, mating disruption was ineffective, with the rate of infestation at picking-time being similar in pheromone-treated and control plots. All the field tests indicated that mating disruption with either Shin-Etsu ropes or TNO squares can control codling moth infestations in apple and pear orchards, at relatively low densities of the pest, as efficiently as commercial insecticide sprays.