Current status of insecticide resistance in Q biotype Bemisia tabaci populations from Crete

BACKGROUND: A major problem of crop protection in Crete, Greece, is the control of Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae) with chemical insecticides owing to the rapid development of resistance. The aim of this study was to investigate the establishment of resistance and the underlying mechanisms to major insecticide classes with classical bioassays and known biochemical resistance markers. RESULTS: During a 2005–2007 survey, 53 Q biotype populations were collected. Application history records showed extensive use of neonicotinoids, organophosphates, carbamates and pyrethroids. High resistance levels were identified in the majority of populations (>80%) for imidacloprid (RF: 38–1958×) and α‐cypermethrin (RF: 30–600×). Low resistance levels (RF < 12) were observed for pirimiphos‐methyl. A strong correlation between resistance to imidacloprid and the number of applications with neonicotinoids was observed. Significant correlations were observed between COE and P450‐dependent monoxygenase activity with resistance to α‐cypermethrin and imidacloprid respectively. A propoxur‐based AChE diagnostic test indicated that iAChE was widespread in most populations. Resistance levels for α‐cypermethrin were increased when compared with a previous survey (2002–2003). Differentiation of LC₅₀ values between localities was observed for imidacloprid only. CONCLUSION: Bemisia tabaci resistance evolved differently in each of the three insecticides studied. Imidacloprid resistance seems less established and less persistent than α‐cypermethrin resistance. The low resistance levels for pirimiphos‐methyl suggest absence of cross‐resistance with other organophosphates or carbamates used. Copyright © 2008 Society of Chemical Industry     

Age-specific expression of resistance to a neonicotinoid insecticide in the whitefly Bemisia tabaci

Neonicotinoid insecticides retain a crucial role within many chemical and integrated control strategies for the tobacco whitefly, Bemisia tabaci Gennadius, in spite of the establishment of potent and widespread resistance in many areas. Metabolic resistance mechanisms mediated by overexpression of P450-dependent monooxygenases have been implicated in neonicotinoid resistance in the two most prevalent B. tabaci biotypes. Further characterisation of resistance to the neonicotinoid imidacloprid in populations of both these B- and Q-types is reported.Expression of resistance to imidacloprid was age specific in B- and Q-type strains of B. tabaci. The highest observed resistance ratio at LC(50) expressed in prepupal nymphs was 13, compared with at least 580 in their adult counterparts. For all strains, resistance expressed in immatures was not sufficiently potent to compromise recommended imidacloprid application rates.Targeting neonicotinoids towards immature life stages of B. tabaci may circumvent the protection conferred by current mechanisms of resistance, simultaneously reducing the selection pressures imposed. However, such tactics may enhance the expression of existing resistance mechanisms in immatures, or promote the establishment of novel ones expressed in all life stages.     

Cotton whitefly (Bemisia tabaci) resistance to organophosphate and pyrethroid insecticides in Pakistan

Resistance to three organophosphate and four pyrethroid insecticides was monitored from 1992 to 2000 in field populations of adult whiteflies, Bemisia tabaci, from Pakistan using a leaf-dip method. There was generally a very high resistance to dimethoate and deltamethrin, and a moderate resistance to monocrotophos during 1992 to 1996. From 1997 to 2000, resistance to these insecticides dropped to low levels because of less reliance on them for whitefly control, and introduction of new chemistries with novel modes of action that had no cross-resistance to conventional insecticides. Concurrently, whitefly resistance to acephate, fenpropathrin, lambda-cyhalothrin and bifenthrin mostly remained low. An insecticide resistance management strategy is recommended that particularly emphasizes the rotation of still-effective insecticides from different chemical classes along with the use of novel chemicals and other tactics of integrated pest management.     

北京地区蔬菜烟粉虱种群动态及其对烟碱类杀虫剂的抗药性监测

为了明确北京地区蔬菜烟粉虱的种群发展动态及对常用烟碱类杀虫剂的抗药性,为烟粉虱防治适期的确定及高效杀虫剂的选择提供基础资料及科学指导,2009年调查了北京地区春茬和秋茬蔬菜烟粉虱的种群动态,并监测了该地区2009-2011年度烟粉虱种群对3种烟碱类杀虫剂的抗药性。结果表明,春茬蔬菜烟粉虱在5月中旬种群快速上升,持续到春茬拉秧;秋茬蔬菜烟粉虱在塑料棚内9月上旬种群数量即开始上升,露地出现时间较晚,10月中旬后种群数量下降;塑料棚内种群数量明显高于露地蔬菜烟粉虱数量。烟粉虱2009年对吡虫啉和噻虫嗪的抗药性很高,2010年和2011年抗性程度显著下降,LC50处于133.94～251.16mg/L,属中等抗性水平;而对新型杀虫剂烯啶虫胺的抗药性在近3年内持续升高,抗性倍数由6.68倍升至83.62倍,即由低抗性水平升至高抗性水平。上述调查结果表明,北京地区蔬菜烟粉虱的防治应掌握春季5月中旬前、秋季9月中旬前种群处于低密度时进行。烟粉虱对新型杀虫剂烯啶虫胺的抗药性在近三年内快速上升,生产中应注意药剂的轮换使用。     

The role of B‐type esterases in conferring insecticide resistance in the tobacco whitefly,Bemisia tabaci (Genn)

Separation of non‐specific esterases on electrophoretic gels has played a key role in distinguishing between races or biotypes of the tobacco whitefly, Bemisia tabaci. One intensively staining esterase in particular (termed E_0.14) has assumed significance as a diagnostic of B‐type whiteflies (aka Bemisia argentifolii), despite any knowledge of its biological function. In this study, a whitefly strain (B‐Null) homozygous for a null allele at the E_0.14 locus that had been isolated from a B‐type population was used to demonstrate a significant role for E_0.14 in resistance of B‐type populations to pyrethroids but not to organophosphates (OPs). Bioassays with pyrethroids, following pre‐treatment with sub‐lethal doses of the OP profenofos (to inhibit esterase activity), coupled with metabolism studies with radiolabelled permethrin, supported the conclusion that pyrethroid resistance in a range of B‐type strains expressing E_0.14 was primarily due to increased ester hydrolysis. In the same strains, OP resistance appeared to be predominantly conferred by a modification to the target‐site enzyme acetylcholinesterase. © 2000 Society of Chemical Industry     

新型双向传导杀虫剂—螺虫乙酯对Q型烟粉虱的防治效果

为了筛选出防治Q型烟粉虱的有效药剂,研究了22.4%螺虫乙酯悬浮剂不同浓度、不同施药时间对烟粉虱卵、若虫的防治效果。螺虫乙酯和对照药阿维菌素均对Q型烟粉虱卵无效,螺虫乙酯对若虫具有非常高的杀虫活性,无论在卵期施药、若虫低龄期施药还是高龄期施药,防效均达95%以上,和对照药阿维菌素的防效无显著性差异。施药后20d接虫,防效大幅降低到21.6%以下,施药30d后接虫对若虫无效。根据烟粉虱的发育历期推算,表明螺虫乙酯持效期可达30d。     

Lack of fitness costs associated with pyriproxyfen resistance in the B biotype of Bemisia tabaci

BACKGROUND: The insect growth regulator pyriproxyfen has provided effective control of the whitefly Bemisia tabaci Gennadius in many countries. Here, whether or not fitness costs were associated with pyriproxyfen resistance in a laboratory‐selected resistant strain (QC02‐R) of the B biotype was determined. RESULTS: Mortality caused by pyriproxyfen and fitness traits over time were measured in unselected and selected hybrid strains, which were created by crossing individuals of the resistant strain with individuals of a susceptible strain. Fitness costs were not associated with resistance in QC02‐R, as mortality caused by pyriproxyfen did not increase over time in unselected hybrid strains and fitness traits were similar in unselected and selected hybrid strains. Using a new method to examine the inheritance of resistance, based on data from fitness cost experiments, it was estimated that pyriproxyfen resistance is controlled by two loci in the QC02‐R strain. CONCLUSION: The lack of fitness costs associated with pyriproxyfen resistance could promote the evolution of resistance in field populations with similar traits to QC02‐R. Copyright © 2008 Society of Chemical Industry     

Development of a low-density DNA microarray for diagnosis of target-site mutations of pyrethroid and organophosphate resistance mutations in the whitefly Bemisia tabaci

BACKGROUND: Rapid and accurate detection of mutations related to insecticide resistance is essential for development of resistance management strategies to support sustainable agriculture. The M918V, L925I and T929V mutations of the voltage‐gated sodium channel gene (vgsc) and the F392W mutation of the acetylcholinesterase I gene (ace1) are reportedly associated with resistance to pyrethroids and organophosphates, respectively, in Bemisia tabaci. In order to detect known base substitutions in the ace1 and vgsc genes, a low‐density microarray with an allele‐specific probe was developed. RESULTS: Specific regions of the ace1 and vgsc gene mutations were amplified by multiplex asymmetrical PCR using Cy3‐labelled primers, and then the PCR products were hybridised on the microarray. After analysing the probe signal data, the microarray containing 12 allele‐specific probes produced a unique pattern of probe signals for field DNA samples of B. tabaci. To determine the optimal cut‐off value of each probe, receiver operating characteristic (ROC) curve analysis was conducted using SPSS. Among 60 individual samples, microarray data for 57 samples were consistent with direct sequencing data. CONCLUSION: Although many molecular detection methods have been employed to monitor insecticide resistance, the present microarray provides rapid and accurate identification of target mutations in B. tabaci for resistance management. Copyright © 2011 Society of Chemical Industry     

6种杀虫剂对Q型烟粉虱的田间防治效果及抗性测定

本文测试了6种常用杀虫剂对黄瓜上Q型烟粉虱的田间防治效果,并进行室内毒力测定,以确定Q型烟粉虱对不同药剂的抗性。结果表明,施药后第3天,阿维菌素防治效果达78.8%,其次为苦参&;#8226;内酯,防效达65.1%,其他药剂防效低于45%;施药7d后,6种供试杀虫剂防效均下降到45%以下,且药剂间的差异不显著。毒力测定显示,Q型烟粉虱对6种常用杀虫剂均产生不同程度的抗性,且抗性水平高于B型。其中,Q型对高效氯氰菊酯抗性（抗性倍数47.69）显著高于B型（抗性倍数7.19）;对阿维菌素抗性则相对较低,抗性指数仅为4.40;对苦参碱、啶虫脒和吡虫啉的抗性居中（抗性倍数4.57~9.44）。     

湖北省棉花主产区烟粉虱生物型分布及系统发育分析

［目的］ 明确武汉、荆州、孝感、随州、武穴5个棉花主产区烟粉虱生物型组成。［方法］ 通过以线粒体细胞色素氧化酶I（mitochondrial cytochrome oxidase I, mtCOI）为基础的PCR RFLP和基因测序的方法,鉴定湖北5个棉花主产区烟粉虱地理种群的生物型组成,并分析5个地理种群与其他省种群的系统发育关系。［结果］ Q型烟粉虱为湖北省棉花主产区的优势种群,而武汉温室种群则均为B型烟粉虱,土著生物型ZHJ1型在大田中略有分布;系统发育分析表明,湖北省棉花主产区B型和Q型烟粉虱种群均与中国其他多个省的烟粉虱种群亲缘关系较近。［结论］ 本文研究结果可以为湖北省5个棉花主产区烟粉虱的有效防治提供参考。     

近10年我国烟粉虱发生为害及防治研究进展

烟粉虱是我国重要农业害虫,近10年来我国烟粉虱发生为害呈现以下特点:烟粉虱的优势生物型/隐种由B型更替为Q型;传播植物病毒(如黄化曲叶病毒、番茄褪绿病毒)成为了烟粉虱重要的为害方式;烟粉虱抗药性问题逐渐突出;化学农药尤其新烟碱类杀虫剂的广泛使用是Q型烟粉虱取代B型烟粉虱的关键因素。烟粉虱的综合防控措施日益完善,抗药性治理与非化学防控措施受到重视,但是依靠农药的现状并未完全改变。本文综述了近10年来我国烟粉虱的发生为害及防治方面的概况,探讨了今后烟粉虱防控方面的研究内容。     

烟粉虱抗、感杀虫剂品系内生菌群落的差异分析

本研究对B型和Q型烟粉虱的吡虫啉或吡丙醚的抗、感品系进行16SrRNA细菌多样性测序分析,以明确内共生菌的种类和数量与烟粉虱生物型和抗性的关系。结果表明Candidatus Nitrososphaera、Bacillus、Sphingobium只在Q型烟粉虱中存在,而Candidatus Hamiltonella、Lactobacillus、Helicobacter等只在B型烟粉虱中存在,说明这些内共生菌可能与烟粉虱B、Q两种生物型的分化有关;内共生菌Rickettsia、Candidatus Portiera、Candidatus Hamiltonella、S24-7、Prevotella等在烟粉虱的新烟碱类杀虫剂吡虫啉抗性品系中明显多于敏感品系,其可能与烟粉虱对吡虫啉的抗性有关;在对保幼激素类似物杀虫剂吡丙醚的抗性品系中,Candidatus Hamiltonella-OTU7、Candidatus Portiera-OTU5、Candidatus Portiera-OTU190、Rickettsia-OTU4、Rickettsia-OTU2的数量明显多于敏感品系,说明这些内共生菌可能与烟粉虱对该类药物的抗性有关。     

Biochemical study of resistance to imidacloprid in B biotype Bemisia tabaci from Guatemala

Systemic uptake bioassays using excised cotton leaves confirmed resistance to imidacloprid in a Guatemalan population of the tobacco whitefly Bemisia tabaci Gennadius. Polyacrylamide gel electrophoresis of naphthyl esterases identified the insects as B-types. Upon collection from the field, resistance was determined to be 58-fold relative to a susceptible strain originating in the Imperial Valley of California. Resistance levels increased to 126-fold in this population during its continuous exposure to systemically treated cotton. In biochemical investigations, there was no detectable NADPH-dependent mixed function oxidase metabolism of 14C-imidacloprid at any time during the selection process. In contrast, microsomal preparations from housefly abdomens readily produced significant amounts of the mono-hydroxy and olefin derivatives of the parent compound. Detoxification of imidacloprid by housefly MFOs may account for reports of lower toxicity of the insecticide towards this insect compared with whiteflies, despite similar binding properties between imidacloprid and the nicotinic acetylcholine receptors in both species.     

宁夏地区烟粉虱对5种常用杀虫剂的抗药性

采用浸叶法分别测定了宁夏8个地区烟粉虱成虫田间种群对4类5种杀虫剂的抗性.结果 表明,供试烟粉虱种群对高效氯氰菊酯已产生了中等至极高水平抗性,抗性倍数为33.31～227.98倍;对烯啶虫胺产生了中等至高水平抗性,抗性倍数为12.14～69.33倍;对毒死蜱和吡虫啉产生了低至中等水平抗性,抗性倍数分别为7.63～38....     

Toxicity of spiromesifen to the developmental stages of Bemisia tabaci biotype B

BACKGROUND: Spiromesifen is a novel insecticidal/acaricidal compound derived from spirocyclic tetronic acids that acts effectively against whiteflies and mites via inhibition of acetyl‐CoA‐carboxylase, a lipid metabolism enzyme. The effects of spiromesifen on the developmental stages of the whitefly Bemisia tabaci (Gennadius) were studied under laboratory conditions to generate baseline action thresholds for field evaluations of the compound. RESULTS: Adult B. tabaci mortality rate after spiromesifen treatment (5 mg L^?1) was 40%. Treatment with 0.5 mg L^?1 reduced fecundity per female by more than 80%, and fertility was almost nil. LC₅₀ for eggs was 2.6 mg L^?1, and for first instar 0.5 mg L^?1. Scanning electron microscopy revealed that eggs laid by treated adult females had an abnormally perforated chorion, and females were unable to complete oviposition. Light and fluorescent microscopy showed significantly smaller eggs following treatment, and smaller, abnormally formed and improperly localized bacteriomes in eggs and nymphs. The number of ovarioles counted in females treated with 5 mg L^?1 was significantly reduced. Spiromesifen showed no cross‐resistance with other commonly used insecticides from different chemical groups, and resistance monitoring in Israel showed no development of field resistance to this insecticide after 1 year of use. CONCLUSION: The strong effect on juvenile stages of B. tabaci with a unique mode of action and the absence of cross‐resistance with major commonly used insecticides from different chemical groups suggest the use of spiromesifen in pest and resistance management programmes. Copyright © 2008 Society of Chemical Industry     

Insecticide resistance in field populations of Bemisia tabaci (Hemiptera: Aleyrodidae) in West Africa

BACKGROUND: The tobacco whitefly, Bemisia tabaci Gennadius (Hemiptera: Aleyrodidae), has developed a high degree of resistance to several chemical classes of insecticides throughout the world. To evaluate the resistance status in West Africa, eight insecticides from different chemical families were tested using the leaf‐dip method on four field populations collected from cotton in Benin, Togo and Burkina Faso. RESULTS: Some field populations showed a significant loss of susceptibility to pyrethroids such as deltamethrin [resistance ratio (RR) 3–5] and bifenthrin (RR 4–36), to organophosphates (OPs) such as dimethoate (RR 8–15) and chlorpyrifos (RR 5–7) and to neonicotinoids such as acetamiprid (RR 7–8) and thiamethoxam (RR 3–7). Bemisia tabaci was also resistant to pymetrozine (RR 3–18) and to endosulfan (RR 14–30). CONCLUSION: The resistance of B. tabaci to pyrethroids and OPs is certainly due to their systematic use in cotton treatments for more than 30 years. Acetamiprid has been recently introduced for the control of whiteflies. Unfortunately, B. tabaci populations from Burkina Faso seem to be already resistant. Because cross‐resistance between these compounds has never been observed elsewhere, resistance to neonicotinoids could be due to the presence of an invasive B. tabaci biotype recently detected in the region. Copyright © 2010 Society of Chemical Industry     

Biochemical and molecular mechanisms of diafenthiuron resistance in the whitefly,Bemisia tabaci

B-biotype Bemisia tabaci has developed high levels of resistance to many insecticides. To investigate the risks and explore possible mechanisms of resistance to diafenthiuron in B. tabaci, a 32.8-fold diafenthiuron-resistant strain (R-DfWf) was established after selection for 36 generations compared with the susceptible strain (S-Lab). Biochemical assays showed that the activity of cytochrome P450 towards p-NA was significantly higher (4.37-fold higher) in the R-DfWf strain than in the S-Lab strain. Similarly, the carboxylesterase (COE) activity and glutathione S-transferase (GST) activity were also significantly higher (3.12- and 1.83-fold higher, respectively) in the R-DfWf strain than in the S-Lab strain. The expression of five of seven P450 genes was significantly higher (>3-fold) in the R-DfWf strain than in the S-Lab strain. The expression of COE2 was significantly higher (>2.5-fold) in the R-DfWf than in the S-Lab strain. The expression of GST and GST2 was significantly higher (>2.3-fold) in the R-DfWf than in the S-Lab. Thus, cytochrome P450, COE and GST may appear to be responsible for the resistance to diafenthiuron in B. tabaci. It is also valuable for usage of insecticides for resistance management and control of this species.     

Gene expression in pyriproxyfen-resistant Bemisia tabaci Q biotype

Pyriproxyfen is a biorational insecticide that acts as a juvenile hormone (JH) analogue and disrupts insect development with an unknown molecular mode of action. Pyriproxyfen is one of the major insecticides used to control the whitefly Bemisia tabaci (Gennadius) and comply with integrated pest management (IPM) programmes, resulting in minimal effects on the environment, humans and beneficial organisms. During the last few years, resistance to pyriproxyfen has been observed in several locations in Israel, sometimes reaching a thousandfold or more. No information exists about the molecular basis underlying this resistance that may lead to understanding the mode of action of pyriproxyfen and developing molecular markers for rapid monitoring of resistance outbreaks. In this communication, a cDNA microarray from B. tabaci was used to monitor changes in gene expression in a resistant B. tabaci population. Based on statistical analysis, 111 expressed sequence tags (ESTs) were identified that were differentially upregulated in the resistant strain after pyriproxyfen treatment. Many of the upregulated ESTs observed in the present study belong to families usually associated with resistance and xenobiotic detoxification such as mitochondrial genes, P450s and oxidative stress, genes associated with protein, lipid and carbohydrate metabolism and others related to JH-associated processes in insects such as oocyte and egg development.     

Susceptibility to insecticides in the Q biotype of Bemisia tabaci is correlated with bacterial symbiont densities

BACKGROUND: The presence of symbiotic microorganisms may influence an insect's ability to tolerate natural and artificial stress agents such as insecticides. The authors have previously shown that Rickettsia in the B biotype of the whitefly Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae) increases this insect's susceptibility to several insecticidal compounds. This communication reports a comparison of the susceptibilities of three isofemale strains of the Q biotype of B. tabaci harbouring different bacterial complements to major insecticides from different chemical groups: one strain harboured only Arsenophonus, one harboured Rickettsia and Arsenophonus and one harboured Arsenophonus and Wolbachia. RESULTS: The presence of different symbiont combinations in the three strains had a significant influence on their susceptibility to most of the insecticides tested. Thiamethoxam, imidacloprid, pyriproxyfen and spiromesifen had a significant influence on strains that had the double infections Rickettsia–Arsenophonus and Wolbachia–Arsenophonus, which also carried higher amounts of symbionts as assessed by quantitative real‐time PCR. No significant differences in mortality rates were observed when the tested strains were treated with diafenthiuron. CONCLUSION: The results suggest a correlation between the presence of high bacterial densities in B. tabaci and the insect's ability to detoxify toxic compounds such as insecticides. Copyright © 2009 Society of Chemical Industry