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1.
We investigated the mechanisms of resistance to α-cypermethrin in a Q biotype, highly resistant Bemisia tabaci strain (GRMAL-RP) isolated from Crete. Cytochrome P450-dependent monoxygenase activity with the substrate ethoxycoumarin, and carboxylesterase activity with the substrates α-naphthyl-acetate, β-naphthyl-acetate, and para-nitrophenol acetate were substantially elevated in the GRMAL-RP, compared to the susceptible SUD-S strain, while glutathione-S-transferase activity with the substrate 1-chloro-2,4-dinitrobenzene was not different. The metabolic inhibitors piperonyl butoxide and S,S,S-tributyl phosphorotrithioate synergised cypermethrin toxicity in the GRMAL-RP strain, however, mortality was still lower than that of the susceptible strain, indicating the presence of an additional resistance mechanism. Analysis of the sequence of the IIS4-IIS6 region of the para sodium channel gene of the GRMAL-RP strain revealed two amino acid replacements compared to that of the SUD-S susceptible strain. One is the leucine to isoleucine substitution at position 925 (L925I) previously implicated in B. tabaci pyrethroid resistance and the other is a novel kdr resistant mutation for B. tabaci, a threonine to valine substitution at position 929 (T929V). Genotype analysis showed that the L925I and T929V were present in all GRMAL-RP males tested, at an approximately 1:1 frequency, but never in combination in the same haplotype.  相似文献   

2.
RNA-seq data analysis of cigarette beetle (Lasioderma serricorne) strains having different sensitivities to pyrethroids identified sodium channel mutations in strains showing pyrethroid resistance: the T929I and F1534S mutations. These results suggest that reduced sensitivity of the sodium channel confers the pyrethroid resistance of L. serricorne. Results also showed that the F1534S mutation mostly occurred concurrently with the T929I mutation. The functional relation between both mutations for pyrethroid resistance is discussed.  相似文献   

3.
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  相似文献   

4.
In addition to the allele frequencies of the L1014F and T929I mutations which are involved in nerve-insensitive resistance to a pyrethroid, those of the M918I mutation were examined using field strains obtained in China, Thailand, and Japan during 2009-2011. Results show that the resistance allele frequencies at the L1014F site were 89-100%, 97-100% and 65-85%, respectively, for strains in China, Thailand, and Japan. The respective allele frequencies at the T929I site were 86-100%, 70-97% and 58-84% for Chinese, Thai, and Japanese strains. With low frequencies up to 27%, M918I was found in Japan and China, but not in Thailand. The strain homozygous for the M918I and L1014F mutations was established and its resistance level to a pyrethroid was examined. The strain lacks a portion of the sodium channel gene corresponding to the 3′ portion of exon 18a, intron 18, and the 5′ portion of exon 18b. Nevertheless, the strain showed a similar level of resistance to that which was homozygous for the T929I and L1014F mutations.  相似文献   

5.
BACKGROUND: Trialeurodes vaporariorum Westwood is an important pest of protected crops in temperate regions of the world. Resistance to pyrethroid insecticides is long established in this species, but the molecular basis of the mechanism(s) responsible has not previously been disclosed. RESULTS: Mortality rates of three European strains of T. vaporariorum to the pyrethroid bifenthrin were calculated, and each possessed significant resistance (up to 662‐fold) when compared with a susceptible reference strain. Direct sequencing revealed three amino acid substitutions in the para‐type voltage‐gated sodium channel (the pyrethroid and DDT target site) of bifenthrin‐resistant T. vaporariorum at positions previously implicated with pyrethroid or DDT resistance (M918L, L925I and T929I) in other related species. CONCLUSION: This study indicates that resistance to bifenthrin in T. vaporariorum is associated with target‐site insensitivity, and that the specific mutations in the sodium channel causing resistance may differ between localities. Copyright © 2012 Society of Chemical Industry  相似文献   

6.
Head lice resistance to permethrin is mainly conferred by the knockdown resistance (kdr) trait, a voltage-sensitive sodium channel (VSSC) insensitivity factor. Three VSSC mutations (M815I, T917I and L920F) have been identified. Functional analysis of the mutations using the house fly VSSC expressed in Xenopus oocytes revealed that the permethrin sensitivity is reduced by the M827I (M815I) and L932F (L920F) mutations when expressed alone but virtually abolished by the T929I (T917I) mutation, either alone or in combination. Thus, the T917I mutation is primarily responsible for permethrin resistance in head lice. Comparison of the expression rates of channel variants indicates that the M815I mutation may play a role in rescuing the decreased expression of channels containing T917I. A step-wise resistance monitoring system has been established based on molecular resistance detection techniques. Quantitative sequencing (QS) has been developed to predict the VSSC mutation frequency in head lice at a population basis. The speed, simplicity and accuracy of QS made it an ideal candidate for a routine primary resistance monitoring tool to screen a large number of wild louse populations as an alternative to conventional bioassay. As a secondary monitoring method, real-time PASA (rtPASA) has been devised for more precise determination of low resistance allele frequencies. To obtain more detailed information on resistance allele zygosity, as well as allele frequency, serial invasive signal amplification reaction (SISAR) has been developed as an individual genotyping method. Our approach of using three tiers of molecular resistance detection should facilitate large-scale routine resistance monitoring of permethrin resistance in head lice using field-collected samples.  相似文献   

7.
BACKGROUND: The tomato red spider mite, Tetranychus evansi (Baker and Pritchard), is a serious pest of solanaceous crops in many African countries. In this study an investigation has been conducted to establish whether mutation of the para‐type sodium channel underlies pyrethroid resistance in T. evansi strains collected in Southern Malawi. RESULTS: Two T. evansi strains from Malawi showed tolerance to the organophosphate chlorpyrifos and resistance (20–40‐fold) to the pyrethroid bifenthrin, but were susceptible to two contemporary acaricides (abamectin and fenpyroximate) in insecticide bioassays. Cloning of a 3.1 kb fragment (domains IIS5 to IVS5) of the T. evansi para gene from pyrethroid‐resistant and pyrethroid‐susceptible strains revealed a single non‐synonymous mutation in the resistant strains that results in an amino acid substitution (M918T) within the domain II region of the channel. Although novel to mites, this mutation confers high levels of resistance to pyrethroids in several insect species where it has always been associated with another mutation (L1014F). This is the first report of the M918T mutation in the absence of L1014F in any arthropod species. Diagnostic tools were developed that allow sensitive detection of this mutation in individual mites. CONCLUSION: This is the first study of pyrethroid resistance in T. evansi and provides contemporary information for resistance management of this pest in Southern Malawi. Copyright © 2011 Society of Chemical Industry  相似文献   

8.
BACKGROUND: The pyrethroid resistance of the diamondback moth Plutella xylostella (L.) is conferred by increased gene expression of cytochrome P450 to detoxify the insecticide and/or through gene mutation of the sodium channel, which makes the individual insensitive to pyrethroids. However, no information is available about the correlation between the increased metabolic detoxification and the target insensitivity in pyrethroid resistance. RESULTS: Frequencies of pyrethroid‐resistant alleles (L1014F, T929I and M918I) and two resistance‐related mutations (A1101T and P1879S) at the sodium channel and expression levels of the cytochrome P450 gene CYP6BG1 were examined individually using laboratory and field strains of P. xylostella. Real‐time quantitative PCR analysis using the laboratory strains revealed that levels of larval expression of the resistant strain, homozygous for the pyrethroid‐resistant alleles other than the M918I, are significantly higher than those of the susceptible strain. In the field strains, the expression levels in insects having the same resistant alleles as those of the resistant strains varied greatly among individuals. The expression levels were not significantly higher than those in the heterozygotes. CONCLUSION: Significant correlation between the target insensitivity and the increased metabolic detoxification in pyrethroid resistance of P. xylostella was observed in the laboratory but not in the field. Copyright © 2010 Society of Chemical Industry  相似文献   

9.
BACKGROUND: Resistance to numerous insecticide classes in Bemisia tabaci Gennadius has impaired field control efficacy in south‐eastern China. The biotype and resistance status of B. tabaci collected from these areas was investigated. RESULTS: Two different biotypes of B. tabaci (B‐biotype and Q‐biotype) were detected in south‐eastern China, and the samples collected from geographical regions showed a prevalence of the Q‐biotype and the coexistence of B‐ and Q‐biotypes in some regions. Moderate to high levels of resistance to two neonicotinoids were established in both biotypes (28–1900‐fold to imidacloprid, 29–1200‐fold to thiamethoxam). Medium to high levels of resistance to alpha‐cypermethrin (22–610‐fold) were also detected in both biotypes. Four out of 12 populations had low to medium levels of resistance to fipronil (10–25‐fold). Four out of 12 populations showed low levels of resistance to spinosad (5.7–6.4‐fold). All populations tested were susceptible to abamectin. CONCLUSION: The Q‐biotype B. tabaci is supplanting the B‐biotype which used to be ubiquitous in China. Field populations of both B‐ and Q‐biotypes of B. tabaci have developed high levels of resistance to imidacloprid and thiamethoxam. Abamectin is the most effective insecticide against adult B. tabaci from all populations. Copyright © 2010 Society of Chemical Industry  相似文献   

10.
BACKGROUND: B and Q biotypes of the whitefly, Bemisia tabaci (Gennadius), are generally regarded as the most significant given their global distribution and strong resistance to insecticides. Since these biotypes can coexist and differ markedly in their insecticide resistance profiles, a rapid but reliable means of discriminating between them would be a valuable complement to resistance monitoring and management programmes. Recently, PCR‐based methods have been developed to determine the biotype status of B. tabaci populations. However, these require post‐amplification procedures, which increase time and labour. RESULTS: The authors have developed an allelic discrimination real‐time PCR assay using fluorescent dye‐labelled probes to distinguish the B and Q biotypes. The assay targets a single nucleotide polymorphism (SNP) in the mitochondrial cytochrome oxidase I (mtCOI) gene. To evaluate the assay, DNA was extracted from individual whiteflies of six known biotype strains, and all scored correctly as either a B or Q biotype. As further validation, 72 individuals from field samples collected in different parts of the world were also tested by the assay. No failed reactions were observed, with all 72 samples scoring clearly as either the B or Q biotype. CONCLUSION: The development of this rapid and high‐throughput assay has important potential for routine monitoring of B and Q biotypes on ornamental plants and for the screening of B. tabaci populations in countries where these biotypes are not yet established. Copyright © 2007 Society of Chemical Industry  相似文献   

11.
Insecticides have been extensively used for house fly control in China, with dichlorvos and deltamethrin being widely used. Knowledge about the current status of insecticide resistance and the underlying genetic changes is crucial for developing effective fly control strategies. The susceptibility to dichlorvos and deltamethrin, and the frequencies of genetic mutations involved in insecticide resistance were studied in five field populations of the house fly collected across China. Bioassay results show that flies exhibit 14- to 28-fold resistance to dichlorvos and 41- to 94-fold resistance to deltamethrin, indicating that dichlorvos and deltamethrin resistance are common in house fly populations in China. Molecular analysis reveals that flies from the five various locations carry resistance alleles at multiple loci and have diverse allelic types, different relative frequencies and combinations of each allele. Four non-synonymous single nucleotide polymorphisms (SNPs) (i.e. V260L, G342A/V, F407Y) in acetylcholinesterase (Ace) and two mutations (W251L/S) in a carboxylesterase (MdαE7) were commonly present in the field house flies. The L1014H rather than L1014F mutation in the voltage sensitive sodium channel gene (Vssc) was widely distributed in Chinese house flies. CYP6D1v1, which confers pyrethroid resistance, was found in all the five tested populations in China, although its frequency in house fly from Shandong province was very low. Our results suggest that resistance monitoring and management of house flies should be customized for a given location.  相似文献   

12.
BACKROUND: In Tetranychus urticae Koch, acetylcholinesterase insensitivity is often involved in organophosphate (OP) and carbamate (CARB) resistance. By combining toxicological, biochemical and molecular data from three reference laboratory and three OP selected strains (OP strains), the AChE1 mutations associated with resistance in T. urticae were characterised. RESULTS: The resistance ratios of the OP strains varied from 9 to 43 for pirimiphos‐methyl, from 78 to 586 for chlorpyrifos, from 8 to 333 for methomyl and from 137 to 4164 for dimethoate. The insecticide concentration needed to inhibit 50% of the AChE1 activity was, in the OP strains, at least 2.7, 55, 58 and 31 times higher for the OP pirimiphos‐methyl, chlorpyrifos oxon, paraoxon and omethoate respectively, and 87 times higher for the CARB carbaryl. By comparing the AChE1 sequence, four amino acid substitutions were detected in the OP strains: (1) F331W (Torpedo numbering) in all the three OP strains; (2) T280A found in the three OP strains but not in all clones; (3) G328A, found in two OP strains; (4) A201S found in only one OP strain. CONCLUSIONS: Four AChE1 mutations were found in resistant strains of T. urticae, and three of them, F331W, G328A and A201S, are possibly involved in resistance to OP and CARB insecticides. Among them, F331W is probably the most important and the most common in T. urticae. It can be easily detected by the diagnostic PCR‐RLFP assay developed in this study. Copyright © 2009 Society of Chemical Industry  相似文献   

13.
Pollen beetle, Meligethes aeneus F. (Coleoptera: Nitidulidae) is a major univoltine pest of oilseed rape in many European countries. Winter oilseed rape is cultivated on several million hectares in Europe and the continuous use of pyrethroid insecticides to control pollen beetle populations has resulted in high selection pressure and subsequent development of resistance. Resistance to pyrethroid insecticides in this pest is now widespread and the levels of resistance are often sufficient to result in field control failures at recommended application rates. Recently, metabolic resistance mediated by cytochrome P450 monooxygenases was implicated in the resistance of several pollen beetle populations from different European regions. Here, we have also investigated the possible occurrence of a target-site mechanism caused by modification of the pollen beetle para-type voltage-gated sodium channel gene. We detected a single nucleotide change that results in an amino acid substitution (L1014F) within the domain IIS6 region of the channel protein. The L1014F mutation, often termed kdr, has been found in several other insect pests and is known to confer moderate levels of resistance to pyrethroids. We developed a pyrosequencing-based diagnostic assay that can detect the L1014F mutation in individual beetles and tested more than 350 populations collected between 2006 and 2010 in 13 European countries. In the majority of populations tested the mutation was absent, and only samples from two countries, Denmark and Sweden, contained pollen beetles heterozygous or homozygous for the L1014F mutation. The mutation was first detected in a sample from Denmark collected in 2007 after reports of field failure using tau-fluvalinate, and has since been detected in 7 out of 11 samples from Denmark and 25 of 33 samples from Sweden. No super-kdr mutations (e.g. M918T) known to cause resistance to pyrethroids were detected. The implications of these results for resistance management strategies of pollen beetle populations in oilseed rape crops are discussed.  相似文献   

14.
The bird cherry-oat aphid (Rhopalosiphum padi L.) is a devastating cereal pest that develops high resistance to organophosphate and carbamate insecticides. Because acetylcholinesterase (ACE) is the target of carbamate and organophosphate insecticides, the resistance mechanism usually involves mutations occurring in ACE-encoding genes, Ace1 and Ace2. Here, we describe a novel polymerase chain reaction (PCR)-based method for the diagnosis of resistance cases associated with the point mutation F368L in the Ace2 gene. We amplified a 127 bp DNA fragment from Ace2 gene using a modified reverse primer, and digested the amplification product using SmaI endonuclease. This procedure enabled a simple and rapid distinction between resistant and susceptible genotypes for F368L mutation. Subsequently, we screened 152 R. padi samples, and found that F368L mutation occurred at low frequency, in both the homozygous (R/R) and heterozygous (R/S) states. Based upon the results of this study, we believe that molecular diagnosis of insecticide resistance should be generalized to genes and mutations involved in this process, toward an optimal accuracy of insecticide applications.  相似文献   

15.
为明确L-阿拉伯糖对B型和Q型烟粉虱毒性及其取食行为的影响,调查了饲喂含有L-阿拉伯糖人工饲料后烟粉虱的死亡率,利用刺吸电位技术(EPG)记录其取食行为,并观察了饲喂后其在人工饲料膜上的刺孔数量及直径。结果表明,B型和Q型烟粉虱的校正死亡率均随L-阿拉伯糖浓度及饲喂时间的增加而升高;在3种浓度下,B型烟粉虱校正死亡率均显著高于Q型烟粉虱;在5%、10%浓度下,Q型烟粉虱校正死亡率分别在第5天和第3天达100%,B型烟粉虱分别在第3天和第2天达到100%;5%L-阿拉伯糖对B型烟粉虱取食行为影响比Q型大;在5%浓度下,B型和Q型烟粉虱在膜上的刺孔数量总体少于对照组。研究表明,L-阿拉伯糖对烟粉虱具有杀虫活性,且对B型和Q型烟粉虱的毒性效果不同。  相似文献   

16.
为有效控制日光温室番茄褪绿病毒病,于2014—2015年通过RT-PCR检测方法研究了济南市日光温室番茄褪绿病毒(Tomato chlorosis virus,ToCV)的发生规律、其与Q型烟粉虱Bemisia tabaci种群动态的关系及防虫网对该病毒病的防控效果。结果表明,春季日光温室番茄植株上Q型烟粉虱成虫数量呈增长趋势,5月下旬最高达到0.10头/叶,秋季日光温室番茄植株上Q型烟粉虱成虫数量9月上旬达最高7.42头/叶,后逐渐下降;日光温室Q型烟粉虱带毒率随着定植时间的延长而逐渐上升,之后维持相对稳定状态,即春季为20.00%~24.14%,秋季为30.00%~40.00%。日光温室ToCV发生与Q型烟粉虱成虫数量和带毒率密切相关,春季番茄最高发病率为12.00%;秋季番茄植株最高发病率为93.02%。番茄育苗和生长期用100目防虫网隔离可显著降低番茄植株带毒率。因此,秋季是日光温室ToCV防控关键期,覆盖防虫网阻隔烟粉虱可有效防治ToCV,推荐在日光温室使用。  相似文献   

17.
为了解我国不同地区棉蚜Aphis gossypii对吡虫啉和氟啶虫胺腈的抗性现状,对代表性棉区棉蚜田间种群进行抗药性监测,同时通过构建具有R81T及V62I单突变和R81T-V62I共同突变的棉蚜烟碱型乙酰胆碱受体（nicotinic acetylcholine receptor,nAChR）蛋白模型,与吡虫啉和氟啶虫胺腈进行分子对接,分析这些突变在吡虫啉和氟啶虫胺腈抗性中的作用,并分析吡虫啉和氟啶虫胺腈之间是否存在交互抗性。结果显示,不同地区棉蚜对吡虫啉产生了高水平抗性,抗性倍数为174.70~56 409.18,对氟啶虫胺腈产生了低至中等水平抗性,抗性倍数为7.35~44.63,说明不同地区的棉蚜对氟啶虫胺腈的敏感度高于吡虫啉,且吡虫啉抗性和氟啶虫胺腈抗性间不存在相关性。R81T、V62I单突变和R81T-V62I共同突变导致吡虫啉与棉蚜nAChR的亲和力降低,对氟啶虫胺腈与棉蚜nAChR的结合无明显影响。R81T及V62I单突变和R81T-V62I共同突变导致棉蚜对吡虫啉产生靶标抗性,但是对氟啶虫胺腈的抗性无明显影响,这些突变不会导致吡虫啉与氟啶虫胺腈产生靶标突变的交互抗性。  相似文献   

18.
Bemisia tabaci adults were collected from pepper and melon at different commercial production greenhouses in Argentina and Uruguay. The biotype status of adults was then established using cytochrome oxidase I gene (mtCOI) as molecular marker. Only the Q biotype was found on all plants sampled. This is the first report of the Q biotype in Argentina and Uruguay.  相似文献   

19.
BACKGROUND: Bemisia tabaci Gennadius Q‐biotype has readily developed resistance to numerous insecticide classes. Studies in the Mediterranean area are needed to clarify the resistance status and cross‐resistance patterns in this invasive whitefly biotype. The levels of resistance in nymphs of seven strains of B. tabaci Q‐biotype from south‐eastern Spain to representative insecticides were determined. RESULTS: Six populations had low to moderate levels of resistance to azadirachtin (0.2‐ to 7‐fold), buprofezin (11‐ to 59‐fold), imidacloprid (1‐ to 15‐fold), methomyl (3‐ to 55‐fold), pyridaben (0.9‐ to 9‐fold), pyriproxyfen (0.7‐ to 15‐fold) and spiromesifen (1‐ to 7‐fold), when compared with a contemporary Spanish Q‐biotype reference population (LC50 = 2.7, 8.7, 15.2, 19.9, 0.34, 20.9 and 1.1 mg L?1 respectively). A single population collected from a greenhouse subject to intensive insecticide use exhibited generally higher resistance levels to the same array of compounds (31‐, 1164‐, 3‐, 52‐, 9‐, 19‐ and 3‐fold respectively). Pyridaben and spiromesifen were extremely effective against nymphs of all strains, with LC50 values significantly below recommended application rates. CONCLUSION: In contrast to previous reports, high rates of efficacy exist for numerous insecticide classes against B. tabaci Q‐biotype populations in these intensive agricultural regions of south‐eastern Spain. This probably reflects the recent and significant reductions in exposure that have resulted from a wider uptake of IPM technologies and strategies. However, the continued presence of resistance genes also suggests that a reversion to levels of high insecticide exposure could result in a rapid selection for resistance. Copyright © 2009 Society of Chemical Industry  相似文献   

20.
为明确烟粉虱Bemisia tabaci取食感染番茄黄化曲叶病毒(Tomato yellow leaf curl virus,TYLCV)的番茄植株后,其体内的芳香基硫酸酯酶B基因(arylsulfatase B,ARSB)是否能够做出应答反应,基于Q型烟粉虱基因组数据克隆得到ARSB基因cDNA全长,采用生物信息学方法分析其序列特征,并通过实时荧光定量PCR技术测定ARSB基因在Q型烟粉虱不同发育阶段、不同组织及携带TYLCV前后的表达量变化情况。结果显示:Q型烟粉虱ARSB基因的cDNA全长为1 731 bp,编码576个氨基酸,分子量为64.89 kD,具有ARSB的保守结构域。ARSB基因在Q型烟粉虱不同发育阶段均有表达,在卵期表达量最高,成虫期表达量最低;该基因在Q型烟粉虱头胸部的表达量显著高于腹部;Q型烟粉虱获取TYLCV 72 h后其体内ARSB基因表达量显著提高。表明ARSB基因在Q型烟粉虱不同龄期、不同组织内存在差异表达,并可能参与Q型烟粉虱对TYLCV的响应和传毒过程。  相似文献   

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