Russell Slater

Mutation of a nicotinic acetylcholine receptor β subunit is associated with resistance to neonicotinoid insecticides in the aphid Myzus persicae

BackgroundMyzus persicae is a globally important aphid pest with a history of developing resistance to insecticides. Unusually, neonicotinoids have remained highly effective as control agents despite nearly two decades of steadily increasing use. In this study, a clone of M. persicae collected from southern France was found, for the first time, to exhibit sufficiently strong resistance to result in loss of the field effectiveness of neonicotinoids.ResultsBioassays, metabolism and gene expression studies implied the presence of two resistance mechanisms in the resistant clone, one based on enhanced detoxification by cytochrome P450 monooxygenases, and another unaffected by a synergist that inhibits detoxifying enzymes. Binding of radiolabeled imidacloprid (a neonicotinoid) to whole body membrane preparations showed that the high affinity [3H]-imidacloprid binding site present in susceptible M. persicae is lost in the resistant clone and the remaining lower affinity site is altered compared to susceptible clones. This confers a significant overall reduction in binding affinity to the neonicotinoid target: the nicotinic acetylcholine receptor (nAChR). Comparison of the nucleotide sequence of six nAChR subunit (Mpα1-5 and Mpβ1) genes from resistant and susceptible aphid clones revealed a single point mutation in the loop D region of the nAChR β1 subunit of the resistant clone, causing an arginine to threonine substitution (R81T).ConclusionPrevious studies have shown that the amino acid at this position within loop D is a key determinant of neonicotinoid binding to nAChRs and this amino acid change confers a vertebrate-like character to the insect nAChR receptor and results in reduced sensitivity to neonicotinoids. The discovery of the mutation at this position and its association with the reduced affinity of the nAChR for imidacloprid is the first example of field-evolved target-site resistance to neonicotinoid insecticides and also provides further validation of exisiting models of neonicotinoid binding and selectivity for insect nAChRs.

Pyrethroid resistance monitoring in European populations of pollen beetle (Meligethes spp.): a coordinated approach through the Insecticide Resistance Action Committee (IRAC)

BACKGROUND Pollen beetle (Meligethes spp.) is a major pest of European oilseed rape crops. Its resistance to pyrethroid insecticides has been recorded in samples of beetles collected in Europe since at least 1999, and problems with the control of the beetle in the field have been widely reported. In 2007, a Pollen Beetle Working Group was formed through the Insecticide Resistance Action Committee (IRAC) in order to coordinate efforts for surveying pyrethroid resistance development. RESULTS The results of the first 3 years of the pollen beetle pyrethroid susceptibility survey using a laboratory test are presented in this paper. Resistant beetle samples were collected from 20 of the 21 countries surveyed, with a general trend of increasing frequency and spread of resistant samples in European oilseed-rape-growing regions. CONCLUSION Pyrethroid-resistant beetles dominate in Western and Central Europe and are becoming established in the North and East, the main oilseed-rape-growing areas of Europe. The development and spread of pyrethroid-resistant pollen beetles highlights the need for effective management strategies for oilseed rape insect pests.

Investigating the mode of action of sulfoxaflor: a fourth-generation neonicotinoid.

BACKGROUND The precise mode of action of sulfoxaflor, a new nicotinic acetylcholine receptor-modulating insecticide, is unclear. A detailed understanding of the mode of action, especially in relation to the neonicotinoids, is essential for recommending effective pest management practices. RESULTS Radiolabel binding experiments using a tritiated analogue of sulfoxaflor ([(3) H]-methyl-SFX) performed on membranes from Myzus persicae demonstrate that sulfoxaflor interacts specifically with the high-affinity imidacloprid binding site present in a subpopulation of the total nAChR pool. In competition studies, imidacloprid-like neonicotinoids displace [(3) H]-methyl-SFX at pM concentrations. The effects of sulfoxaflor on the exposed aphid nervous system in situ are analogous to those of imidacloprid and nitenpyram, and finally the high-affinity sulfoxaflor binding site is absent in a Myzus persicae strain (clone FRC) possessing a single amino acid point mutation (R81T) in the β-nAChR, a region critical for neonicotinoid interaction. CONCLUSION The nicotinic acetylcholine receptor pharmacological profile of sulfoxaflor in aphids is consistent with that of imidacloprid. Additionally, the insecticidal activity of sulfoxaflor and the current commercialised neonicotinoids is affected by the point mutation in FRC Myzus persicae. Therefore, it is suggested that sulfoxalfor be considered a neonicotinoid, and that this be taken into account when recommending insecticide rotation partnering for effective resistance management programmes.

Susceptibility of standard clones and European field populations of the green peach aphid, Myzus persicae, and the cotton aphid, Aphis gossypii (Hemiptera: Aphididae), to the novel anthranilic diamide insecticide cyantraniliprole

BACKGROUND Parthenogenetic clones of the green peach aphid, Myzus persicae (Sulzer), and the cotton aphid, Aphis gossypii Glover, were tested with the anthranilic diamide insecticide cyantraniliprole (i.e. DuPont(™) Cyazypyr(™) ) in systemic-uptake bioassays to investigate potential for cross-resistance conferred by mechanisms of insecticide resistance to organophosphates, carbamates and pyrethroids and, in the case of M. persicae, reduced sensitivity to neonicotinoids. These data were compared with the response of field samples of M. persicae and A. gossypii collected from around Europe. RESULTS Cyantraniliprole was not cross-resisted by any of the known insecticide resistance mechanisms present in M. persicae or A. gossypii. The compound was equally active against resistant and susceptible aphid strains. The responses of the M. persicae field samples were very consistent with a maximum response ratio of 2.9 compared with a standard laboratory clone. The responses of the A. gossypii field samples were more variable, although a majority of the responses were not statistically different. CONCLUSION Cyantraniliprole is currently the only anthranilic diamide (IRAC MoA 28) insecticide targeting aphid species such as M. persicae and A. gossypii. There is no evidence to suggest that the performance of this compound is affected by commonly occurring mechanisms that confer resistance to other insecticide chemistries. Cyantraniliprole is therefore a valuable tool for managing insecticide resistance in these globally important pests.

A mutation (L1014F) in the voltage-gated sodium channel of the grain aphid, Sitobion avenae, is associated with resistance to pyrethroid insecticides

BACKGROUND The grain aphid, Sitobion avenae Fabricius (Hemiptera: Aphididae), is an important pest of cereal crops. Pesticides are the main method for control but carry the risk of selecting for resistance. In response to reports of reduced efficacy of pyrethroid sprays applied to S. avenae, field samples were collected and screened for mutations in the voltage-gated sodium channel, the primary target site for pyrethroids. Aphid mobility and mortality to lambda-cyhalothrin were measured in coated glass vial bioassays. RESULTS A single amino acid substitution (L1014F) was identified in the domain IIS6 segment of the sodium channel from the S. avenae samples exhibiting reduced pyrethroid efficacy. Bioassays on aphids heterozygous for the kdr mutation (SR) or homozygous for the wild-type allele (SS) showed that those carrying the mutation had significantly lower susceptibility to lambda-cyhalothrin. CONCLUSION The L1014F (kdr) mutation, known to confer pyrethroid resistance in many insect pests, has been identified for the first time in S. avenae. Clonal lines heterozygous for the mutation showed 35-40-fold resistance to lambda-cyhalothrin in laboratory bioassays, consistent with the reported effect of this mutation on pyrethroid sensitivity in other aphid species.

Characterising metabolic resistance in pyrethroid-insensitive pollen beetle (Meligethes aeneus F.) from Poland and Switzerland.

BACKGROUND Pollen beetle (Meligethes aeneus F.) has become the most important pest of oilseed rape in Europe, but its control has been greatly hindered by pyrethroid resistance. Target-site resistance has been implicated previously, and, whilst synergism has been found with piperonyl butoxide (PBO), the exact nature of metabolic resistance has remained unknown. The use of PBO, in conjunction with its analogue EN 16/5-1, has allowed the characterisation of metabolic resistance. RESULTS In vitro assays in combination with in vivo studies using PBO and EN 16/5-1 showed that high synergism of pyrethroids was primarily correlated with an oxidative mechanism, although a limited contribution by esterases was implicated in one population. CONCLUSION Differential synergism has enabled the characterisation of pyrethroid resistance in populations of M. aeneus. It was found to be principally due to an oxidative-based mechanism, and, if a synergist were to be used to inhibit this enzyme group, renewed control against resistant pests could be achieved.

An Individual-Based Model of the Evolution of Pesticide Resistance in Heterogeneous Environments: Control of Meligethes aeneus Population in Oilseed Rape Crops

Preventing a pest population from damaging an agricultural crop and, at the same time, preventing the development of pesticide resistance is a major challenge in crop protection. Understanding how farming practices and environmental factors interact with pest characteristics to influence the spread of resistance is a difficult and complex task. It is extremely challenging to investigate such interactions experimentally at realistic spatial and temporal scales. Mathematical modelling and computer simulation have, therefore, been used to analyse resistance evolution and to evaluate potential resistance management tactics. Of the many modelling approaches available, individual-based modelling of a pest population offers most flexibility to include and analyse numerous factors and their interactions. Here, a pollen beetle (Meligethes aeneus) population was modelled as an aggregate of individual insects inhabiting a spatially heterogeneous landscape. The development of the pest and host crop (oilseed rape) was driven by climatic variables. The agricultural land of the landscape was managed by farmers applying a specific rotation and crop protection strategy. The evolution of a single resistance allele to the pyrethroid lambda cyhalothrin was analysed for different combinations of crop management practices and for a recessive, intermediate and dominant resistance allele. While the spread of a recessive resistance allele was severely constrained, intermediate or dominant resistance alleles showed a similar response to the management regime imposed. Calendar treatments applied irrespective of pest density accelerated the development of resistance compared to ones applied in response to prescribed pest density thresholds. A greater proportion of spring-sown oilseed rape was also found to increase the speed of resistance as it increased the period of insecticide exposure. Our study demonstrates the flexibility and power of an individual-based model to simulate how farming practices affect pest population dynamics, and the consequent impact of different control strategies on the risk and speed of resistance development.

Development of a high-throughput real-time PCR assay for the detection of the R81T mutation in the nicotinic acetylcholine receptor of neonicotinoid-resistant Myzus persicae

BACKGROUND Myzus persicae is a globally important aphid pest that is mainly controlled through the application of chemical insecticides. Recently, a clone of M. persicae exhibiting control-compromising levels of resistance to neonicotinoid insecticides was described. The resistance of this clone was associated with reduced affinity of imidacloprid for the target site (the nicotinic acetylcholine receptor) as a result of mutation of a key amino acid residue (R81T) in the loop D region of a nAChR β1 subunit. The potent levels of resistance conferred by this mechanism are cause for considerable concern, and the frequency and distribution of the mutation in worldwide populations of M. persicae require careful monitoring. In this study, a high-throughput assay has been developed that allows detection of the mutation in individual aphids. RESULTS A real-time TaqMan assay to detect the R81T substitution was developed that proved to be sensitive and specific in tests of analytical sensitivity and in a blind genotyping trial of DNA extracted from individual aphids comprising the three possible genotypes. The assay was then used to examine the frequency of the R81T mutation in aphids collected and stored in ethanol from peach orchards in southern France. The R81T frequency varied from 33 to 100% in seven populations from the department of Gard, France. CONCLUSIONS This study describes a rapid and sensitive assay that very effectively detects the R81T mutation in individual aphids. The results also have practical significance for the control of M. persicae in southern France and provide contemporary data to inform current resistance management strategies.

Behavioural avoidance and enhanced dispersal in neonicotinoid-resistant Myzus persicae (Sulzer)

BACKGROUND The peach potato aphid Myzus persicae is a major agricultural pest capable of transmitting over 100 plant viruses to a wide range of crops. Control relies largely upon treatment with neonicotinoid insecticides such as thiamethoxam (TMX). In 2009, a strain denoted FRC, which exhibits between 255- and 1679-fold resistance to current neonicotinoids previously linked to metabolic and target site resistance, was discovered in France. Dispersal behaviour may potentially further enhance the resistance of this strain. This study investigated this possibility and is the first to compare the dispersal behaviour of aphid clones of the same species with differing levels of neonicotinoid resistance. RESULTS Comparing the dispersal behaviour of the FRC strain with that of a clone of lower neonicotinoid resistance (5191A), and a susceptible clone (US1L) highlighted several differences. Most importantly, the FRC strain exhibited an increased ability to locate untreated areas when presented with an environment consisting of both TMX-treated and untreated plant tissue. CONCLUSION The altered dispersal behaviour of the FRC may partially account for the high level of neonicotinoid resistance exhibited by this strain in the field. Since the dispersal of aphid vectors is key to the transmission of viruses across crop fields this has implications for current crop protection practice.

Use of an individual‐based simulation model to explore and evaluate potential insecticide resistance management strategies

BACKGROUND Tools with the potential to predict risks of insecticide resistance and aid the evaluation and design of resistance management tactics are of value to all sectors of the pest management community. Here we describe use of a versatile individual-based model of resistance evolution to simulate how strategies employing single and multiple insecticides influence resistance development in the pollen beetle, Meligethes aeneus. RESULTS Under repeated exposure to a single insecticide, resistance evolved faster to a pyrethroid (lambda-cyhalothrin) than to a pyridine azomethane (pymetrozine), due to difference in initial efficacy. A mixture of these compounds delayed resistance compared to use of single products. The effectiveness of rotations depended on the sequence in which compounds were applied in response to pest density thresholds. Effectiveness of a mixture strategy declined with reductions in grower compliance. At least 50% compliance was needed to cause some delay in resistance development. CONCLUSION No single strategy meets all requirements for managing resistance. It is important to evaluate factors that prevail under particular pest management scenarios. The model used here provides operators with a valuable means for evaluating and extending sound resistance management advice, as well as understanding needs and opportunities offered by new control techniques.