Field-simulator study of insecticide resistance conferred by esterase-, MACE- and kdr-based mechanisms in the peach-potato aphid,Myzus persicae (Sulzer)

Insecticide sprays were applied to Myzus persicae (Sulzer) populations carrying various combinations of three insecticide resistance mechanisms (esterase-based metabolic resistance and two target site mechanisms, known as MACE and kdr), supported on host plants growing in field simulator cages. The study showed that MACE confers extreme resistance to pirimicarb and triazamate (carbamate insecticides) but not to deltamethrin + heptenophos (16 + 1) (Decisquick) or dimethoate (an organophosphorus insecticide). Resistance to dimethoate depends solely on levels of esterase-based resistance, while resistance to Decisquick depends on kdr and esterase. None of the four insecticides is effective against aphids carrying MACE combined with extreme esterase-based resistance. This knowledge, in association with current monitoring of the mechanisms, will play an important role in making decisions on insecticide use against M persicae in the UK. © 1999 Society of Chemical Industry     

The effect of insecticide application sequences on the control and insecticide resistance status of the peach-potato aphid, Myzus persicae (Hemiptera:Aphididae), on field crops of potato

Experiments were done on commercial potato crops in the UK to investigate the effect of different insecticide sequences on the control and insecticide resistance status of Myzus persicae (Sulzer). The work was done to provide field validation of similar laboratory studies done in 'field simulators'. To ensure adequate aphid populations and to influence the initial resistance status of the aphid population, cultured M. persicae from a clone of known resistance status (esterase R1, kdr heterozygote, non-MACE (modified acetylcholinesterase)) were inoculated into both experiments. Two-spray programmes starting with lambda-cyhalothrin (a pyrethroid insecticide) gave poor control in comparison with programmes starting with pirimicarb (a carbamate insecticide) or pirimicarb-containing mixtures. This concurred closely with the results obtained from single applications in field simulator studies. Treatment sequences containing pymetrozine (a pyridine azomethine insecticide) were also effective, though slower-acting. This again concurs with field simulator studies. The proportions of aphids carrying different resistance mechanisms were largely unaffected by treatment in these experiments. The implications of these results for field control strategies are discussed.     

Temporal and spatial dynamics of insecticide resistance in Myzus persicae (Hemiptera: Aphididae)

Advances in understanding insecticide resistance in the peach-potato aphid, Myzus persicae (Sulzer), at the genotypic, biochemical and molecular levels have led to rapid and precise methods for the detection of several resistance mechanisms (elevated carboxylesterase, modified acetylcholinesterase or MACE, and knockdown resistance or kdr) in individual insects, and for monitoring their frequencies over space and time. This paper summarises the results of two long-term surveys of resistance dynamics in M persicae in England, based on samples collected directly from field and glasshouse crops or from four 12.2-m suction traps. The study showed marked fluctuations in resistance frequencies that probably reflect the counteracting forces of selection imposed by insecticides for aphids possessing more copies of esterase resistance genes, and selection against these forms when insecticide use is relaxed. There is growing evidence that several different resistance mechanisms in M persicae have associated fitness costs. In the case of esterase and MACE, these costs are apparently strong enough to effect a decline in resistance frequency over winter, and a more prolonged decline over successive cropping seasons when aphid numbers are insufficient to trigger intensive chemical applications. Changes in the overall frequency of resistance genotypes may also be influenced by the predominance of year-round parthenogenesis in M persicae in the UK, leading to non-random associations between mechanisms and selection operating on clonal lineages rather than individual genotypes.     

Variation in response to neonicotinoid insecticides in peach-potato aphids,Myzus persicae (Hemiptera: Aphididae)

Laboratory bioassays applying the neonicotinoid insecticides imidacloprid, acetamiprid and nitenpyram against clones of the peach-potato aphid Myzus persicae (Sulzer) demonstrated that these compounds effectively circumvent the known carboxylesterase, modified acetylcholinesterase (MACE) and knock-down (kdr) insecticide resistance mechanisms in this species. However, some clones showed cross-tolerance (up to 18-fold) of these compounds relative to susceptible standards. A survey assessing the frequency of neonicotinoid tolerance in M persicae in the UK, based on samples collected from the field and glasshouses between 1997 and 2000, showed that such tolerance is still rare. Experiments on neonicotinoid-susceptible and -tolerant populations of M persicae under simulated field conditions in the laboratory showed that, although the latter were well controlled by imidacloprid applied at recommended application rates, they were more likely to survive and reproduce when this compound was applied at lower concentrations. Such conditions are probably periodically present in imidacloprid-treated field and glasshouse crops. Selection favouring tolerant forms of M persicae could lead to increases in their frequency and the evolution of more potent resistance to neonicotinoids.     

Bioassay and field-simulator studies of the efficacy of pymetrozine against peach-potato aphids,Myzus persicae (Hemiptera: Aphididae), possessing different mechanisms of insecticide resistance

Twenty-one clones of the peach-potato aphid, Myzus persicae (Sulzer), carrying various combinations of known mechanisms of insecticide resistance were assessed for their response to the new pyridine azomethine compound, pymetrozine, in leaf-dip bioassays. Pymetrozine was also applied as a foliar spray to populations of four different UK M persicae clones on potato plants in field-simulator chambers. Neither study showed any evidence of cross-resistance to this compound. Pymetrozine, used in conjunction with other effective aphicides, such as pirimicarb and imidacloprid, can therefore play an important role in insecticide resistance-management strategies for M persicae.     

Use of the RFLP-PCR diagnostic test for characterizing MACE and kdr insecticide resistance in the peach potato aphid Myzus persicae

The peach-potato aphid Myzus persicae (Sulzer) has developed a number of insecticide resistance mechanisms owing to the high selective pressure produced by world-wide insecticide treatments. Knowledge of the geographical distribution and the temporal evolution of these resistant phenotypes helps to develop suitable pest-management programs. Current understanding of the major mechanisms of resistance at the molecular level makes it possible to diagnose the presence of modified acetylcholinesterase (MACE) or knockdown resistance (kdr). This paper describes a rapid method for the identification of both resistance mechanisms in a single molecular assay by using restriction fragment length polymorphism of PCR products (RFLP-PCR) in individual as well as pooled aphids.     

Insecticide resistance status of Myzus persicae (Hemiptera: Aphididae) populations from peach and tobacco in mainland Greece

The susceptibility of 88 and 38 field samples of Myzus persicae (Sulzer) to imidacloprid and deltamethrin respectively was examined using the FAO dip test bioassay. The field samples were collected from tobacco and peach from various regions of Greece in the period from 2004 to 2006. In addition, 497, 349 and 370 clones originating from peach and tobacco were screened for the three known resistance mechanisms, elevated esterases, modified acetylcholinesterase (MACE) and knockdown resistance (kdr) respectively, using biochemical and DNA diagnostics. Most of the samples assayed with imidacloprid showed low resistance factors (RFs)-39% below 5 and 21% between 5 and 10. However, 9% of the samples (all from tobacco) showed relatively high RF values (24-73). Differences were found between crops, with higher RF values recorded in samples from tobacco than in those from peach. Bioassays with deltamethrin revealed the development of strong resistance in the populations examined. The RFs were mostly higher than 23, and in 29% of the samples they were extremely high (152-436). Finally, the three known resistance mechanisms were found in high frequencies in the populations examined, although some differences between crops and years were detected. The implications of the study for management schemes against M. persicae are discussed.     

Amplified esterase genes and their relationship with other insecticide resistance mechanisms in English field populations of the aphid,Myzus persicae (Sulzer)

Myzus persicae samples were collected from populations present on a range of field crops between 1997 and 2000. A combination of biochemical, DNA-based diagnostics and bioassays was used to assess the presence of three insecticide resistance mechanisms: elevated carboxylesterase (E4 or FE4), insensitive acetylcholinesterase and insensitive sodium channels (knockdown resistance, kdr). For the carboxylesterases, both the levels of enzyme and the type of gene present (E4 or FE4) were determined. The results showed that during the time period studied there was a dramatic reduction in the proportion of aphids with very high levels of E4 and an increase in those with lower levels of FE4. There was also a slightly different E4 gene present in a limited number of samples. The change in esterase genes was accompanied by a virtual loss of the insensitive AChE variant and a maintenance of aphids with kdr. The selection pressures and other factors leading to these changes in field populations of M persicae are discussed.     

Biochemical evidence that an S431F mutation in acetylcholinesterase-1 of Aphis gossypii mediates resistance to pirimicarb and omethoate

Molecular changes in acetylcholinesterase (AChE) leading to target-site resistance to carbamate and organophosphate insecticides have recently been identified. Of particular interest is the S431F mutation in ace2 and its orthologue ace1 of Myzus persicae Sulzer and Aphis gossypii Glover, respectively. This mutation has been correlated with resistance to pirimicarb, but biochemical evidence has not yet been provided. Here, we describe for the first time that recombinantly expressed AChE1 from A gossypii carrying the S431F mutation is insensitive to pirimicarb and omethoate, but sensitive to demeton-S-methyl and hypersensitive to carbofuran. Furthermore, site-directed mutagenesis of the serine residue at position 431 in ace1 from a pirimicarb-susceptible clone of A gossypii conferred insensitivity to pirimicarb. We conclude that AChE1 of A gossypii is the target of toxicological relevance of carbamates and organophosphates.     

Association of MACE-based insecticide resistance in Myzus persicae with reproductive rate, response to alarm pheromone and vulnerability to attack by Aphidius colemani

Reproductive success and response to alarm pheromone, both potentially important components of fitness, were assessed using clones of Myzus persicae (Sulzer) to establish associations with insecticide resistance conferred by insensitive modified acetylcholinesterase (MACE). Both traits showed significant trends that were apparently related to this mechanism. MACE forms appeared to reproduce at slower rates than non-MACE forms expressing moderate (R1) levels of another resistance mechanism based on elevated carboxylesterase. However, MACE forms were more responsive to alarm pheromone than their non-MACE counterparts. The potential implications for parasitoid performance were tested using two clones showing clear differences in alarm response. The level of parasitism of M persicae by the parasitoid Aphidius colemani (Viereck) was significantly lower in MACE forms on pepper crops compared to non-MACE forms. In addition, the distribution of MACE and non-MACE forms differed on the pepper plants, with more MACE forms being found on the growing points. The presence of the parasitoid A colemani did not alter this change in distribution.     

Diversity of insecticide resistance mechanisms and spectrum in European populations of the codling moth, Cydia pomonella

Only a few of the registered insecticides against Cydia pomonella L. are still effective in areas where insecticide resistance has emerged in this pest. Resistance mechanisms are multiple, and their lone or cumulative effects in a single population are not completely understood. A detailed estimation of resistance spectrum is still required to define the suitable insecticides to use against a given population. The efficacy of ten insecticides was therefore investigated together with the resistance mechanisms expressed in four laboratory strains and 47 field populations of C. pomonella from five countries. Bioassays were performed using topical applications of diagnostic concentrations on diapausing larvae, and resistance mechanisms were analysed on adults emerging from control insects. All populations exhibited a reduced susceptibility to at least one insecticide when compared with the susceptible laboratory strain. Cross-resistances were observed between azinphos-methyl or phosalone and more recent compounds such as spinosad and thiacloprid. Resistances to azinphos-methyl, diflubenzuron, spinosad, tebufenozide and thiacloprid were significantly correlated with mixed-function oxidase activity, while increased glutathione-S-transferase and reduced non-specific esterase activities were correlated with resistance to azinphos-methyl and emamectin, respectively. Conversely, resistances to azinphos-methyl, tebufenozide and thiacloprid were negatively correlated with increased esterase activity. None of the observed mechanisms explained the loss of susceptibility of populations to chlorpyrifos-ethyl, and no significant correlation was detected between resistance to deltamethrin and the presence of the kdr mutation. The suitability of such non-target instars to monitor insecticide resistance in field populations is discussed.     

Temporal synergism can enhance carbamate and neonicotinoid insecticidal activity against resistant crop pests

BACKGROUND: Piperonyl butoxide (PBO) effectively synergises synthetic pyrethroids, rendering even very resistant insect pests susceptible, provided a temporal element is included between exposure to synergist and insecticide. This concept is now applied to carbamates and neonicotinoids. RESULTS: A microencapsulated formulation of PBO and pirimicarb reduced the resistance factor in a clone of Myzus persicae (Sulzer) from >19 000- to 100-fold and in Aphis gossypii (Glover) from >48 000- to 30-fold. Similar results were obtained for a strain of Bemisia tabaci Gennadius resistant to imidacloprid and acetamiprid, although a second resistant strain did not exhibit such a dramatic reduction, presumably owing to the presence of target-site insensitivity and the absence of metabolic resistance. Synergism was also observed in laboratory susceptible insects, suggesting that, even when detoxification is not enhanced, there is degradation of insecticides by the background enzymes. Use of an analogue of PBO, which inhibits esterases but has reduced potency against microsomal oxidases, suggests that acetamiprid resistance in whiteflies is largely oxidase based. CONCLUSION: Temporal synergism can effectively enhance the activity of carbamates and neonicotinoids against resistant insect pests. Although the extent of this enhancement is dependent upon the resistance mechanisms present, inhibition of background enzymes can confer increased sensitivity against target-site resistance as well as increased metabolism. .     

抗吡虫啉桃蚜种群的选育及其交互抗性研究

采用浸渍法,在室内用吡虫啉对桃蚜敏感种群进行抗性选育,经过15代的连续汰选,获得抗性指数为14.34倍的抗吡虫啉桃蚜种群。对9种常用杀虫剂的交互抗性测定结果表明,该种群对拟除虫菊酯类如高效氯氟氰菊酯(抗性指数12.76倍)和溴氰菊酯(10.24倍),有机磷类如氧乐果(7.95倍)、辛硫磷(5.44倍)和甲胺磷(5.32倍)以及吡虫啉·高效氯氰菊酯混剂(8.90倍)均产生了不同程度的交互抗性,而对氨基甲酸酯类如灭多威(3.15倍)、有机氯类如硫丹(1.64倍)以及阿维菌素·辛硫磷混剂(2.31倍)等无明显的交互抗性。     

草地贪夜蛾对农药主要抗性机制的概述

草地贪夜蛾Spodoptera frugiperda是一种迁飞性害虫,源自美洲,已入侵非洲和亚洲部分国家并暴发成灾,2019年初入侵我国,严重威胁着我国玉米、小麦等粮食作物。目前草地贪夜蛾的防治主要依赖于化学农药和种植转基因作物。因长期使用农药,草地贪夜蛾已经对氨基甲酸酯类、有机磷酸酯类和拟除虫菊酯类等多种杀虫剂产生了不同程度的抗性。草地贪夜蛾对不同种类杀虫剂表现的抗性机制不同,主要有代谢抗性和靶标抗性2种。本文主要从代谢抗性和靶标抗性2个方面分别综述了草地贪夜蛾对不同种类杀虫剂的抗性机制,分析草地贪夜蛾对不同杀虫剂的抗性作用方式,并对草地贪夜蛾抗药性管理和监测进行展望。     

Communication to the Editor Insecticide Resistance in a Strain of Aphis gossypii from Southern France

A strain (R) of Aphis gossypii from Southern France was found to be resistant to several insecticides, particularly to pirimicarb, as compared to a susceptible strain (S). Resistance levels were determined by biological tests, and the highest resistance factor (1350) was for pirimicarb. Resistance was mainly restricted to anticholinesterase inhibitors. Use of synergists, DEF and PB, suggested that resistance mechanisms based on detoxification were involved to a minor extent, since a good correlation was observed between I₅₀ values and k_i values of AChE and in-vivo bioassay data. The two strains differed in esterase activity, with a 27·7-fold increase in the R strain. Resolution of esterases by polyacrylamide gel electrophoresis showed different patterns in the S and R strains, and two isozymes were less sensitive to pirimicarb in the S strain; however, no in-vitro degradation of [¹⁴C]pirimicarb was observed. These data suggest that the main mechanism of resistance was through a decrease in the sensitivity of the target, AChE, to the insecticides. © 1997 SCI.     

A biochemical mechanism of resistance to pirimicarb in two glasshouse clones of Aphis gossypii

The susceptibility of three clones of Aphis gossypii Glover to 15 insecticides was established by bioassay. A high level of resistance towards pirimicarb was confirmed for a clone from Holland (Dutch R) and a clone from Japan (Jap R), while the susceptible clone (S) was killed by very low doses of the insecticide. However, only limited cross-resistance was shown towards other carbamates and organophosphates, and no marked resistance to the pyrethroids tested. The acetylcholinesterase (AChE) of both resistant clones hydrolysed acetyl-choline faster than that of susceptible aphids, with greatest enzyme activity shown by the Dutch R clone. Inter-clone differences in these rates were consistent with differences in catalytic centre activities. Inhibition (I₅₀) of AChE by pirimicarb was approximately 900-fold higher for the resistant clones than for the S clone. First-order kinetics revealed that resistance to pirimicarb in Dutch R and Jap R involved a modified AChE which had a reduced (approximately 350-fold) affinity (K_d) for pirimicarb. The marked change in AChE affinity for pirimicarb was not repeated with the other carbamates tested, ethiofencarb and aldicarb. It was considered that the resistant aphids would not require mechanisms in addition to insensitive AChE in order to show the high level of resistance to pirimicarb shown in the bioassay.     

Analysis of insecticide-resistant Myzus persicae (Sulzer) populations collected in Italian peach orchards

Several populations of the green peach aphid, Myzus persicae (Sulzer), were collected in the most important Italian peach-growing areas. In these populations, collected as primary colonies or as secondary ones, total esterase content and acetylcholinesterase sensitivity towards pirimicarb was assessed using biochemical assays. Most of the individuals analysed generally exhibited an elevated esterase content, and in some cases acetylcholinesterase insensitivity. The implications of resistance levels and distribution on M persicae control strategies are discussed.     

我国西南烟区桃蚜Myzus persicae (Sulzer)的抗药性水平

采用毛细管点滴法测定了我国西南烟区云南和贵州省的10个桃蚜Myzus persicae（Sulzer）种群对氰戊菊酯、氧化乐果、灭多威和吡虫啉的抗性。结果表明，云南楚雄种群对氰戊菊酯的抗性高达26．87倍，对氧化乐果、灭多威、吡虫啉的抗性分别为敏感种群的10．02、6．00和5．83倍。云南大理、丽江、石林和曲靖4个种群对氰戊菊酯的抗性分别达24．98、14．94、11．11和10．33倍，抗性水平也较高，但对氧乐果、灭多威和吡虫啉仍较为敏感，应尽早实施抗性治理措施。贵州省的5个种群仍然处在敏感阶段。     

Involvement of a sodium channel mutation in pyrethroid resistance in Cydia pomonella L, and development of a diagnostic test

Populations of the codling moth, Cydia pomonella L (Lepidoptera, Tortricidae) have developed resistance to several classes of insecticide such as benzoylureas, juvenile hormone analogues, ecdysone agonists and pyrethroids, but the corresponding resistance mechanisms have not been extensively studied. Knockdown resistance (kdr) to pyrethroid insecticides has been associated with point mutations in the para sodium channel gene in a great variety of insect pest species. We have studied two susceptible strains (S and Sv) and two resistant strains (Rt and Rv) of C pomonella that exhibited 4- and 80-fold resistance ratios to deltamethrin, respectively. The region of the voltage-dependent sodium channel gene which includes the position where kdr and super-kdr mutations have been found in Musca domestica L was amplified. The kdr mutation, a leucine-to-phenylalanine replacement at position 1014, was found only in the Rv strain. In contrast, the super-kdr mutation, a methionine-to-threonine replacement at position 918, was not detected in any C pomonella strain. These data allowed us to develop a PCR-based diagnostic test (PASA) to monitor the frequency of the kdr mutation in natural populations of C pomonella in order to define appropriate insecticide treatments in orchards.     

Biochemical and molecular diagnosis of insecticide resistance conferred by esterase, MACE, kdr and super-kdr based mechanisms in Italian strains of the peach potato aphid, Myzus persicae (Sulzer)

In this paper we analysed the basis of insecticide resistance in 59 Italian strains of the peach potato aphid Myzus persicae using both molecular and biochemical assays. Our data as a whole clearly indicate that most M. persicae strains (76.3%) have high or extremely high production of an esterase enzyme which sequester and detoxify insecticides with esteric group. Kdr genotypes conferring resistance towards pyrethoids are present in 57.7% of the analysed populations. Moreover, 26.5% of the kdr positive strains possess also the M918T mutation conferring super-kdr phenotype. Strains with modified AChE (MACE) are not so numerous (27.1%), although they can be found almost everywhere in Italy. Considering all the strains analysed, both MACE and kdr phenotypes are associated with high levels of esterase activity. In Central–Southern regions, kdr and MACE resistance mechanisms resulted in linkage disequilibrium. Bioassays performed in order to evaluate the efficacy of a pyrethroid insecticide against a strain possessing a F979S mutation within its para-type sodium channel gene suggests that this amino acid substitution could affect the sodium channel responsivity to pyrethroids.