Use of biochemical and DNA diagnostics for characterising multiple mechanisms of insecticide resistance in the peach-potato aphid,Myzus persicae (Sulzer)

The peach-potato aphid Myzus persicae (Sulzer) can resist a range of insecticides by over-producing detoxifying esterase and having mutant-insensitive forms of the target proteins, acetylcholinesterase (AChE), and the sodium channel. Using a combination of bioassays, biochemical and DNA diagnostics, it is now possible to diagnose all three mechanisms in individual aphids, and thereby establish their spatial distributions and temporal dynamics. A survey of 58 samples of wide geographic origin showed that all 46 resistant clones had amplified esterase genes (E4 or FE4) conferring broad-spectrum resistance to pyrethroids, organophosphates and carbamates. These occurred in combination with insensitive AChE (11 clones), conferring resistance to pirimicarb and triazamate, and/or mutant sodium channel genes (25 clones), conferring knockdown (kdr) resistance to pyrethroids and DDT. Amplified esterase genes were in linkage disequilibrium with both insensitive AChE and the kdr mutation, reflecting tight physical linkage, heavy selection favouring aphids with multiple mechanisms, and/or the prominence of parthenogenesis in many M. persicae populations. An ability to monitor individual mechanisms with contrasting cross-resistance profiles has important implications for the development of resistance management recommendations. ©1997 SCI     

The detection and distribution of organophosphorus and carbamate insecticide-resistant Myzus persicae (Sulz.) in Britain in 1976

Examination of the enzyme that determines the level of resistance to organophosphorus insecticides and carbamates in Myzus persicae (Sulz.) and bioassays were used to establish the frequency and resistance levels of resistant aphids on outdoor crops in Britain in 1976. The biochemical tests, staining esterase-4 after electrophoresis and total esterase determination, were more sensitive than bioassays. However the dip-test, a simple, rapid and inexpensive bioassay designed to detect resistance and its different levels gave satisfactory results which warrant its use where biochemical detection of resistance is not possible. Carboxylesterase activities of M. persicae collected in 1976 fell into three groups: low, moderate and high, and these were correlated with differences in tolerance to dimethoate, demeton-S-methyl and pirimicarb. Aphids with low esterase activity were susceptible(S). Those with the moderately active enzyme (R₁) had five- to seven-fold resistance to the two organophosphorus insecticides and were marginally resistant (about two-fold) to pirimicarb. The insects with the most active esterase (R₂) were strongly resistant to dimethoate (resistance factor, RF × 126) and moderately resistant to demeton-S-methyl (RF × 17) and pirimicarb (RF × 8). Some R₁, but no S aphids survived the recommended dose of demeton-S-methyl on field crops probably because they were under the lowest leaves and therefore protected from direct contact with the spray. Laboratory tests demonstrated that these R₁ aphids tolerated the residual deposit and systemic dose present in the leaves of the treated potato-plants. This enabled their numbers to recover in treated fields much faster than the susceptible insects which could do so by immigration only when the residual dose in the plant was no longer toxic. R₁ aphids were common throughout the country particularly in eastern England where susceptible aphids were rare, but in the Shardlow area of Derbyshire susceptible aphids were in the majority throughout the summer. R₂ aphids were found only in samples from the west of Scotland and northern England. The implications of the presence of aphids with different levels of resistance for aphid control are discussed.     

辛氰和乐氰混剂对甘蓝桃蚜抗性演化的影响

用辛硫磷与氰戊菊酯,乐果与氰戊菊酯的最佳配比及其相应单剂对甘蓝桃蚜进行抗性选育,汰洗２０代时桃蚜对两种混剂的抗性,分别为４．２０和２．６８倍,比相应单剂的抗性发展速度慢,尤其比氰戊菊酯单剂（２８９倍）更慢,表明上述两种混剂都可延缓桃蚜的抗性发展。用酶抑制剂进行增效应用测定结果表明：Ｋ１和ＴＰＰ对辛硫磷和乐果都有增效作用,尤以Ｋ１的增效作用更显著,其增效比分中辊为１０５３和３８,说明桃蚜对辛硫磷和乐     

Lethal and Sublethal Effects of Imidacloprid on Nicotine-Tolerant Myzus nicotianae and Myzus persicae

Bioassays of nicotine and imidacloprid against clones of Myzus persicae (Sulzer) and Myzus nicotianae (Blackman) from around the world demonstrated that some had low levels of resistance to both compounds. This was expressed not only as a reduced mortality, but more markedly as differential inhibition of feeding by imidacloprid concentrations in the parts per billion range. Such concentrations also reduced aphid fecundity by inhibiting the production and viability of nymphs, and this effect was more marked for susceptible aphids than for those showing reduced direct lethal and antifeedant effects.     

Further studies on the genetics of the carboxylesterase regulatory system involved in resistance to organophosphorus insecticides in Myzus persicae (Sulzer)

Levels of carboxylesterase activity in F₁ clones of Myzus persicae, obtained by crossing sexuales from a resistant, high esterase clone with those from a susceptible, low esterase clone, fell into two distinct groups intermediate between the levels of carboxylesterase in the parent clones. When sexuales of F₁ clones of the lower of these two intermediate levels of carboxylesterase activity were crossed, segregation ratios in the F₂ generation indicated that this lower intermediate activity level (about 0.4 μmol mg^?1 h^?1). which is about twice the level in susceptible clones, is due to mutation at a single regulatory locus. The results obtained with backcrosses, between sexuales of an F₁ clone having the higher intermediate level of carboxylesterase activity and a parent susceptible, low esterase clone, suggest that a second locus may be involved in the expression of higher levels of esterase activity.     

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     

Virus transmission by aphids in potato crops

This paper reviews the contribution of vector activity and plant age to virus spread in potato crops. Determining which aphid species are vectors is particularly important for timing haulm destruction to minimize tuber infection by potato virus Y (PVY). Alate aphids of more than 30 species transmit PVY, and aphids such asRhopalosiphum padi, that migrate in large numbers before flights of the more efficient vector,Myzus persicae, appear to be important vectors. Differences in methodology, aphid biotypes and virus strains prevent direct comparisons between estimates of vector efficiencies obtained for aphids in different countries in north western Europe. M. persicae is also the most efficient vector of potato leafroll virus (PLRV), but some clones ofMacrosiphum euphorbiae transmit PLRV efficiently toNicotiana clevelandii and potato test plants. The removal of infected plants early in the season prevents the spread of PLRV in cool regions with limited vector activity. The proportion of aphids acquiring PLRV from infected potato plants decreases with plant age, and healthy potato plants are more resistant to infection later in the season. Severe symptoms of secondary leafroll developed on progeny plants of cv. Maris Piper derived from mother plants inoculated with PLRV in June or July of the previous year. Progeny plants derived from mother plants inoculated in August showed only mild symptoms, but the concentration of PLRV in these plants was as high as that in the plants with severe symptoms.     

基于EPG技术分析杭菊两主栽品种对三种菊蚜抗性及其相关抗性物质

为比较杭白菊主栽品种早、晚小洋菊对菊小长管蚜Macrosiphoniella sanborni、棉蚜Aphis gossypii、桃蚜Myzus persicae的抗性,通过刺吸电位技术(electrical penetration graph,EPG)研究菊蚜在菊顶叶上刺吸行为,并检测了菊顶叶主要化合物。结果表明:在晚小洋菊上,菊小长管蚜的E1、E2波平均持续时间4.31、3.47 min,分别短于在早小洋菊上的4.63、3.75 min;棉蚜、桃蚜的E1、E2波平均持续时间分别为4.32、4.72 min和4.92、4.64 min;3种蚜虫的平均刺探次数均大于在早小洋菊上的平均刺探次数。聚类分析结果显示,2个主栽品种之间抗性存在差异。早小洋菊顶叶平均可溶性糖和可溶性蛋白含量分别为2.71 mg/g和25.36 mg/g,均高于晚小洋菊;早小洋菊顶叶总酚含量为0.24 mg/g,显著低于晚小洋菊;早、晚小洋菊总黄酮含量分别为3.46 mg/g和3.37 mg/g。早小洋菊顶叶可溶性糖、可溶性蛋白含量基本上与每种菊蚜的E1、E2波持续时间显著正相关,总酚含量、总黄酮含量与每种菊蚜的E1或E2波持续时间显著负相关。推测早小洋菊对于菊长管蚜的抗性稍弱于晚小洋菊,2种杭白菊对棉蚜或桃蚜的抗性相当,且菊叶中的这4种物质含量与抗蚜性相关。     

Virus transmission by aphids in potato crops

This paper reviews the contribution of vector activity and plant age to virus spread in potato crops. Determining which aphid species are vectors is particularly important for timing haulm destruction to minimize tuber infection by potato virus Y (PVY). Alate aphids of more than 30 species transmit PVY, and aphids such asRhopalosiphum padi, that migrate in large numbers before flights of the more efficient vector,Myzus persicae, appear to be important vectors. Differences in methodology, aphid biotypes and virus strains prevent direct comparisons between estimates of vector efficiencies obtained for aphids in different countries in north western Europe.M. persicae is also the most efficient vector of potato leafroll virus (PLRV), but some clones ofMacrosiphum euphorbiae transmit PLRV efficiently toNicotiana clevelandii and potato test plants. The removal of infected plants early in the season prevents the spread of PLRV in cool regions with limited vector activity. The proportion of aphids acquiring PLRV from infected potato plants decreases with plant age, and healthy potato plants are more resistant to infection later in the season. Severe symptoms of secondary leafroll developed on progeny plants of cv. Maris Piper derived from mother plants inoculated with PLRV in June or July of the previous year. Progeny plants derived from mother plants inoculated in August showed only mild symptoms, but the concentration of PLRV in these plants was as high as that in the plants with severe symptoms.     

Using bioassays with the green peach aphid (Myzus persicae) to determine residual activity of two systemically soil-applied neonicotinoid insecticides in field-grown tobacco

Duration of systemic pesticide activity under field conditions has wide implications for pest management. Our aim was to determine the duration of activity of systemic insecticides commonly used in cultivated tobacco (Nicotiana tabacum) by measuring the levels of insect infestations on field plots and effects on reproduction and survival of the green peach aphid (Myzus persicae) in controlled bioassays using field grown leaves. Plants were treated with different concentrations of two systemic neonicotinoid pesticides, imidacloprid and thiamethoxam, and grown in small field plots. Our results show that these materials are effective under field conditions against aphids for at least 13 weeks after transplant. Pesticides also affected aphid reproduction and nymph survival in bioassays, although some aphids survived on pesticide-treated leaves. We also observed that leaf age affected aphid survival. We showed that neonicotinoids were very effective against M. persicae, aphids are a useful organism to assess pesticide efficacy early in the growing season, but plant characteristics are more important than pesticide concentration in the second half of the growing season.     

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. Copyright © 2011 Society of Chemical Industry     

A carboxylesterase with broad substrate specificity causes organophosphorus,carbamate and pyrethroid resistance in peach-potato aphids (Myzus persicae)

Carboxylesterase E4, the enzyme previously shown to cause resistance to organophosphorus insecticides in peach-potato aphids, Myzus persicae, was purified and characterized by electrophoretic and enzyme kinetic techniques. Its insecticidal substrate specificity, determined by following the rate of recovery of esterase activity after inhibition by a range of acylating inhibitors, included a variety of carbamates and organophosphates, although the catalytic center activity for these substrates was low. Radiometric measurement of hydrolysis of the pyrethroid, permethrin, showed that E4, whether purified or in crude aphid homogenates, hydrolyzed the (1S)trans enantiomer rapidly but hydrolysis of the other three isomers could not be detected. Such absolute specificity for one enantiomer of a pyrethroid is rare. The rates of hydrolysis of the various insecticidal classes correlated well with the relative degrees of resistance to them, and no other resistance mechanisms have been detected. Although the enzyme is relatively inefficient in degrading insecticidal esters, it is produced in very large quantity, accounting for approximately 3% of the total protein in very resistant aphids. Its effect is thus mediated not only by hydrolysis but also by sequestering a substantial proportion of a toxic dose of insecticide. It is effective in this respect because the molar amount present is similar to that of a lethal dose of insecticide. These results support earlier indirect evidence for “overproduction” of E4, probably because of structural gene duplication or amplification and have direct implications for strategies to delay the buildup of resistance or for developing synergists to overcome resistance.     

The symbiotes of Myzus persicae (Sulz.) in strains resistant and susceptible to demeton-S-methyl

The lengths of individual symbiotes and the numbers of these per unit area in ultrathin sections of the mycetomes did not differ significantly between strains of Myzus persicae (Sulz.) that were resistant or susceptible to demeton-S-methyl in Britain. This is contrary to observations made in Germany on the symbiotes of similar aphids resistant or susceptible to parathion.     

The fate of some organophosphorus compounds applied topically to peach-potato aphids (Myzus persicae (Sulz.)) resistant and susceptible to insecticides

The fate of [¹⁴C]-dimethoate and [¹⁴C]-parathion after topical application to strains of Myzus persicae resistant and susceptible to organophosphorus compounds was investigated. In 4 h approximately 65% of the dimethoate or para-thion applied was lost from the aphids by evaporation, and this limited the amount available for penetration into the insect. Only approximately 25% of the dose applied penetrated, of which a very small proportion was excreted as metabolites. There was no difference in the amounts penetrating into the resistant and susceptible strains, indicating that penetration was not a factor contributing to the observed resistance. These findings emphasise that evaporation from the cuticle can greatly influence results when insecticides are applied topically.     

Response of susceptible and resistant peach-potato aphids Myzus persicae (Sulz.) to insecticides in leaf-dip bioassays

The response of susceptible (S), moderately resistant (R₁) and strongly resistant (R₂) peach-potato aphids, Myzus persicae (Sulz.) to organophosphorus, carbamate and pyrethroid insecticides was tested by a leaf-dip bioassay. The aphids were placed on potato leaves (dipped in insecticide solutions 1–2 or 24 h before infestation) and their mortality examined 48 h later. R₁ aphids were virtually susceptible to most of the carbamates, demephion and acephate, but were slightly to moderately resistant (2.1–9.4 times) to permethrin, cypermethrin and (S)-α-cyano-3-phenoxybenzyl (1R)-cis-3-(2,2-dibromovinyl)-2,2-dimethylcyclopropanecarboxylate (I), (NRDC 161), to 5,6,7,8-tetrahydro-2-methylquinolin-4-yl dimethylcarbamate (II), (Hoechst 25 682) and demeton-S-methyl. R₂ aphids resisted more strongly or very strongly (between 65 and 1280 times) the pyrethroids, demeton-S-methyl (×94), II (×83) and demephion (×9), and were slightly to moderately (2–5 times) resistant to acephate, pirimicarb, ethiofencarb and 2-(dimethylcarbamoyloxyimino)-3-methoxyimino-N,N- dimethylbutyramide (III), (DPX 3853). Both resistant strains were susceptible to nitrilacarb [4,4-dimethyl-5-(methylcarbamoyloxyimino)pentanenitrile] complex (1:1) with zinc chloride (IV), (AC 85 258). The implications of these results in terms of practical aphid control are discussed.     

释放异色瓢虫对北京温室甜椒和圆茄上桃蚜的控害效果

为评价异色瓢虫在温室条件下对桃蚜的控害能力及效益,在甜椒和圆茄生产温室中以生物农药防治为对照,开展释放异色瓢虫控制桃蚜的示范试验,分析天敌害虫的种群动态变化,并计算防治成本。结果显示:异色瓢虫能够持续控制甜椒温室中桃蚜种群密度,其定殖率在蚜虫暴发高峰期时最高,为64%;且甜椒产量及经济效益高于生物农药防治。在圆茄温室中,前期释放的异色瓢虫使桃蚜高峰延缓1周出现;在增加瓢虫释放量后,1周内桃蚜种群密度下降了79%,且瓢虫定殖率达到86%,控害效果较好。表明通过生产期全程监测天敌-害虫的种群动态,在植株定植15 d后每周确定益害比,通过2~3个月持续释放异色瓢虫,可有效、持续控制整个生产期桃蚜为害。     

Evaluation of semiochemical toxicity to houseflies and stable flies (Diptera: Muscidae)

BACKGROUND: The housefly, Musca domestica L., and stable fly, Stomoxys calcitrans (L.) are cosmopolitan pests of both farm and home environments. Houseflies have been shown to be resistant to a variety of insecticides, and new chemistries are slow to emerge on the market. Toxicities of selected semiochemicals with molecular structures indicative of insecticidal activity were determined against adults from an insecticide‐susceptible laboratory strain of houseflies. The three most active semiochemicals were also evaluated against recently colonized housefly and stable fly strains. RESULTS: Nineteen semiochemicals classified as aliphatic alcohols, terpenoids, ketones and carboxylic esters showed toxicity to houseflies and stable flies. Rosalva (LC₅₀ = 25.98 µg cm^?2) followed by geranyl acetone and citronellol (LC₅₀ = 49.97 and 50.02 µg cm^?2) were identified as the most toxic compounds to houseflies. Permethrin was up to 144‐fold more toxic than rosalva on the susceptible strain. However, it was only 35‐fold more toxic to the insecticide‐tolerant field strain. The compounds generated high toxicity to stable flies, with LC₅₀ values ranging from 16.30 to 40.41 µg cm^?2. CONCLUSION: Quantification of LC₅₀ values of rosalva, citronellol and geranyl acetone against susceptible housefly and field‐collected housefly and stable fly strains showed that semiochemicals could serve as potent insecticides for fly control programs. Copyright © 2010 Society of Chemical Industry     

Stability of insecticide resistance in the peach-potato aphid,Myzus persicae (Sulzer)

Anholocyclic Myzus persicae (Sulz.) from the glasshouse and field can be classified into six variants, each with a characteristic carboxylesterase activity and consequent resistance to carbamate and organophosphorus insecticides. Overlapping activity distributions prevent unequivocal identification of an individual from measurements of its enzyme activity, but this may be done by cloning and measuring the frequency distribution of the enzyme activity in its progeny. Although each of the four lowest esterase variants has a frequency distribution with a single peak, only the susceptible (USLL) is best described by a single Normal distribution. Those of the other three clones (MSIG, French R and T1V) correspond better with two Normally-distributed components. However, these are not well separated and the lower component accounts for only 10-25% of the individuals. The very resistant clones from glasshouses have higher mean activities but with very broad distributions covering the whole range observed in this aphid. These broad distributions have peaks corresponding to those of the less resistant variants and arise from the appearance at each generation of a small proportion of individuals with much less active enzyme than their parents. Careful monitoring of the changes in activity between parents and offspring of the most resistant variant (clone G6) showed that complete loss of resistance can occur either in a single step or over several generations, and that below a threshold value, reversion to higher levels is very rare. Such reversion could only be detected by selection, with insecticide, of large clonal populations from an individual that had lost activity. High esterase activity was not stabilised by breeding for 16 generations only from individuals with high esterase activity, and is probably maintained in glasshouses by continuous exposure to insecticides. Spontaneous loss of esterase activity and resistance was observed only in glasshouse populations.     

Release of Aphidius matricariae for control of Myzus persicae in glasshouses1

Aphidius matricariae, the most effective antagonist of Myzus persicae, has been released in experiments to control this aphid on capsicum, Anthurium andreanum and Ficus lyrata. In all cases, effective short-term as well as long-term control has been achieved through an infestation-linked application of the parasite, at parasitoid-host ratios from 1:5 to 1:50. Control experiments in capsicum over 31 weeks showed effective control of M. persicae by A. matricariae, even at a high infestation level. Chemical insecticides were not needed against aphids until week 26, when it became necessary to avoid damage due to mass propagation of naturally occurring Aphis gossypii and Macrosiphum euphorbiae. The results show that, with increasing complexity of biological control methods, the species diversity of aphids increases their significance as pests. In particular, biocontrol of aphids seems to necessitate the use of several antagonists.     

Insecticidal spider venom toxin fused to snowdrop lectin is toxic to the peach-potato aphid, Myzus persicae (Hemiptera: Aphididae) and the rice brown planthopper, Nilaparvata lugens (Hemiptera: Delphacidae)

The SFI1/GNA fusion protein, comprising of snowdrop lectin (Galanthus nivalis agglutinin, GNA) fused to an insecticidal spider venom neurotoxin (Segestria florentina toxin 1, SFI1) was tested for toxicity against the rice brown planthopper Nilaparvata lugens (Stål) and the peach‐potato aphid Myzus persicae (Sulzer) by incorporation into artificial diets. Significant effects on the mortality of N. lugens were observed, with 100% of the insects fed on the SFI1/GNA fusion protein diet dead by day 7. The survival of the aphid M. persicae was also reduced when fed on the SFI1/GNA fusion protein. After 14 days, only 49% of the aphids that were fed on the fusion protein were still alive compared with approximately 90% of the aphids fed on the control diet or on diet containing GNA only. The SFI1/GNA fusion protein also slowed the development of M. persicae, and the reproductive capacity of the aphids fed on the SFI1/GNA fusion protein was severely reduced. The ability of GNA to act as a carrier protein, and deliver the SFI1 neurotoxin to the haemolymph of N. lugens, following oral ingestion, was investigated. The successful delivery of intact SFI1/GNA fusion protein to the haemolymph of these insects was shown by western blotting. Haemolymph taken from the insects that were fed on the fusion protein contained two GNA‐immunoreactive proteins of molecular weights corresponding to GNA and to the SFI1/GNA fusion protein. © 2005 British Crown Copyright. Published for SCI by John Wiley & Sons, Ltd.