Spatial and temporal dynamics of Myzus persicae clones in fields and suction traps

1 The population of peach‐potato aphid Myzus persicae in Scotland is comprised almost entirely of long‐term asexual clones. 2 Over a ten year period, M. persicae from Scottish fields and suction traps were analysed with six microsatellite markers. 3 Out of 1497 individuals analysed, 14 clones (denoted A–N) comprised over 98% of the collection. 4 Some clones were particularly abundant but most clones had a widespread distribution on all available plants. 5 Clones E and L had distinct features in their distributions as clone L was geographically totally restricted to the north east of Scotland and clone E showed a marked preference for brassica crops. 6 Clones E and L provide direct evidence of a role for local adaptations in the distribution of M. persicae clones.     

Efficacy of pymetrozine against Myzus persicae and in reducing potato virus Y transmission on tobacco plants

Forty‐four parthenogenetic lineages of Myzus persicae s.l. (Sulzer) from tobacco crops and peach orchards located in various regions of Greece were examined to determine their response to the insecticide pymetrozine using leaf‐dip bio‐assays. The results show that the aphid has not developed resistance, as all lineages exhibited resistance factors bellow 6.0. In transmission experiments of potato virus Y (PVY) using a lineage of the tobacco‐adapted subspecies M. persicae nicotianae Blackman on tobacco plants, one foliar application with pymetrozine provided adequate protection for 7 days. Pymetrozine significantly reduced both virus acquisition and inoculation compared with the untreated control and the reduction was comparable to a mineral oil application. These results are discussed in terms of the advantage of incorporating pymetrozine as a compound of pest management strategies against M. persicae s.l. and for control of non‐persistent viruses, especially in crops such as tobacco because of the high selection pressure from neonicotinoids resulting in potential of resistance developing in aphid populations.     

Incidence of insecticide resistance alleles in sexually-reproducing populations of the peach-potato aphid Myzus persicae (Hemiptera: Aphididae) from southern France

Intensive chemical treatments have led to the development of a number of insecticide resistance mechanisms in the peach-potato aphid Myzus persicae (Sulzer). Some of these mechanisms are known to be associated with negative pleiotropic effects (resistance costs). Molecular and biochemical methods were used to determine the genotypes or phenotypes associated with four insecticide resistance mechanisms in single aphids from sexually-reproducing populations in southern France. The mechanisms considered were E4 and FE4 carboxylesterase overproduction, modified acetycholinesterase, and kdr and rdl resistance-associated mutations. A new method for determining individual kdr genotypes is presented. Almost all resistant individuals overproduced FE4 carboxylesterase, whereas modified acetylcholinesterase was rare. Both the kdr and rdl resistance mutations were present at high frequencies in French sexually-reproducing populations. The frequencies of insecticide resistance genes were compared before and after sexual reproduction in one peach orchard at Avignon to evaluate the potential impact of selection on the persistence of resistance alleles in the over-wintering phase. The frequencies of the kdr and rdl mutations varied significantly between autumn and spring sampling periods. The frequency of the kdr mutation increased, probably due to pyrethroid treatments at the end of the winter. Conversely, the frequency of the rdl mutation decreased significantly during winter, probably because of a fitness cost associated with this mutation.     

Reduced response of insecticide-resistant aphids and attraction of parasitoids to aphid alarm pheromone; a potential fitness trade-off

Response to the alarm pheromone, (E)-beta-farnesene, produced by many species of aphids, was assessed in laboratory bioassays using an aphid pest, Myzus persicae (Sulzer), and its primary endoparasitoid, Diaeretiella rapae (McIntosh). This was done in three separate studies, the first investigating responses of a large number of M. persicae clones carrying different combinations of metabolic (carboxylesterase) and target site (kdr) insecticide resistance mechanisms, and the other two investigating the responses of young virgin female adult parasitoids. In M. persicae, both insecticide resistance mechanisms were associated with reduced repellence suggesting that each has a pleiotropic effect on aphid behaviour. In contrast, D. rapae females were attracted to the alarm pheromone source. The implications of this apparent fitness trade-off for the evolution and dynamics of insecticide resistance, and the potential for using beneficial insects to combat resistance development are discussed.     

Performance of Myzus persicae (Hemiptera: Aphididae) clones on different host-plants and their host preference

The performance of eighteen clones of Myzus persicae (Sulzer) on pepper and tobacco plants at 20 degrees C and L16:D8 and the choice of young adult apterae between tobacco and pepper leaf-discs were examined. The clones were collected from weeds and peach in two tobacco-growing regions: Katerini, northern Greece and Karditsa, central Greece (only from weeds) and from Lehonia, central eastern Greece where tobacco is not cultivated. All clones did well on both hosts. However, the analysis of data revealed a significant effect of "region / host plant origin" on aphid performance. The mean values of adult weight, intrinsic rate of increase and fecundity of the clones collected in Lehonia and reared on tobacco were significantly lower than the observed values for clones from Katerini and Karditsa. Aphids from Lehonia had significantly higher mean values for developmental time on tobacco than clones from the other regions whereas the opposite was observed when aphids were reared on pepper. Aphids collected in Lehonia performed better on pepper than those originating from the tobacco-growing regions. A choice test revealed differences among the clones originating from different regions. Fifty three percent and 43% of aphids from weeds and peach from Lehonia, respectively, chose pepper. By comparison 41.5% and 40.0% of aphids from peach and weeds from Katerini, respectively and 49.5% of aphids from Karditsa preferred tobacco. The results are discussed in relation to host specialization in M. persicae.     

Amplification and methylation of an esterase gene associated with insecticide-resistance in greenbugs, Schizaphis graminum (Rondani) (Homoptera: Aphididae)

The greenbug aphid, Schizaphis graminum (Rondani) has developed resistance to organophosphorus insecticides by the over-production of esterases that have been classified as Type I and Type II. The first twenty N-terminal amino acids of the Type I esterase were determined and used to design an oligonucleotide, which in conjunction with an active site primer derived from conserved sequences of other insect esterases and two internal primers specific for esterases from another aphid species resulted in a 0.85 kb genomic DNA fragment from resistant greenbugs. This was extended by 5′ RACE which provided approximately 1.2 kb of the 5′ end of the esterase gene. The 5′ DNA sequence corresponded to 19 of the 20 known amino acids of the Type I esterase, with the last needing only a one base change (probably resulting from a PCR artifact). Furthermore, the sequence showed very close similarity to the amplified E4/FE4 esterase genes of Myzus persicae (Sulzer). A comparison of sequences suggested that the S. graminum gene has introns in the same positions as the first two introns of E4/FE4, with the second intron being considerably larger in S. graminum. Probing of Southern blots with the 0.85 kb esterase fragment showed that the gene encoding the Type I esterase is amplified 4- to 8-fold in resistant S. graminum and that the amplified sequences contain 5-methylcytosine at MspI/HpaII sites, again in agreement with previous findings for M. persicae genes.     

不同品种烤烟对烟蚜羧酸酯酶和谷胱甘肽转移酶活性的影响

为明确不同烤烟品种对烟蚜Myzus persicae(Sulzer)解毒酶活力的影响,比较了9个品种烤烟上的绿色型烟蚜的解毒酶活力,发现烤烟品种对烟蚜α-乙酸萘酯羧酸酯酶、β-乙酸萘酯羧酸酯酶、谷胱甘肽转移酶活力有显著影响,不同品种烤烟上烟蚜的α-乙酸萘酯羧酸酯酶、β-乙酸萘酯羧酸酯酶、谷胱甘肽转移酶均有差异。烤烟品种V2、云烟87和红大上的烟蚜羧酸酯酶活力较高,云烟317、云烟85和K326居中,K346、K358和G28则较低。9个品种烤烟对烟蚜谷胱甘肽转移酶活力有所影响,但差异不大。     

寄主植物对烟蚜药剂敏感性及相关酶活性的影响

以甘蓝、烟草和桃树上的饲养的烟蚜Myzus persicae（Sulzer）种群为研究材料,分别测试了其对吡虫啉、丁硫克百威和氰戊菊酯这3种杀虫剂的敏感性。结果表明,桃树上的烟蚜种群敏感性最高,其次为烟草种群,甘蓝种群最低。测试了在3种不同寄主上饲养的烟蚜种群的羧酸酯酶、乙酰胆碱酯酶和谷光甘肽-s-转移酶的比活力,这3种酶均表现为甘蓝种群最高,其次为烟草种群,桃树种群最低。     

Identification of novel aphid-killing bacteria to protect plants

Aphids, including the peach-potato aphid, Myzus persicae, are major insect pests of agriculture and horticulture, and aphid control measures are limited. There is therefore an urgent need to develop alternative and more sustainable means of control. Recent studies have shown that environmental microbes have varying abilities to kill insects. We screened a range of environmental bacteria isolates for their abilities to kill target aphid species. Tests demonstrated the killing aptitude of these bacteria against six aphid genera (including Myzus persicae). No single bacterial strain was identified that was consistently toxic to insecticide-resistant aphid clones than susceptible clones, suggesting resistance to chemicals is not strongly correlated with bacterial challenge. Pseudomonas fluorescens PpR24 proved the most toxic to almost all aphid clones whilst exhibiting the ability to survive for over three weeks on three plant species at populations of 5–6 log CFU cm⁻² leaf. Application of PpR24 to plants immediately prior to introducing aphids onto the plants led to a 68%, 57% and 69% reduction in aphid populations, after 21 days, on Capsicum annuum, Arabidopsis thaliana and Beta vulgaris respectively. Together, these findings provide new insights into aphid susceptibility to bacterial infection with the aim of utilizing bacteria as effective biocontrol agents.     

Genetic variation in Myzus persicae populations associated with host-plant and life cycle category

Random amplified polymorphic DNA (RAPD) analysis was applied on 96 clones of Myzus persicae (Sulzer) (Homoptera: Aphididae) representing seven populations collected from different host-plants and regions of Greece. Ten decamer random primers were used to evaluate genetic variation among the examined samples. Despite the variability found between clones, no specific RAPD marker was detected to discriminate the different populations. A significant finding was that aphids from peach and pepper, which were collected far away from tobacco-growing regions, especially those from peach, showed genetic divergence from the tobacco-feeding clones. Moreover, data analysis revealed a significant genetic divergence between holocyclic and anholocyclic populations from tobacco. Lastly, holocyclic clones showed higher level of estimated heterozygosity than the nonholocyclic (anholocyclic, androcyclic and intermediate) ones.     

A multiple homogenizer for rapid sample preparation in immunoassays and electrophoresis

A multiple homogenizer is described for preparing samples of small invertebrates or tissue in a flat-bottom immunoplate. Its efficiency was evaluated by immunoassay of a carboxylesterase (E4), the enzyme conferring insecticide resistance in the peach potato aphid (Myzus persicae). This equipment was shown to release more enzyme, with less variability, than homogenizing individual aphids and its efficiency allows one person to analyze up to 3000 individual insects per day. It is also suitable for preparing samples for electrophoretic analysis. In the present study samples were loaded onto electrophoresis gels rapidly and accurately by using an eight-channel multi-pipette.R. H. ffrench-Constant was supported by an AFRC studentship.     

Poor survival on an artificial diet of two genotypes of the aphid Myzus persicae: a fitness cost of insecticide resistance?

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Fitness and predation potential of Macrolophus pygmaeus reared under artificial conditions

Abstract The biological parameters of Macrolophus pygmaeus Rambur after prolonged rearing in the absence of plant materials were compared with those of conventionally plant‐reared predators. When eggs of Ephestia kuehniella Zeller were provided as food, developmental and reproductive fitness of M. pygmaeus reared for over 30 consecutive generations using artificial living and oviposition substrates was similar to that of predators kept on tobacco leaves. Plantless‐reared fifth instars of the predator also had similar predation rates on second instars of the tobacco aphid, Myzus persicae nicotianae Blackman, as their peers maintained on plant materials. In a further experiment, predation on aphid prey by fifth instar M. pygmaeus fed one of two egg yolk‐based artificial diets was compared with that of nymphs fed E. kuehniella eggs. Despite their lower body weights, predators produced on either artificial diet killed similar numbers of prey as their counterparts reared on lepidopteran eggs. Our study indicates that artificial rearing systems may be useful to further rationalize the production of this economically important biological control agent.     

Variation in the life cycle and morphology of the tobacco host-race of Myzus persicae (Hemiptera: Aphididae) in relation to its geographical distribution

Morphological variation and life cycle category were examined in 121 clones of Myzus persicae (Sulzer). The clones were collected from tobacco from three localities in Greece (Xanthi, Nea Efessos and Naphplion), one in Germany (Rheinstetten), one in France (Bergerac) and one in Spain (Madrid). Before morphometrics, all aphids were laboratory-reared on potato. The morphological variation was investigated using both canonical variates analysis and a novel non-parametric classification tree method. The life cycle category was examined by rearing the clones for three generations under short day conditions. In Nea Efessos a relative high proportion of clones was found to overwinter as eggs on the primary host. In the other regions all collected clones were non-holocyclic. Intermediate genotypes were found in all regions at percentages ranging from 4.0 to 24.0%. Androcyclic clones were found only in Xanthi, Greece (4.0%) and Rheinstetten, Germany (16.7%). The canonical variates analysis and the tree classification method revealed important intrapopulation polymorphisms in clones from Bergerac, Nea Efessos and Madrid. Both methods separated the populations originating from Greece from those collected elsewhere in western Europe. The observed morphological variation was probably due to genetic differences, since all clones were reared in a common environment. The results are discussed in relation to factors responsible for genetic divergence in M. persicae populations.     

Life cycle variation of Myzus persicae (Hemiptera: Aphididae) in Greece

During the years 1995-1999 the life cycle category of 2797 clones of Myzus persicae (Sulzer) was examined. The clones originated from primary and secondary hosts from different localities of North and Central Greece and the island of Crete in the south. Four different overwintering life cycle strategies were found that have also been described for M. persicae and other heteroecious species previously. A geographical variation was found in the proportion of holocyclic clones from tobacco and other secondary hosts associated with the abundance of the primary host in the sampling regions. In Central Macedonia, around the main peach-growing regions, the proportion of holocyclic clones was mostly above 50% and in some cases reached 100%. In localities of East Macedonia, holocyclic clones were also frequent. On the other hand, further south or in north-eastern Greece, where peach is not common, the proportion of holocyclic clones varied between 0 and 33%. Fifty seven percent of examined anholocyclic clones produced males under short day conditions, suggesting that androcyclic clones in Greece represent an important factor of genetic variability. Intermediate clones were sampled from all host-plants but at low frequencies (3.6% of total examined clones and 6.9% of non-holocyclic ones). Moreover, a regional variation was found in different colour forms feeding on tobacco plants. Red clones were predominant in regions where aphids overwinter parthenogenetically on weeds or winter crops. However, almost all clones from the primary host were green. The ecological aspects of life cycle variation are discussed.     

Distribution of common genotypes of Myzus persicae (Hemiptera: Aphididae) in Greece, in relation to life cycle and host plant

Microsatellite genotyping was used to identify common clones in populations of the Myzus persicae group from various hosts and regions in mainland Greece and southern Italy and to compare their distribution and occurrence on tobacco and other crops. Common clones were defined as genotypes collected at more than one time or in more than one population; and, therefore, unlikely to be participating in the annual sexual phase on peach. Sixteen common genotypes were found, accounting for 49.0% of the 482 clonal lineages examined. Eight of these genotypes were subjected, in the laboratory, to short days and found to continue parthenogenetic reproduction, i.e. they were anholocyclic. Four of the six commonest genotypes were red, and one of these accounted for 29.6% of the samples from tobacco and 29.4% of those from overwintering populations on weeds. All six commonest genotypes were found on weeds and five of them both on tobacco and on other field crops. In mainland Greece, the distribution of common clones corresponded closely with that of anholocyclic lineages reported in a previous study of life cycle variation. Common genotypes were in the minority in the commercial peach-growing areas in the north, except on weeds in winter and in tobacco seedbeds in early spring, but predominated further south, away from peach trees. This contrasts with the situation in southern Italy, reported in a previous paper, where peaches were available for the sexual phase, yet all samples from tobacco were of common genotypes.     

Within-species variability of the response to 20-hydroxyecdysone in peach-potato aphid (Myzus persicae Sulzer)

Phytoecdysteroids have been proposed as new tools for controlling crop pests because of their endocrine disruption and deterrent effects on insects and nematodes. There is increasing evidence of variability between taxa in sensitivity to phytoecdysteroids, but the genetic variability of this sensitivity within species is unknown. However, knowledge about this intraspecies variability is required for predicting evolution of the pest's response to new control methods. We assessed the variability of the response of the aphid Myzus persicae Sulzer, a major agricultural pest, to 20-hydroxyecdysone (20E). We determined the number of nymphs produced by six clones of M. persicae exposed to various concentrations of 20E and the capacity of these clones to detect 20E in choice experiments. High concentrations of 20E significantly decreased the number of nymphs produced for two clones and both increases and decreases in the number of offspring were detected at low concentrations. Two clones significantly avoided food with 20E, while one significantly preferred it, suggesting that 20E does not always act as a deterrent in this species. We conclude that genetic variability in the response to 20E exists in natural populations of M. persicae. The consequences of this finding on the sustainability of control methods using 20E are discussed.     

Genetic, biochemical, and behavioral uniformity among populations of Myzus nicotianae and Myzus persicae

Prior to designation as distinct species, an appellation presently in question, the tobacco aphid, Myzus nicotianae Blackman (Homoptera: Aphididae), was classified as a tobacco-feeding form of the green peach aphid, Myzus persicae (Sulzer). In this study, RAPD polymorphisms distinguished members of the Myzus persicae complex (M. persicae and M. nicotianae) from three outgroup Myzus species (M. cerasi (F.), M. hemerocallis Takahashi, and M. varians Davidson). Polymorphisms within the complex did not separate populations on the basis of host association (tobacco versus other host plants) or geographic origin (collections from the United States, Europe, and Japan). Similarly, while GC-MS analysis of cuticular hydrocarbon profiles revealed both developmental and inter-populational differences within the M. persicae complex, it did not separate populations of tobacco feeding aphids from those collected off non-tobacco hosts. Finally, with the exception of their responses to a choice between lettuce and collards, the host preference behavior of a green peach aphid population, a red tobacco aphid population, and a green tobacco aphid population was indistinguishable in host preference experiments. These results add to a growing body of evidence suggesting M. nicotianae and M. persicae are conspecific.