Clonal turnover of MACE-carrying peach-potato aphids (Myzus persicae (Sulzer), Homoptera: Aphididae) colonizing Scotland

Peach-potato aphids, Myzus persicae (Sulzer), collected in Scotland in the years 1995 and 2002-2004 were characterized using four microsatellite loci and three insecticide resistance mechanisms. From 868 samples, 14 multilocus genotypes were defined (designated clones A-N). Five of these (denoted A, B, H, M and N) carried modified acetylcholinesterase (MACE) resistance, the most recent resistance mechanism to have evolved in M. persicae. The current paper shows that the continued presence of MACE aphids is due to turnover, as clones A and B were replaced in field samples by clones H, M and N in later seasons. Thus, insecticide-resistant populations in Scotland can be attributed to multiple waves of rapid clone colonisations and not to the continued presence of stable resistant clones or mutation or sexual recombination in local populations. The MACE clones carried varying levels of the other insecticide resistance mechanisms, kdr and esterase. The presence of these mechanisms could alter the clones success in the field depending on insecticide spraying (positive selection) and resistance fitness costs (negative selection).     

The attack of the clones: tracking the movement of insecticide-resistant peach-potato aphids Myzus persicae (Hemiptera: Aphididae)

Myzus persicae (Sulzer) collected in Scotland were characterized for four microsatellite loci, intergenic spacer fingerprints and the resistance mechanisms modified acetylcholinesterase (MACE), overproduced carboxylesterase and knockdown resistance (kdr). Microsatellite polymorphisms were used to define a limited number of clones that were either fully susceptible to insecticides or possessed characteristic combinations of resistance mechanisms. Within these clones, intergenic spacer fingerprints could either be very consistent or variable, with the latter indicating ongoing evolution within lineages, most likely derived from the same zygote. Two clones (termed A and B) possessed all three resistance mechanisms and predominated at sites treated with insecticides. Their appearance on seed potatoes and oilseed rape in Scotland in 2001 coincided with extensive insecticide use and severe control failures. Clones C, I and J, with no or fewer resistance mechanisms, were found in samples from 1995 and were dominant at untreated sites in 2001. A comparison of Scottish collections with those from other UK and non-UK sites provides insight into the likely origins, distribution and dynamics of M. persicae clones in a region where asexual (anholocyclic) reproduction predominates, but is vulnerable to migration by novel genotypes from areas of Europe where sexual (holocyclic) reproduction occurs.     

Probiotic effects of beta-glucuronidase on the peach-potato aphid Myzus persicae (Aphididae)

beta-glucuronidase (GUS) is a reporter protein commonly expressed in transgenic plants allowing the visualization of the transformed individuals. In our recent work, we showed that consumption of transformed potato plants expressing this GUS enzyme improves performance of the phloem feeding aphid Myzus persicae. Those results led us to the conclusion that the expression of GUS in potato plants might be responsible for the probiotic effect measured in feeding aphids. In the present paper, artificial diets were used to provide active GUS (10 and 500 microg ml(-1)), inactivated heated GUS (500 microg ml(-1)), glucuronic acid (10, 100 and 500 microg ml(-1)), and bovine serum albumin (500 microg ml(-1)) to M. persicae. Our results reveal that these chemicals provided as food intake might influence the biological parameters of this aphid. Experiments showed a probiotic effect of 500 microg ml(-1) GUS diet, resulting in reduced larval mortality, and increased adult reproduction period and fecundity, which led to an increased population growth potential (r(m)=0.17+/-0.01 versus r(m)=0.12+/-0.03 for aphids fed on control diet). A lower amount of added GUS led to fewer variations, biological parameters being only slightly altered (r(m)=0.14+/-0.03). Statistically similar alterations of the biological parameters were obtained when comparing aphids fed on the diet added with inactivated GUS or the non-structural bovine serum albumin protein (r(m)=0.15+/-0.02 and 0.14+/-0.03, respectively). Feeding assays conducted with glucuronic acid supplemented diets enhanced longevity and nymph production of the adult aphids and reduced larval mortality, resulting in r(m)=0.15+/-0.02 for the highest dose (500 microg ml(-1)). Although 100 microg ml(-1) glucuronate diet did not induce any effect on M. persicae (r(m)=0.12+/-0.03), aphids fed on 10 microg ml(-1) glucuronate diet exhibited unexpected reduced demographic parameters (r(m)=0.10+/-0.03). Immuno-histological analysis showed GUS labeling along the whole digestive epithelium of adults and in various tissues including embryos and bacteriocytes. These results suggest that GUS crosses through the digestive tract. Western blots performed with protein extracts of transformed potato plants expressing the gus gene showed a unique band of molecular weight 76 kDa. On the contrary, in extracts from aphids fed on transgenic potato plants or bred on GUS 500 microg ml(-1) artificial diet, several proteins of lower molecular weight were hybridized, revealing proteolysis of ingested GUS. It is concluded that GUS protein, and more precisely GUS activity, is responsible for the probiotic effects on aphid feeding. The possible pathways of induction of such physiological alterations by GUS are discussed.     

Microsatellite isolation, linkage group identification and determination of recombination frequency in the peach-potato aphid, Myzus persicae (Sulzer) (Hemiptera: Aphididae)

Fifteen polymorphic microsatellite markers were used to establish linkage groups and relative rates of recombination in male and female Myzus persicae (Sulzer) (Hemiptera: Aphididae) (peach-potato aphid). We cloned nine markers from M. persicae and for these we report primer sequences and levels of allelic diversity and heterozygosity in four Australian M. persicae populations. Of the remaining six loci, four loci, also cloned from M. persicae, were obtained from G. Malarky (Natural History Museum, London) and two loci from Sitobion miscanthi were used. Additionally, the primer sequences of locus M77, a locus monomorphic in M. persicae but polymorphic in the closely related Myzus antirrhinii, are presented. Eleven of the 15 polymorphic markers were autosomal and four were X-linked. A linkage analysis was performed on a European pedigree of aphids containing five families with between seven and 11 offspring each. There was no linkage between any loci in females. In males, several pairwise comparisons yielded no recombinant offspring. With the exception of locus M40, these observations were supported in a linkage analysis performed on larger families produced from Australian M. persicae crosses. Locus M40 showed segregation consistent with involvement in a translocation between autosomes 1 and 3 in European samples but not in the Australian samples. From the Australian crosses we report an absence of recombination in males but high recombination rates in females. One X chromosome and four autosomal linkage groups were identified and tentatively assigned to chromosomes. The relevance of achiasmate meiosis to the evolution of sex is discussed.     

Clonal composition of the peach-potato aphid Myzus persicae (Homoptera: Aphididae) in France and Scotland: Comparative analysis with IGS fingerprinting and microsatellite markers

Fourteen colonies of the peach‐potato aphid, Myzus persicae, were taken either from French peach trees or weeds in 2001. Thirty five apomictic parthenogenetic lineages (APLs) were established. Ribosomal DNA intergenic spacer (IGS) fingerprinting was used to characterise these and 28 fingerprints were duly obtained. Those lineages with different fingerprints were considered different genotypes and those with the same fingerprint as the same. The genetic identity of APLs was further tested using four microsatellite loci. APLs that differed by IGS fingerprint had distinct microsatellite allele combinations and those that had the same IGS fingerprint had the same microsatellite allele combinations. The results confirmed that IGS types corresponded to different aphid genotypes. Independent APLs with identical IGS and microsatellite genotype were therefore considered different representatives of the same clone. APLs from M. persicae found on Scottish crops in 1995, 1996 and 2001, as well as a long‐term laboratory line were also examined by the same methods. Their IGS fingerprints were similar or identical suggesting that they all belonged to the same clone. Microsatellite markers also suggested that these lineages were derived from a single clone. Some field lineages exhibited slight modifications to their IGS fingerprints confirming that the IGS evolves more rapidly than these microsatellite alleles. Thus, IGS will continue to provide a useful marker for aphid fieldwork.     

Segregation of autosomes during spermatogenesis in the peach-potato aphid (Myzus persicae) (Sulzer) (Hemiptera: Aphididae)

Most aphids are cyclic parthenogens, so are ideal models in studies of the mechanisms and consequences of sex and recombination. However, owing to a shortage of physical and genetic markers, there have been few studies of the most fundamental genetic processes in these organisms. For example, it is not known whether autosomal segregation during male spermatogenesis is in Mendelian proportions: we address that question here. The aphid Myzus persicae has a typical karyotype of 2n = 12 in females (XX), while males are XO (2n = 11). During male meiosis, only the spermatocytes with an X chromosome are viable. We hypothesized that assortment of autosomes might be non-random because chromosomal imprinting leading to elimination of the paternal autosomes is seen in the closely related coccoids. In other aphid models, we have observed segregation distortions at single microsatellite loci (Wilson, 2000). Such distortions may have nothing to do with 'selfish' behaviour, but may be caused by mutation accumulation causing fitness differentials. Thus single-locus distortions might be predicted to be more likely to be detected via the male lines of clones that have lost the ability to reproduce sexually (male-producing obligate parthenogenesis (androcyclic)). Using microsatellites we show that genetic imprinting or selfish autosome behaviour does not occur in male M. persicae. Generally, loci segregated in Mendelian proportions in both sexes of cyclically parthenogenetic (holocyclic) clones. However, in androcyclic clones, segregation distortions consistently involved the same two autosomes. This is consistent with linkage of markers to deleterious mutations associated with a loss of sexual reproduction.     

Studies on the morphometry of the green peach aphid, Myzus persicae (Sulzer) (Homoptera: Aphididae)

Aphids are one of the most important groups of phytophagous insects because of their polyphenism, host alternation, heteroecious behavior and reproductive habits. Laboratory culture of the aphid Myzus persicae on the host plant Brassica pekinensis Hubner was maintained seasonally for biometrical studies. The metric analysis revealed gradual developmental changes from the first instar nymph to the adult stage in size, shape, and a number of characters. The transitional changes were sufficiently distinct to allow differentiation of each of the life stages irrespective of the season. A key to the nymphal stages of the pest is provided.     

An interval timer controls the production of sexual morphs in Myzus persicae (Homoptera: Aphididae)

Abstract The interval timer mechanism was examined in two clones of Myzus persicae (Sulzer) originating from peach in Naousa, North Greece. Overwintering eggs were collected in April 1997 and after egg hatch the morphs of progeny were recorded at LD 10 : 14 h and 17 °C until the appearance of sexuals. In one clone, observations were continued until the 21st generation after egg hatch. In addition, the production of sexual morphs was examined in two other clones originating from Lehonia, Central Greece, when apterae (wingless females) were transferred from long to short days and 17 °C in the second and eighth generation of parthenogenetic rearing after their collection from peach trees. In clones from Naousa, the first gynoparae appear and comprise 10% of alate (winged) females in the second half of the third generation after egg hatch. The percentage of gynoparae increased in each consecutive generation, reaching 100% in fifth and seventh generations in clones Nb and Na, respectively. First males appeared later than gynoparae, in fifth and sixth generation in clones Nb and Na, respectively. In clone Nb, the number of males, after their appearance in fifth generation, was relatively constant from the sixth to the ninth generation (10.8–12.3). Then a significant increase was observed in the 10th generation, thereafter remaining relatively constant until the 21st generation (18.1–19.8). A similar trend in the number of males produced was observed in clones deriving from Lehonia.     

Separation of Myzus (Nectarosiphon) antirrhinii (Macchiati) from Myzus (N.) persicae (Sulzer) and related species in Europe (Homoptera: Aphididae)

Abstract. Multivariate morphometric analysis (canonical variates) was used to discriminate between closely related taxa within the Myzus persicae group. It is demonstrated that dark green, anholocyclic populations with a 2n=13 or 2n=14 karyotype in Europe, hitherto treated as a form or biotype of M.persicae, all conform to a discrete morphometric grouping and should therefore be treated as a separate taxonomic entity, permanently isolated from the M.persicae gene pool. It is suggested that this taxon was first described as Siphonophora antirrhinii by Macchiati in 1883. A discriminant function is provided to separate most individual apterae of antirrhinii from those of persicae. Myzus icelandicus Blackman sp.n., on Caryophyllaceae and other plants in Iceland, is distinguished from M.polaris H.R.L. and M.certus (Walker), and a key is given to apterous virginoparae of the species of the M.persicae group in Europe.     

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.     

Pathogenicity of hyphomycet fungi to aphids Aphis gossypii Glover and Myzus persicae (Sulzer) (Hemiptera: Aphididae)

The aphids Aphis gossypii and Myzus persicae are cosmopolitan, poliphagous and damage cultivated plants. The effects of the entomopathogenic fungi Beauveria bassiana (isolate IBCB 66), Metarhizium anisopliae (isolate IBCB 121), Paecilomyces fumosoroseus (isolate IBCB 141) and Lecanicillium (= Verticillium) lecanii (isolate JAB 02) on third instar nymphs of A. gossypii and M. persicae were evaluated in the laboratory at 25 degrees C, 70 +/- 10% RH and 12h photophase. The aphids were transferred to petri dishes with a foliar disk (cotton or pepper) with a layer of 1 cm tick of agar-water. The fungi were applied in a suspension containing 1.0 x 106 to 1.0 x 108 conidia/ml. In the control treatment 1 ml of sterilized water was added to the foliar disks. The mortality of aphids was evaluated daily. B. bassiana and M. anisopliae caused 100% mortality at the seventh day after inoculation, for both species. L. lecanii was the fungus that provided mortality later in the aphids and M. persicae was more susceptible to both fungi than A. gossypii.     

The resistance of three Solanum species to Myzus persicae, Macrosiphum euphorbiae and Aulacorthum solani (Aphididae: Homoptera)

A study was made of the resistance of three wild potato species, Solanum chacoense, S. stoloniferum and S. demissum, to three aphid species, namely Myzus persicae (Sulz.), Aulacorthum solani (Kalt.) and Macrosiphum euphorbiae (Thomas), which all commonly infest the cultivated potato, S. tuberosum. The resistance of each Solanum species differed with the species of aphid, with the part of the plant and with the physiological condition of the leaf. As a result, it may be difficult to utilize such resistance, since it is ineffective against some of the pest aphid species and may break down as the physiological condition of the plant changes.     

Biological performance of Myzus persicae and Macrosiphum euphorbiae (Homoptera: Aphididae) on seven wild Solanum species

Resistance level of seven accessions of wild Solanum species (Solanaceae) to Myzus persicae and Macrosiphum euphorbiae (Homoptera: Aphididae) was evaluated by measuring survival and fecundity during sleeve cage experiments and population growth on whole plants in a controlled environment. The survival was lowest on the Solanum circaeifolium spp. capsicibaccatum, Solanum pinnatisectum and Solanum trifidum accessions for M. persicae and on the S. circaeifolium spp. capsicibaccatum, Solanum okadae and S. trifidum, accessions with M. euphorbiae. Plant species significantly influenced the fecundity of both aphid species. Aphid population growth on whole plants was negatively affected by the age of the plant, but generally followed the levels of net reproductive rate on different plant species observed during the sleeve cage experiment. The population of M. persicae varied among the seven wild potato accessions with the lowest growth rates on S. circaeifolium spp. capsicibaccatum, Solanum polyadenium, Solanum tarijense and S. trifidum. The number of M. euphorbiae also varied among accessions but the smallest population was collected from S. polyadenium. The results can be used to identify sources of resistance to aphids within those accessions already known to be resistant to the Colorado potato beetle. This study highlights the difficulties involved in developing a high-throughput screening test for aphid resistance compatible with a potato breeding programme.     

Mass Rearing of Aphidius gifuensis (Hymenoptera: Aphidiidae) for Biological Control of Myzus persicae (Homoptera: Aphididae)

The green peach aphid, Myzus persicae, is a major pest of tobacco, Nicotiana tabacum, in Yunnan province, China, where its control still depends on the use of insecticides. In recent years, the local government and farmers have sought to improve the biological control of this tobacco pest. In this paper, we present methods for mass rearing Aphidius gifuensis, a dominant endoparasitoid of M. persicae on tobacco plants in this region. The tobacco cultivar K326 (N. tabacum) was used as the host plant and M. persicae as the host insect. In the greenhouse, we collected tobacco seedlings for about 35 days (i.e., until the six-true-leaf stage), transferred them to 7.5-cm diameter pots, and kept these plants in the greenhouse for another 18 days. These pots were then transferred to an insectary-greenhouse, where the tobacco seedlings were inoculated with five to seven wingless adult M. persicae per pot. After 3 days, the infested seedlings were moved to a second greenhouse to allow the aphid population to increase, and after an additional 4±1 days when 182±4.25 aphid adults and nymphs were produced per pot, they were inoculated with A. gifuensis. With this rearing system, we were able to produce 256±8.8 aphid mummies per pot, with an emergence rate of 95.6±2.45%; 69% were females. The daily cost of parasite production (recurring costs only) was US$ 0.06 per 1000 aphid mummies. With this technique, we released 109 800 parasitoids in 1998, 196 000 in 1999, 780 000 in 2000, and 5 600 000 in 2001 during a 2-month period each year. This production method is discussed with respect to countrywide usage in biological control and integrated control of M. persicae.     

Two new species of aphids (Homoptera: Aphididae) from North-west India

Two new aphids viz. Prociphilus (Stagona) himalayaensis sp. nov. infesting the roots of Pinus excelsa and an unidentified plant and Capitophorus tricholepidis sp. nov. infesting Tricholepis furcata, collected from Simla Himalayas and Kumaon Himalayas, respectively, of the North-west India are described.     

Volatiles from potato plants infected with potato leafroll virus attract and arrest the virus vector, Myzus persicae (Homoptera: Aphididae)

The influence of viral disease symptoms on the behaviour of virus vectors has implications for disease epidemiology. Here we show that previously reported preferential colonization of potatoes infected by potato leafroll virus (genus Polerovirus) (luteovirus) (PLRV) by alatae of Myzus persicae, the principal aphid vector of PLRV, is influenced by volatile emissions from PLRV-infected plants. First, in our bioassays both differential immigration and emigration were involved in preferential colonization by aphids of PLRV-infected plants. Second, M. persicae apterae aggregated preferentially, on screening above leaflets of PLRV-infected potatoes as compared with leaflets from uninfected plants, or from plants infected with potato virus X (PVX) or potato virus Y (PVY). Third, the aphids aggregated preferentially on screening over leaflet models treated with volatiles collected from PLRV-infected plants as compared with those collected from uninfected plants. The specific cues eliciting the aphid responses were not determined, but differences between headspace volatiles of infected and uninfected plants suggest possible ones.     

Rapid method to screen resistance of potato plants against Myzus persicae (Homoptera: Aphididae) in the laboratory

With the objective to develop a potato, Solanum tuberosum L., resistance program against aphids, we propose a rapid screening method with Myzus persicae (Sulzer) in the laboratory. We aimed to optimize the duration of the whole procedure and to decrease the frequency of measurements. In a first experiment, intrinsic rate of natural increase (r(m)) values were compared between adult aphids reared throughout their entire life and adults reared only during a period equivalent to their prereproductive period. No significant differences were observed. In a second experiment, four groups of aphids were distinguished according to the sampling frequency, i.e., those whose biological parameters were evaluated every single, second, third, and fourth day. Except for the fourth-day experiment, the r(m) values estimated on aphids reared on the three potato lines were not significantly different whatever sampling frequency of single, second, or third day used to check aphids. Thus, screening efforts in laboratory can be largely optimized by evaluating adult aphids only during a period equivalent to their prereproductive period and assessing M. persicae populations every third day. Our method is reliable and adapted to screen a large number of potato plants against M. persicae because it allows an average 70% reduction in the time required for the whole experimental process.     

Microsatellite DNA and behavioural studies provide evidence of host-mediated speciation in Myzus persicae (Hemiptera: Aphididae)

The fossil record of mammals records a major interchange of northern and southern faunas in the New World, upon closure of the Panamanian isthmus approximately 3 Mya, termed the Great American Biotic Interchange (GABI). Due to their poor preservation in the fossil record, the degree of participation of birds in this interchange remains largely unknown. A phylogeny for wrens of the genus Campylorhynchus (Aves: Passeriformes) was reconstructed using DNA sequences from the mitochondrial control region and cytochrome b gene. This phylogeny, in combination with biogeographical inference and molecular clock methods, allows estimates of the importance of Late Pliocene interchange to the history of the group. Biogeographical reconstructions and divergence date estimates suggest that the genus began diversification in North America prior to closure of the Panamanian isthmus, consistent with a hypothesized North American origin for the family Troglodytidae. These reconstructions are consistent with pre-GABI dispersal of at most a single Campylorhynchus lineage into South America, with subsequent dispersal of additional lineages, probably across the fully formed isthmus. Increased sampling of avian taxa with widespread New World distributions will continue to clarify the timing and direction of continental interchange.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 90 , 687–702.