Fitness effects of two facultative endosymbiotic bacteria on the pea aphid,Acyrthosiphon pisum,and the blue alfalfa aphid,A. kondoi

The effects of two bacterial endosymbionts, designated PASS and PAR, were evaluated on the pea aphid, Acyrthosiphon pisum (Harris) (Hemiptera:Aphididae), in which they occur facultatively, and on the blue alfalfa aphid, A. kondoi Shinji, in which these bacteria have not been found in natural populations. Subclones of pea aphids and blue alfalfa aphids, derived from parent aphid clones that did not contain PASS or PAR, were infected with one or both bacteria, generating PASS- and/or PAR-positive subclones with minimal genetic differences from the parent clones. Under laboratory conditions at 20 °C, PAR consistently reduced the fecundity (by between 19 and 60%) of subclones derived from three different parent pea aphid clones on bur clover, Medicago hispida Gaertn. PAR had intermediate effects on pea aphids reared on sweet pea, Lathyrus odoratus L., and had no significant effect on pea aphids on alfalfa, Medicago sativa L. The effect of PASS was either neutral or negative, depending on parent clone as well as host plant. Also at 20 °C, PASS reduced fecundity (70–77%) and longevity (40–48%), and increased the age of first reproduction (by up to 1.5 days) of blue alfalfa aphid reared on alfalfa and clover. PAR had a less dramatic effect (e.g., 30–39% reduction in fecundity) on these traits of blue alfalfa aphid. In contrast, PAR and PASS increased the fitness of pea aphid subclones of one parent clone reared for three generations at 25 °C on each of the three test plants. Without facultative bacteria, fecundity of the parent clone was reduced to a mean total of < 6 offspring per adult at this elevated temperature, but with PASS or PAR, mean total fecundity of its subclones was > 35. However, this ameliorative effect of facultative bacteria at 25 °C was not found for two other sets of parent clones and their derived subclones. Alate production in pea aphids was significantly increased in large populations of two PASS- and PAR-positive subclones relative to their parent clones. Attempts to transmit PASS or PAR horizontally, i.e., from aphid to aphid via feeding on host plants (bur clover), were unsuccessful.     

Aphid genotypes vary in their response to the presence of fungal endosymbionts in host plants

Genetic variation for fitness‐relevant traits may be maintained in natural populations by fitness differences that depend on environmental conditions. For herbivores, plant quality and variation in chemical plant defences can maintain genetic variation in performance. Apart from plant secondary compounds, symbiosis between plants and endosymbiotic fungi (endophytes) can produce herbivore‐toxic compounds. We show that there is significant variation among aphid genotypes in response to endophytes by comparing life‐history traits of 37 clones of the bird cherry‐oat aphid Rhopalosiphum padi feeding on endophyte‐free and endophyte‐infected tall fescue Lolium arundinaceum. Clonal variation for life‐history traits was large, and most clones performed better on endophyte‐free plants. However, the clones differed in the relative performance across the two environments, resulting in significant genotype × environment interactions for all reproductive traits. These findings suggest that natural variation in prevalence of endophyte infection can contribute to the maintenance of genetic diversity in aphid populations.     

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.     

Intraspecific variation of Myzus persicae on sugar beet (Beta vulgaris)

Differences in inherited resistance among seven sugar-beet stocks had similar effects on Myzus persicae clones representing the range of variation in aphid response to resistant and susceptible sugar beet observed in fifty-eight clones collected between 1969 and 1971. Three sugar-beet stocks were consistently resistant. Statistically significant interactions between beet stocks and aphid clones did not indicate the existence of biotypes with specific abilities to overcome resistance. M. persicae clones differed in their vigour of colonizing sugar beet, irrespective of the differences between beet stocks. The readiness of adult aphids to settle determined the size of aphid population produced and included a component related to the response of the aphid clone to sugar beet as a host, and a component related to the resistance ranking of the beet stock. Breeding sugar beet with resistance to aphids will be simplified, as the results indicate that, at present, differences between aphid biotypes need not be considered a problem.     

Recent changes in the distribution of carboxylesterase genes and associated chromosomal rearrangements in Greek populations of the tobacco aphid Myzus persicae nicotianae

We present data on the frequency of amplified E4 and FE4 carboxylesterase genes in Myzus persicae s.l. clones collected during the years 2002–2007 and 2012 in Greece. Most clones were of the tobacco aphid, Myzus persicae nicotianae. Samples from 2012 were genotyped with microsatellite DNA markers and a number of them were karyotyped. Aphid clones with amplified FE4 genes predominated in all years, whereas E4 was present in only 3.5% of all samples and always occurred in clones with FE4. Most of the clones examined showed high carboxylesterase activity levels (R2 resistant category). The results showed marked changes in the frequencies of the two carboxylesterase genes in the tobacco aphid populations compared to published data that were collected in Greece in the mid 1990s, when E4 was recorded on its own in 20% of all samples and in 32% of samples from tobacco. A parallel change in karyotype was also observed because the A1,3 translocation, which had a worldwide association with amplified E4 genes in the 1990s, was not detected in the clones analyzed in 2012. Possible causes for these changes are discussed, although selection as a result of pest management practices appears to be the major one. Novel chromosomal rearrangements were also found in M. persicae nicotianae clones. These rearrangements could be a result of clastogenic effects of nicotine, which could persist because of the holocentric nature of aphid chromosomes. The results are discussed in relation to rapid evolution events that have taken place in the tobacco aphid in Greece during the last two decades. © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 113 , 455–470.     

Genetic variation in the effect of a facultative symbiont on host-plant use by pea aphids

Ecological specialisation on different host plants occurs frequently among phytophagous insects and is normally assumed to have a genetic basis. However, insects often carry microbial symbionts, which may play a role in the evolution of specialisation. The bacterium Regiella insecticola is a facultative symbiont of pea aphids (Acyrthosiphon pisum) where it is found most frequently in aphid clones feeding on Trifolium giving rise to the hypothesis that it may improve aphid performance on this plant. A study in which R. insecticola was eliminated from a single naturally infected aphid clone supported the hypothesis, but a second involving two aphid clones did not find the same effect. We created a series of new pea aphid–R. insecticola associations by injecting different strains of bacteria into five aphid clones uninfected by symbionts. For all aphid clones, the bacteria decreased the rate at which aphids accepted Vicia faba as a food plant and reduced performance on this plant. Their effect on aphids given Trifolium pratense was more complex: R. insecticola negatively affected acceptance by all aphid clones, had no effect on the performance of four aphid clones, but increased performance of a fifth, thus demonstrating genetic variation in the effect of R. insecticola on pea aphid host use. We discuss how these results may explain the distribution and frequency of this symbiont across different aphid populations. Julia Ferrari and Claire L. Scarborough contributed equally to the work.     

Bottom‐up effects mediated by an organic soil amendment on the cabbage aphid pests Myzus persicae and Brevicoryne brassicae

Earthworm‐produced compost or vermicompost has been shown to increase resistance of plants to a variety of insect pests, but it is still unclear whether this resistance is dose dependent and whether the mechanisms responsible are the same for insect species with differing feeding habits and preferences. Therefore, we tested the effects of plants grown in various vermicompost concentrations (0, 20, 40, and 60%) on the preference and performance of generalist, Myzus persicae L., and specialist, Brevicoryne brassicae L. (both Hemiptera: Aphididae), aphid pests. Preference was evaluated with leaf disk (apterous) and whole plant (alate) choice assays. After 24 h of feeding, there was no significant negative effect on the feeding preference noted for apterae of either species of any of the treatments tested. To the contrary, apterae B. brassicae showed a significant preference for vermicompost treatments over control leaf disks. Alate M. persicae preferred alighting on control plants over vermicompost‐grown plants, but B. brassicae showed no preference toward any of the treatments tested. Both aphid species deposited significantly more nymphs on control plants than on those grown in 20% vermicompost. Furthermore, plants grown in soil amended with 20% vermicompost significantly suppressed mass accumulation, as well as numbers of adults and nymphs of both aphid species compared to controls. These data clearly show that vermicompost soil amendments can significantly influence pest aphid preference and performance on plants and that these effects are not dose dependent, but rather species and morph dependent.     

Opposing effects of organic and conventional fertilizers on the performance of a generalist and a specialist aphid species

• 1 Sustainable and conventional farming systems use fertilizers that differ in the availability of nitrogen, which may affect plant quality to alter the abundance and performance of potential pest species.
• 2 We grew brassica plants in several types of fertilizer, including those commonly used in conventional and sustainable farming systems, and an unfertilized control. The effects of fertilizer type on the performance of two aphid species and foliar glucosinolate content were investigated.
• 3 Both aphid species performed poorly (with reduced fecundity) on the unfertilized treatment compared with those feeding on fertilized host plants.
• 4 Brevicoryne brassicae, the brassica specialist, performed best on Brassica oleracea plants fertilized with an organic animal manure, with a 72% increase in fecundity and an 18% increase in intrinsic rate of increase compared with plants fertilized with ammonium nitrate.
• 5 By contrast, the generalist Myzus persicae had an intrinsic rate of increase that was reduced by 15% on plants growing in the animal manure compared with those growing in ammonium nitrate.
• 6 These results may explain earlier findings on the effects of fertilizer type on aphid populations in the field, and are discussed in the context of pest species' responses to sustainable and conventional agricultural systems.

     

Performance of clones and morphs of two cereal aphids on wheat plants with high and low nitrogen content

The great variability of the aphid life cycle and their tendency for host alternation gives rise to aphid clones and morphs. Inter‐ and intraclonal variability may be observed in the responses of aphids to various environmental factors. In this study we aimed to evaluate the influence of intrinsic factors (clone and morph type) on the intrinsic rate of increase (r_m) of the English grain aphid, Sitobion avenae (Fabricius), and the bird cherry‐oat aphid, Rhopalosiphum padi (Linnaeus). For each species four apterous clones were collected from established laboratory colonies and compared to assess their relative fitness on high‐ and low‐nitrogen wheat plants under laboratory conditions. The clones had significantly different intrinsic rates of increase on high‐ and low‐nitrogen plants. All R. padi clones had a higher intrinsic rate of increase and mean relative growth rate than S. avenae. Experiments were also conducted to compare the mean fecundity of apterous and alate morphs of S. avenae and R. padi clones on high‐ and low‐nitrogen wheat plants. On high‐nitrogen plants the apterous morphs of S. avenae clones had significantly higher mean fecundity than alate morphs. There were no significant differences between the mean fecundity of alate morphs of the same species on high‐ and low‐nitrogen plants. The results support the idea of better fitness of specific clones/morphs on certain host plants due to higher and lower intrinsic rates of increase.     

Unusual bacterial populations observed in a full-scale municipal sludge digester affected by intermittent seawater inputs

This study investigated the bacterial community of a full-scale anaerobic digester, which suffers from intermittent seawater contaminations, using 16S rRNA gene clone analysis over different seasons. Bacteroidetes, Proteobacteria, and unclassifiable bacteria were the three major bacterial groups within the clone library (a total of 290 clones). A significant portion of the total clones (29.3%) was not affiliated to any previously reported phylum, and 55.3% of the unclassifiable clones (16.9% of the total clones) showed potential relations to the species of Thermotogae, rarely present under normal mesophilic anaerobic conditions. These results suggested that the novel populations may have the potential to play an important role in anaerobic processes, particularly under abnormal environmental conditions. Additionally, statistical analysis supported that seasonal variations in influent characteristics, and potential competitions among different populations, may be related to the unusual bacterial diversity and community dynamics observed over the study period.     

Toxicity of plant essential oil vapours to aphid pests and their coccinellid predators

The comparative toxicity of five essential oil vapours was tested against four aphid species, the black bean aphid Aphis fabae, the pea aphid Acyrthosiphon pisum, the chrysanthemum aphid Macrosiphoniella sanborni, the green peach aphid Myzus persicae and on two of the most common coccinellid predators, the seven-spotted ladybird Coccinella septempunctata and the two-spotted ladybird Adalia bipunctata. All essential oils were highly toxic to the aphid species tested with LC₅₀ and LC₉₉ values ranging between 0.17 and 1.92 and 0.44 and 4.83 µL/L air, respectively, depending on the aphid species and on the essential oil. Coccinellid predators were also highly susceptible to the essential oil vapours and the selective toxicity ratio varied depending on aphid species, coccinellid predator and essential oil. The possibilities for the utililization of essential oils as aphicides, especially in IPM programmes in glasshouses are discussed with regard to the present findings.     

Transmission Comparisons of Cucumber Mosaic Virus Subgroup I and II Isolates by Different Aphid Species

To investigate the transmission differences between Cucumber mosaic virus (CMV) subgroup isolates, we carried out a comparative study with five aphid species Myzus persicae, Aphis gossypii, Lipaphis erysimi, Aphis craccivora and Megoura viciae in laboratory and field experiments to evaluate spread of CMV Subgroup I NX and subgroup II AG isolates in tobacco. Both NX and AG varied in transmission efficiency by the five aphids, and our transmission results revealed important differences in transmission efficiency of two isolates by Myzus persicae and Aphis gossypii. In contrast, significant transmission differences were not detected with Lipaphis erysimi, Aphis craccivora or Megoura viciae. Interestingly, the overall transmission efficiencies of the two different subgroup strains were almost equal when field transmissions were tested with mixed populations of the five aphid species. Our results together with our previously reported experiments on competition of CMV subgroup isolates in tobacco suggest that variations in aphid vector populations contribute substantially to the epidemic potential of CMV subgroup isolates.     

Spread of zucchini yellow mosaic potyvirus in squash in Hungary

The temporal and spatial distribution of zucchini yellow mosaic potyvirus (ZYMV) was studied in a 3000‐m² zucchini squash field. The first infected plants were found 4 weeks after the field was exposed to virus source plants. The infection increased to nearly 74% by the end of the study. Alate aphids were active from the beginning of the study and 43 species were trapped in the field. Flights of vector species Acyrthosiphon pisum and Myzus persicae peaked during the fourth week which resulted in high virus incidence 4 weeks later. There was a significant correlation between the number of vectors caught in yellow pan traps and the number of infected plants in the field. In laboratory studies evaluating 11 aphid species, Aphis pomi de Geer was identified as a new vector species of ZYMV. Although this aphid was not caught in our field studies, it may be an important contributor in other areas where cucurbits are grown in close proximity to apple or other hosts of this aphid.     

A strain of the bacterial symbiont Regiella insecticola protects aphids against parasitoids

Aphids commonly harbour facultative bacterial endosymbionts and may benefit from their presence through increased resistance to parasitoids. This has been demonstrated for Hamiltonella defensa and Serratia symbiotica, while a third common endosymbiont, Regiella insecticola, did not provide such protection. However, this symbiont was recently detected in a highly resistant clone of the peach-potato aphid, Myzus persicae, from Australia. To test if resistance was indeed conferred by the endosymbiont, we eliminated it from this clone with antibiotics, and we transferred it to two other clones of the same and one clone of a different aphid species (Aphis fabae). Exposing these lines to the parasitoid Aphidius colemani showed clearly that unlike other strains of this bacterium, this specific isolate of R. insecticola provides strong protection against parasitic wasps, suggesting that the ability to protect their host against natural enemies may evolve readily in multiple species of endosymbiotic bacteria.     

Temporal and Spatial Development of Papaya Ringspot in Veracruz, Mexico

The temporal progress and spatial distribution of papaya ringspot virus (PRV) and populations of aphid vectors were monitored in two papaya (Carica papaya) plantations in the state of Veracruz, México. The incidence of PRV had a typical sigmoidal curve and the logistic model was more acceptable for describing the disease progress than the Gompertz model. The rate (r^L of increase in disease incidence (0.034 and 0.023/unit/day in Plot A and B, respectively) differed (P = 0.05) between the plots; differences in number of aphids trapped could account for the difference. The initial determination of an apparent regular spatial pattern early in the epidemic, obtained with a quadrat size determined by Greig-Smith's method (n = 8 plants/quadrat) at 50 % disease incidence, was inconsistent with our biological observations in the field. An arbitrarily selected, square quadrat size (n = 9 plants/quadrat), was more consistent with visual observation for describing the spatial pattern in the field. With 9 plants/quadrat a random pattern was found. Aphid populations had a bimodal distribution at both sites with the highest population peak in December–February and a secondary peak in August–September and change in disease incidence was generally related to the aphid population level in the previous month. None of the five potential aphid vectors (Myzus persicae, Aphis gossypii A. nerii, A. citricola and Macrosiphon euphorbiae) of PRV in Mexico colonized the papaya plants, however, which may explain the absence of clustering of disease.     

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.     

Quantifying aphid behavioral responses to environmental change

Aphids are the most common vector of plant viruses, and their feeding behavior is an important determinant of virus transmission. Positive effects of global change on aphid performance have been documented, but effects on aphid behavior are not known. We assessed the plant‐mediated behavioral responses of a generalist aphid, Myzus persicae (Sulzer) (Hemiptera: Aphididae), to increased CO₂ and nitrogen when feeding on each of three host species: Amaranthus viridis L. (Amaranthaceae), Polygonum persicaria L. (= Persicaria maculosa Gray) (Polygonaceae), and Solanum dulcamara L. (Solanaceae). Via a family of constrained Markov models, we tested the degree to which aphid movements demonstrate preference among host species or plants grown under varying environmental conditions. Entropy rates of the estimated Markov chains were used to further quantify aphid behavior. Our statistical methods provide a general tool for assessing choice and quantitatively comparing animal behavior under different conditions. Aphids displayed strong preferences for the same host species under all growth conditions, indicating that CO₂‐ and N‐induced changes in plant chemistry have minimal effects on host preference. However, entropy rates increased in the presence of non‐preferred hosts, even when preferred hosts were available. We conclude that the presence of a non‐preferred host species affected aphid‐feeding behavior more than changes in plant leaf chemistry when plants were grown under elevated CO₂ and increased N availability.     

Aphid populations showing differential levels of virulence on Capsicum accessions

The green peach aphid,Myzus persicae,is one of the most threatening pests in pepper cultivation and growers would benefit from resistant varietices.Previously,we identified two Capsicum acessions as susceptible and three as resistant to M.persicae using an aphid population originating from the Netherlands(NL).Later on we identified an aphid population originating from a diferent gcographical region(Switserland,SW)that was virulent on all tested Capsicum acessions.The objeetive of the current work is to describe in detail diferent aspects of the interaction between two aphid populations and two sclected Capsicum acessions(one that was susceptible[PB2013046]and one that was resistant[PB2013071]to population NL),including biochemical processes involved.Electrical penetration graph(EPG)recordings showed similar feeding activities for both aphid populations on PB2013046.On acession PB2013071 the aphid population sw was able to devote significantly more time to phloem ingestion than population NL.We also studied plant defense response and found that plants of acession PB2013046 could not induce an accumulation of reactive oxygen species and callose formation after infestation with either aphid population.However,plants of PB2013071 induced a stronger defense response after infestation by population NL than after infestation by population SW.Based on these results,population SW of M.persicae seems to have overcome the resistance of PB2013071 that prevented feeding of aphids from NL population.The potential mechanism by which SW population overcomes the resistance is discussed.     

Horizontally transferred genes as RNA interference targets for aphid and whitefly control

RNA interference (RNAi)-based technologies are starting to be commercialized as a new approach for agricultural pest control. Horizontally transferred genes (HTGs), which have been transferred into insect genomes from viruses, bacteria, fungi or plants, are attractive targets for RNAi-mediated pest control. HTGs are often unique to a specific insect family or even genus, making it unlikely that RNAi constructs targeting such genes will have negative effects on ladybugs, lacewings and other beneficial predatory insect species. In this study, we sequenced the genome of a red, tobacco-adapted isolate of Myzus persicae (green peach aphid) and bioinformatically identified 30 HTGs. We then used plant-mediated virus-induced gene silencing (VIGS) to show that several HTGs of bacterial and plant origin are important for aphid growth and/or survival. Silencing the expression of fungal-origin HTGs did not affect aphid survivorship but decreased aphid reproduction. Importantly, although there was uptake of plant-expressed RNA by Coccinella septempunctata (seven-spotted ladybugs) via the aphids that they consumed, we did not observe negative effects on ladybugs from aphid-targeted VIGS constructs. To demonstrate that this approach is more broadly applicable, we also targeted five Bemisia tabaci (whitefly) HTGs using VIGS and demonstrated that knockdown of some of these genes affected whitefly survival. As functional HTGs have been identified in the genomes of numerous pest species, we propose that these HTGs should be explored further as efficient and safe targets for control of insect pests using plant-mediated RNA interference.     

Strong parasitoid-mediated selection in experimental populations of aphids

Clonal diversity in asexual populations may be maintained if different clones are favoured under different environmental conditions. For aphids, parasitoids are an important variable of the biotic environment. To test whether parasitoids can mediate selection among host clones, we used experimental populations consisting of 10 clones of the peach-potato aphid, Myzus persicae, and allowed them to evolve for several generations either without parasitoids or in the presence of two species of parasitoid wasps. In the absence of parasitoids, strong shifts in clonal frequencies occurred, mostly in favour of clones with high rates of increase. The parasitoid Diaeretiella rapae hardly affected aphid densities but changed the outcome of competition by favouring one entirely resistant clone and disfavouring a highly susceptible clone. Aphidius colemani, the more infective parasitoid, strongly reduced aphid densities and dramatically changed host clonal frequencies. The most resistant clone, not a successful clone without parasitoids, became totally dominant. These results highlight the potential of temporal or spatial variation in parasitoid densities to maintain clonal diversity in their aphid hosts.