Uninfected Aphids Research Articles

Physiological costs of facultative endosymbionts in aphids assessed from energy metabolism

Abstract Many insect species harbour heritable bacterial endosymbionts. Some facultative endosymbionts provide benefits to their hosts under certain environmental conditions. Facultative endosymbionts are expected to impose additional energetic expenditures to their host, reducing host fitness. While there is accumulating evidence in plant sucking insects that facultative endosymbionts reduce the fitness of their host under permissive conditions, no direct energy costs associated with facultative endosymbionts have been identified. Using the standard metabolic rate (SMR) as a measure of the energy cost of self‐maintenance, we investigated whether two common facultative endosymbionts Hamiltonella defensa or Regiella insecticola increase the maintenance cost of the pea aphid Acyrthosiphon pisum which could translate into host fitness reduction (‘compensation hypothesis’). In addition, we tested if there was a link between SMR and the aphid fitness and whether it depended on endosymbiont density and aphid energetic reserves. Finally, we measured SMR at different temperatures to assess the impact of suboptimal thermal conditions on physiological cost of endosymbionts. In the presence of facultative endosymbionts, aphids expressed generally a lower fitness and a higher SMR compared to uninfected ones, in accordance with the ‘compensation hypothesis’. However, the SMR difference between infected and uninfected aphids tended to decrease with increasing temperature. Complex host genotype‐by‐symbiont genotype‐by‐temperature interactions on SMR were also revealed. Energetic budget of adult aphids appeared weakly influenced by the aphid genotype and endosymbiont species, suggesting that facultative endosymbionts primarily impact the consumption of energy resources rather than their acquisition. Density of facultative endosymbionts varied largely among aphid lines but was not associated with the fitness nor metabolic rate of aphids. This work supports the energy basis of facultative endosymbiont associated fitness costs and raises new questions about the effect of facultative endosymbionts on the energy metabolism of their host. Read the free Plain Language Summary for this article on the Journal blog.

Urbanization and plant pathogen infection interact to affect the outcome of ecological interactions in an experimental multitrophic system

Abstract Urbanization can change interactions in insect communities, and the few studies of tritrophic interactions in urban settings focus on interactions between plants, herbivorous insects and their mutualists and natural enemies. Plant pathogen infection is also widespread and common, and infection may also alter such interactions, but we have no understanding of whether the ecological consequences of pathogen infection vary with urbanization. Using replicated aphid colonies on experimental plants, we investigated how infection by the plant pathogen Botrytis cinerea influences interactions between plants, aphids and the aphid natural enemies and ant mutualists in highly urbanized, suburban and rural study sites. Aphid and natural enemy abundance were highest in the suburban site, while mutualist ants were most abundant in the urban site, reversing the usual positive density-dependent relationship between natural enemies and aphids. The effect of pathogen infection varied with trait and site, mediated by natural enemy preference for hosts or prey on uninfected plants. The effect of infection on aphid abundance was only seen in the suburban site, where natural enemies were most abundant on uninfected plants and aphid numbers were greatest on infected plants. In the urban site, there was no effect of infection, while in the rural site, aphid numbers were lower on infected plants. Uninfected plants were smaller than infected plants and differed between locations. This study suggests that the effects of urbanization on ecological interactions may become more complex and difficult to predict as we study ecological assemblages and communities at greater levels of structural complexity.

Similar cost of Hamiltonella defensa in experimental and natural aphid-endosymbiont associations.

Endosymbiont‐conferred resistance to parasitoids is common in aphids, but comes at a cost to the host in the absence of parasitoids. In black bean aphids (Aphis fabae), costs in terms of reduced lifespan and lifetime reproduction were demonstrated by introducing 11 isolates of the protective symbiont Hamiltonella defensa into previously uninfected aphid clones. Transfection of H. defensa isolates into a common genetic background allows to compare the costs of different endosymbiont isolates unconfounded by host genetic variation, but has been suggested to overestimate the realized costs of the endosymbiont in natural populations, because transfection creates new and potentially maladapted host–symbiont combinations that would be eliminated by natural selection in the field. In this experiment, we show that removing H. defensa isolates from their natural host clones with antibiotics results in a fitness gain that is comparable to the fitness loss from their introduction into two new clones. This suggests that estimating cost by transfecting endosymbiont isolates into a shared host genotype does not lead to gross overestimates of their realized costs, at least not in the two recipient genotypes used here. By comparing our data with data reported in previous publications using the same lines, we show that symbiont‐induced costs may fluctuate over time. Thus, costs estimated after extended culture in the laboratory may not always be representative of the costs at the time of collection in the field. Finally, we report the accidental observation that two isolates from a distinct haplotype of H. defensa could not be removed by cefotaxime treatment, while all isolates from two other haplotypes were readily eliminated, which is suggestive of variation in susceptibility to this antibiotic in H. defensa.

Aphids harbouring different endosymbionts exhibit differences in cuticular hydrocarbon profiles that can be recognized by ant mutualists

Cuticular hydrocarbons (CHCs) have important communicative functions for ants, which use CHC profiles to recognize mutualistic aphid partners. Aphid endosymbionts can influence the quality of their hosts as ant mutualists, via effects on honeydew composition, and might also affect CHC profiles, suggesting that ants could potentially use CHC cues to discriminate among aphid lines harbouring different endosymbionts. We explored how several strains of Hamiltonella defensa and Regiella insecticola influence the CHC profiles of host aphids (Aphis fabae) and the ability of aphid-tending ants (Lasius niger) to distinguish the profiles of aphids hosting different endosymbionts. We found significant compositional differences between the CHCs of aphids with different infections. Some endosymbionts changed the proportions of odd-chain linear alkanes, while others changed primarily methyl-branched compounds, which may be particularly important for communication. Behavioural assays, in which we trained ants to associate CHC profiles of endosymbiont infected or uninfected aphids with food rewards, revealed that ants readily learned to distinguish differences in aphid CHC profiles associated with variation in endosymbiont strains. While previous work has documented endosymbiont effects on aphid interactions with antagonists, the current findings support the hypothesis that endosymbionts also alter traits that influence communicative interactions with ant mutualists.

Aphid Facultative Symbionts Aid Recovery of Their Obligate Symbiont and Their Host After Heat Stress

Environmental conditions affect insect fitness, with many species constrained by specific temperature ranges. Aphids are limited to temperate climates and it is hypothesized that this is partly due to their heat-susceptible obligate nutritional symbiont Buchnera. Aphids often carry additional facultative symbionts which can increase the host’s fitness after heat stress. Here we used the pea aphid (Acyrthosiphon pisum) and three of its facultative endosymbionts (Candidatus Regiella insecticola, Candidatus Fukatsuia symbiotica (X-type; PAXS), and Candidatus Hamiltonella defensa) to investigate how these species respond to heat stress and whether their presence affects the fitness of the host or the obligate symbiont. We exposed aphid lines to a single high temperature event and measured lifetime fecundity and population densities of both obligate and facultative symbionts. Heat shock reduced aphid fecundity, but for aphids infected with two of the facultative symbionts (Regiella or Fukatsuia), this reduction was less than in uninfected aphids. The population density of Buchnera was also reduced after heat shock, and only recovered in aphids infected with Regiella or Fukatsuia but not in uninfected aphids or those with Hamiltonella. Although heat shock initially reduced the densities of two of the facultative symbionts (Hamiltonella and Fukatsuia), all facultative symbiont densities recovered by adulthood. Two of the facultative symbionts tested therefore aided the recovery of the obligate symbiont and the host, and we discuss possible underlying mechanisms. Our work highlights the beneficial effects of protective symbionts on obligate symbiont recovery after heat stress and how facultative symbionts may affect the wider ecological community.

Can aphids be controlled by fungus? A mathematical model

Abstract The control of insect pests in agriculture is essential for food security. Chemical controls typically damage the environment and harm beneficial insects such as pollinators, so it is advantageous to identify targetted biological controls. Since predators are often generalists, pathogens or parasitoids are more likely to serve the purpose. Here, we model a fungal pathogen of aphids as a potential means to control of these important pests in cereal crops. Typical plant herbivore pathogen models are set up on two trophic levels, with dynamic variables the plant biomass and the uninfected and infected herbivore populations. Our model is unusual in that (i) it has to be set up on three trophic levels to take account of fungal spores in the environment, but (ii) the aphid feeding mechanism leads to the plant biomass equation becoming uncoupled from the system. The dynamical variables are therefore the uninfected and infected aphid population and the environmental fungal concentration. We carry out an analysis of the dynamics of the system. Assuming that the aphid population can survive in the absence of disease, the fungus can only persist (and control is only possible) if (i) the host grows sufficiently strongly in the absence of infection, and (ii) the pathogen transmission parameters are sufficiently large. If it does persist the fungus does not drive the aphid population to extinction, but controls it below its disease-free steady state value, either at a new coexistence steady state or through oscillations. Whether this control is sufficient for agricultural purposes will depend on the detailed parameter values for the system.

Circulation of the Cultivable Symbiont Serratia symbiotica in Aphids Is Mediated by Plants.

Symbiosis is a common phenomenon in nature that substantially affects organismal ecology and evolution. Fundamental questions regarding how mutualistic associations arise and evolve in nature remain, however, poorly studied. The aphid-Serratia symbiotica bacterium interaction represents a valuable model to study mechanisms shaping these symbiotic interspecific interactions. S. symbiotica strains capable of living independently of aphid hosts have recently been isolated. These strains probably resulted from horizontal transfers and could be an evolutionary link to an intra-organismal symbiosis. In this context, we used the tripartite interaction between the aphid Aphis fabae, a cultivable S. symbiotica bacterium, and the host plant Vicia faba to evaluate the bacterium ability to circulate in this system, exploring its environmental acquisition by aphids and horizontal transmission between aphids via the host plant. Using molecular analyses and fluorescence techniques, we showed that the cultivable S. symbiotica can enter the plants and induce new bacterial infections in aphids feeding on these new infected plants. Remarkably, we also found that the bacterium can have positive effects on the host plant, mainly at the root level. Furthermore, our results demonstrated that cultivable S. symbiotica can be horizontally transferred from infected to uninfected aphids sharing the same plant, providing first direct evidence that plants can mediate horizontal transmission of certain strains of this symbiont species. These findings highlight the importance of considering symbiotic associations in complex systems where microorganisms can circulate between different compartments. Our study can thus have major implications for understanding the multifaceted interactions between microbes, insects and plants.

A fitness cost resulting from Hamiltonella defensa infection is associated with altered probing and feeding behaviour in Rhopalosiphum padi.

Many herbivorous arthropods, including aphids, frequently associate with facultative endosymbiotic bacteria, which influence arthropod physiology and fitness. In aphids, endosymbionts can increase resistance against natural enemies, enhance aphid virulence and alter aphid fitness. Here, we used the electrical penetration graph technique to uncover physiological processes at the insect-plant interface affected by endosymbiont infection. We monitored the feeding and probing behaviour of four independent clonal lines of the cereal-feeding aphid Rhopalosiphum padi derived from the same multilocus genotype containing differential infection (+/-) with a common facultative endosymbiont, Hamiltonella defensa Aphid feeding was examined on a partially resistant wild relative of barley known to impair aphid fitness and a susceptible commercial barley cultivar. Compared with uninfected aphids, endosymbiont-infected aphids on both plant species exhibited a twofold increase in the number of plant cell punctures, a 50% reduction in the duration of each cellular puncture and a twofold higher probability of achieving sustained phloem ingestion. Feeding behaviour was also altered by host plant identity: endosymbiont-infected aphids spent less time probing plant tissue, required twice as many probes to reach the phloem and showed a 44% reduction in phloem ingestion when feeding on the wild barley relative compared with the susceptible commercial cultivar. Reduced feeding success could explain the 22% reduction in growth of H. defensa-infected aphids measured on the wild barley relative. This study provides the first demonstration of mechanisms at the aphid-plant interface contributing to physiological effects of endosymbiont infection on aphid fitness, through altered feeding processes on different quality host plants.

Use of electrical penetration graphs (EPG) and quantitative PCR to evaluate the relationship between feeding behaviour and Pandora neoaphidis infection levels in green peach aphid, Myzus persicae

A real-time qPCR method was developed, validated, and used to quantity the fungal pathogen, P. neoaphidis, within aphids at different times during infection; colonization rate fitted the Gompertz model well (R2 = 0.9356). Feeding behaviour of P. neoaphidis-infected and uninfected M. persicae were investigated, for the first time, using DC-electrical penetration graphs (DC-EPG) that characterized the waveforms made during different aphid stylet probing periods corresponding to epidermis penetration, salivation and ingestion. In the 6 h following the 12-h incubation period (to achieve infection), there were significant differences in the number of events of Np (non-probing) and C (stylet pathway) between infected and uninfected aphids. However, the difference between total duration of Np and C were not significantly different between infected and uninfected aphids. There were no significant differences in the number of events or total duration of E1 (phloem salivation) or E2 (phloem ingestion) between infected and uninfected aphids. There were significant differences in mean number of events and total duration of the pd waveform (intracellular punctures) in infected and uninfected aphids. In the 16 h prior to death, the same differences in behaviour were observed but they were even more obvious. Furthermore, the total duration time of E2 was significantly greater in uninfected aphids than infected aphids, a change that had not been observed in the first 6 h observation period. In conclusion, qPCR quantification demonstrated ‘molecular’ colonization levels throughout infection, and EPG data analysis during the two periods (during early infection and then during late infection just prior to death) demonstrated the actual physical effects of fungal infection on feeding behaviour of M. persicae; this has the potential to decrease the aphid’s capacity of transmission and dispersal. These studies increase our understanding of the interaction between P. neoaphidis and its host aphid.

The defensive aphid symbiont Hamiltonella defensa affects host quality differently for Aphelinus glycinis versus Aphelinus atriplicis

Endosymbiont interactions with hosts have important effects on fitness, including the fitness of many pest and beneficial species. Among these interactions, facultative endosymbiotic bacteria can protect aphids from parasitoids. Aphis craccivora and Acyrthosiphon pisum can harbor the symbiotic bacteria Hamiltonella defensa and its bacteriophage APSE. Infection by H. defensa defends these aphids against some but not all parasitoid species in the hymenopteran family Braconidae. Here, we report results on the effect of H. defensa on parasitism of these aphids by species in the other major lineage of aphid parasitoids, Aphelinus species in the family Aphelinidae. Parasitism of aphids infected with H. defensa/APSE by two Aphelinus species did not differ from that of uninfected aphids. While Aphelinus atriplicis showed no difference in fitness components between infected and uninfected aphids, Aphelinus glycinis actually produced more adult progeny and larger female progeny on infected than on uninfected aphids. Aphelinus glycinis may increase host quality for itself by changing the titer of nutritional versus protective bacteria in such a way that aphids infected with H. defensa can be made more suitable for parasitoid development than uninfected aphids. Our results and reasoning suggest that these Aphelinus species may be less prone to harm by H. defensa/APSE that affect eggs because they have anhydropic, heavily chorionated eggs, which may not absorb toxins during embryogenesis.

The influence of facultative endosymbionts on honeydew carbohydrate and amino acid composition of the black bean aphid Aphis fabae

AbstractThe facultative endosymbionts Hamiltonella defensa and Regiella insecticola are commonly found in aphids. They are linked with various ecological benefits but generally occur at low prevalence, which indicates a possible harbouring cost. Little is known about how the presence of facultative endosymbionts is reflected in honeydew composition. Honeydew is the key mediator of the mutualism between aphids and their tending ants. The present study examines whether endosymbionts have an influence on aphid honeydew quality by comparing the amino acid and carbohydrate concentrations between infected and uninfected aphids. To this end, two genetic lines of the aphid Aphis fabae Scopoli are experimentally infected with different strains of Hamiltonella and Regiella. Infected aphids are shown to have reduced concentrations of amino acids in the honeydew compared with uninfected aphids. However, the presence of endosymbionts has no effect on the absolute amount of carbohydrates produced. Nevertheless, interclonal variation in honeydew composition between aphid genotypes is observed for both carbohydrate and amino acid production. These results imply that the nutritional value of honeydew depends on aphid genotype, as well as on the presence of secondary bacterial endosymbionts, which suggests that there is a physiological cost of harbouring endosymbionts and which could also impact aphid attractiveness to tending ants.

Conditional Reduction of Predation Risk Associated with a Facultative Symbiont in an Insect

Symbionts are widespread among eukaryotes and their impacts on the ecology and evolution of their hosts are meaningful. Most insects harbour obligate and facultative symbiotic bacteria that can influence their phenotype. In the pea aphid Acyrthosiphon pisum, an astounding symbiotic-mediated phenotype has been recently observed: when infected with the symbiotic bacteria Rickettsiella viridis, young red aphid larvae become greener at adulthood and even darker green when co-infected with Rickettsiella viridis and Hamiltonella defensa. As body colour affects the susceptibility towards natural enemies in aphids, the influence of the colour change due to these facultative symbionts on the host survival in presence of predators was tested. Our results suggested that the Rickettsiella viridis infection may impact positively host survival by reducing predation risk. Due to results from uninfected aphids (i.e., more green ones attacked), the main assumption is that this symbiotic infection would deter the predatory ladybird feeding by reducing the profitability of their hosts rather than decreasing host detection through body colour change. Aphids co-infected with Rickettsiella viridis and Hamiltonella defensa were, however, more exposed to predation suggesting an ecological cost associated with multiple infections. The underlying mechanisms and ecological consequences of these symbiotic effects are discussed.

Development and Reproduction of Chrysoperla externa (Hagen) (Neuroptera: Chrysopidae) Fed on Myzus persicae (Sulzer) (Hemiptera: Aphididae) Vectoring Potato leafroll virus (PLRV).

The aim of this study was to evaluate the biological parameters of Chrysoperla externa (Hagen) while feeding on Myzus persicae (Sulzer) acting as a vector of potato leafroll virus (PLRV). In laboratory experiments, three different diets were offered ad libitum to C. externa during the larval period: M. persicae fed on PLRV-infected potato leaves, M. persicae fed on uninfected potato leaves, and eggs of the factitious prey Anagasta kuehniella (Zeller) as the control. The following parameters were studied: the developmental time and survival rate of the larval and pupal stages, the sex ratio, the proportion of fertile females, female fecundity and egg viability, and the survival curve of the first 30 days after adult emergence. PLRV-infected aphids influenced the C. externa larval developmental time and survival compared to PLRV-uninfected aphids and A. kuehniella eggs. The pupal developmental time of C. externa was shorter when fed on aphids compared to A. kuehniella eggs, but no differences were recorded between the PLRV-infected and uninfected aphid diets. Additionally, no differences were observed for pupal survival and reproduction. However, adult survival was affected by the prey type, as 75% of C. externa control adults remained alive at an age of 30 days compared to 51 and 54% for those fed on PLRV-uninfected and infected aphids, respectively.

Peroxiredoxin 1 protects the pea aphid Acyrthosiphon pisum from oxidative stress induced by Micrococcus luteus infection

Reactive oxygen species (ROSs) are generated in organisms in response to infections caused by invading microbes. However, excessive ROSs will inflict oxidative damage on the host. Peroxiredoxins (Prxs) are antioxidative enzymes that may eliminate ROSs efficiently. In this study, ApPrx1 from the pea aphid Acyrthosiphon pisum was cloned, and its function was investigated in vitro and in vivo. In the presence of DTT, recombinant ApPrx1 protein from Escherichia coli showed antioxidative activity by eliminating H2O2 effectively. The H2O2 levels were significantly higher in Micrococcus luteus-infected aphids than in uninfected aphids, and ApPrx1 expression was remarkably up-regulated when the aphids were infected with M. luteus or injected with H2O2. When ApPrx1 expression was reduced by dsRNA injection, the survival of the aphids decreased significantly after M. luteus infection. Knockdown of ApPrx1 decreased M. luteus loads inside the aphids 48h post-infection. While under infection conditions, the H2O2 levels were much higher in ApPrx1 knockdown aphids than in dsGFP-injected aphids, indicating that the decreased survival of the aphids was caused by increased oxidative stress. Taken together, our results reveal that ApPrx1 plays a protective role in oxidative stress caused by bacterial infection.

Aphid behavioral responses to virus‐infected plants are similar despite divergent fitness effects

AbstractWe compared the settling preferences and reproductive potential of an oligophagous herbivore, the pea aphid, Acyrthosiphon pisum Harris (Hemiptera: Aphididae), in response to pea plants, Pisum sativum L. cv. ‘Aragorn’ (Fabaceae), infected with two persistently transmitted viruses, Pea enation mosaic virus (PEMV) and Bean leaf roll virus (BLRV), that differ in their distribution within an infected plant. Aphids preferentially oriented toward and settled on plants infected with PEMV or BLRV in comparison with sham‐inoculated plants (plants exposed to herbivory by uninfected aphids), but aphids did not discriminate between plants infected with the two viruses. Analysis of plant volatiles indicated that plants inoculated with either virus had significantly higher green leaf volatile‐to‐monoterpene ratios. Time until reproductive maturity was marginally influenced by plant infection status, with a trend toward earlier nymph production on infected plants. There were consistent age‐specific effects of plant infection status on aphid fecundity: reproduction was significantly enhanced for aphids on BLRV‐infected plants across most time intervals, though mean aphid fecundity did not differ between sham and PEMV‐infected plants. There was no clear pattern of age‐specific survivorship; however, mean aphid lifespan was reduced on plants infected with PEMV. Our results are consistent with predictions of the host manipulation hypothesis, extended to include plant viruses: non‐viruliferous A. pisum preferentially orient to virus‐infected host plants, potentially facilitating pathogen transmission. These studies extend the scope of the host manipulation hypothesis by demonstrating that divergent fitness effects on vectors arise relative to the mode of virus transmission.

Virulence of Verticillium lecanii (Z.) against cereal aphids; does timing of infection affect the performance of parasitoids and predators?

Entomopathogenic fungi such as Verticillium lecanii (Z.) (Mycotal(®)) are used for pest control as an alternative to chemical control. In this study, the effect of V. lecanii on cereal aphids is assessed. In addition, an investigation is carried out to determine whether the use of V. lecanii affects the performance of two natural enemies of aphids, the predator Harmonia axyridis (P.) and the parasitoid Aphidius colemani (V.), in no-choice experiments under laboratory conditions. The number of Rhopalosiphum padi (L.) and Sitobion avenae (F.) killed was increased by increasing the concentration of V. lecanii. The timing of application of fungus to aphids affected the efficacy of other biocontrol agents, a parasitoid and a predator. Parasitation by A. colemani (V.) in both cereal aphids (S. avenae and R. padi) was not affected by V. lecanii when aphids were first treated with V. lecanii and then exposed to A. colemani. The emergence of adults from parasitised mummies was, however, lower in infected aphids than in uninfected aphids when the aphids were first exposed to the parasitoids and then treated with fungus. The female sex ratio in the emerging adults was lower in V. lecanii-treated aphids in both species. When aphids were first treated with V. lecanii, 72 h before predation, fewer aphids of both species were consumed by H. axyridis (P.). Use of entomopathogenic fungus as a biological control agent could be a complementary strategy in an integrated pest management programme against cereal aphids, but it can reduce the efficiency of other biocontrol agents (parasitoids and predators) when applied simultaneously.

Parasitic wasp responses to symbiont-based defense in aphids

BackgroundRecent findings indicate that several insect lineages receive protection against particular natural enemies through infection with heritable symbionts, but little is yet known about whether enemies are able to discriminate and respond to symbiont-based defense. The pea aphid, Acyrthosiphon pisum, receives protection against the parasitic wasp, Aphidius ervi, when infected with the bacterial symbiont Hamiltonella defensa and its associated bacteriophage APSE (Acyrthosiphon pisum secondary endosymbiont). Internally developing parasitoid wasps, such as A. ervi, use maternal and embryonic factors to create an environment suitable for developing wasps. If more than one parasitoid egg is deposited into a single aphid host (superparasitism), then additional complements of these factors may contribute to the successful development of the single parasitoid that emerges.ResultsWe performed experiments to determine if superparasitism is a tactic allowing wasps to overcome symbiont-mediated defense. We found that the deposition of two eggs into symbiont-protected aphids significantly increased rates of successful parasitism relative to singly parasitized aphids. We then conducted behavioral assays to determine whether A. ervi selectively superparasitizes H. defensa-infected aphids. In choice tests, we found that A. ervi tends to deposit a single egg in uninfected aphids, but two or more eggs in H. defensa-infected aphids, indicating that oviposition choices may be largely determined by infection status. Finally, we identified differences in the quantity of the trans-β-farnesene, the major component of aphid alarm pheromone, between H. defensa-infected and uninfected aphids, which may form the basis for discrimination.ConclusionsHere we show that the parasitic wasp A. ervi discriminates among symbiont-infected and uninfected aphids, and changes its oviposition behavior in a way that increases the likelihood of overcoming symbiont-based defense. More generally, our results indicate that natural enemies are not passive victims of defensive symbionts, and that an evolutionary arms race between A. pisum and the parasitoid A. ervi may be mediated by a bacterial symbiosis.

Entomopathogenic fungi stimulate transgenerational wing induction in pea aphids, Acyrthosiphon pisum (Hemiptera: Aphididae)

1. Aphid natural enemies include not only predators and parasitoids but also pathogens, of which fungi are the most studied for biological control. While wing formation in aphids is induced by abiotic conditions, it is also affected by biotic interactions with their arthropod natural enemies. Wing induction via interactions with arthropod natural enemies is mediated by the increase in their physical contact when alarmed (pseudo‐crowding). Pathogenic fungi do not trigger this alarm behaviour in aphids and, therefore, no pseudo‐crowding occurs.2. We hypothesise that, while pathogenic fungi will stimulate maternally induced wing formation, the mechanism is different and is influenced by pathogen specificity. We tested this hypothesis using two entomopathogenic fungi, Pandora neoaphidis and Beauveria bassiana, an aphid specialist and a generalist respectively, on the pea aphid, Acyrthosiphon pisum Harris.3. We first demonstrate that pea aphids infected with either pathogen and maintained in groups on broad bean plants produced a higher proportion of winged morphs than uninfected control aphids. We then show that, when maintained in isolation, aphids infected with either pathogen also produced higher proportions of winged offspring than control aphids. There was no difference between P. neoaphidis and B. bassiana in their effects on wing induction in either experiment.4. Unlike the effect of predators and parasitoids on pea aphid wing induction, the effect of pathogens is independent of physical contact with other aphids, suggesting that physiological cues induce wing formation in infected aphids. It is possible that aphids benefit from wing induction by escaping infected patches whilst pathogens may benefit through dispersion. Possible mechanisms of wing induction are discussed.

Parasitic wasp responses to symbiont-based defense in aphids

BackgroundRecent findings indicate that several insect lineages receive protection against particular natural enemies through infection with heritable symbionts, but little is yet known about whether enemies are able to discriminate and respond to symbiont-based defense. The pea aphid, Acyrthosiphon pisum, receives protection against the parasitic wasp, Aphidius ervi, when infected with the bacterial symbiont Hamiltonella defensa and its associated bacteriophage APSE (Acyrthosiphon pisum secondary endosymbiont). Internally developing parasitoid wasps, such as A. ervi, use maternal and embryonic factors to create an environment suitable for developing wasps. If more than one parasitoid egg is deposited into a single aphid host (superparasitism), then additional complements of these factors may contribute to the successful development of the single parasitoid that emerges.ResultsWe performed experiments to determine if superparasitism is a tactic allowing wasps to overcome symbiont-mediated defense. We found that the deposition of two eggs into symbiont-protected aphids significantly increased rates of successful parasitism relative to singly parasitized aphids. We then conducted behavioral assays to determine whether A. ervi selectively superparasitizes H. defensa-infected aphids. In choice tests, we found that A. ervi tends to deposit a single egg in uninfected aphids, but two or more eggs in H. defensa-infected aphids, indicating that oviposition choices may be largely determined by infection status. Finally, we identified differences in the quantity of the trans-β-farnesene, the major component of aphid alarm pheromone, between H. defensa-infected and uninfected aphids, which may form the basis for discrimination.ConclusionsHere we show that the parasitic wasp A. ervi discriminates among symbiont-infected and uninfected aphids, and changes its oviposition behavior in a way that increases the likelihood of overcoming symbiont-based defense. More generally, our results indicate that natural enemies are not passive victims of defensive symbionts, and that an evolutionary arms race between A. pisum and the parasitoid A. ervi may be mediated by a bacterial symbiosis.

Development, specificity and sublethal effects of symbiont‐conferred resistance to parasitoids in aphids

Summary1. One of the most exciting recent discoveries in the field of ecological immunology has been that insects employ the help of heritable symbionts as a defence against parasitoids and pathogens. Aphids commonly harbour the facultative bacterial endosymbiont Hamiltonella defensa, which is known to increase their resistance to parasitoids. It is unknown how this resistance develops during the aphids’ ontogeny, following the transmission bottleneck between mother and offspring, and how specific symbiont‐conferred defences are.2. We addressed these issues in the black bean aphid, Aphis fabae, by exposing aphids of different age classes to the parasitoid Lysiphlebus fabarum. The susceptibility of aphids that were either naturally or experimentally infected with H. defensa was compared with that of uninfected aphids.3. Susceptibility to parasitoids decreased with aphid age, but aphids harbouring H. defensa showed an earlier and/or steeper decline to lower levels of susceptibility than aphids without this symbiont. This is consistent with the hypothesis that during aphid development, symbiont‐conferred resistance builds up with bacterial population growth, which we documented using quantitative polymerase chain reaction (qPCR).4. Parasitoids that successfully overcame the symbiont‐conferred resistance still suffered from sublethal effects of H. defensa. They exhibited lower emergence, delayed development and reduced size compared with parasitoids developing in aphids without H. defensa.5. The most striking result was a strong interaction on the rates of parasitism between aphid sublines infected with different isolates of H. defensa and the parasitoid lines they were exposed to, suggesting a high specificity of symbiont‐conferred resistance.6. Based on these results, we conclude that when faced with hosts possessing H. defensa, aphid parasitoids are under selection to preferentially attack the youngest host stages and/or to discriminate against symbiont‐protected aphids. Furthermore, the specificity induced by H. defensa in the interaction between host and parasitoid is likely to have important consequences for co‐evolution. It may result in negative frequency‐dependent selection and thus promote genotypic variation.