Pea Aphid Acyrthosiphon Pisum Research Articles

Deep Conservation of Hid-Like RHG Gene Family Homologs in Winged Insects Revealed by "Taxon Hopping" BLAST.

Together with sickle (skl), the Drosophila paralogs reaper (rpr), head involution defective (hid), and grim (RHG) control a critical switch in the induction of programmed cell death. RHG homologs have been identified in other dipteran and lepidopteran species but not beyond. Revisiting this issue with a "taxon hopping" BLAST search strategy in current genome and transcriptome resources, I detected high confidence RHG homologs in Coleoptera, Hymenoptera, Hemiptera, and Dictyoptera. Analyses of gene structure and protein sequence conservation revealed aconserved splicing pattern and highly conserved amino acid residues at both the N- and C-terminal ends that identify hid as the most ancestrally organized RHG gene family member in Drosophila. hid-like RHG homologs were also detected in mosquitoes, redefining their michelob_x (mx) genes as an expansion of derived RHG homologs. Only singleton homologs were detected in the large majority of other insect clades. Lepidopteran RHG homologs, however, stand out by producing an evolutionarily-derived splice isoform, identified in previous work, in addition to the newly detected hid-like isoform. Exceptional sequence diversification of select RHG homologs at the family- and genus-level explain their previous elusiveness in important insect genome model species like the red flour beetle Tribolium castaneum and the pea aphid Acyrthosiphon pisum. Combined, these findings expand the minimal age of the RHG gene family by about 100 million years and open new avenues for molecular cell death studies in insects.

Effects of Aqueous Extracts of Endophyte-Infected Grass Achnatherum inebrians on Growth and Development of Pea Aphid Acyrthosiphon pisum.

Simple SummaryDrunken horse grass Achnatherum inebrians is a widely distributed perennial poisonous grass on the grasslands of Northern and Northwestern China. The present study focused on contact toxicity of aqueous extracts of endophyte-infected (E+) and endophyte-free (E−) plants of A. inebrians in different growth periods on pea aphids. These results show that extracts from endophyte-containing plants may contain compounds that may be used to control insects and lay a foundation for deeper chemical analysis to identify insecticidal compounds.The pea aphid Acyrthosiphon pisum has a worldwide distribution and causes serious losses for agricultural production. Drunken horse grass Achnatherum inebrians is a widely distributed perennial poisonous grass on the grasslands of Northern and Northwestern China. The present study focused on contact toxicity activity of aqueous extracts of endophyte-infected (E+) and endophyte-free (E−) A. inebrians in different growth periods of pea aphids, and the growth and development of two color morphs of F1 generation nymphs. Both of the color morphs had development durations in E+ treatments that tended to be longer at 1st, 2nd, 3rd, and 4th instars than E− and control (CK). The E+ treated aphids also showed decreased weights at maturity with over all lower mean relative growth rates (MRGR). Aphid survival of E+ treated aphids was lower than that of E− and CK at all growth periods. Seeding stage E+ extracts showed a greater propensity for negatively affecting aphids than did E+ extract at maturity and the yellowing stage. These results show that extracts from endophyte-containing plants may contain compounds that may be used to control insects.

Evolutionary diversification of insulin-related peptides (IRPs) in aphids and spatiotemporal distribution in Acyrthosiphon pisum

Members of the insulin superfamily activate the evolutionarily highly conserved insulin/insulin-like growth factor signaling pathway, involved in regulation of growth, energy homeostasis, and longevity. In the current study we focus on aphids to gain more insight into the evolution of the IRPs and how they may contribute to regulation of the insulin-signaling pathway. Using the latest annotation of the pea aphid (Acyrthosiphon pisum) genome, and combining sequence alignments and phylogenetic analyses, we identified seven putative IRP encoding-genes, with IRP1-IRP4 resembling the classical insulin and insulin-like protein structures, and IRP5 and IRP6 bearing insulin-like growth factor (IGF) features. We also identified IRP11 as a new and structurally divergent IRP present in at least eight aphid genomes. Globally the ten aphid genomes analyzed in this work contain four to 15 IRPs, while only three IRPs were found in the genome of the grape phylloxera, a hemipteran insect representing an earlier evolutionary branch of the aphid group. Expression analyses revealed spatial and temporal variation in the expression patterns of the different A. pisum IRPs. IRP1 and IRP4 are expressed throughout all developmental stages and morphs in neuroendocrine cells of the brain, while IRP5 and IRP6 are expressed in the fat body. IRP2 is expressed in specific cells of the gut in aphids in non-crowded conditions and in the head of aphids under crowded conditions, IRP3 in salivary glands, and both IRP2 and IRP3 in the male morph. IRP11 expression is enriched in the carcass. This complex spatiotemporal expression pattern suggests functional diversification of the IRPs.

Aphids Facing Their Parasitoids: A First Look at How Chemical Signals May Make Higher Densities of the Pea Aphid Acyrthosiphon pisum Less Attractive to the Parasitoid Aphidius ervi.

Simple SummarySuccessful foraging behavior of parasitoids depends on specific organic information emitted by host-infested plants. For instance, the emission of volatile compounds increases in infested plants, and these are the first indicator of host presence. Parasitoids are attracted by these volatiles in a quite specific way. By combining behavioral and chemical studies, we showed bottom-up effects in a broad bean Vicia faba (Fabaceae)–pea aphid Acyrthosiphon pisum (Homoptera: Aphididae)–parasitoid Aphidius ervi (Hymenoptera: Braconidae) model system. We found that behavioral selection of parasitoid females toward plants with a high density of aphid infestation was reduced, and this can be linked to reduced emission of volatile compounds. In practice, if parasitoids are less attracted to plants with high-density aphid infestations, there may be potential negative impacts on biological control. Therefore, the common recommendation in biological control is to release parasitoids early in the season when aphid density on crop plants is still low.Herbivore-induced plant volatiles constitute the first indicators of insect host presence, and these can affect the foraging behavior of their natural enemies. The density of insect hosts may affect the nature and concentration of these plant-induced volatiles. We tested the impact of infestation density (low, intermediate, and high) of the pea aphid, Acyrthosiphon pisum (Homoptera: Aphididae), feeding on the broad bean Vicia faba, on the attractiveness of the parasitoid Aphidius ervi (Hymenoptera: Braconidae), using a Y-tube olfactometer (infested vs. non-infested plants). The emitted volatile compounds from both infested and non-infested plants were collected and identified. In addition, two series of experiments were carried out to test the impact of the presence of a conspecific female parasitoid within the aphid/plant complex on the attractiveness to other females. Parasitoids were significantly more attracted to the plants with low and intermediate aphid infestation levels. The volatile blend composition of the infested plants changed in relation to aphid density and may explain the low attraction of parasitoids toward high aphid density. The presence of conspecific females on the aphid patch had no apparent impact on the behavioral choices of other parasitoid females. Our study adds a new aspect to understanding plant–aphid–parasitoid interactions, including the possibility that aphids may manipulate chemical cues of host plants affecting the orientation of parasitoids.

Frequent Drivers, Occasional Passengers: Signals of Symbiont-Driven Seasonal Adaptation and Hitchhiking in the Pea Aphid, Acyrthosiphon pisum.

Simple SummaryRapid adaptation has been observed for several insects, including a number of agricultural pests. In these instances, it is expected that variation in insect-encoded genes encodes the variable phenotypes being acted on by natural selection. However, in addition to the thousands of genes encoded in their genomes, many insects harbor maternally transmitted, symbiotic bacteria encoding hundreds to thousands of genes of their own. Variation in the genes or presence and absence of such bacteria may, thus, also cause phenotypic variation, providing further raw material for natural selection. Here we studied symbiotic bacteria of the pea aphid, demonstrating that several change in prevalence within a single growing season. The frequencies of some bacteria shifted in concert with environmental factors predicted to determine their costs and benefits. Interpreting these correlations as plausible signals of symbiont-mediated insect adaptation, we found little evidence in support of alternative hypotheses, and that defense against fungal pathogens was possibly the most common beneficial symbiont service. Yet we also found that the particular combinations of bacteria living within an aphid may sometimes shape how aphids respond to natural selection. Our results have implications for the management of crop pests and for understanding the nature of rapid insect adaptation.Insects harbor a variety of maternally inherited bacterial symbionts. As such, variation in symbiont presence/absence, in the combinations of harbored symbionts, and in the genotypes of harbored symbiont species provide heritable genetic variation of potential use in the insects’ adaptive repertoires. Understanding the natural importance of symbionts is challenging but studying their dynamics over time can help to elucidate the potential for such symbiont-driven insect adaptation. Toward this end, we studied the seasonal dynamics of six maternally transferred bacterial symbiont species in the multivoltine pea aphid (Acyrthosiphon pisum). Our sampling focused on six alfalfa fields in southeastern Pennsylvania, and spanned 14 timepoints within the 2012 growing season, in addition to two overwintering periods. To test and generate hypotheses on the natural relevance of these non-essential symbionts, we examined whether symbiont dynamics correlated with any of ten measured environmental variables from the 2012 growing season, including some of known importance in the lab. We found that five symbionts changed prevalence across one or both overwintering periods, and that the same five species underwent such frequency shifts across the 2012 growing season. Intriguingly, the frequencies of these dynamic symbionts showed robust correlations with a subset of our measured environmental variables. Several of these trends supported the natural relevance of lab-discovered symbiont roles, including anti-pathogen defense. For a seventh symbiont—Hamiltonella defensa—studied previously across the same study periods, we tested whether a reported correlation between prevalence and temperature stemmed not from thermally varying host-level fitness effects, but from selection on co-infecting symbionts or on aphid-encoded alleles associated with this bacterium. In general, such “hitchhiking” effects were not evident during times with strongly correlated Hamiltonella and temperature shifts. However, we did identify at least one time period in which Hamiltonella spread was likely driven by selection on a co-infecting symbiont—Rickettsiella viridis. Recognizing the broader potential for such hitchhiking, we explored selection on co-infecting symbionts as a possible driver behind the dynamics of the remaining six species. Out of twelve examined instances of symbiont dynamics unfolding across 2-week periods or overwintering spans, we found eight in which the focal symbiont underwent parallel frequency shifts under single infection and one or more co-infection contexts. This supported the idea that phenotypic variation created by the presence/absence of individual symbionts is a direct target for selection, and that symbiont effects can be robust under co-habitation with other symbionts. Contrastingly, in two cases, we found that selection may target phenotypes emerging from symbiont co-infections, with specific species combinations driving overall trends for the focal dynamic symbionts, without correlated change under single infection. Finally, in three cases—including the one described above for Hamiltonella—our data suggested that incidental co-infection with a (dis)favored symbiont could lead to large frequency shifts for “passenger” symbionts, conferring no apparent cost or benefit. Such hitchhiking has rarely been studied in heritable symbiont systems. We propose that it is more common than appreciated, given the widespread nature of maternally inherited bacteria, and the frequency of multi-species symbiotic communities across insects.

Horizontal Transfer of Microbial Toxin Genes to Gall Midge Genomes.

A growing body of evidence has underscored the role of horizontal gene transfer (HGT) in animal evolution. Previously, we discovered the horizontal transfer of the gene encoding the eukaryotic genotoxin cytolethal distending toxin B (cdtB) from the pea aphid Acyrthosiphon pisum secondary endosymbiont (APSE) phages to drosophilid and aphid nuclear genomes. Here, we report cdtB in the nuclear genome of the gall-forming “swede midge” Contarinia nasturtii (Diptera: Cecidomyiidae) via HGT. We searched all available gall midge genome sequences for evidence of APSE-to-insect HGT events and found five toxin genes (aip56, cdtB, lysozyme, rhs, and sltxB) transferred horizontally to cecidomyiid nuclear genomes. Surprisingly, phylogenetic analyses of HGT candidates indicated APSE phages were often not the ancestral donor lineage of the toxin gene to cecidomyiids. We used a phylogenetic signal statistic to test a transfer-by-proximity hypothesis for animal HGT, which suggested that microbe-to-insect HGT was more likely between taxa that share environments than those from different environments. Many of the toxins we found in midge genomes target eukaryotic cells, and catalytic residues important for toxin function are conserved in insect copies. This class of horizontally transferred, eukaryotic cell-targeting genes is potentially important in insect adaptation.

Effect of Soy Leaf Flavonoids on Pea Aphid Probing Behavior.

Simple SummaryFlavonoids are plant phenolic compounds whose biological activities include participation in plant responses to various stresses of biological and environmental origins, including protection against insect herbivore attack. We were interested in whether the specific flavonoids detected in soybean leaves have the potential to discourage the pea aphid from infesting other leguminous plants, peas in particular. We immersed the pea leaves in ethanolic solutions of the flavonoids apigenin, daidzein, genistein and kaempferol, offered them to the pea aphids and observed their behavior when they probed plant tissues with their piercing–sucking mouthparts. We discovered that aphids readily probed the pea leaves whether they were treated with flavonoids or not. However, later on, the behavior of the aphids changed depending on the flavonoid applied. Apigenin, daidzein and kaempferol caused a decrease in the intensity of plant sap ingestion. In addition, daidzein and kaempferol made the finding of sap-transporting vessels more difficult for aphids. In contrast, genistein did not affect the pea aphids’ feeding activity. Our findings provide the plant breeders and plant protection services with information on what direction their efforts should take to protect leguminous plants against aphids in a sustainable and environmentally friendly way.Flavonoids detected in soybean Glycine max (L.) Merr. (Fabaceae) cause various alterations in the metabolism, behavior, and development of insect herbivores. The pea aphid Acyrthosiphon pisum (Harris) (Hemiptera: Aphididae) poses potential threat to soybeans, but the effect of individual flavonoids on its feeding-associated behavior is relatively unknown. We monitored probing behavior (stylet penetration activities) of A. pisum on its preferred host plant, Pisum sativum L. untreated (control) and treated with 0.1% ethanolic solutions of flavonoids apigenin, daidzein, genistein, and kaempferol. We applied the electrical penetration graph (electropenetrography, EPG) technique, which visualizes the movements of aphid stylets within plant tissues. None of the applied flavonoids affected the propensity to probe the plants by A. pisum. However, apigenin enhanced the duration of probes in non-phloem tissues, which caused an increase in the frequency and duration of stylet mechanics derailment and xylem sap ingestion but limited the ingestion of phloem sap. Daidzein caused a delay in reaching phloem vessels and limited sap ingestion. Kaempferol caused a reduction in the frequency and duration of the phloem phase. Genistein did not affect aphid probing behavior. Our findings provide information for selective breeding programs of resistant plant cultivars to A. pisum.

Diversity and geographic variation of endosymbiotic bacteria in natural populations of the pea aphid (Acyrthosiphon pisum) in China

• We firstly characterized symbiont communities and diversity of the pea aphid in China. • The obligate symbiont, Buchnera , was present in all aphid samples. • Four facultative symbionts ( Serratia , Hamiltonella , Regiella , and Rickettsia ) showed a significant geographic variation. • Bacterial communities of the Chinese pea aphid populations were mainly influenced by environmental factors. Bacterial symbionts in aphids are known to benefit the insect host and associated with aphid’s ecological adaptation. The pea aphid ( Acyrthosiphon pisum ), an important legume pest worldwide, carries at least eight endosymbionts, providing a model system to study insect–bacteria interactions. However, species diversity and geographic variations of endosymbionts are unknown in Chinese populations; therefore, we characterized symbiont communities and diversity of 27 pea aphid samples from 13 geographic populations of China. Via amplicon high-throughput sequencing and diagnostic PCR, we found that bacterial communities of Chinese populations were dominated by Proteobacteria and Firmicutes. Among eight known endosymbionts, five ( Buchnera , Serratia , Hamiltonella , Regiella , and Rickettsia ) were detected by both methods, with a specific geographical distribution. The obligate symbiont, Buchnera , was present in all aphid samples, while the four facultative symbionts showed a significant geographic variation. Each population was randomly infected with distinct endosymbionts, ranging from three to five species. Serratia and Rickettsia showed relatively higher abundance in central regions of China, Regiella was predominant in eastern and western China, whereas Hamiltonella showed an extremely low abundance and was absent in four populations. Samples grouped by altitudes showed a significant diversity difference, whereas there was no significant difference between red and green body colors. Bacterial community structures of the Chinese pea aphid populations were mainly influenced by environmental factors, other than body colors. These data can guide the development of potential biocontrol techniques against this aphid.

Antixenosis in Glycine max (L.) Merr against Acyrthosiphon pisum (Harris)

To reveal the antixenosis potential against the pea aphid Acyrthosiphon pisum (Harris) (Hemiptera: Aphididae) we analyzed the pea aphid survival and probing behavior, and the quantitative and qualitative variation of flavonoids in the leaves of selected soybean Glycine max (L.) Merr (Fabaceae) cultivars ‘Aldana’, ‘Annushka’, ‘Augusta’, ‘Madlen’, ‘Mavka’, ‘Simona’, ‘Violetta’, and ‘Viorica’. Aphid survival was drastically impeded on all cultivars. The electronic monitoring of aphid probing using the Electrical Penetration Graph (EPG) technique revealed that on all soybean cultivars, A. pisum readily probed into leaf tissues but the probes were usually terminated before reaching vascular tissues, which demonstrates the activity of antixenosis mechanisms in peripheral tissues epidermis and/or mesophyll in soybean leaves. The potency of antixenosis factors differed among soybean cultivars, which was reflected in differences in aphid survival and frequency and duration of phloem sap ingestion. Seven flavonoids were found: apigenin, daidzein, genistein, glycitein, isorhamnetin, kaempferol, and rutin, which occurred in different amount and proportion in individual cultivars. The content of apigenin and genistein in all soybean cultivars studied probably made them relatively unacceptable to A. pisum. Kaempferol in ‘Aldana’ might be responsible for the observed strong antixenosis resistance of this cultivar to A. pisum. The results of our survey provide the first detailed data that can be used for future studies.

Responses to predation risk cues and alarm pheromones affect plant virus transmission by an aphid vector.

Herbivores assess predation risk in their environment by identifying visual, chemical, and tactile predator cues. Detection of predator cues can induce risk-avoidance behaviors in herbivores that affect feeding, dispersal, and host selection in ways that minimize mortality and reproductive costs. For herbivores that transmit plant pathogens, including many aphids, changes in herbivore behavior in response to predator cues may also affect pathogen spread. However, few studies have assessed how aphid behavioral responses to different types of predator cues affect pathogen transmission. Here, we conducted greenhouse experiments to assess whether responses of pea aphids (Acyrthosiphon pisum) to predation risk and alarm pheromone (E-β-Farnesene), an aphid alarm signal released in response to predation risk, affected transmission of Pea enation mosaic virus (PEMV). We exposed A. pisum individuals to risk cues, and quantified viral titer in aphids and pea (Pisum sativum) host plants across several time periods. We also assessed how A. pisum responses to risk cues affected aphid nutrition, reproduction, and host selection. We show that exposure to predator cues and alarm pheromone significantly reduced PEMV acquisition and inoculation. Although vectors avoided hosts with predator cues, predator cues did not alter vector reproduction or reduce nutrient acquisition. Overall, these results suggest that non-consumptive effects of predators may indirectly decrease the spread of plant pathogens by altering vector behavior in ways that reduce vector competence and pathogen transmission efficiency.

Aphids show interspecific and intraspecific variation in life history responses to host plant infection by the fungal pathogen Botrytis cinerea

AbstractThe life histories of insect herbivores are affected by variation in host plant quality, with poor quality typically being associated with reduced herbivore fecundity, size and longevity. Plant pathogens are ubiquitous in nature and can alter host plant quality as experienced by insect herbivores. We asked how host plant infection by the widespread and economically important fungal pathogen Botrytis cinerea affected the life history traits of two aphid species. We found that the life history traits of the black bean aphid Aphis fabae were negatively affected by being reared on infected host plants, showing reduced fecundity, population growth rate, size, off‐plant survival time and development rate. In contrast, we found that pea aphids Acyrthosiphon pisum benefitted from being reared on infected plants, and that the degree of benefit varied between pea aphid clonal lines. This work suggests that the ecological and economic consequences of plant pathogen infection on the dynamics of aphid pests could be difficult to predict.

Male-Biased microRNA Discovery in the Pea Aphid.

Simple SummaryMales and females develop from a genome that is largely similar, yet the sexes have dramatically different traits. How this happens has driven interest in the role of epigenetic mechanisms—i.e., changes in gene function that are not due to changes in the DNA—in regulating sexual dimorphisms. Our focus here is on differences between the sexes in one epigenetic mechanism, the downregulation of gene expression by small RNAs called microRNAs. Typical genomes code for hundreds of microRNAs and each one can target many protein-coding RNAs, ultimately causing them to produce fewer protein copies. Here, we focus on microRNAs in male versus female pea aphids. We use small RNA sequence data to identify all the microRNAs in the pea aphid genome and to find microRNAs that are highly biased towards expression in males. This work is foundational for future studies of the epigenetic basis of sex differences in the pea aphid.Epigenetic mechanisms modulate gene expression levels during development, shaping how a single genome produces a diversity of phenotypes. Here, we begin to explore the epigenetic regulation of sexual dimorphism in pea aphids (Acyrthosiphon pisum) by focusing on microRNAs. Previous analyses of microRNAs in aphids have focused solely on females, so we performed deep sequencing of a sample containing early-stage males. We used this sample, plus samples from Genbank, to find 207 novel pea aphid microRNA coding loci. We localized microRNA loci to a chromosome-level assembly of the pea aphid genome and found that those on the X chromosome have lower overall expression compared to those on autosomes. We then identified a set of 19 putative male-biased microRNAs and found them enriched on the X chromosome. Finally, we performed protein-coding RNA-Seq of first instar female and male pea aphids to identify genes with lower expression in males. 10 of these genes were predicted targets of the 19 male-biased microRNAs. Our study provides the most complete set of microRNAs in the pea aphid to date and serves as foundational work for future studies on the epigenetic control of sexual dimorphism.

Long-Term Effect of Elevated CO2 on the Development and Nutrition Contents of the Pea Aphid (Acyrthosiphon pisum).

It is predicted that the current atmospheric CO2 level will be doubled by the end of this century. Here, we investigate the impacts of elevated CO2 (550 and 750 μL/L) on the development and nutrition status of the green pea aphid for six generations, which is longer than previous studies. All seven examined physiological parameters were not affected over six generations under the ambient CO2 level (380 μL/L). However, the elevated CO2 levels (550 and 750 μL/L) prolonged nymph duration, decreased adult longevity, female fecundity and protein content, and increased the contents of total lipid, soluble sugar and glycogen. There was a significant interaction between the effect of CO2 levels and the effect of generations on nymph duration, female fecundity and adult longevity. The elevated CO2 had immediate effects on the female fecundity and the contents of total protein, total lipid and soluble sugar, starting within F0 generation. The adult longevity decreased, and the glycogen content increased from the F1 generation. However, the significant effect on the nymph development was only observed after three generations. Our study indicates that the elevated CO2 levels first influence the reproduction, the nutrition and the energy supply, then initiate aphid emergency responses by shortening lifespan and increasing glucose metabolism, and finally result in the slow development under further persistent elevated CO2 conditions after three generations, possibly leading to population decline under elevated CO2 conditions. Our results will guide further field experiments under climate change conditions to evaluate the effects of elevated CO2 on the development of the pea aphids and other insects, and to predict the population dynamics of the green pea aphid.

Elucidation of the whole carotenoid biosynthetic pathway of aphids at the gene level and arthropodal food chain involving aphids and the red dragonfly

BackgroundAphids can be positioned as robust pest insects in farming and as ones of the model organisms for arthropods in molecular biology. Carotenoids are pigments that protect organisms from photooxidative damage caused by excessive light. Aphids were shown to possess genes of fungal origin for carotenoid biosynthesis, whereas a little knowledge was available about the functions of the genes and the biosynthetic pathway. Even carotenoid species contained in aphids were not enough understood. Main purpose of this study is to clarify these insufficient findings.ResultsThe whole carotenoid biosynthetic pathway of the pea aphid (Acyrthosiphon pisum) was elucidated at the gene level, through comprehensive functional analysis of its carotenogenic genes, using Escherichia coli that synthesized carotenoid substrates, along with structural and quantitative analysis of carotenoids from various aphid species. Four genes were needed to synthesize all carotenoids accumulated in aphids from geranylgeranyl diphosphate. The tor gene mediated desaturation reaction from phytoene to 3,4-didehydrolycopene. It was revealed that a gene designated ApCrtYB3, which was considered to have functionally evolved in aphids, can convert lycopene into uncommon carotenoids with the γ-ring such as (6′S)-β,γ-carotene and γ,γ-carotene. We further demonstrated that the atypical carotenoids work as ecological indicators for estimating the food chain from aphids to predatory arthropods, and showed that aphids contributed with significant levels to the food chain from insect herbivores to several predatory arthropods, i.e., the red dragonfly (Sympetrum frequens; adults), seven-spotted ladybird (Coccinella septempunctata), and two spiders, Oxyopes sertatus and Nephila clavata. Gut microflora of the dragonfly (mature adults) was also found to include endosymbiotic bacteria such as Serratia symbiotica specific to the black bean aphid (Aphis fabae).ConclusionsWe revealed the whole carotenoid biosynthetic pathway of aphids, including functional identification of the corresponding genes. Subsequently, we showed that arthropodal food chain can be estimated using the uncommon carotenoids of aphids as ecological indicators. This result indicated that aphids made significant contributions to the food chain of several predatory arthropods including the red-dragonfly adults. Aphids are likely to be positioned as an important “phytochemicals” source for some predatory insects and arachnids, which are often active under bright sunlight.

Luteoviruses affected energy reserves and fatty acid composition of their aphid vectors

AbstractLuteoviruses are plant‐infecting RNA viruses that are transmitted by aphids in a persistent circulative mode. Luteoviruses have been shown to positively influence their aphid vectors fitness. To find out the physiological responses of aphid vectors to luteoviruses, fatty acid (FA) composition and energy components (EC) of aphid vectors including pea aphid (Acyrthosiphon pisum Harris), black bean aphid (Aphis fabae Scopoli) and bird cherry‐oat aphid (Rhopalosiphum padi L.) were evaluated in response to Pea enation mosaic virus 1 (PEMV1), Bean leafroll virus (BLRV) and Barley yellow dwarf virus‐PAV (BYDV‐PAV), respectively. Our results showed that the number and the proportion of FA of total lipid are altered in aphid vectors in response to the luteoviruses. Compared to non‐viruliferous (NV) aphids, BLRV and PEMV1 elevated the amount of myristic acid (1.13 and 1.03‐fold increase) in A. fabae and A. pisum, respectively. V R. padi individuals exhibited a 1.11‐fold increase in palmitic acid content compared to the NV individuals. In contrast, other V aphids showed a reduced proportion of FA in response to the viruses. These changes in FA profile enhanced the ratio of saturated to unsaturated FA in V aphids (4.09‐fold increase). BLRV decreased protein and lipid contents in its aphid vector, A. fabae, while carbohydrate content was found to increase in V R. padi and A. pisum. Thus, our results demonstrate that luteoviruses change the content of some FA and EC in their aphid vectors highlighting a specific interaction among virus species, plant host and aphid vector.

Intraspecific variation in immune gene expression and heritable symbiont density.

Host genetic variation plays an important role in the structure and function of heritable microbial communities. Recent studies have shown that insects use immune mechanisms to regulate heritable symbionts. Here we test the hypothesis that variation in symbiont density among hosts is linked to intraspecific differences in the immune response to harboring symbionts. We show that pea aphids (Acyrthosiphon pisum) harboring the bacterial endosymbiont Regiella insecticola (but not all other species of symbionts) downregulate expression of key immune genes. We then functionally link immune expression with symbiont density using RNAi. The pea aphid species complex is comprised of multiple reproductively-isolated host plant-adapted populations. These 'biotypes' have distinct patterns of symbiont infections: for example, aphids from the Trifolium biotype are strongly associated with Regiella. Using RNAseq, we compare patterns of gene expression in response to Regiella in aphid genotypes from multiple biotypes, and we show that Trifolium aphids experience no downregulation of immune gene expression while hosting Regiella and harbor symbionts at lower densities. Using F1 hybrids between two biotypes, we find that symbiont density and immune gene expression are both intermediate in hybrids. We propose that in this system, Regiella symbionts are suppressing aphid immune mechanisms to increase their density, but that some hosts have adapted to prevent immune suppression in order to control symbiont numbers. This work therefore suggests that antagonistic coevolution can play a role in host-microbe interactions even when symbionts are transmitted vertically and provide a clear benefit to their hosts. The specific immune mechanisms that we find are downregulated in the presence of Regiella have been previously shown to combat pathogens in aphids, and thus this work also highlights the immune system's complex dual role in interacting with both beneficial and harmful microbes.

Does getting defensive get you anywhere?-Seasonal balancing selection, temperature, and parasitoids shape real-world, protective endosymbiont dynamics in the pea aphid.

Facultative, heritable endosymbionts are found at intermediate prevalence within most insect species, playing frequent roles in their hosts' defence against environmental pressures. Focusing on Hamiltonella defensa, a common bacterial endosymbiont of aphids, we tested the hypothesis that such pressures impose seasonal balancing selection, shaping a widespread infection polymorphism. In our studied pea aphid (Acyrthosiphon pisum) population, Hamiltonella frequencies ranged from 23.2% to 68.1% across a six-month longitudinal survey. Rapid spikes and declines were often consistent across fields, and we estimated that selection coefficients for Hamiltonella-infected aphids changed sign within this field season. Prior laboratory research suggested antiparasitoid defence as the major Hamiltonella benefit, and costs under parasitoid absence. While a prior field study suggested these forces can sometimes act as counter-weights in a regime of seasonal balancing selection, our present survey showed no significant relationship between parasitoid wasps and Hamiltonella prevalence. Field cage experiments provided some explanation: parasitoids drove modest ~10% boosts to Hamiltonella frequencies that would be hard to detect under less controlled conditions. They also showed that Hamiltonella was not always costly under parasitoid exclusion, contradicting another prediction. Instead, our longitudinal survey - and two overwintering studies - showed temperature to be the strongest predictor of Hamiltonella prevalence. Matching some prior lab discoveries, this suggested that thermally sensitive costs and benefits, unrelated to parasitism, can shape Hamiltonella dynamics. These results add to a growing body of evidence for rapid, seasonal adaptation in multivoltine organisms, suggesting that such adaptation can be mediated through the diverse impacts of heritable bacterial endosymbionts.

Drop when the stakes are high: adaptive, flexible use of dropping behaviour by aphids

Abstract For herbivorous insects, dropping from the host plant is a commonly-observed antipredator defence. The use of dropping compared to other behaviours and its timing in relation to contact with a predator was explored in both pea aphids (Acyrthosiphon pisum) and potato aphids (Macrosiphum euphorbiae). Pea aphids dropped more frequently in response to ladybird adults (Adalia bipunctata) than lacewing larvae (Chrysoperla carnea). Potato aphids mainly walked away or backed-up in response to both predator types; but they dropped more frequently relative to other non-walking defences when faced with ladybird adults. Contact with a predator was an important influencer of dropping for both species, and most drops occurred from adjacent to the predator. Dropping appears to be a defence adaptively deployed only when the risk of imminent predation is high; factors that increase dropping likelihood include presence of faster-foraging predators such as adult ladybirds, predator proximity, and contact between aphid and predator.

Secondary Symbionts Affect Foraging Capacities of Plant-Specialized Genotypes of the Pea Aphid.

Ecological specialization is widespread in animals, especially in phytophagous insects, which have often a limited range of host plant species. This host plant specialization results from divergent selection on insect populations, which differ consequently in traits like behaviors involved in plant use. Although recent studies highlighted the influence of symbionts on dietary breadth of their insect hosts, whether these microbial partners influence the foraging capacities of plant-specialized insects has received little attention. In this study, we used the pea aphid Acyrthosiphon pisum, which presents distinct plant-specialized lineages and several secondary bacterial symbionts, to examine the possible effects of symbionts on the different foraging steps from plant searching to host plant selection. In particular, we tested the effect of secondary symbionts on the aphid capacity (1) to explore habitat at long distance (estimated through the production of winged offspring), (2) to explore habitat at short distance, and (3) to select its host plant. We found that secondary symbionts had a variable influence on the production of winged offspring in some genotypes, with potential consequences on dispersal and survival. By contrast, symbionts influenced both short-distance exploration and host plant selection only marginally. The implication of symbionts' influence on insect foraging capacities is discussed.

Limited influence of gain and loss of symbionts on host plant selection in specialized pea aphid genotypes

Recent advances in insect-microbe interactions show that microbial associates can modulate the behaviors of their hosts, such as defensive or food choice behaviors. For phytophagous insects, host plant choice is of utmost importance, because most of them are specialized on a few plant species, which represent not only their food source but often their habitat and reproduction site. Therefore, identifying how microbial symbionts could influence host plant selection provides valuable information on their potential role in the evolution of the diet breath of phytophagous insects. In this study, we investigated whether bacterial symbiont loss or acquisition modulate host plant selection in the pea aphid Acyrthosiphon pisum Harris (Hemiptera: Aphididae). Several aphid lineages were considered, originating from three plant-specialized populations, each adapted to a different plant species. The symbiotic status of aphid lineages was experimentally manipulated to mimic natural events of symbiont loss and acquisition through horizontal transfer. Host plant selection was then tested by releasing aphids in a cage containing three plant species (1 host and 2 non-hosts) and determining the proportion of aphids on each plant. Our results show minor effects of the symbiotic status on aphid host plant choice, indicating that this trait is determined primarily by the aphid genome.