Juvenile hormone effects on polymorphism in the pea aphid, Acyrthosiphon pisum

When reared in short days (LD 12:12) at 15°C, apterous Acyrthosiphon pisum gave birth to sexual females (oviparae) exclusively for the first eight days of larviposition. After this time they switched to the production of parthenogenetic females (viviparae). Topical application of juvenile hormones I, II and III to fourth instar or adult ovipara-producers induced the precocious appearance of parthenogenetic females in the progeny sequence. Various forms intermediate between oviparae and viviparae were also produced and repetitive JH-I treatments resulted in a few alatiform progeny. However, many of the JH induced apterous, parthenogenetic females appeared to be normal viviparae and were capble of reproduction. Thus, prenatal treatment of oviparous embryos with JH diverts development towards the viviparous form. JH-I treatment of long-day reared A. pisum had no effect on the type of progeny produced.

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Effets de l'hormone juvénile sur le polymorphisme d'Acythosiphon pisum

Résumé Quand il est élevé sous jours courts (LN 12/12) à 15°C, le type anglais vert d'Acyrthosiphon pisum ne donne naissance qu'à des femelles sexuées (ovipares) pendant la première partie de sa période de reproduction. Ensuite quelques types intermédiaires ovipares/vivipares peuvent apparaitre avant que les pucerons ne bifurquent spontanément vers la production de femelles parthénogénétiques (vivipares). L'application cutanée d'hormones juvéniles (JH I, II, et III) aux larves de quatrième stade ou à des adultes producteurs d'ovipares provoque l'apparition prématurée d'intermédiaires et de vivipares dans la descendance. Les différentes formes intermédiaires produites par des applications répétées de J.H. comprenaient des types ailés ou partiellement ailés. Cependant, les vivipares aptères induits par J.H. étaient morphologiquement normaux et beaucoup étaient capables de se reproduire. Des traitements semblables aux J.H. de vivipares élevées en jours longs (LN 16/8) n'ont pas eu d'effets sur le type de la descendance.On ne sait pas si l'action de JH exogène sur l'induction des vivipares est direct ou indirect. La reprogrammation des embryons, autrement destinés à se développer comme ovipares, est examinée en relation avec notre connaissance du contrôle endocrine du polymorphisme des pucerons.

     

Microsatellite DNA markers for the pea aphid Acyrthosiphon pisum

Microsatellite loci were isolated from enriched partial genomic libraries of Acyrthosiphon loti and Acyrthosiphon pisum. Twenty of those loci were characterized in A. pisum. Fifteen of those loci were polymorphic. Genetic diversity varied across loci, allele repeat number ranging from two to 15, and observed heterozygosity from 0.1 and 0.96. An additional eight microsatellite loci originally isolated from other aphids but cross‐priming with A. pisum showed polymorphism as well. Allele size ranged from three to 9 and observed heterozygosity from 0.43 to 0.84. Overall, we present 23 microsatellite loci that can be used to reveal polymorphism in pea aphids.     

Instar-specific defense of the pea aphid,Acyrthosiphon pisum: Influence on oviposition success of the parasiteAphelinus asychis (Hymenoptera: Aphelmidae)

The parasite Aphelinus asychisWalker (Hymenoptera: Aphelinidae) oviposits in all four instars of the pea aphid, Acyrthosiphon pisum(Harris) (Homoptera: Aphididae). Searching females display a highly stereotyped sequence of behaviors when encountering a host. Once recognized, an aphid is examined and probed by the wasp with the everted ovipositor prior to oviposition. Oviposition success is influenced by aphid behavior that is related to aphid size and expressed through instarspecific escape and defense reactions. Being smaller and less able to defend themselves, first and early-second instars of pea aphid are more susceptible to successful parasitism than third and fourth instars, in that order. Observed patterns of preference by Aphelinus females for particular aphid species and instars reflect the outcome of behavioral interactions between the hosts and the parasites, rather than preference in the strict sense.     

Influence of rearing temperature on triacylglycerol storage in the pea aphid,Acyrthosiphon pisum

Total fatty acids in the pea aphid reared at low temperatures increased significantly compared to that at high rearing temperatures. This change is reflected in a large increase of myristic acid, which occurs exclusively in triacylglycerols. When aphids were moved from 25°C to a lower rearing temperature at 10°C, saturated fatty acids accumulated over time, reaching a maximum at 16th day. When aphids were moved to 4°C, a temperature below the developmental threshold, those aphids did not accumulate saturated fatty acids. Similar results were observed when aphids were exposed to sequential decrease in rearing temperature. However, both total fatty acids and myristic acid in the aphids from the treatments of sequential decreasing rearing temperature were significantly higher compared to those in the aphids from the treatments of sudden decreasing rearing temperature. This result, therefore, supports the hypothesis that cold‐adapted aphids can survive under threshold temperature for a longer period of time than noncold‐adapted aphids. Acetyl‐CoA carboxylase activity in the aphids at 25°C was twofold higher than that in the aphids at 10°C, whereas fatty acid synthase activities in the aphids reared at 25 and 10°C are similar. Aphids reared at 10°C showed a threefold reduction in reproduction rates. This reduced production of new nymphs reduces energy demand and would allow for accumulation of energy in the form of triacylglycerols. Therefore, the increased level of saturated fatty acids in aphids reared at low temperature is probably related to lower utilization of fatty acids rather than increased rates of biosynthesis.     

Temporal changes in host adaptation in the pea aphid, Acyrthosiphon pisum

Abstract.

• 1 Temporal changes in host adaptation were followed in a local population of the pea aphid, Acyrthosiphon pisum. Aphid clones were collected in one alfalfa and one clover field at three different times. In the spring, first-generation females were collected. Later, in the autumn, females belonging to the last parthenogenetic generation were collected. Lastly, sexual females were collected after mating in autumn and allowed to produce eggs which were hatched. The performance was evaluated on alfalfa and clover. The spring-collected individuals were also assessed on peas.
• 2 On the overwintering hosts clover and alfalfa, the clones performed best on the plant of origin, i.e. negative correlations in performance. Correlations between performance on the temporary summer host, pea, and that on clover/alfalfa were weak or nonsignificant.
• 3 Significant variation in host performance was found within both host fields at spring, which is a prerequisite for changes in clone composition due to selection/migration.
• 4 The clones from alfalfa showed an increase in mean performance on alfalfa between spring and autumn, whereas no changes among the clones from the clover field were observed. This difference in seasonal response between the two fields could have been the result of larger variation in performance among the alfalfa clones and/or a differential tendency to migrate among clones in both fields.
• 5 After sexual recombination in the autumn, mean performance in the alfalfa field returned to the spring level, probably as a result of emergence of new genetic combinations. In the clover field, mean performance did not change significantly over time.

     

Lipophorin receptor regulates the cuticular hydrocarbon accumulation and adult fecundity of the pea aphid Acyrthosiphon pisum

Cuticular hydrocarbons form a barrier that protects terrestrial insects from water loss via the epicuticle. Lipophorin loads and transports lipids, including hydrocarbons, from one tissue to another. In some insects, the lipophorin receptor (LpR), which binds to lipophorin and accepts its lipid cargo, is essential for female fecundity because it mediates the incorporation of lipophorin by developing oocytes. However, it is unclear whether LpR is involved in the accumulation of cuticular hydrocarbons and its precise role in aphid reproduction remains unknown. We herein present the results of our molecular characterization, phylogenetic analysis, and functional annotation of the pea aphid (Acyrthosiphon pisum) LpR gene (ApLpR). This gene was transcribed throughout the A. pisum life cycle, but especially during the embryonic stage and in the abdominal cuticle. Furthermore, we optimized the RHA interference (RNAi) parameters by determining the ideal dose and duration for gene silencing in the pea aphid. We observed that the RNAi-based ApLpR suppression significantly decreased the internal and cuticular hydrocarbon contents as well as adult fecundity. Additionally, a deficiency in cuticular hydrocarbons increased the susceptibility of aphids to desiccation stress, with decreased survival rates under simulated drought conditions. Moreover, ApLpR expression levels significantly increased in response to the desiccation treatment. These results confirm that ApLpR is involved in transporting hydrocarbons and protecting aphids from desiccation stress. Furthermore, this gene is vital for aphid reproduction. Therefore, the ApLpR gene of A. pisum may be a novel RNAi target relevant for insect pest management.     

Latitudinal trend in the reproductive mode of the pea aphid Acyrthosiphon pisum invading a wide climatic range

The maintenance of sexuality is a puzzling phenomenon in evolutionary biology. Many universal hypotheses have been proposed to explain the prevalence of sex despite its costs, but it has been hypothesized that sex could be also retained by lineage‐specific mechanisms that would confer some short‐term advantage. Aphids are good models to study the maintenance of sex because they exhibit coexistence of both sexual and asexual populations within the same species and because they invade a large variety of ecosystems. Sex in aphids is thought to be maintained because only sexually produced eggs can persist in cold climates, but whether sex is obligate or facultative depending on climatic conditions remains to be elucidated. In this study, we have inferred the reproductive mode of introduced populations of the pea aphid Acyrthosiphon pisum in Chile along a climatic gradient using phenotypic assays and genetic‐based criteria to test the ecological short‐term advantage of sex in cold environments. Our results showed a latitudinal trend in the reproductive mode of Chilean pea aphid population from obligate parthenogenesis in the north to an intermediate life cycle producing both parthenogenetic and sexual progeny in the southernmost locality, where harsh winters are usual. These findings are congruent with the hypothesis of the ecological short‐term advantage of sex in aphids.     

The economics of escape behaviour in the pea aphid,Acyrthosiphon pisum

Summary Pea aphids have several alternative responses to the detection of alarm pheromone produced by conspecifics. One of these, dropping from the feeding site to the ground, is potentially costly owing to the risk of desiccation-induced mortality on the ground before another host plant can be reached. Both dropping and walking from the feeding site incur a cost due to lost feeding opportunity. The aphids' decision as to which anti-predator tactic to use should be sensitive to the costs of their behaviour. Consequently, aphids should be less likely to drop when the risk of desiccation is higher, and less likely to drop or walk when the lost opportunity cost is higher. We tested these predictions by manipulating climatic severity (temperature and humidity) and host quality, respectively. As predicted, aphids are less likely to drop or walk in response to pheromone when feeding on high quality than on low quality hosts, and less likely to drop when the environment is hot and dry than when it is more benign. The latter is true whether the aphids are feeding on real or simulated leaves. Since all aphids were of the same clone, these results show that individual aphid genotypes possess the ability to adaptively modify their escape behaviour with changes in prevailing conditions. A number of other behavioural observations in the aphid literature may be interpreted in an economic or cost-benefit framework. The approach holds considerable promise for understanding many aspects of the anti-predator behaviour of aphids and other animals.     

The effects of two species of hymenopterous parasitoid on the reproductive system of the pea aphid, Acyrthosiphon pisum

Two cultures of first instar pea aphids, Acyrthosiphon pisum (Harris) (Homoptera: Aphididae), were subjected to parasitization by Ephedrus plagiator (Nees) and Aphidius ervi Haliday (Hymenoptera: Braconidae) respectively, and a third culture, of fourth instar pea aphids, was subjected to parasitization by E. plagiator. Significant parasitogenic effects on the reproductive system occurred within 24 h of exposure to parasitoids in the form of reductions in both embryo number and the size of the largest embryo, and degenerative changes in the embryos themselves. No evidence was found of direct feeding by the larvae on the host's tissues, until just before larval pupation. The largest embryos of some fourth instar aphids escaped the effects of parasitization probably because they had developed a resistant cuticle by the time of oviposition by the parasitoid. Results suggested that embryonic degeneration was an indirect effect of parasitization due to starvation, interference with the aphid's hormone levels, or both.

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Résumé Trois élevages de 100 à 200 premiers stades d'A. pisum, nés à 24 heures les uns des autres, ont été divisés chacun en un lot témoin et un lot expérimental. Deux des élevages expérimentaux ont alors été soumis au parasitisme d'Ephedrus plagiator ou de Aphidius ervi, le troisième élevage a pu se développer jusqu'au 4è stade avant d'être exposé à E. plagiator. Des échantillons de 10 pucerons ont été retirés de chaque lot après 24 heures et ensuite toutes les 48 heures pour être disséqués dans du liquide physiologique. Les nombres d'ovarioles et d'embryons et la taille du plus gros embryon ont été notés. Des cellules développées, issue de la séreuse de l'oeuf de l'hyménoptère se sont disséminées dans l'hémolymphe de l'hôte et ont servi d'aliment aux larves en développement. Seul le dernier stade larvaire des parasitoïdes s'est alimenté sur les tissus de l'hôte et ceci avait lieu juste avant la nymphose. Des effets significatifs se sont traduits après 24 heures par la réduction du nombre des embryons, par la diminution de la taille du plus grand de ceux-ci et par des altérations dans les embryons eux-mêmes.

     

Identification and characterisation of antixenosis and antibiosis to pea aphid (Acyrthosiphon pisum) in Pisum spp. germplasm

Acyrthosiphon pisum is a polyphagous aphid of major importance on the pea crop to which few resistant cultivars are available. In this study, we screened a germplasm collection of Pisum spp. under field conditions over two seasons yielding the identification of a number of accessions with intermediate levels of resistance. Selected accessions were further studied under semi‐controlled and controlled conditions in no choice and choice assays to validate the responses, and to further characterise the mechanisms of resistance operative. Results elucidated the valuable resistance of accession P40 (Pisum sativum ssp. sativum) and P665 (P. sativum ssp. syriacum), with the combination of both antixenosis, by reducing aphid preference, and antibiosis, by diminishing aphid proliferation.     

Erwinia aphidicola,a new species isolated from pea aphid,Acyrthosiphon pisum

Five strains of Gram-negative, oxidase-negative, facultatively anaerobic, fermentative, motile, rod-shaped bacterium with the general characteristics of the family Enterobacteriaceae were isolated from the gut of multiple specimens of the pea aphid. All the strains caused aphid mortality when ingested by insects via a synthetic diet. The results of biochemical tests showed that these strains are most related to Erwinia herbicola and Pantoea agglomerans. According to DNA-DNA hybridization, the five strains showed more than 96% relatedness to each other, indicating that these organisms are members of a single species. These strains were most closely related to Erwinia herbicola (22% DNA relatedness). Phenotypic differentiation of these strains from Erwinia herbicola, which was also detected from aphid gut, was based on negative reactions in tests of yellow pigment production, gelatin liquefaction, acid production from inulin, starch and dulcitol, and positive acid production from melibiose, inositol, cellobiose and glycerol. On the basis of these data, the name Erwinia aphidicola is proposed for the new organism. The type strain is strain X 001 (=IAM 14479).     

Allelic and genotypic diversity in long-term asexual populations of the pea aphid, Acyrthosiphon pisum in comparison with sexual populations

Many aphid species exhibit geographical variation in the mode of reproduction that ranges from cyclical parthenogenesis with a sexual phase to obligate parthenogenesis (asexual reproduction). Theoretical studies predict that organisms reproducing asexually should maintain higher allelic diversity per locus but lower genotypic diversity than organisms reproducing sexually. To corroborate this hypothesis, we evaluated genotypic and allelic diversities in the sexual and asexual populations of the pea aphid, Acyrthosiphon pisum (Harris). Microsatellite analysis revealed that populations in central Japan are asexual, whereas populations in northern Japan are obligatorily sexual. No mixed populations were detected in our study sites. Phylogenetic analysis using microsatellite data and mitochondrial cytochrome oxidase subunit I (COI) gene sequences revealed a long history of asexuality in central Japan and negated the possibility of the recent origin of the asexual populations from the sexual populations. Asexual populations exhibited much lower genotypic diversity but higher allelic richness per locus than did sexual populations. Asexual populations consisted of a few predominant clones that were considerably differentiated from one another. Sexual populations on alfalfa, an exotic plant in Japan, were most closely related to asexual populations associated with Vicia sativa L. The alfalfa-associated sexual populations harboured one COI haplotype that was included in the haplotype clade of the asexual populations. Available evidence suggests that the sexuality of the alfalfa-associated populations has recently been restored through the northward migration and colonization of alfalfa by V. sativa- associated lineages. Therefore, our results support the theoretical predictions and provide a new perspective on the origin of sexual populations.     

Attachment forces of pea aphids (Acyrthosiphon pisum) on different legume species

1. Sympatric populations of insects adapted to different host plants are good model systems not only to study how they adapt to the chemistry of their food plant, but also to investigate whether morphological modifications evolved enabling them to live successfully on a certain plant species. 2. The pea aphid, Acyrthosiphon pisum (Harris) encompasses at least 11 genetically distinct sympatric host races, each showing a preference for a certain legume species. The leaflet surfaces of these legumes differ considerably in their wax coverage. 3. It was investigated whether the attachment structures of three pea aphid genotypes from different host races are adapted to the different surface properties of their host plants and whether they show differences in their attachment ability on the respective host and non‐host plants. 4. The surface morphology of plants and aphid tarsi was examined using SEM (scanning electron microscopy). The ability of the aphids to walk on specific surfaces was tested using traction force measurements. 5. The presence of wax blooms on the leaflets lowers the aphids' attachment ability considerably and diminishes their subsequent attachment on ‘neutral’ surfaces like glass. The pea aphid host races differ in their ability to walk on certain surfaces. However, the genotype from the adapted aphid host race was not necessarily the one with the best walking performance on their host plant. All aphids, regardless of the original host plant, were most efficient on the neutral control surface glass. The general host plant Vicia faba was the plant with the most favourable surface for all aphid host races.     

Cannibalism in the pea aphid,Acyrthosiphon pisum

Previous observations of cannibalism have been made in the aphid Acyrthosiphon pisum （L.）： this article seeks to quantify factors contributing to such be- haviors. We observed and quantified the responses of a number of clones and life stages to varying levels of starvation, in the form of increasingly desiccated Vicafaba L. plants （receiving 50, 25, or 10 mL every second day） or a complete absence of host plant. We found that, while the longest incidences of cannibalism are carried out by juveniles （F = 3.45, P = 0.019, df = 3） and targeted at adults, the starvation treatments had the most significant effect on the prevalence of cannibalism in mature A. pisum （F ---- 2.24, P = 0.025, df = 9）. Furthermore, there was no difference between the prevalence or dura- tion of cannibalistic activities within and between different clones （P 〉 0.05 in all cases）, though juveniles were more likely to target unrelated aphids （V = 6 112, P = 0.011）, and spent more time feeding on aphids from the same culture （V = 6 062, P = 0.018）.     

Facultative 'secondary' bacterial symbionts and the nutrition of the pea aphid, Acyrthosiphon pisum

Abstract.  Facultative 'secondary' bacterial symbionts influence various traits of aphids, including plant utilization patterns and resistance to parasitoids. The present study is designed to test the hypothesis that these multiple effects are underlain by symbiont-mediated changes to the aphid requirement for the dominant dietary nutrients, sucrose and amino acids. The performance of pea aphids ( Acyrthosiphon pisum ) on chemically defined diets of systematically altered sucrose and amino acid content varies among eight parthenogenetic clones, with a pattern that does not match the aphid complements of secondary symbionts, Hamiltonella defensa , Regiella insecticola and Serratia symbiotica . Aphid performance is reduced, increased and unaffected by elimination of S. symbiotica , R. insecticola and H. defensa , respectively, but with no significant effect on the range of diets on which aphids performed well. It is concluded that the impact of secondary symbionts on aphid traits is most unlikely to have a purely nutritional basis.     

Effects of parasitism by Praon pequodorum on age-specific fecundity and population growth of the pea aphid, Acyrthosiphon pisum

The effects of parasitism by Praon pequodorum Viereck (Hymenoptera: Aphidiidae) on the survival, development and reproduction of apterous pea aphids, Acyrthosiphon pisum (Harris) (Homoptera: Aphididae), were studied in the laboratory. Ten cohorts of aphids differing in age from 0.5 h to 9 days were tested. At 20°C, pea aphids survived an average of 7 days after parasitization. Parasitized first and second instars died before reaching the adult stage. Third instars died as adults but without producing any offspring. Parasitized fourth instars and adult aphids produced a variable number of offspring; their mean total fecundity increased as an exponential function of the age at parasitization. Host age did not significantly affect the period from the beginning of parasitism to the end of reproduction. Equations are provided to estimate the length of the reproductive period, the total fecundity, the intrinsic rate of increase, and the doubling time as functions of aphid age at parasitization. The data are discussed with regard to host population growth. It is shown that a parasite's potential impact on an aphid population is critically dependent on its preference for and successful oviposition in early host instars.

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Résumé Les effets du parasitisme par P. pequodorum Viereck (Hym.: Aphidiidae), sur la survie, le développement et la reproduction des aptères de A. pisum, ont été examinés au laboratoire. L'étude a porté sur 10 cohortes de pucerons dont les âges différaient de 0,5 heure à 9 jours. A 20°C, les pucerons parasités ont survécu en moyenne pendant 7 jours après avoir été parasités. Leur évolution ultérieure dépend au stade auquel ils ont été parasités: au premier et second stades, ils sont morts avant de devenir adultes; au troisième stade, ils sont morts adultes avant de se reproduire; au quatrième stade et adultes, ils ont donné un nombre variable de descendants. La fécondité totale moyenne a crû comme une fonction exponentielle de l'âge au moment où ils avaient été parasités. L'âge de l'hôte n'a pas modifié significativement la durée de la période entre le moment où ils ont été parasités et la fin de la phase de reproduction. Des équations ont été établies en fonction de l'âge des pucerons au moment où ils ont été parasités, pour estimer la longueur de la période de reproduction, la fécondité totale, le taux intrinsèque d'accroissement et le temps de doublement des cohortes de pucerons. Les résultats sont discutés sous l'angle de la dynamique de la population. On en déduit que l'impact potentiel d'un parasite sur une population de pucerons dépend d'abord de ses préférences pour les premiers stades de l'hôte et du succès de sa ponte sur ces stades.

     

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.