Effect of entomopathogenic Aspergillus strains against the pea aphid,Acyrthosiphon pisum (Hemiptera: Aphididae)

Aphids (Homoptera: Aphididae) are sap-sucking insect pests that feed on several plants of agronomical importance. Entomopathogenic fungi are valuable tools for potential aphid control. As part of a selection process, laboratory bioassays were carried with five different concentrations of Aspergillus clavatus (Desmazières), Aspergillus flavus (Link) and Metarhizium anisopliae ((Metschnikoff) Sorokin) spores against the pea aphid, Acyrthosiphon pisum (Harris). Aspergillus isolates induced higher mortalities than M. anisopliae, which is a well-known entomopathogen in the literature. Lethal concentrations (LC₅₀ and LC₉₀) were 1.23 × 10³ and 1.34 × 10⁷ spores/ml for A. flavus, 4.95 × 10² and 5.65 × 10⁷ spores/ml for A. clavatus, and 3.67 × 10³ and 9.71 × 10⁷ spores/ml for M. anisopliae 5 days after treatment. Mycelia development and sporulation on adult cadavers were observed 48 h after incubation. The intrinsic growth rate of A. pisum decreased with increased spore concentration for all fungal strains, suggesting an increase in pathogen fitness related to a consumption of host resources. In conclusion, Aspergillus species could be useful in aphid control as pest control agents despite their saprophytic lifestyle. This is also to our knowledge the first report of A. clavatus and A. flavus strains pathogenic to aphids.     

Influence of temperature on pea aphid Acyrthosiphon pisum (Hemiptera: Aphididae) resistance to natural enemy attack

The ability to resist or avoid natural enemy attack is a critically important insect life history trait, yet little is understood of how these traits may be affected by temperature. This study investigated how different genotypes of the pea aphid Acyrthosiphon pisum Harris, a pest of leguminous crops, varied in resistance to three different natural enemies (a fungal pathogen, two species of parasitoid wasp and a coccinellid beetle), and whether expression of resistance was influenced by temperature. Substantial clonal variation in resistance to the three natural enemies was found. Temperature influenced the number of aphids succumbing to the fungal pathogen Erynia neoaphidis Remaudière & Hennebert, with resistance increasing at higher temperatures (18 vs. 28 degrees C). A temperature difference of 5 degrees C (18 vs. 23 degrees C) did not affect the ability of A. pisum to resist attack by the parasitoids Aphidius ervi Haliday and A. eadyi Stary, González & Hall. Escape behaviour from foraging coccinellid beetles (Hippodamia convergens Guerin-Meneville) was not directly influenced by aphid clone or temperature (16 vs. 21 degrees C). However, there were significant interactions between clone and temperature (while most clones did not respond to temperature, one was less likely to escape at 16 degrees C), and between aphid clone and ladybird presence (some clones showed greater changes in escape behaviour in response to the presence of foraging coccinellids than others). Therefore, while larger temperature differences may alter interactions between Acyrthosiphon pisum and an entomopathogen, there is little evidence to suggest that smaller changes in temperature will alter pea aphid-natural enemy interactions.     

Oral activity of FMRFamide-related peptides on the pea aphid Acyrthosiphon pisum (Hemiptera: Aphididae) and degradation by enzymes from the aphid gut

Insect myosuppressins and myosuppressin analogues were tested for oral toxicity against the pea aphid Acyrthosiphon pisum (Harris) by incorporation into an artificial diet. Acyrthosiphon pisum myosuppressin (Acypi-MS) and leucomyosuppressin (LMS) had significant dose-dependent effects (0.1-0.5μg peptide/μl diet) on feeding suppression, mortality, reduced growth and fecundity compared with control insects, but Acypi-MS was more potent than LMS. One hundred percent of aphids had died after 10days of feeding on 0.5μg Acypi-MS/μl diet whereas 40% of aphids feeding on 0.5μg LMS/μl diet were still alive after 13days. Myosuppressins were degraded by aphid gut enzymes; degradation was most likely due to a carboxypeptidase-like protease, an aminopeptidase and a cathepsin L cysteine protease. The estimated half-life of Acypi-MS in a gut extract was 30min, whereas LMS was degraded more slowly (t?=54min). No toxicity was observed when the analogues δR(9) LMS and citrolline(9) Acypi-MS or FMRFamide were fed to the pea aphid. These findings not only help to better understand the biological effects of myosuppressins in aphids but also demonstrate the potential use of myosuppressins in a strategy to control aphid pests.     

Bacteria from the alimentary canal of the pea aphid,Acyrthosiphon pisum (Harr.) (Homoptera,Aphididae)

Bacteria isolated from the alimentary canal of the pea aphid, Acyrthosiphon pisum, were identified mainly as species of the genera Sarcina, Micrococcus, Achromobacter, and Flavobacterium. Starch was rapidly hydrolysed by all species of Sarcina and Flavobacterium and by most species of Achromobacter and slowly by some species of Micrococcus. All of the types of bacteria found in the alimentary tract of the pea aphid were represented in the bacterial flora on the leaves of the pea plants on which the aphids were reared. It is suggested that these organisms become established in the gut after being picked up from the leaf surface while the aphid is probing for the phloem bundles.     

Clones of pea aphid, Acyrthosiphon pisum (Hemiptera: aphididae) distinguished using genetic markers, differ in their damaging effect on a resistant alfalfa cultivar

CUF 101, a resistant cultivar of alfalfa, was exposed to 15 clones of Acyrthosiphon pisum Harris collected from alfalfa fields in three regions of France (east, south, central west) to determine whether the level of resistance varied across the different clones. The survival of alfalfa seedlings infested at the cotyledon stage was assessed using a standardized method. Although no difference in seedling mortality was detected between clones grouped by region, there was a significant variation among the 15 pea aphid clones. In particular, two clones of southern origin were more aggressive. In addition, the different pea aphid clones were characterized using allozyme and RAPD-PCR markers. Among the 15 clones, seven allozyme genotypes (plus one when adding colour polymorphism) and 12 RAPD-PCR genotypes were distinguished. The two southern clones differing by their aggressiveness on the resistant alfalfa belonged to the same allozyme and RAPD genotype which was distinct from the other pea aphid clones. Our results reinforce the need to take into account aphid genetic diversity in breeding programmes for resistance in cultivated plants.     

Spiroplasma symbiont of the pea aphid, Acyrthosiphon pisum (Insecta: Homoptera)

From a laboratory strain of the pea aphid, Acyrthosiphon pisum, we discovered a previously unknown facultative endosymbiotic bacterium. Molecular phylogenetic analysis based on 16S ribosomal DNA revealed that the bacterium is a member of the genus Spiroplasma. The Spiroplasma organism showed stable vertical transmission through successive generations of the host. Injection of hemolymph from infected insects into uninfected insects established a stable infection in the recipients. The Spiroplasma symbiont exhibited negative effects on growth, reproduction, and longevity of the host, particularly in older adults. Of 58 clonal strains of A. pisum established from natural populations in central Japan, 4 strains possessed the Spiroplasma organism.     

Time-specific life tables for the pea aphid,Acyrthosiphon pisum (Harris), on alfalfa

Time-specific life tables were constructed for three pea aphid, Acyrthosiphon pisum (Harris ) (Homoptera: Aphididae), populations using a modification ofHughes' analytical procedure. All populations were studied on second-growth alfalfa (mid-June to mid-July) in south central Wisconsin; data for two populations were collected during 1980, and data for the third population were collected during 1982. The intrinsic rate of increase (r_m) estimated on a physiological time (day-degree) scale under field conditions but in the absence of natural enemies, provided a reliable estimate of potential population growth rate and was used in preference toHughes' approach of estimating potential population growth rates directly from stage structure data. Emigration by adult alatae and fungal disease were the major sources of A. pisum mortality in each of the three populations studied. These factors were most important because of their impact on reducing birth rates within the local population. Parasitism was never greater than 9 percent. Mortality attributable to predation ranged from 0.0 to about 30.0%; however, even at the highest predator densities A. pisum populations increased exponentially.     

Intracellular symbiotes of the pea aphid,Acyrthosiphon pisum

Histological and ultrastructural studies on the mycetome of the pea aphid, Acyrthosiphon pisum, disclosed two types of symbiotes. The more common primary symbiotes were oval in shape and were found in large mycetocytes making up the bulk of the mycetome. The secondary symbiotes were smaller, rod-shaped, and were restricted to an apparently syncytial sheath partially enclosing the primary mycetocytes. Extensive rough endoplasmic reticulum and Golgi occurred in the sheath but not in the primary mycetocytes. Lysosomal breakdown occurred in both primary and secondary symbiotes but the two processes differed markedly. In the primary mycetocytes, a small number of symbiotes were broken down individually to form small, compact residual bodies. In the sheath, breakdown of secondary symbiotes was more extensive: large numbers were broken down within cytolysomes.     

Evaluation of four co-occurring ladybirds for use as biocontrol agents of the pea aphid,Acyrthosiphon pisum (Homoptera: Aphididae)

The potential of four co-occurring ladybirds, Coccinella septempunctata (L.), Coccinella transversalis (F.), Cheilomenes sexmaculata (F.), and Propylea dissecta (Mulsant), as predators of the pea aphid, Acyrthosiphon pisum (Harris) was investigated. Larger ladybirds were more voracious while smaller ones had higher consumption indices and were more efficient converters of food.     

Lipid composition of the pea aphid,Acyrthosiphon pisum (Harris) (homoptera: Aphididae), reared on host plant and on artificial media

Polar and neutral lipids and their constitutive fatty acids were quantified in the pea aphid, Acyrthosiphon pisum (Harris), grown on host plant or on a lipid free artificial diet. The results were compared to determine if lipids were involved in the suitability of the diet for continuous rearing of this A. pisum biotype. For apterous adults grown on plants, the lipids were characterized by a low amount of neutral lipids (2.5% weight/fresh weight) almost entirely (96.4%) composed of hexanoyl and sorboyl dimyristin. These storage lipids were higher in the alatae (3.8%), probably correlated with potential flight activity. The phospholipid amounts were identical in these two morphs (1.3–1.4% weight/fresh weight), comprised mainly of phosphatidylethanolamines (52%) and phosphatidylcholines (40.6%). These phospholipids contained a still unidentified fatty acid, with a retention time close to that of linolenic acid and synthesized by the aphid or its bacterial symbionts (not found in plants). The apterous adult aphids reared on an artificial diet showed an accumulation of neutral lipids (8.9% for the first generation); this increase was shown to be slightly greater for the hexanoyl and sorboyl triglycerides. In contrast, the phospholipids decreased in aphids reared on an artificial diet (1.1% and 0.9%, respectively, for first and second generation), correlated with a phospholipid fatty acid profile significantly deficient in C18:3 and in the unidentified peculiar fatty acid. These phospholipids are essential components of biological membranes and a diet-driven deficient synthesis in some of their components may result in the observed symptoms. © 1992 Wiley-Liss, Inc.     

Application of DL-beta-aminobutyric acid (BABA) as a root drench to legumes inhibits the growth and reproduction of the pea aphid Acyrthosiphon pisum (Hemiptera: Aphididae)

DL-beta-aminobutyric acid (BABA) is a non-protein amino acid that is an effective inducer of resistance against a variety of plant pathogens. However, examples of BABA-induced resistance against insect herbivores have not been reported. We applied BABA as a soil drench to legumes and monitored its effects on the pea aphid Acyrthosiphon pisum (Harris). On tic bean (Vicia faba var. minor), BABA increased aphid mortality, caused a reduction in the mean relative growth rate of individual insects and lessened the intrinsic rate of population increase (rm). BABA also caused significant reductions in the growth rate of A. pisum on pea (Pisum sativa), broad bean (Vicia faba var. major), runner bean (Phaseolus coccineus), red clover (Trifolium pratense) and alfalfa (Medicago sativa). No direct toxic effects of BABA against A. pisum were found, and no phytotoxic effects that may have caused a reduction in aphid performance were detected. Possible mechanisms behind this BABA-induced inhibition of aphid performance are discussed.     

Sex versus parthenogenesis: a transcriptomic approach of photoperiod response in the model aphid Acyrthosiphon pisum (Hemiptera: Aphididae)

Most aphids develop a cyclic parthenogenesis life-cycle. After several generations of viviparous parthenogenetic females, it follows a single annual generation of sexual individuals, usually in autumn, that mate and lay the sexual eggs. Shortening of photoperiod at the end of the summer is a key factor inducing the sexual response. With the survey here reported we aimed at identifying a collection of candidate genes to participate at some point in the cascade of events that lead to the sexual phenotypes. Following a suppression subtractive hybridization methodology (SSH) on the model aphid Acyrthosiphon pisum, we built and characterised two reciprocal cDNA libraries (SDU and SDD) enriched respectively in genes up-regulated or down-regulated by short photoperiod conditions that lead to the sexual response in this aphid species. A total of 557 ESTs were obtained altogether representing 223 non-overlapping contigs. 29% of these were new sequences not present in previous aphid EST libraries. BLAST searches allowed putative identification of about 54% of the contigs present in both libraries. Relative quantification of expression through real-time quantitative PCR demonstrated the differential expression in relation with the photoperiod of 6 genes (3 up-regulated and 3 down-regulated by shortening the day length). Among these, expression of a tubulin gene, two cuticular proteins and a yet unidentified sequence along the day-night cycle was further investigated. Implications for current studies on gene regulation of the dichotomy sex vs. parthenogenesis in aphids are discussed.     

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.

     

Characterisation of immune responses in the pea aphid, Acyrthosiphon pisum

The innate immune system of insects provides effective defence against a range of parasites and pathogens. The pea aphid, Acyrthosiphon pisum, is a novel study system for investigating host-parasite interactions due to its complex associations with both well-characterised bacterial symbionts and a diversity of pathogens and parasites, including several important biological control agents. However, little is known about the cellular and humoral immune responses of aphids. Here we identify three morphologically distinct types of haemocytes in circulation that we name prohemocytes, granulocytes and oenocytoids. Granulocytes avidly phagocytose Gram negative Escherechia coli and Gram positive Micrococcus luteus while oenocytoids exhibit melanotic activity. Prohaemocytes increase in abundance immediately following an immune challenge, irrespective of the source of stimulus. Pea aphids form melanotic capsules around Sephadex beads but do not form cellular capsules. We also did not detect any antimicrobial peptide activity in the haemolymph using zone of inhibition assays. We discuss these results in relation to recent findings from the pea aphid genome annotation project that suggest that aphids have a reduced immune gene repertoire compared to other insects.     

The host plant range of the pea aphid subspecies Acyrthosiphon pisum ssp. destructor (Johnson) (Hom., Aphididae)

Abstract:  In order to establish the host range of the pea aphid subspecies, Acyrthosiphon pisum ssp. destructor , and hence from which plant species pea crops are likely to become infested, the performance of this aphid on different leguminous plants was assessed. The plant species used were: Lotus uliginosus , Medicago sativa , Melilotus officinalis , Ononis repens , Sarothamnus scoparius , Trifolium hybridum , Trifolium pratense , Trifolium repens , Vicia cracca and Vicia faba . Vicia faba and Trifolium hybridum were the plants on which aphids reached the greatest size, took the least time to reach maturity, and experienced the lowest mortality. The time taken for the aphids to develop to maturity was negatively correlated with adult size, whereas survival to maturity was positively correlated with adult size. The host preference of the aphids was also assessed. The plant species selected as hosts by alatae were those on which their offspring performed best.     

Rapid response to intraclonal selection in the pea aphid (Acyrthosiphon pisum)

Summary Pea aphids show intraclonal variability in antipredator behaviour. Among the offspring of a single parthenogenetically reproducing female, some individuals drop from the plant in response to alarm pheromone while others remain on the plant. We demonstrate that this intraclonal behavioural variability can be altered by selection. The proportion of aphids dropping in response to alarm pheromone was significantly greater in lines in which this behaviour was selected than in clonally identical lines in which the opposite phenotype was favoured. This change occurred within one generation and could not be attributed to grand-maternal effects, nor to environmental effects. These results demonstrate the ability of clonal aphids to adapt to changes in the environment within a single generation.