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.     

Expression pattern analysis of odorant‐binding proteins in the pea aphid Acyrthosiphon pisum

Odorant‐binding proteins (OBPs) are soluble proteins mediating chemoreception in insects. In previous research, we investigated the molecular mechanisms adopted by aphids to detect the alarm pheromone (E)‐β‐farnesene and we found that the recognition of this and structurally related molecules is mediated by OBP3 and OBP7. Here, we show the differential expression patterns of 5 selected OBPs (OBP1, OBP3, OBP6, OBP7, OBP8) obtained performing quantitative RT‐PCR and immunolocalization experiments in different body parts of adults and in the 5 developmental instars, including winged and unwinged morphs, of the pea aphid Acyrthosiphon pisum. The results provide an overall picture that allows us to speculate on the relationship between the differential expression of OBPs and their putative function. The expression of OBP3, OBP6, and OBP7 in the antennal sensilla suggests a chemosensory function for these proteins, whereas the constant expression level of OBP8 in all instars could suggest a conserved role. Moreover, OBP1 and OBP3 are also expressed in nonsensory organs. A light and scanning electron microscopy study of sensilla on different body parts of aphid, in particular antennae, legs, mouthparts, and cornicles‐cauda, completes this research providing a guide to facilitate the mapping of OBP expression profiles.     

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.

     

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.     

Seasonal monitoring of Drosophila suzukii and its non-crop hosts in Wuhu,Eastern China

Drosophila suzukii (Matsumura) (Diptera: Drosophilidae) is a serious pest that prefers fresh fruits and is native to Southeast Asia. In our study, apple cider vinegar bait traps were used to capture and monitor the population dynamics of this native pest in Wuhu City, China, from May/June 2017 to May 2018. The research was conducted at 15 locations in two fruit orchards in Wuhu. Traps caught more adults in general in a Meiling blueberry orchard than in a Xicun mixed orchard, and the highest trap counts occurred near harvest time (October). Females had more mature eggs from September to November, and the number of mature eggs declined thereafter. We found several non-crop hosts, which can provide food and reproductive resources for D. suzukii and are common in forests and field margins. By comparing the number of captured adults in the Meiling and Xicun orchards, we found that blueberry was preferred by D. suzukii among the fruits in our search. Fruit ripening times differed among crops; therefore, fly populations moved between crop and non-crop habitats during the year or had varying population dynamics on different crops in different seasons. The D. suzukii population and the number of mature eggs decreased in summer and winter but increased in spring and autumn. Drosophila suzukii had higher survival rates with blueberry than those with other fruits, and D. suzukii could use four non-crop species growing around the orchards as host plants.     

European yellow lupine, Lupinus luteus, and narrow-leaf lupine, Lupinus angustifolius, as hosts for the pea aphid, Acyrthosiphon pisum

The pea aphid, Acyrthosiphon pisum Harris (Homoptera: Aphididae), fed, developed, and reproduced on yellow lupine, Lupinus luteus L. (Fabaceae: Genisteae). No clear preferences for any variety within L. luteus were found. Acyrthosiphon pisum showed negative values of relative growth rate and no aphid completed development on any variety of narrow-leaf lupine Lupinus angustifolius L. Aphids did not ingest phloem sap while probing on L. angustifolius and the probes were very short. All varieties of L. angustifolius were rejected by aphids during an early stage of probing in peripheral tissues, that is, epidermis or mesophyll. There were qualitative and quantitative differences in alkaloid and soluble sugar content between the two lupine species. Within species, the relative content of individual compounds differed among the varieties. Lupinus angustifolius contained four quinolizidine alkaloids (13-hydroxylupanine, dehydrolupanine, lupanine, and angustifoline), while L. luteus contained two (lupanine and sparteine). Lupanine occurred in all varieties of both lupine species. The total content of soluble carbohydrates was similar in L. luteus and L. angustifolius . The following cyclitols were found in both lupine species: myo -inositol, D-ononitol, and D-pinitol. Lupinus angustifolius also contained D- chiro -inositol. The study of aphid probing behaviour, development, and reproduction demonstrated that L. luteus is a suitable host plant for A. pisum while L. angustifolius is not. It is likely that the rejection of L. angustifolius by A. pisum was caused by chemical factors detected by aphids at the epidermis and mesophyll level.     

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.     

Recognition of host-specific chemical stimulants in two sympatric host races of the pea aphid Acyrthosiphon pisum

Abstract.  1. In ecological speciation , adaptation to variation in the external environment provides the crucial push that starts the process of genetic divergence and eventually leads to speciation. This emphasis on the role of ecological specialisation in speciation events has brought with it a renewed interest in its proximate mechanisms in recently diverged groups such as host races. Here, the proximate mechanisms of feeding specialisation are investigated in two host races of the pea aphid Acyrthosiphon pisum .
2. Using alfalfa and clover extracts, enclosed in diet chambers or applied on whole plants, it is shown that feeding specialisation depends on recognition of stimulants specific to the host plant, not on deterrents or toxins specific to the non-host plants.
3. Because pea aphids mate on their host plant, feeding specialisation leads to de facto assortative mating. This study suggests that behavioural recognition of host-specific chemicals, rather than avoidance of deterrents or/and plant toxins, contributes to gene flow restriction between the alfalfa and clover host races.     

Synthesis of proteins and nucleic acids by the pea aphid Acyrthosiphon pisum (aphididae) in the absence of a full complement of dietary amino acids

Protein, nucleic acids, and nucleotide syntheses were studied in pea aphids, Acyrthosiphon pisum (Harris), by feeding them labeled ¹⁴C-amino acids and [5-³H]-orotic acid in sucrose. It was demonstrated that in the absence of dietary essential amino acids, aphids were capable of synthesizing nucleic acids, nucleotides, and proteins when provided with a single dietary amino acid in sucrose. It is suggested that other required amino acids were possibly supplied by the symbionts present in the pea aphid and/or were obtained from the amino acid pool in the hemolymph or glucose, one of the end products of sucrose digestion. Of the various amino acids tested, synthesis of measurable amounts of protein or other compounds occurred when alanine, aspartic acid, glutamic acid, glycine, proline, or serine were provided, but no synthesis occurred with cysteine.     

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.     

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.     

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.     

The significance of gut sucrase activity for osmoregulation in the pea aphid, Acyrthosiphon pisum

The osmotic pressure of the body fluids of aphids is lower than in their diet of plant phloem sap. It is hypothesised that aphids reduce the osmotic pressure of ingested food by sucrase-mediated hydrolysis of dietary sucrose to glucose and fructose, and the polymerisation of glucose into oligosaccharides of low osmotic pressure per hexose unit. To test this hypothesis, the impact of the alpha-glucosidase inhibitor acarbose on the sugar relations and osmoregulation of aphids was explored. Acarbose inhibited sucrase activity in gut homogenates and the production of monosaccharides and oligosaccharides in the honeydew of live aphids. Acarbose caused an increase in the haemolymph osmotic pressure for aphids reared on a diet (containing 0.75 M sucrose) hyperosmotic to the haemolymph and not on the isoosmotic diet containing 0.2 M sucrose. It did not affect aphid feeding rate over 2 days, except at high concentrations on 0.75 M sucrose diet, and this may have been a secondary consequence of osmotic dysfunction. Acarbose-treated aphids died prematurely. With 5 microM dietary acarbose, mean survivorship on 0.2 M sucrose diet was 4.2 days, not significantly different from starved aphids, indicating that, although these aphids fed, they were deprived of utilisable carbon; and on 0.75 M sucrose diet, mean survivorship was just 2.8 days, probably as a consequence of osmotic failure. It is concluded that the aphid gut sucrase activity is essential for osmoregulation of aphids ingesting food hyperosmotic to their body fluids.     

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.     

The secreted salivary proteome of the pea aphid Acyrthosiphon pisum characterised by mass spectrometry

Nine proteins secreted in the saliva of the pea aphid Acyrthosiphon pisum were identified by a proteomics approach using GE‐LC‐MS/MS and LC‐MS/MS, with reference to EST and genomic sequence data for A. pisum. Four proteins were identified by their sequences: a homolog of angiotensin‐converting enzyme (an M2 metalloprotease), an M1 zinc‐dependant metalloprotease, a glucose‐methanol‐choline (GMC)‐oxidoreductase and a homolog to regucalcin (also known as senescence marker protein 30). The other five proteins are not homologous to any previously described sequence and included an abundant salivary protein (represented by ACYPI009881), with a predicted length of 1161 amino acids and high serine, tyrosine and cysteine content. A. pisum feeds on plant phloem sap and the metalloproteases and regucalcin (a putative calcium‐binding protein) are predicted determinants of sustained feeding, by inactivation of plant protein defences and inhibition of calcium‐mediated occlusion of phloem sieve elements, respectively. The amino acid composition of ACYPI009881 suggests a role in the aphid salivary sheath that protects the aphid mouthparts from plant defences, and the oxidoreductase may promote gelling of the sheath protein or mediate oxidative detoxification of plant allelochemicals. Further salivary proteins are expected to be identified as more sensitive MS technologies are developed.     

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.     

The photoperiodic responses and phenology of an English strain of the pea aphid Acyrthosiphon pisum

Abstract. 1. The response curves for the photoperiodic induction of the sexual forms (oviparae and males) differ significantly in an English clone of the pea aphid Acyrthosiphon pisum Harris. Male production is sharply peaked. The late summer scotophases (dark periods) that initiate male production are 1.0–1.5 h shorter than those that initiate ovipara production; the induction of males, but not oviparae, virtually ceases at scotophases longer than 12 h. This disparity suggests that there are two photoperiodic clocks.
2. All ovipara-producing aphids switch spontaneously to virginopara production part-way through the progeny sequence, irrespective of photoperiod. This may confer the ability to overwinter partheno-genetically under favourable climatic conditions. Since males are always born last in the progeny sequence this possibility would be pre-empted unless the terminal male sequences were suppressed by long winter scotophases.
3. The role of the photoperiodic response in determining the seasonal phenology was tested in the field by exposing a succession of laboratory-reared aphids to natural photoperiods, in late summer and autumn. Adult males and oviparae developed synchronously in early October, the difference in critical scotophase compensating for the late appearance of males in the progeny sequence. A large part of the 'civil twilight' is photoperiodically active.
4. Clones propagated outdoors by isolating aphids in each generation from the terminal sequence of virginoparae did not survive the whole winter of 1985/86. The chances of survival were reduced by severe weather and by the inability of the photoperiodic system to 'turn off' the production of early born oviparae which are, therefore, 'wasted'.     

Identification and functional characterization of an odorant receptor in pea aphid,Acyrthosiphon pisum

The sensitive olfactory system is necessary for survival of insects.Odorant receptors (ORs)are located on the dendrites of olfactory receptor neurons and play a critical role in odor detection.Insect ORs are functionally analyzed via heterologous expression in a Xenopus oocyte system using a two-electrode voltage-clamp (TEVC)electrophysiological recording.Here,we have identified a novel OR in the pea aphid,Acyrthosiphon pisum,then we cloned and named it ApisOR4.We analyzed the ApisOR4 tissue expression patterns and found expression only in antennae tissues.Further functional analysis using TEVC revealed that ApisOR4 is broadly tuned to eight volatiles,which elicit electrophysiological response in pea aphid antennae.This study provides an initial functional analysis of aphid ORs and identifies candidate volatiles to be used in developing new strategies for aphid control.