Isolation of the primary intracellular symbiote of the pea aphid, Acyrthosiphon pisum

A method for isolating the primary intracellular symbiote of the pea aphid, Acyrthosiphon pisum, is presented. The purity of the preparation, based on an area symbiote to area contaminant and a number to number basis, averages 93%.     

Sexual morph determination in the aphid, Acyrthosiphon pisum

The effects of two temperatures and a series of photoperiods on sexual morph determination in an aphid are examined. Photoperiodic response curves are presented and analysed. Female morph determination and male production were influenced by both these environmental factors. Sexuals were produced only at shorter photoperiods and the critical photoperiods for sexual production were shorter at the higher temperature. The critical photoperiod for male production differed from that for female morph determination and was more sensitive to temperature. These results are discussed in relation to the ‘maternal switching mechanism’ and sex determination. The significance of the temperature effect and also the variation in photoperiodic response are discussed.     

Phase contrast and electron microscopy of the mycetocytes and symbiotes of the pea aphid, Acyrthosiphon pisum

Phase contrast, scanning electron, and transmission electron microscopy of the symbiotes of Acyrthosiphon pisum was undertaken. Some staining properties of the symbiotes were also studied.The symbiotes of the pea aphid were found to be coccoid bodies 2 to 5 μ in diameter, gram negative, stained slightly by Fuelgen's, and stained blue by Machiavello's. The symbiotes appear to be surrounded by three membranes. Ribosomes may occur within the cytoplasm of the symbiotes. The cytoplasm of the mycetocytes contains large numbers of mitochondria, endoplasmic reticulum, ribosomes, and a large nucleus, and nucleolus.A discussion of the classification of the symbiotes is also presented.     

The reproductive sequence and sex determination in the aphid, Acyrthosiphon pisum

The effects of photoperiod and temperature on birth-rates and the pattern of female and male production in the aphid A. pisum are described. Females only are produced first and are followed, after a brief reproductive pause, by the production of males. The birth-rate of males is lower than that of females. The onset of male production is affected by photoperiod over a narrow range near the critical photoperiod. The characteristics of the reproductive sequence are discussed in relation to them echanism of sex determination.     

Biostable multi-Aib analogs of tachykinin-related peptides demonstrate potent oral aphicidal activity in the pea aphid Acyrthosiphon pisum (Hemiptera: Aphidae)

The tachykinin-related peptides (TRPs) are multifunctional neuropeptides found in a variety of arthropod species, including the pea aphid Acyrthosiphon pisum (Hemiptera: Aphidae). Two new biostable TRP analogs containing multiple, sterically hindered Aib residues were synthesized and found to exhibit significantly enhanced resistance to hydrolysis by angiotensin converting enzyme and neprilysin, membrane-bound enzymes that degrade and inactivate natural TRPs. The two biostable analogs were also found to retain significant myostimulatory activity in an isolated cockroach hindgut preparation, the bioassay used to isolate and identify the first members of the TRP family. Indeed one of the analogs (Leuma-TRP-Aib-1) matched the potency and efficacy of the natural, parent TRP peptide in this myotropic bioassay. The two biostable TRP analogs were further fed in solutions of artificial diet to the pea aphid over a period of 3 days and evaluated for antifeedant and aphicidal activity and compared with the effect of treatment with three natural, unmodified TRPs. The two biostable multi-Aib TRP analogs were observed to elicit aphicidal effects within the first 24 h. In contrast natural, unmodified TRPs, including two that are native to the pea aphid, demonstrated little or no activity. The most active analog, double-Aib analog Leuma-TRP-Aib-1 (pEA[Aib]SGFL[Aib]VR-NH₂), featured aphicidal activity calculated at an LC₅₀ of 0.0083 nmol/μl (0.0087 μg/μl) and an LT₅₀ of 1.4 days, matching or exceeding the potency of commercially available aphicides. The mechanism of this activity has yet to be established. The aphicidal activity of the biostable TRP analogs may result from disruption of digestive processes by interfering with gut motility patterns and/or with fluid cycling in the gut; processes shown to be regulated by the TRPs in other insects. These active TRP analogs and/or second generation analogs offer potential as environmentally friendly pest aphid control agents.     

Maintenance of the primary symbiote of the pea aphid Acyrthosiphon pisum in liquid media

The primary symbiote of the pea aphid was maintained in liquid culture media for 12 hr for the purpose of optimizing certain culture parameters. The optimum temperature for protein and nucleic acid synthesis was found to be 28–30°C. There was no dependence of this microorganism on fetal calf serum added to the growth medium. Grace's insect tissue culture medium and Grace's insect tissue culture medium (modified by Tokumitsu and Maramorosch) were found to be best suited for the maintenance of this organism.The suitability of the growth medium for the microorganism was directly related to the age of the culture medium.     

The elimination of intracellular microorganisms from insects: an analysis of antibiotic-treatment in the pea aphid (Acyrthosiphon pisum)

Antibiotics are routinely used to eliminate intracellular prokaryotic microorganisms from a wide range of insect species, but concerns about deleterious effects of antibiotic therapy on the insect host are seldom addressed. Here, the impact of antibiotic therapy in the symbiosis between the pea aphid Acyrthosiphon pisum and bacteria of the genus Buchnera is reviewed. Antibiotic-treatment produces aposymbiotic (i.e. symbiont-free) aphids, but does not depress the mitochondrial complement, the assimilation of dietary amino acids or the incorporation of amino acids into protein in these insects and does not impair osmoregulation, feeding rate and the capacity to penetrate plant tissues. It is concluded that the general malaise associated with aposymbiotic aphids is not attributable to a direct effect of the antibiotic. However, an important implication of this study is that aposymbiotic insects exhibit substantial metabolic adjustments to loss of the symbiosis; they are not simply aphids from which the symbiotic bacteria have been removed.     

Aggregation behavior of pea aphids, Acyrthosiphon pisum

First and second instar pea aphid nymphs were allowed to respond in multiple choice test chambers having five parafilm feeding stations, each of which contained 20% sucrose solution. Under light conditions, the aphids assumed a negative bionomial distribution. This same phenomenon occurred in darkness, but with a lesser degree of aggregation. The ability to see other aphids was not a prerequisite to aggregate formation. The formation of aggregates on certain stations was not enhanced by aphid aggregations that had previously occurred but were removed from those stations. Aggregate formation was not in response to a humidity gradient.

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Zusammenfassung Erbsenblattläuse im ersten und zweiten Larvenstadium wurden in Testkammern gehalten, in denen sie nach Belieben fünf mit Parafilm überzogene Futterplätze besuchen durften. Jeder Futterplatz enthielt eine 20% ige Rohrzuckerlösung. Nach zwei Stunden wurde die Zahl der Blattläuse auf jeder Membran festgestellt und ein Häufigkeitsdiagramm angelegt. Unter Lichtbedingungen war die Verteilung der Blattläuse negativ binomisch. Die gleiche Erscheinung zeigte sich im Dunklen. Die Blattläuse traten jedoch unter Lichtbedingungen dichter gehäuft auf. Diese Fähigkeit, Ansammlungen zu bilden, lässt eine Art Verständigung vermuten. Der Sehfaktor ist für diese Verständigung nicht verantwortlich. Die Bildung von Ansammlungen auf mehreren Futterplätzen wurde nicht gefördert von früheren Ansammlungen von Aphiden, die zuvor von diesen Plätzen entfernt waren. Die Anhäufungen standen nicht in Beziehung zum Feuchtigkeitsgehalt.

     

光周期对两种色型豌豆蚜种群参数的影响

为探讨光周期对红绿两种色型豌豆蚜种群增长的影响,于室内模拟光周期22L∶2D、16L∶8D、10L∶14D和4L∶20D四个水平,研究了红绿两种色型豌豆蚜的发育、繁殖和生命表。结果表明:相对于其它光周期,在光周期22L∶2D时,红绿两种色型豌豆蚜若蚜存活率最高,繁殖力最强;其红色型豌豆蚜种群增长内禀增长率(r_m=0.3263)高于绿色型豌豆(r_m=0.2863)。光周期16L∶8D时,红色型豌豆蚜的若蚜存活率(72.22%)高于绿色型豌豆蚜(69.45%),而成蚜寿命(6.58d)较短;但绿色型豌豆蚜内禀增长率(r_m=0.2648)高于红色型豌豆蚜(r_m=0.2249),其种群繁殖力强。光周期10L∶14D时,红色型豌豆蚜与绿色型豌豆蚜相比,若蚜存活率(80.55%)高,内禀增长率(r_m=0.2490)高,种群增长力强,而绿色型豌豆蚜成蚜寿命(12.71d)较长。光周期4L∶20D时,两种色型豌豆蚜若蚜存活率最低,繁殖力最弱,成蚜寿命显著延长;红色型豌豆蚜若蚜存活率(64.67%)高于绿色型(35.50%),成蚜没有产生后代;绿色型豌豆蚜内禀增长率(r_m=-0.0366),其种群为负增长。综上,短光照条件(4L∶20D)延长了两种色型豌豆蚜的若蚜发育历期及成蚜寿命,降低了若蚜存活率、繁殖力及生命表各项参数;长光照条件(22L∶2D)缩短了两种色型豌豆蚜的若蚜发育历期及成蚜寿命,若蚜存活率高、繁殖力强;红色型豌豆蚜更适宜光周期10L∶14D,而绿色型豌豆蚜更适宜光周期16L∶8D。     

High variation in host adaptation among clones of the pea aphid,Acyrthosiphon pisum on peas,Pisum sativum

The performance of one clone of the pea aphid,Acyrthosiphon pisum (Harris), was assessed on 37 different cultivars and species ofPisum L. In addition, random samples of 36 pea aphid clones collected on alfalfa and clover were tested on a selection of fivePisum sativum L. cultivars. Aphid performance was evaluated in terms of the mean relative growth rate (MRGR) during the first five days of life or other life history variables. The MRGR of the first-mentioned pea aphid clone differed little between cultivars. No significant differences in MRGR were found between wild and cultivatedPisum species or between modern and oldP. sativum cultivars. There was considerable variation in host adaptation among the 36 pea aphid clones within each sampled field. The pea aphid clones showed no consistent pattern in performance on four of the five pea cultivars i.e. there was a significant pea aphid genotype —pea genotype interaction. On one of the cultivars all clones performed well. Pea aphid clones collected from red clover generally performed relatively poorly on pea cultivars, in contrast to the pea aphid clones collected on alfalfa. There was no difference in performance between the two pea aphid colour forms tested. Possible reasons for the high variation and the observed adaptation patterns are discussed. The fact that all clones were collected in two adjacent fields indicates thatA. pisum shows high local intraspecific variability in terms of host adaptation.     

Distribution in time and space of resistance to the pathogenic fungus Erynia neoaphidis in the pea aphid Acyrthosiphon pisum

Two biotypes of pea aphid, Acyrthosiphon pisum, have been recognized in Australia, by their susceptibility (S) or resistance (R) to certain isolates of the fungal pathogen Erynia neoaphidis. The responses of these two biotypes to 40 isolates of the fungus have shown that the R form is largely confined to two States, New South Wales and Victoria, but appears to have recently spread into Queensland and Tasmania. There is no evidence to suggest it occurs outside Australia. Sequential sampling of two field populations of pea aphids during 1981 and 1982 showed that the proportion of R form remained stable at 10.7±3.0 and 14.6±2.6% (mean±standard error) for the two populations. Glasshouse competition experiments run at the comparatively high temperature of 25°C resulted in the R form becoming dominant even when the initial ratio was 4:1 in favour of the S form. The ecological and genetical implications are discussed.

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Résumé Deux types biologiques d'Acyrthosiphon pisum ont été définis, en Australie, suivant leur sensibilité (S) ou leur résistance (R) à certains isolats du champignon pathogène Erynia neoaphidis. Les réponses des deux biotypes à 40 isolats du champignon ont montré que la forme R est essentiellement cantonnée à deux états: Nouvelle Galle due Sud et Victoria, mais a récemment gagné le Queensland et la Tasmanie. Aucun élément ne fait dire qu'elle existe hors d'Australie. Des échantillonnages séquentiels des deux populations de pucerons dans la nature en 1981 et 1982 ont montré que la proportion de la forme R est restée stable à 10,7 et 14,62 dans les deux populations. Des expériences de compétition en serre à la température relativement haute de 25°C ont rendu la forme R dominante, même quand le rapport initial était de 4/1 en faveur de la forme S. La discussion porte sur les conséquences écologiques et génétiques.

     

醉马草水浸液对豌豆蚜触杀活性及种群增长的影响

为探讨醉马草水浸液对豌豆蚜触杀活性及种群增长的影响,采用带虫浸叶法比较不同生育期醉马草带菌（E+）和不带菌（E-）水浸液对豌豆蚜触杀活性及种群生命表,测定了豌豆蚜的死亡率及触杀后对其生殖期,平均繁殖力,繁殖率及生命表参数的影响。结果表明,不同生育期醉马草带菌（E+）水浸液触杀豌豆蚜后对其各项指标均有显著影响。在苗期时,E+水浸液触杀豌豆蚜后校正死亡率最高,繁殖力最低,内禀增长率（r_m=0.145 d^-1）和净生殖率（R₀=4.802头）均为最小值。在成熟期时,E+水浸液触杀豌豆蚜24 h、48 h和72 h后校正死亡率分别为26.15%,19.01%,9.07%;繁殖期（3.87 d）,平均繁殖力（8.80头）,繁殖率（1.40%）,内禀增长率（r_m=0.208 d^-1）和净生殖率（R₀=8.820头）。在枯黄期时,E+水浸液触杀豌豆蚜后校正死亡率最低,繁殖力最强,内禀增长率（r_m=0.247 d^-1）和净生殖率（R₀=13.647头）均为最大值。不同生育期醉马草E-水浸液触杀豌豆蚜后对其种群繁殖无显著影响,与对照差异不显著（P>0.05）。综上,苗期醉马草E+水浸液对豌豆蚜有较好的触杀效果,校正死亡率高,且触杀后当代繁殖力减弱,种群扩建时间延长,不利于其种群繁殖和增长;故苗期醉马草E+水浸液具有很好的杀虫潜力,所采用水浸液方法制备简单,成本低,可为新型植物源农药研发提供重要理论依据。     

The susceptibility of the pea aphid to intrahemocoelic infection by Serratia marcescens

The bacterium Serratia marcescens is pathogenic for the pea aphid, Acyrthosiphon pisum. When S. marcescens cells are inoculated into the hemocoel of the aphid, there is a variable lag period of 2–3 hr duration. This phase is followed by a rapid increase in pathogen numbers causing a general septicemia and the death of the aphid. The LD₅₀ for fourth-instar pea aphid nymphs was estimated as 190 viable S. marcescens cells (95% confidence limits, 123–288). It is suggested that being sucking insects aphids are protected against infection by bacterial pathogens in the field.     

Feeding and excretion by the pea aphid, Acyrthosiphon pisum (Harr.) (Homoptera: Aphididae), reared on different varieties of peas

The amount of honeydew excreted by the pea aphid, Acyrthosiphon pisum (Harr.), reared singly and in groups on susceptible resistant varieties of peas, Pisum sativum L., grown under greenhouse conditions, was measured. Excretion by isolated aphids was interrupted at moulting for mean periods of 12 and 16 hr, and between moults and in the adult stage for mean periods of 3 to 15 hr, although in many cases the aphids remained in the feeding position. Droplet volume, frequency, and rate of excretion were generally proportional to the susceptibility of the host variety on which the aphids were feeding. The rate of feeding on susceptible varieties was higher and contributed therefore to the faster rate of aphid growth; a greater proportion of the material ingested was excreted and a lesser proportion assimilated than on resistant varieties. Honeydew from aphids feeding on susceptible varieties usually contained a slightly higher concentration of free amino acids and amides.

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Résumê La production de miellat par le puceron du pois, Acyrthosiphon pisum (Harr.), élevé isolément et en groupes a été mesurée sur des variétés sensibles et résistantes de pois, Pisum sativum L., cultivées en serre. La production de miellat par les pucerons isolés fut interrompue au moment de la mue durant les périodes moyennes de 12 et 16 hr; elle fut interrompue entre les mues et ausi au stade adulte durant des périodes moyennes de 3 á 15 hr, même lorsque les aphides en observation paraissaient se nourrir, le bout de leur rostre étant placé sur les nervures des feuilles. Le volume moyen des gouttelettes, la fréquence et le taux d'excrétion furent en général proportionnels au degré de sensibilité de la variétéhôte. Le taux d'ingestion de ces insectes fut plus élevé lorsqu'ils se nourrissaient sur les variétés sensibles, contribuant ainsi à accélérer leur croissance. Une plus grande proportion de la matière ingérée fut excrétée et une proportion moindre assimilée sur les plantes sensibles. Le miellat des pucerons se nourrissant sur les variétés sensibles avait une teneur légèrement plus élevée en amides et acides aminés.

     

Population level effects of cadmium and the insecticide imidacloprid to the parasitoid, Aphidius ervi after exposure through its host, the pea aphid, Acyrthosiphon pisum (Harris)

The objective of this study was to assess the influence of toxic substances with different modes of action on a two-species system: an aphid-specific parasitoid, Aphidius ervi Haliday, feeding on the pea aphid, Acyrthosiphon pisum (Harris). The instantaneous rate of population increase (r_i) was used as a measure of population level toxic effect in this study. The toxicants evaluated were imidacloprid, a nonpersistent neurotoxic insecticide, and cadmium, a chronic pollutant with a tendency to accumulate. We evaluated the effects of cadmium and imidacloprid on A. pisum and A. ervi because both toxicants can occur together in polluted areas where crops are grown. Cadmium (200 or 400 mg kg⁻¹ dry weight soil) and imidacloprid (4 or 40 g a.i. ha⁻¹) were applied to soil contained in plastic pots in which broad bean plants, Vicia faba L., were grown. Results of this study indicated that cadmium at the concentrations tested, reduced population growth rate of the pea aphid. Imidacloprid also reduced aphid growth rate, but only at the highest concentration tested (40 g a.i. ha⁻¹). Combinations of cadmium and imidacloprid had the greatest impact on aphid growth rate. Imidacloprid alone had no effect on population growth rate of the parasitoid. However, cadmium alone or in combination with imidacloprid had a negative impact on A. ervi by reducing population growth rate 77%. These results indicate that negative impacts on parasitoids may occur in areas where cadmium contamination is present and imidacloprid is used to control aphids.     

Toxicity of lectins and processing of ingested proteins in the pea aphidAcyrthosiphon pisum

Acute toxicity of thirty lectins was tested against the pea aphidAcyrthosiphon pisum (Harris) (Homoptera, Aphididae: Macrosiphini). Activity was measured on artificial diets containing moderate concentrations of lectins (10–250 μg/ml) by scoring mortality and growth inhibition over the whole nymphal period (7 days at 20°C). Most of the proteins tested exhibited low toxicity, but some induced significant mortality; these included the lectins from jackbean (Concanavalin A), amaranth, lentil and snowdrop. There was no direct correlation between toxicity and sugar specificity of the lectin; however, many mannose-binding lectins were toxic towardsA. pisum. Concanavalin A was also tested on five other aphid species (Aphis gossypii, Aulacortum solani, Macrosiphum euphorbiae, Macrosiphum albifrons andMyzus persicae) at concentrations between 10–1500 μg/ml. Mortality was very variable from one species to another. Strong growth inhibition invariably occurred within this concentration range, although dose-response curves differed substantially between aphid species. The peptidase complement ofA. pisum’s digestive tract was also investigated, as well as the oral toxicity of some protease inhibitors (PIs) to this aphid. Most protein PIs were inactive, and no part of the digestive tract contained detectable amounts of endo-protease activity. This is in contrast to the strong amino-peptidase activity which was shown to occur predominantly in the midgut and crop portions of the digestive tract. The potential of lectins in transgenic crops to confer Host-Plant Resistance to aphids is discussed.     

Performance of pea aphid (Acyrthosiphon pisum) clones on host plants and synthetic diets mimicking the same plants phloem amino acid composition

The performances of three clones of pea aphids, with different host affiliations, were evaluated on four host plants species and on four artificial diets. The amino acid compositions of the diets mimicked those of the phloem sap of the respective host plants. The total concentration of amino acids was the same in all the diets. The pea aphid clones performance were significantly affected by amino acid composition of the diets in different ways, implying physiological and/or behavioural differences among coexisting pea aphid clones in response to amino acids in artificial diets. The observed differences in performance on diets between clones were not related to host plant affiliations. Thus, even if the variation in amino acid composition in phloem sap among the host plants affects the pea aphid clones when tested on artificial diets, this variation has no observable effect on pea aphid performance on natural host plants.