Isolation of microsatellite markers from the rosy apple aphid Dysaphis plantaginea

We report the development of five polymorphic microsatellite loci from the aphid Dysaphis plantaginea, a serious pest on apple. Using a multimotif enrichment protocol, we have isolated loci that are proving to be valuable tools for studying aphid movement at both between‐county and within‐orchard scales.     

The potential of three native insect predators to control the rosy apple aphid, Dysaphis plantaginea

The potential of three aphidophagous predators, Adalia bipunctata, Aphidoletes aphidimyza, and Episyrphus balteatus to control the rosy apple aphid, Dysaphis plantaginea Pass., a major pest on apple in Europe, was assessed by means of laboratory and field cage experiments in Northern Switzerland. Under laboratory conditions, all three predators efficiently preyed upon D. plantaginea on apple seedlings. The searching success of larvae of A. bipunctata for individual aphids was not dependent on the size of branches of apple trees varying in leaf surface area from 150 cm² to 960 cm². Fifty and 70% of individual aphids were found and killed 6 hours and 48 hours, respectively, after release of single second instar larva of A. bipunctata. In a first field cage experiment in 1996, A. bipunctata, and to a lesser extent E. balteatus, proved to be effective and consistent predators of D. plantaginea during spring conditions, being little affected by cool temperatures and wet weather. In a subsequent field cage experiment in 1997, larvae of A. bipunctata and E. balteatus were released singly and in combination on aphid infested apple seedlings to study interactions between these two promising control agents. Both species had a significant negative effect on aphid population increase. The two species did not significantly interact and thus, their joint effect is best explained by an additive model. Combined releases of the two predator species reduced aphid densities to 5% of the control. This indicates the potential for augmentative releases of these native aphid predators to control D. plantaginea.     

Weak spatial and temporal population genetic structure in the rosy apple aphid, Dysaphis plantaginea, in French apple orchards

We used eight microsatellite loci and a set of 20 aphid samples to investigate the spatial and temporal genetic structure of rosy apple aphid populations from 13 apple orchards situated in four different regions in France. Genetic variability was very similar between orchard populations and between winged populations collected before sexual reproduction in the fall and populations collected from colonies in the spring. A very small proportion of individuals (～2%) had identical multilocus genotypes. Genetic differentiation between orchards was low (F(ST)<0.026), with significant differentiation observed only between orchards from different regions, but no isolation by distance was detected. These results are consistent with high levels of genetic mixing in holocyclic Dysaphis plantaginae populations (host alternation through migration and sexual reproduction). These findings concerning the adaptation of the rosy apple aphid have potential consequences for pest management.     

Conservation biological control of rosy apple aphid, Dysaphis plantaginea (Passerini), in eastern North America

Because of the potentially serious damage rosy apple aphid, Dysaphis plantaginea (Passerini) (Homoptera: Aphididae), can cause to apple fruit and branch development, prophylactic insecticides are often used for control. If biological control could be relied on, the amount of pesticide applied in orchards could be reduced. This study examined biological control of rosy apple aphid in eastern West Virginia and the potential for enhancement through conservation biological control, in particular, the effect of interplanting extrafloral nectar-bearing peach trees. By 20 d after first bloom, only 2% of fundatrices initially present survived to form colonies based on regression of data from 687 colonies. Exclusion studies showed that many of the early colonies were probably destroyed by predation; the major predator responsible seemed to be adult Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae). Mortality before apple bloom was most important in controlling rosy apple aphid population growth but by itself is not sufficiently reliable to prevent economic injury. Interplanting of extrafloral nectar-bearing trees did not increase biological control, and interplanting with 50% trees with extrafloral nectar glands reduced biological control. The number of leaf curl colonies in the 50% interplanted orchards was lower than in monoculture orchards, suggesting a preference of alate oviparae for more diverse habitats, supporting the resource concentration hypothesis but not at a level sufficient to prevent injury. Predation and parasitism after the formation of leaf curl colonies was not adequate to control rosy apple aphid populations.     

In-field production of parasitoids of Dysaphis plantaginea by using the rowan aphid Dysaphis sorbi as substitute host

A system was developed to provide the parasitic wasp Ephedrus persicae Froggatt (Hymenoptera: Braconidae: Aphidiinae), which attacks the rosy apple aphid Dysaphis plantaginea (Passerini) (Homoptera: Aphididae), with the alternative host Dysaphis sorbi Kaltenbach (Homoptera: Aphididae) in apple orchards. Rowan trees (Sorbus aucuparia L.) arranged along the side of an unsprayed orchard were artificially infested in late February 2002 with eggs of D. sorbi. Colonies of D. sorbi successfully developed from the introduced eggs and persisted on several trees until the end of June. The only primary parasitoid species emerging from a sample of mummified aphids collected in spring from the infested rowan trees was the braconid wasp species E. persicae. In a host-switching experiment, nymphs of D. plantaginea proved suitable for female parasitoids originating from mummified D. sorbi. A series of mummies collected from the rowan trees in early summer contained diapausing parasitoids and hyperparasitoids that only hatched in April of the following spring. These observations suggest the possibility of establishing a local population of E. persicae in apple orchards, so that D. plantaginea can be readily attacked by diapause-emerging parasitoids in early spring.     

Seasonal coincidence between rosy apple aphid fundatrices and their parasitoids: implications for an early biological control of Dysaphis plantaginea

The potential contribution of the aphid parasitoid Ephedrus persicae Froggatt (Hymenoptera: Braconidae, Aphidiinae) in regulating stem mothers of the rosy apple aphid Dysaphis plantaginea (Passerini) (Homoptera: Aphididae) was investigated in culture conditions using both species' phenological emergence data in spring and host stage suitability trials. In 2003, emergence of E. persicae started on April 1st, i.e., 108.2 degree-days (base: 4.5 degrees C; start date: January 1st) later than its host on March 10th. When less than 3% of parasitoids had emerged on April 12th, more than 97% of D. plantaginea stem mothers had moulted beyond their 1st instar. The latest parasitoids left their diapause mummies in late April as the majority of fundatrices had reached the adult stage. Parasitisation trials demonstrated the suitability for E. persicae of all developmental stages of rosy apple aphid fundatrices, including mature individuals. No offspring were deposited by fundatrices parasitised as Ist-instar nymphs but later instars did produce a progeny whose size was substantially reduced compared with unparasitised individuals, and scaled against their age at the time of parasitisation. Although the temperature accumulation required for emergence in spring is reached later for E. persicae than for its host aphids, thus allowing part of the stem mother population to temporarily escape parasitisation and reach maturity, the marked reduction in fecundity of individuals parasitised even as late-instar nymphs does confer to E. persicae a non-negligible role in potentially regulating D. plantaginea.     

Effects of autumn kaolin treatments on the rosy apple aphid, Dysaphis plantaginea (Pass.) and possible modes of action

Abstract:  Autumn applications of the repellent processed-kaolin particle film (Surround® WP) might be an alternative to the insecticides commonly used in spring to control the rosy apple aphid, Dysaphis plantaginea (Pass.) (Hom., Aphididae). To assess the mode of action and the impact of kaolin on autumn forms of D. plantaginea , trials were conducted in field cages and in open-fields in autumn 2003. Choice and no-choice experiments in field cages showed that winged aphids landed in significantly lower numbers and gave birth to significantly fewer females on kaolin-treated branches compared with the untreated control. In a first open-field trial, single applications at 10 different dates and with two different concentrations of kaolin were conducted after harvest to assess the influence of the kaolin concentration on the autumn forms of D. plantaginea . No differences were found between the different concentrations and spraying dates. In a second open-field trial, single and multiple applications of kaolin were tested at different dates after harvest. Repeated applications of kaolin significantly reduced females in autumn and fundatrices in spring, whereas single kaolin treatments had no significant effect on D. plantaginea . None of the kaolin treatments reduced aphids below the economic threshold. In conclusion, kaolin showed promising results to control autumn forms of D. plantaginea. However, with a more detailed forecasting model for the autumnal flight of this aphid, treatments could be conducted more precisely and more effectively, especially in years with exceptional climatic conditions, such as in 2003.     

The induction of leaf-roll galls by the apple aphids Dysaphis devecta and D. plantaginea

Dysaphis devecta causes the leaves of its host plant to roll laterally whereas D. plantaginea causes them to roll longitudinally. Both species of aphid are phloem feeders. D. devecta prefers to feed on the smaller veins in the lamina whereas D.plantaginea chooses the midrib. However, groups of D. devecta or D. plantaginea confined to the stem of an apple seedling induced young leaves several centimetres away to develop leaf rolls characteristic of each species. A single larva of D. devecta or D. plantaginea can induce a leaf-roll or a stem-bend on an apple seedling within 24 h. It is suggested that characteristic abnormalities in leaves attacked by D. devecta and D. plantaginea are caused by specific substances in the saliva of each aphid.     

Resistance of apple cultivars to Dysaphis plantaginea (Hemiptera: Aphididae): role of tree phenology in infestation avoidance

To test the importance of flowering phenology in damage caused to apple cultivars by rosy apple aphid, Dysaphis plantaginea (Passerini) (Hemiptera: Aphididae), susceptibility of 16 cultivars was compared in greenhouse conditions after infestation with aptera and in the field after natural tree colonization. Flowering phenology was recorded in the field for all the cultivars. In both greenhouse and field trials, there were differences among cultivars with respect to aphid infestation and damage. However, damage in the greenhouse and field was not significantly correlated. Flowering phenology was also different among cultivars. Flowering order among cultivars was significantly negatively correlated with aphid infestation and damage in field, i.e., early leafing cultivars showed higher infestation and damage than late-leafing cultivars. If egg hatching occurs before bud bursting, neonate larvae will suffer a high mortality because they cannot feed on these late cultivars. A later recolonization of these trees is hampered because (1) winged aphids cannot live on apple but only on the secondary host, (2) apterous forms have a limited dispersal capability, and (3) aphid predators progressively increase in the orchard. Therefore, synchronization between egg hatching and bud bursting is of critical importance in the success or failure of infestation.     

Effects of autumn kaolin and pyrethrin treatments on the spring population of Dysaphis plantaginea in apple orchards

The impact of treatments with the pyrethrin insecticide Pyrethrum FS and the repellent kaolin product Surround^® WP to control the autumn forms of Dysaphis plantaginea Pass. (Hom., Aphididae), a major insect pest of apple, was assessed in 1‐year field experiments in Switzerland. Single and multiple applications of pyrethrin and kaolin were tested at different dates after apple harvesting in autumn 2002 when sexuales of the rosy apple aphid were present. Repeated applications of Surround^® WP in autumn significantly reduced the number of females in autumn and, consequently, the number of hatched fundatrices in spring. Single kaolin treatments were less effective. Unexpectedly, neither single nor multiple applications of the contact insecticide Pyrethrum FS had a knock‐down effect on females in autumn. However, pyrethrin significantly reduced the number of hatched fundatrices in spring. Neither pesticide completely controlled the rosy apple aphid but with a more detailed analysis of factors influencing the efficacy of autumn treatments a new approach to control this serious pest could be achieved.     

Effects of augmentative releases of eggs and larvae of the ladybird beetle, Adalia bipunctata, on the abundance of the rosy apple aphid, Dysaphis plantaginea, in organic apple orchards

The impact of augmentative releases of larvae and eggs of the indigenous ladybird beetle Adalia bipunctata (L.) (Coleoptera: Coccinellidae) against the rosy apple aphid Dysaphis plantaginea Pass. (Homoptera: Aphididae), a major pest insect on apple trees, was assessed in field experiments in Switzerland, during 1997. In a first experiment, eggs and larvae were released on 3-year old apple trees infested with five aphids at four different predator-prey ratios (0:5, 1:5, 1:1, 5:1). In a second experiment, eggs and larvae were released at a predator-prey ratio of 5:1 on branches of apple trees naturally infested with aphids. In both experiments, the interaction with ants was taken into account and the releases were done at two different times in spring. The results showed that an augmentative release of larvae significantly prevented the build-up of colonies of D. plantaginea. Significant reductions in aphid numbers were recorded at the two highest predator-prey ratios, 1:1 and 5:1. Larvae were efficient just before flowering of apple trees at a time when growers normally have to spray their trees. On trees where ants were present the larvae of A. bipunctata were significantly less efficient. Effects of eggs of A. bipunctata, however, were less reliable. At the first date of release (5 April), they did not hatch, probably as a consequence of bad weather conditions.     

DL‐β‐Aminobutyric acid application negatively affects reproduction and larval development of the rosy apple aphid,Dysaphis plantaginea,on apple

The rosy apple aphid, Dysaphis plantaginea (Passerini) (Hemiptera: Aphididae), is one of the major pests of European apple orchards, commonly controlled by the use of synthetic insecticides. In the present work, the non‐protein amino acid DL‐β‐aminobutyric acid (BABA), known to induce plant resistance against a wide range of abiotic and biotic stresses, has been tested for its protective effect against this pest on apple. We first verified the lack of any contact effect of BABA on the insect itself. Next we applied BABA as a soil drench to apple and monitored its effect on the population development of aphids after artificial infestation. We demonstrated that BABA strongly reduced the population growth and that this compound severely affected various life‐history characteristics of the aphid such as female longevity and fecundity, nymph mortality, and larval development.     

The biology and energetics of the potato aphid Macrosiphum euphorbiae, living in galls of the apple aphids Dysaphis devecta and Aphis pomi

A non-gall-forming aphid, Macrosiphum euphorbiae, was reared within the galls of Dysaphis devecta and Aphis pomi which had been induced in apple seedlings. Similar aphids were also reared on ungalled seedling leaves of a similar age and also on mature leaves and in old galls.There is a weak positive linear relationship between lipid content and pre-reproductive adult dry weight, but aphids living on young galled or ungalled plant tissue have 30% lipid compared to 11% for aphids living on old plant tissue. Adult aphids gradually decline in weight, losing between 24% and 38% of their pre-reproductive biomass before death. The energy content of this loss subsidieses reproduction. Reproduction of aphids feeding in old galls or on mature leaves was negligible, but on younger tissues average fecundity was 31, there being little difference between aphids living on galled and ungalled tissue. There is no simple relationship between pre-reproductive adult embryo content and fecundity.The principal factor mediating honeydew production was aphid size and this factor overrides any variation caused by different feeding sites. Honeydew production averaged 0.43 mg during a life of 30 days. Aphids successfully completing their life cycle have a life-time energy consumption of 30.58 joules of which 18% is lost as heat during respiration (R), 30% is contained in honeydew (F+U), 1% is contained in exuviae (Pe). Larval growth accounts for 17% (Pg), and adult reproduction for 34% (Pr).The P/C ratio for M. euphorbiae is 53% and is broadly comparable with aphids living on herbaceous plants.

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Résumé Un puceron ne formant pas de galles, Macrosiphum euphorbiae, a été élevé dans des galles de Dysaphis devecta et Aphis pomi formées sur semis de pommiers. D'autres M. euphorbiae ont été élevés sur feuilles sans galles du même âge provenent de semis identiques, sur feuilles mûres et sur vieilles galles.Il y a une faible relation positive linéaire entre la teneur en lipides et le poids sec des adultes avant la reproduction, mais les pucerons élevés sur tissus végétaux jeunes avec ou sans galles contiennent 30% de lipides contre 11% pour ceux élevés sur tissus âgés. Le poids des pucerons adultes diminue progressivement, la perte avant la mort se situant entre 24 et 38% de la biomasse avant la reproduction. Le contenu énergétique de ces pertes alimente la reproduction. La reproduction de pucerons consommant les vieilles galles ou les feuilles mûres était négligeable, mais sur jeunes tissus la fécondité moyenne était de 31-avec peu de différences entre pucerons sur galles ou feuilles-. Il n'y a pas de corrélation simple entre le contenu en embryon des adultes avant la reproduction et la fécondité.Le principal paramètre conditionnant la production de miellat est la taille du puceron; ce paramètre surpasse toute variation provoquée par les divers lieux d'alimentation. La production moyenne de miellat est de 0,43-mg pendant une vie de 30 jours. Les pucerons accomplissant leur cycle avec succès utilisent pendant leur vie 30,58 joules, dont 18% sont dissipés sous forme de chaleur pendant la respiration (R); le contenu du miellat (F+U) correspond à 30%, et 1% est contenu dans l'exuvie (Pe). La croissance larvaire (Pg) utilise 17%, et l'activité reproductrice des adultes (Pr) 34% de l'énergie. Le rapport P/C-53% pour M. euphorbiae-est globalement comparable à celui des pucerons vivant sur plantes herbacées.

     

Molecular interactions between rosy apple aphids, Dysaphis plantaginea, and resistant and susceptible cultivars of its primary host Malus domestica

The use of crop varieties resistant or tolerant to insect pests or other stress factors is one approach in non‐chemical crop‐protection. Knowledge of the biochemical and molecular background of insect–plant interactions is a prerequisite for optimizing breeding for resistance. However, the resistance genes involved in plant–aphid interactions have so far only been identified and characterized in very few plant species. Our work aims to elucidate the molecular and biochemical mechanisms involved in resistance of apple trees, Malus domestica L. (Rosaceae), against its primary aphid pest, the rosy apple aphid, Dysaphis plantaginea (Passerini) (Homoptera: Aphididae), which is considered a serious economic pest of apple. Gene expression in both resistant and susceptible apple cultivars after infestation with rosy apple aphids was investigated by employing the cDNA‐AFLP method (cDNA–Amplified Fragment Length Polymorphism). From approximately 12 500 cDNA fragments detected on polyacrylamide gels, 21 bands were apparently up‐ or down‐regulated only in the resistant cultivar ‘Florina’ after aphid infestation compared to the susceptible cultivar ‘Topaz’ and/or mechanically wounded or non‐infested leaves. These fragments were cloned, sequenced, and the pattern of gene expression for six fragments was subsequently verified by virtual Northern blots. Sequence comparisons of these fragments to GenBank accessions revealed homologies to already known genes, most of them isolated from Arabidopsis thaliana L. Among them, a putative RNase‐L‐inhibitor‐like protein, a pectinacetylesterase, an inositol‐phosphatase‐like protein, a precursor of the large chain of the ribulose‐1,5‐biphosphate‐carboxylase, and defence‐related genes such as a vacuolar H(+)‐ATPase subunit‐like protein and an ADP‐ribosylating enzyme were identified. The results are discussed in relation to a putative role of these genes in conferring aphid resistance in apple trees.     

Resistance genes in apple and biotypes of Dysaphis devecta

Apple cultivars and young seedlings were repeatedly inoculated with Dysaphis devecta from six locations in South East England. Four genes governing resistance were associated with three aphid biotypes. The gene for resistance to biotypes 1 and 2 from Cox's Orange Pippin was given the symbol Sd₁ while that from Northern Spy, for resistance to biotype 1 only, was designated Sd₂. A single gene (Sd₃) for resistance to biotype 3 was located in Malus robusta MAL 59/9 and M. zumi MAL 68/5. Worcester Pearmain, Cox, Mcintosh, Lane's Prince Albert and MAL 68/5, are heterozygous for a precursor gene Sd_pr without which Sd₁Sd₂, and Sd₃ are ineffective.     

Susceptibilities of apple aphid and spirea aphid collected from apple in the Pacific Northwest to selected insecticides

Laboratory bioassays using leaf disks of apple dipped in test solutions of insecticides demonstrated that the apple aphid, Aphis pomi De Geer, and the spirea aphid, Aphis spiraecola Patch, differed significantly in susceptibility to a number of insecticides registered for control of aphids on apple (Malus spp.). Compared with A. pomi, A. spiraecola was approximately four- and three-fold less susceptible to pirimicarb and lambda-cyhalothrin, respectively, whereas there was little difference in response to dimethoate. Pymetrozine is thought to act on aphids primarily as a feeding inhibitor. Exposure of aphids to this material generated data that fit the probit model for only half the tested clones. However, the LC50 value for one clone of A. spiraecola was nearly 1,000 times higher than the value for one clone of A. pomi. Although the results from these trials did not indicate that either species had developed significant levels of resistance to the test materials, differences in LC50 levels of > 10-fold suggest insecticide tolerances and the possibility of control failures in the future. The demonstrated differences in susceptibility to insecticides between these two morphologically similar species also should be considered during the evaluation of new products for use on apple.     

Baseline susceptibilities to imidacloprid for green apple aphid and spirea aphid (Homoptera: Aphididae) collected from apple in the Pacific northwest

Susceptibilities to the neonicotinyl insecticide imidacloprid were determined for clones of apple aphid, Aphis pomi De Geer, and spirea aphid, Aphis spiraecola Patch, collected from conventional and organic apple orchards and from crab apple and wild apple in Washington state and British Columbia over a period of 6 yr. For aphids collected during 1996--1998, adults were dipped in test solutions by using the Food and Agriculture Organization protocol, and third instars and adults were reared on treated apple leaf disks. During the final 3 yr of study, bioassays involved only third instars on treated leaf material. Tests showed that A. spiraecola was significantly more tolerant to imidacloprid compared with A. pomi. Depending on the bioassay method and aphid developmental stage, average LC50 values for A. spiraecola were 4.4 -5.7 times higher than those for A. pomi established under the same test conditions. Clones of both species from Washington were marginally more tolerant to imidacloprid than clones from British Columbia, but the differences were generally not significant. Average measures of susceptibility for clones from organic orchards or unsprayed trees also did not differ from those for clones from conventional orchards, and there was no evidence for increasing LC50 values over the 6 yr of study. Differences in susceptibility to insecticides between these two anatomically similar species should be considered during the testing of new products for use on apple.     

Phytohormonal signaling in plant responses to aphid feeding

Aphid feeding induces various defense signaling mechanisms in plants. The recognition of feeding activities by plants occurs through the use of transmembrane pattern recognition receptors (PRRS) or, acting largely inside the cell, polymorphic nucleotide-binding leucine-rich-repeat (NB-LRR) protein products, encoded by most R genes. Activation may induce defensive reactions which are the result of highly coordinated sequential changes at the cellular level comprising, among other changes, the synthesis of signaling molecules. The ensuing plant responses are followed by the transmission of defense response signal cascades. Signals are mediated by bioactive endogenous molecules, i.e. phytohormones, such as jasmonic acid (JA), salicylic acid (SA), ethylene (ET), abscisic acid (ABA), gibberellic acid (GA) and free radicals such as hydrogen peroxide (H₂O₂) and nitric oxide (NO) which independently provide direct chemical resistance. Plant-induced defenses are also regulated by a network of inter-connecting signaling pathways, in which JA, SA, and ET play dominant roles. Both synergistic and inhibitory aspects of the cross-talk among these pathways have been reported. This paper presents molecular mechanisms of plant response to aphid feeding, the precise activation of various endogenous bioactive molecules signaling in the response of many plant species and their participation in the regulation of numerous defense genes, which lead to a specific metabolic effect. Selected important points in signal transduction pathways were also discussed in studies on plant response to aphid feeding.