Infected Aphids Research Articles

Belowground effects of organic and conventional farming on aboveground plant–herbivore and plant–pathogen interactions

Soil organisms may significantly affect the aboveground system. However, the influence of farming practices in modifying the effects of soil organisms on aboveground systems is poorly understood. The aim of our study was to investigate: (1) How important are soil organisms for plant growth and the development of herbivores and pathogens above the ground? (2) How do agricultural management practices affect interactions between soil organisms, plants and their aboveground herbivores and pathogens? To answer these questions we investigated the effect of experimental defaunation of soils from organic versus conventional farms on growth of wheat, abundance of aphids and infection of wheat by Septoria fungi. Plant biomass in soil from conventional farms exceeded that of soils from organic farms, presumably due to the higher nutrient input in the conventional farming system. Soil defaunation likely mobilized nutrients that increased plant growth. Aphid abundance and Septoria infection was reduced by defaunation but only in organic soils. This suggests that soil organisms in organic farming systems are more important for aphid performance and the infection rate by Septoria than in conventional systems. Hence, changes in the soil animal food web caused by farming practice feed back on aboveground organisms, and this appears to be more pronounced in organic farming systems. Further, the results indicate that soil organisms may modify higher trophic levels (aphid and pathogen infection) without significantly affecting lower trophic levels (plant growth).

The players in a mutualistic symbiosis: insects, bacteria, viruses, and virulence genes.

Aphids maintain mutualistic symbioses involving consortia of coinherited organisms. All possess a primary endosymbiont, Buchnera, which compensates for dietary deficiencies; many also contain secondary symbionts, such as Hamiltonella defensa, which confers defense against natural enemies. Genome sequences of uncultivable secondary symbionts have been refractory to analysis due to the difficulties of isolating adequate DNA samples. By amplifying DNA from hemolymph of infected pea aphids, we obtained a set of genomic sequences of H. defensa and an associated bacteriophage. H. defensa harbors two type III secretion systems, related to those that mediate host cell entry by enteric pathogens. The phage, called APSE-2, is a close relative of the previously sequenced APSE-1 but contains intact homologs of the gene encoding cytolethal distending toxin (cdtB), which interrupts the eukaryotic cell cycle and which is known from a variety of mammalian pathogens. The cdtB homolog is highly expressed, and its genomic position corresponds to that of a homolog of stx (encoding Shiga-toxin) within APSE-1. APSE-2 genomes were consistently abundant in infected pea aphids, and related phages were found in all tested isolates of H. defensa, from numerous insect species. Based on their ubiquity and abundance, these phages appear to be an obligate component of the H. defensa life cycle. We propose that, in these mutualistic symbionts, phage-borne toxin genes provide defense to the aphid host and are a basis for the observed protection against eukaryotic parasites.

Wheat containing snowdrop lectin (GNA) does not affect infection of the cereal aphid Metopolophium dirhodum by the fungal natural enemyPandora neoaphidis

Studies were carried out to determine if susceptibility of the cereal aphid Metopolophium dirhodum to the fungus Pandora neoaphidis was affected by wheat expressing snowdrop lectin (GNA). Aphid infection did not differ significantly between the transgenic GNA and non-transformed lines (91 and 82%, respectively). Fecundity also did not differ between aphids on the two lines, and was ca. 18 nymphs adult(-1). Time to infection was ca. 5 days for M. dirhodum on both lines in two of three assays. Our results indicate that wheat expressing GNA would not compromise the efficacy of P. neoaphidis as a biocontrol agent.

Mass harvesting of the entomopathogenic fungus, Neozygites fresenii, from natural field epizootics in the cotton aphid, Aphis gossypii

Epizootics caused by insect pathogens sometimes occur over wide areas and result in millions of infected insects. Naturally infected insects can be considered a natural resource for harvesting insect pathogens. We developed methods to mass harvest the fungal pathogen Neozygites fresenii (Entomophthorales: Neozygitaceae) from epizootics in Aphis gossypii (Homoptera: Aphididae) in a commercial cotton field in Arkansas. A total of 30,722 aphids infected with N. fresenii in the mature hyphal body or early conidiophore stages were harvested, dried, and frozen. Three desiccants were compared: silica gel, dry rock salt, and wet rock salt. Silica gel was the superior material because it more rapidly and thoroughly dried cotton leaves and aphids. Using this method a mean of 193.4 infected aphids could be harvested per hour from cotton leaves dried over silica gel. The quality of harvested infected aphids was high and resulted in a mean of 70.4% sporulation from infected aphids harvested from the silica gel desiccant.

Can plants use an entomopathogenic virus as a defense against herbivores?

It is by now well established that plants use various strategies to defend themselves against herbivores. Besides conventional weapons such as spines and stinging hairs and sophisticated chemical defenses, plants can also involve the enemies of the herbivores in their defense. It has been suggested that plants could even use entomopathogens as part of their defense strategies. In this paper, we show that Brassica oleraceae plants that are attacked by Myzus persicae aphids infected with an entomopathogenic parvovirus (M. persicae densovirus) transport the virus through the phloem locally and systematically. Moreover, healthy aphids that fed on the same leaf, but separated from infected aphids were infected via the plant. Hence, this is proof of the principle that plants can be vectors of an insect virus and can possibly use this virus as a defense against herbivores.

Further studies on Carrot virus Y: hosts, symptomatology, search for resistance, and tests for seed transmissibility

Carrot virus Y (CarVY) was studied to provide information on its host range and symptoms, identify any alternative natural hosts and sources of host resistance in carrot germplasm, and determine whether it is seed-borne. Twenty-two species belonging to the Apiaceae were inoculated with CarVY by viruliferous aphids in the glasshouse. Systemic infection with CarVY developed in carrot itself, 4 other Daucus species, 5 herbs, 1 naturalised weed, and 2 Australian native plants. When 7 of these host species were exposed to infection in the field, all became infected systemically. In both glasshouse and field, the types of symptoms that developed in infected plants and their severity varied widely from host to host. Following inoculation with infective sap, the virus was detected in inoculated leaves of 1 additional species in the Apiacaeae, and 2 species of Chenopodiaceae. A field survey did not reveal any alternative hosts likely to be important as CarVY infection reservoirs. When 34 accessions of wild carrot germplasm and 16 of other Daucus spp. were inoculated with infective aphids, symptom severity varied widely among accessions but no source of extreme resistance to CarVY was found. Tests on seedlings grown from seed collected from individual infected plants or field plantings (most with CarVY incidences of >92%) of cultivated carrot (34 135 seeds), wild carrot (20 978 seeds), Anethum graveolens (22 921 seeds), and 3 other host species (3304 seeds) did not detect any seed transmission of CarVY. The implications of these results for control of the virus in carrot crops, minimising the losses it causes, and avoiding its introduction to new locations are discussed.

Plant waxy bloom on peas affects infection of pea aphids by Pandora neoaphidis

This study examined the effects of the surface wax bloom of pea plants, Pisum sativum, on infection of pea aphids, Acyrthosiphon pisum, by the fungal pathogen Pandora neoaphidis. In prior field surveys, a higher proportion of P. neoaphidis-killed pea aphids (cadavers) had been observed on a pea line with reduced wax bloom, as compared with a sister line with normal surface wax bloom. Laboratory bioassays were conducted in order to examine the mechanisms. After plants of each line infested with aphids were exposed to similar densities of conidia, the rate of accumulation of cadavers on the reduced wax line was significantly greater than on the normal wax bloom line; at the end of the experiment (13 d), the proportion of aphid cadavers on the reduced wax line was approximately four times that on the normal wax bloom line. When plants were exposed to conidia first and then infested with aphids, the rate of accumulation of cadavers was slightly but significantly greater on the reduced wax line, and infection at the end of the experiment (16 d) did not differ between the lines. When aphids were exposed first and then released onto the plants, no differences in the proportion of aphid cadavers were observed between the pea lines. Greater infection of pea aphid on reduced wax peas appears to depend upon plants being exposed to inoculum while aphids are settled in typical feeding positions on the plant. Additional experiments demonstrated increased adhesion and germination by P. neoaphidis conidia to leaf surfaces of the reduced wax line as compared with normal wax line, and this could help explain the higher infection rate by P. neoaphidis on the reduced wax line. In bioassays using surface waxes extracted from the two lines, there was no effect of wax source on germination of P. neoaphidis conidia.

Glutenin and gliadin contents of flour derived from wheat infested with different aphid species.

Aphid feeding in cereals results in both quantitative and qualitative yield losses; moreover aphid-transmitted viruses are responsible for other quantitative and qualitative damage so that direct or indirect effects of aphid infection are always of interest. At the beginning of stem elongation, wheat plants were caged regardless of the presence or absence of indigenous cereal aphids. Half of the caged plants were artificially infected with Diuraphis noxia. The number of D noxia and Rhopalosiphum padi individuals on plants destined for flour preparation was approximately 150 aphids per tiller at peak population. The number of Sitobion avenae individuals was ca 25 aphids per tiller. Bread-making quality of wheat flour is determined primarily by the protein content, the gluten proteins (gliadins, glutenins) being the prime factors. The absolute amounts and/or the relative proportion of gliadins and glutenins are very important in dough making and in determining baking quality. Flour from grains originating from plants infected naturally with cereal aphids indigenous to Hungary, and flour infected artificially with D noxia, were analysed for glutenin and gliadin content using size exclusion HPLC. It was found that aphid infection had a significant effect on the gliadin/glutenin ratio, which was significantly lower in wheat flour prepared from aphid-infected plants than in that from uninfected control. The most significant decrease in gliadin/glutenin ratio was caused by D noxia infection, followed by R padi and then S avenae. As the gliadin/glutenin ratio was significantly lower in flour made from aphid-infected wheat seeds, it can be supposed that aphid feeding results in decreased bread making quality of wheat flour.

Effect of spatial heterogeneity on the role of Coccinella septempunctata as an intra-guild predator of the aphid pathogen Pandora neoaphidis.

The foraging behavior of starved and non-starved adult and larval Coccinella septempunctata on groups of plants in the presence of Pandora neoaphidis-infected Acyrthosiphon pisum, uninfected aphids or a mixture of these two prey types was compared. In general results of these studies confirmed the results of previous work comparing foraging behavior on a smaller spatial scale in Petri dishes. However, behaviors were modified in response to spatial complexity, prey quality, and the host plant. Starved C. septempunctata adults and larvae fed for longer and consumed more aphids than non-starved coccinellids. Both larvae and adults fed on infected aphids and in some cases entirely consumed them. This was thought to be due to the ease of capture of infected (dead) aphids and the feeding stimuli provided by the presence of the host plant and, where there was a choice of prey, uninfected aphids in the environment. Both larvae and adults spent the majority of the time foraging in the upper regions of plants and visited more plants when they were not starved or when they were in the presence of less suitable, infected aphid prey.

Facultative bacterial symbionts in aphids confer resistance to parasitic wasps.

Symbiotic relationships between animals and microorganisms are common in nature, yet the factors controlling the abundance and distributions of symbionts are mostly unknown. Aphids have an obligate association with the bacterium Buchnera aphidicola (the primary symbiont) that has been shown to contribute directly to aphid fitness. In addition, aphids sometimes harbor other vertically transmitted bacteria (secondary symbionts), for which few benefits of infection have been previously documented. We carried out experiments to determine the consequences of these facultative symbioses in Acyrthosiphon pisum (the pea aphid) for vulnerability of the aphid host to a hymenopteran parasitoid, Aphidius ervi, a major natural enemy in field populations. Our results show that, in a controlled genetic background, infection confers resistance to parasitoid attack by causing high mortality of developing parasitoid larvae. Compared with uninfected controls, experimentally infected aphids were as likely to be attacked by ovipositing parasitoids but less likely to support parasitoid development. This strong interaction between a symbiotic bacterium and a host natural enemy provides a mechanism for the persistence and spread of symbiotic bacteria.

Spore germination and appressorium formation in the entomopathogenic Alternaria alternata

The entomopathogenic fungus Alternaria alternata grew equally well on potato dextrose agar (PDA) and also on potato extract agar (PEA). The latter was excellent for sporulation. The fungus grew at a wide temperature range of 15–35°C with an optimum at 25°. On PDA sporulation was much reduced as compared to that on PEA. The spores of the fungus germinated only at 100% relative humidity (rh). Darkness significantly increased the germination percentages in the early stages of germination at all temperatures tested. Differences in germination percentages between light and dark conditions were mostly levelled 24 h post incubation. The germ tubes formed distinct appressoria, both on glass slides and on the aphid host Anuraphis nerii on which appressoria were formed on all parts of the insect bodies, even on the hardest ones, such as the head and thorax. Most appressoria were single globose and formed terminally on hyphal branches. Few were subapical and still fewer compound and multilobed. Detachment of appressoria from their original position revealed an exceedingly smoothened appression area, around which a mucilagenous material was secreted, apparently cementing them onto the host epicuticle. Optimum temperature for appressorium formation was 20–25°. At these temperatures more than 80 % of the emerging germ tubes formed appressoria. The factors affecting spore germination and appressorium formation are discussed in relation to the prevailing environmental conditions in the mediterranean area as well as their role in influencing the spread and activities of the pathogen and the infection of aphids under field conditions.

A simple test for evaluation of transmission efficiency of barley yellow dwarf virus by aphids

Barley yellow dwarf virus (BYDV) transmission test systems involve the use of clip-cages or of whole plants in cages, which are both labor-intensive methods and require large controlled environment units. Employing detached leaves for assessment of the inoculation efficiency of aphids proved reliable for assessing transmission of a BYDV PAV-like isolate byRhopalosiphum padi. One use of the system could be for the rapid determination of the infectivity of field-collected aphids, an essential part of any epidemiological study of BYDV.

Effects of constant and fluctuating temperatures on sporulation and infection by the aphid‐pathogenic fungus Pandora neoaphidis

AbstractLaboratory studies were performed to assess the importance of temperature on sporulation and infection by the aphid‐pathogenic fungus Pandora neoaphidis (Remaudière and Hennebert) Humber. Numbers of primary conidia discharged from mycelium formulated as alginate granules and unformulated mycelial mats were assessed, as well as infection of the potato aphid, Macrosiphum euphorbiae (Thomas) (Homoptera, Hemiptera, Aphididae), using culture plugs as inoculum sources. Sporulation from experiments at constant temperatures indicated the optimum temperature range was 10–20 °C for both mycelial preparations and there was no or very little sporulation at 30 °C. Infection of aphids kept at 15 °C was 34–50%, while infection at 25 °C was 11–44%. At 20 °C, 77–79% of aphids were infected. Under fluctuating temperature cycles, conidia numbers did not differ when mycelial preparations were maintained at 18–25 °C compared with 18–20 °C, but fewer conidia were recorded when preparations were exposed continuously to 18–30 °C. Infections of inoculated aphids kept for varying numbers of days at 18–25 °C varied between 24–47%, but only 3–32% of aphids were infected when exposed to a cycle of 18–30 °C for various times. Unformulated mycelial mats of P. neoaphidis appear to be superior to forumlated alginate granules for use in experimental greenhouse and field trials, since temperature stability is similar for both materials but mycelial mats are much easier to produce.

Detection of Strawberry crinkle virus in plants and aphids by RT‐PCR using conserved L gene sequences

About 10% of the large (L) protein gene of Strawberry crinkle virus (SCV) was sequenced after amplification with degenerate primers designed to conserved regions of the rhabdovirus L protein. The virus sequence was extended to 1362 nucleotides through rapid amplification of cDNA ends. One pair of degenerate L gene primers amplified a 683‐bp fragment from four different isolates of SCV cultured in the experimental host Physalis pubescens; the nucleotide sequences of these fragments differed by < 1% to 10% indicating the suitability of this region as a diagnostic target. This information enabled the development of a reverse transcription polymerase chain reaction (RT‐PCR) detection method for SCV using primers designed to the L gene sequence. SCV was amplified from infected P. pubescens (573 bp fragment) and from infected Chaetosiphon fragaefolii aphids (770 bp fragment). SCV was also detected by RT‐PCR in total RNA extracts from three strawberry plants showing symptoms typical of SCV infection but failed when the intensity of the disease symptoms decreased. However, both SCV positive‐sense RNA, and negative‐sense genomic RNA, were detected by nested PCR in chronically infected strawberry plants sampled in September.

Conidial discharge in the aphid pathogen Erynia neoaphidis

The patterns of conidial discharge of Erynia neoaphidis were measured from three species of aphids: nettle aphid (Microlophium carnosum), grain aphid (Sitobion avenae) and pea aphid (Acyrthosiphon pisum). The effects of release height, morph of aphid and temperature on the horizontal and vertical discharge of conidia were studied. Numbers of conidia deposited in the dorsal and lateral directions were distributed with distance in truncated bell shaped patterns. Discharge distances ranged from 2 to 11 mm and half the conidia caught travelled further than 5 mm. There was little difference in the horizontal dispersal patterns of conidia when ejected from cadavers of different morphs of the same species, or from different species. The weight of infected aphids showed little correlation with discharge patterns. Values of maximum discharge distance, Dm, were generally between 6 and 9 mm. Temperature had a significant effect on conidia discharged from the dorsal surface of apterous A. pisum cadavers. Dm was greater at 18 °C than at 10 or 25 °. Vertical discharge distances for conidia released from A. pisum cadavers ranged between about 2 and 8 mm. The maximum height to which conidia were projected vertically was not affected by temperature. However, the average height tended to be greater at 18 °. Half of the conidia reached heights of about 3.5 mm while the maximum heights reached were about 8 mm. The initial speeds of conidia were estimated from measured discharge distances. The results suggest that conidia of E. neoaphidis may be ejected at speeds of about 8 ms−1.

Targeted Dispersal of the Aphid Pathogenic Fungus Erynia neoaphidis by the Aphid Predator Coccinella septempunctata

The potential of adult and larval C. septempunctata to vector the aphid-specific entomopathogenic fungus E. neoaphidis was assessed through a series of laboratory and field experiments. The ability of coccinellids to vector conidia from a colony of E. neoaphidis -infected pea aphids, Acyrthosiphon pisum, to a colony of uninfected A. pisum was demonstrated in a laboratory study. Adult coccinellids which had previously foraged on plants infested with different densities of sporulating cadavers (1, 5, 15, 30 cadavers per plant) initiated infection in a proportion of uninfected pea aphids (4, 0, 2 and 8%, respectively) when subsequently allowed to forage on A. pisum infested bean plants. Further laboratory studies demonstrated that fourth instar larvae and adult coccinellids artificially inoculated with conidia initiated infection in 11 and 13% of an A. pisum population in which they foraged, respectively. Furthermore, a proportion of A. pisum placed on bean plants which had previously been foraged on by inoculated larval and adult coccinellids also died from infection (3 and 10% of A. pisum, respectively). However, although coccinellid adults inoculated with conidia initiated infection in 19% of A. pisum, cereal aphids, S. avenae , exposed to the inoculated coccinellids did not become infected. A further laboratory study demonstrated that infection of A. pisum only occurred if inoculated coccinellids were transferred to A. pisum populations immediately post inoculation. However, a proportion of A. pisum placed on bean plants which had been foraged on by inoculated coccinellids transferred 0, 4 and 24 h post inoculation died from infection (9, 3 and 7%, respectively). A field study further demonstrated the potential of coccinellids to vector E. neoaphidis. Single spring sown field bean plants (Long Hoos Experimental Plots, IACRRothamsted Farm) were enclosed within nylon mesh bags and 25 adult A. pisum were added to each bag with one of the following treatments: no further addition (control), coccinellid adult (control), inoculated coccinellid adult, inoculated A. pisum or sporulating A. pisum cadavers. No aphids died of E. neoaphidis in the control treatments; 5, 16 and 33% of aphids were infected with E. neoaphidis on the other treatments, respectively.

Altered behavior and distribution of pea aphids, Acyrthosiphon pisum (Homoptera: Aphididae), infected with Pandora neoaphidis (Zygomycetes: Entomophthorales)

The distribution and mobility of infected aphid hosts can have a great effect on the ability of a pathogen to spread throughout a population. The distribution of dead and living pea aphids (Acyrthosiphon pisum) infected with Pandora neoaphidis was compared with that of their healthy conspecifics. Infected aphids were significantly more likely to be found on the undersides of alfalfa leaves and off of the plants than were healthy aphids. These two shifts in microhabitat location have potential costs and benefits for both the host and the pathogen.

Seasonal dynamics and entomophthoralean infection of the pea aphid, Acyrthosiphon pisum Harris

The seasonal dynamics of the pea aphid, Acyrthosiphon pisum Harris, and entomopathogenic fungi from the order Entomophthorales attacking the aphid were analysed in alfalfa and field pea crops during the growing seasons of 1998 and 1999 at Nitra-Malanta. In both years, pea aphid populations on pea showed a pattern with one peak, culminated at flowering and pod formation. The infestation level on alfalfa was low in both years. Entomopathogenic fungi attacking the pea aphid were identified as Erynia neoaphidis Remaudičre and Hennebert and Conidiobolus obscurus (Hall and Dunn) Remaudičre and Keller. Both pathogens infected the aphid on pea, but only E. neoaphidis was found in the alfalfa plots. Infected aphids were not found on alfalfa during 1999. The maximum levels of infected aphids on pea were 10.30% and 48.39% in 1998 and 1999, respectively. During both years alate aphids were more frequently attacked than apteral ones. Correlation coefficients indicated a positive relationship between the number of infected aphids and precipitation, but this relationship was weak or moderately strong. A strong correlation was found between the number of dead aphids and number of alate aphids counted 5 to 10 days earlier.

Alternaria alternata, a new pathotype pathogenic to aphids

A fungus, identified as Alternaria alternata, was isolated from dying or dead aphids and proved to be pathogenic. It was isolated from different parts of Greece from aphid specimens on cultivated plants, ornamentals and weeds. In the laboratory, disease development started with the germination of spores on the insect integument and the subsequent growth of mycelium. The fungus formed apical and intercalary, globose or lobate appressoria which were firmly attached onto the host exoskeleton and facilitated entrance of the mycelium into the insect body. Under favorable conditions of temperature (15–35 °C) and relative humidity (100%), infected aphids died in 2–4 days. A characteristic brown discoloration accompanied the death of the insects. Both mycelial growth and sporulation were profuse on dead specimens. The pathogen infected all 26 aphid species tested but was unable to infect other insects (Drosophila melanogaster and Ceratitis capitata) or aphid host plants. There were significant differences in mortality rate among aphid species only during the first two days after inoculation. It is suggested that A. alternata may be a good candidate to be exploited for the biological control of aphids.

The potential of some adjuvants in promoting infection with Verticillium lecanii: laboratory bioassays with Myzus persicae

Summary The potential of 14 adjuvants, including mineral oils, vegetable oils, stickers, spreaders, surfactants and nutrients, in promoting the virulence of the entomopathogenic fungus Verticillium lecanii were compared relative to a commercial formulation of the pathogen. The rate of infection over time with each adjuvant relative to the commercial formulation was tested using the peach potato aphid Myzus persicae. The proportion of infected aphids was modelled binomially with a logistic rate function assumed for the proportion infected over the time period of the experiment. Four of the adjuvant treatments, Codacide, skimmed milk, glycerol and Cutinol together with the commercial control had similar logistic slope parameters and were modelled using parallel regressions. The parallel regressions model showed significant differences between the intercepts with both glycerol and Cutinol significantly better than V. lecanii alone. Glycerol had a more positive intercept than Cutinol but the difference was not statistically significant. Although highly variable, the numbers of progeny produced by the treated aphids were significantly reduced by all the treatments relative to untreated control aphids. Glycerol and Cutinol were the only treatments that significantly reduced the number of progeny relative to the V. lecanii control.