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Vector specificity of barley yellow dwarf virus (BYDV) transmission: identification of potential cellular receptors binding BYDV-MAV in the aphid, Sitobion avenae.

Two proteins (SaM35 and SaM50) isolated from head tissues of the aphid vector, Sitobion avenae, were identified as potential receptors for barley yellow dwarf virus MAV isolate (Luteoviridae) based on MAV virus overlay assays and immunoblots of urea SDS 2-D gels. An anti-idiotypic antibody (MAV4 anti-ID) that mimics an epitope on MAV virions and competes with MAV in antibody binding assays also bound to SaM50 and SaM35 and to six additional proteins including a GroEL homolog. No MAV-binding proteins were detected from the nonvector aphid, Rhopalosiphum maidis, although MAV4 anti-ID did react with four proteins from R. maidis. It is hypothesized that SaM35 and SaM50 may be MAV receptors involved in MAV transmission based on their high affinity for MAV and their unique association with the vector, S. avenae. The additional aphid proteins binding the MAV4 anti-ID may represent less specific virus-binding proteins facilitating transmission through different aphid tissues.

Animals↗

Discovery and molecular characterization of a plasmid localized in Buchnera sp. bacterial endosymbiont of the aphid Rhopalosiphum padi.

We have identified and completely sequenced a novel plasmid isolated from the aphid Rhopalosiphum padi. Evidence which suggests that the plasmid occurs localized within the bacterial endosymbionts is presented. The plasmid contains the four genes that constitute the entire leucine operon. This fact makes it really unique since most plasmids are dispensable and lack genes that encode essential anabolic functions. Four more phloem-feeding aphid species also seem to contain homologous plasmids. Although further work is necessary, we hypothesize that this plasmid has appeared during the evolution of the symbiotic association between the aphid and the bacterial endosymbiont. The fact that this plasmid contains the entire leucine operon can be related to physiological evidence showing that the aphid host's diet of plant phloem is deficient in essential amino acids.

Amino Acid Sequence↗

Effect of dietary cholesterol on the pattern of osmium deposition in the symbiote-containing cells of the pea aphid.

Pea aphids left for 48 h in unbuffered osmium tetroxide show heavy staining of many organelles in the symbiote-containing cells (mycetocytes and sheath), embryos and oenocytes very similar to that characteristic of mammalian sterol-synthesizing cells. However, the staining of the per-aphid cells is, to a large extent, dependent on the presence of cholesterol benzoate, or free cholesterol, in the aphid's diet. In aphids cultured in vitro with 3H mevalonate in the presence of added cholesterol, the incorporation of label into the cholesterol and lanosterol fractions is significantly reduced. If the dietary cholesterol effects a similar inhibition in vivo, the cholesterol-dependent osmium staining could be due to precursors(s) of cholesterol accumulating in the intracellular sites described. There is also osmium staining of large (normally electron-transparent) vacuoles in mycetocytes, gut and fat body, irrespective of dietary cholesterol.

Animals↗

Ribosomal DNA is frequently concentrated on only one X chromosome in permanently apomictic aphids, but this does not inhibit male determination.

The ribosomal DNA arrays in the nucleolar organizer regions (NORs) of aphids are generally-located in a telomeric or subtelomeric position on the X chromosomes. In aphid populations or species that have lost the sexual part of their life cycle and become permanent apomicts, multiple rDNA copies are often concentrated on only one of the original X chromosomes. This situation has been found in apomictic members of three aphid subfamilies, although not as yet in the tribe Aphidini, which includes several permanent apomicts that are important pests. Some clonal cultures of Acyrthosiphon pisum and Myzus persicae reared for many years in conditions that prolong apomixis and inhibit sexual reproduction also have their rDNA arrays concentrated on one X chromosome. A 50-year-old clone of Aphis fabae, however, still retains a pair of similar-sized rDNA arrays on its X chromosomes. Although pairing of the X chromosomes by their NORs during prophase of the maturation division seems to be required for determination of XO males in aphids, a clone of A. pisum with one rDNA array was nevertheless able to produce males when subjected to appropriate environmental conditions.

Animals↗

Enhancement of resistance to aphids by introducing the snowdrop lectin gene gna into maize plants.

In order to enhance the resistance to pests, transgenic maize (Zea mays L.) plants from elite inbred lines containing the gene encoding snowdrop lectin (Galanthus nivalis L. agglutinin; GNA) under control of a phloem-specific promoter were generated through the Agrobacterium tumefaciens-mediated method. The toxicity of GNA-expressing plants to aphids has also been studied. The independently derived plants were subjected to molecular analyses. Polymerase chain reaction (PCR) and Southern blot analyses confirmed that the gna gene was integrated into maize genome and inherited to the following generations. The typical Mendelian patterns of inheritance occurred in most cases. The level of GNA expression at 0.13%-0.28% of total soluble protein was observed in different transgenic plants. The progeny of nine GNA-expressing independent transformants that were derived separately from the elite inbred lines DH4866, DH9942, and 8902, were selected for examination of resistance to aphids. These plants synthesized GNA at levels above 0.22% total soluble protein, and enhanced resistance to aphids was demonstrated by exposing the plants to corn leaf aphid (Rhopalosiphum maidis Fitch) under greenhouse conditions. The nymph production was significantly reduced by 46.9% on GNA-expressing plants. Field evaluation of the transgenic plants supported the results from the inoculation trial. After a series of artificial self-crosses, some homozygous transgenic maize lines expressing GNA were obtained. In the present study, we have obtained new insect-resistant maize material for further breeding work.

Agrobacterium tumefaciens↗

Ribosomal protein S1 (RpsA) of Buchnera aphidicola, the endosymbiont of aphids: characterization of the gene and detection of the product.

Buchnera aphidicola is a prokaryotic endosymbiont found in specialized cells of the aphid Schizaphis graminum. Many of the previously cloned B. aphidicola genes are preceded by a poor ribosome-binding site. Ribosomal protein S1 (RpsA) allows the translation of messenger RNAs that lack or have a poor ribosome binding site. We have cloned and sequenced a 4.5-kilobase (kb) B. aphidicola DNA fragment containing four open reading frames corresponding to aroA-rpsA-himD-tpiA. The deduced amino acid sequence of B. aphidicola RpsA was 75% identical to that of the Escherichia coli protein. The major difference was in the number of basic amino acids, which were present in higher numbers in B. aphidicola RpsA. Antiserum to E. coli RpsA was prepared and used to detect B. aphidicola RpsA in cell-free extracts of aphids. During the first 12 days of aphid growth there is a slight decrease in the amount of RpsA per unit of aphid weight. The three additional genes found on the 4.5-kb DNA fragment encoded for proteins involved in aromatic amino acid biosynthesis (aroA), DNA bending (himD), and carbohydrate metabolism (tpiA). The presence of these genes in B. aphidicola is additional evidence of its similarity to free-living bacteria.

Amino Acid Sequence↗

Molecular characterization of the leucine plasmid from Buchnera aphidicola, primary endosymbiont of the aphid Acyrthosiphon pisum.

The complete sequence of the leucine plasmid of Buchnera aphidicola from the aphid Acyrthosiphon pisum (pLeu-BAp) is reported. Its gene organization was concordant with those of other leucine plasmids of Buchnera from aphids of the Aphidini and Macrosiphini tribes. Three inverted repeats are present in pLeu-BAp. Two of them are also present in pLeu from the family Aphididae: (i) SIR1, located downstream the leucine operon, resembles a rho-independent terminator of transcription, and (ii) LIR1, located upstream of the leucine operon, is suggested to be involved in transcription termination or messenger stability. The third, located near the putative ATGC repeats involved in the origin of replication, is specific in aphids of the Macrosiphini tribe. Phylogenetic analyses based on sequences of leuA, leuB, leuC, leuD, repA1 and ORF1 showed a closer relationship between Buchnera (A. pisum) and Buchnera (Diuraphis noxia). However, tree topologies indicate that the split between both aphid species took place soon after the formation of the Macrosiphini lineage.

Animals↗

Predator-induced transgenerational phenotypic plasticity in the cotton aphid.

Transgenerational phenotypic changes, whereby offspring have an altered trait or a distinct alternate phenotype, frequently occur in response to increased maternal predation risk. The cotton aphid, Aphis gossypii (Glover), is unique, however, as offspring consist of four distinct phenotypes ("normal" light green apterae, "normal" dark green apterae, "dwarf" yellow apterae, and alatae), all with divergent life history patterns and resulting population dynamics. Here, we show that increased predation risk induces transgenerational phenotypic changes in cotton aphids. When exposed to search tracks from larval or adult convergent ladybird beetles, Hippodamia convergens Guerin-Meneville, cotton aphids produced greater numbers of winged offspring. In a subsequent experiment, apterous and alate individuals on clean plants were found to have primarily normal and dwarf offspring, respectively. We suggest that elevated predation risk may cause phenotypic changes in aphids over multiple generations, resulting in a more precipitous decline in herbivore populations than could be explained solely by increased predation rates.

Analysis of Variance↗

Doppler radar detection of exceptional mass-migration of aphids into Finland.

Our objective was to detect mass migrations of insects of economic significance by insect traps and a Doppler weather radar. Migrants were sampled by suction traps, tow nets and light traps in the Helsinki region. We used radar to observe the migrating insects, and trajectories to backtrack mass migrations of aphids (Homoptera, Aphididae) in spring 1988. The aphid migrations were clearly observed in trap catches and by radar. The first migration, mainly involving Euceraphis betulae, occurred on 18 May and was tracked back to northern Poland. The second migration, mainly of Rhopalosiphum padi (a serious pest of small-grain cereals), occurred 3 days later and was tracked back to a large area covering Latvia and western Russia south of St Petersburg. The third migration included both E. betulae and R. padi, and took place on 30 May. It originated from Estonia. Neither trap nor radar data provide exact quantitative information on migrations. Trapping efficiency depends strongly on wind speed and insect size. Radar echo intensity is very strongly related to the sizes of insects in the large volume of air measured, and the sizes are not known accurately. Weather data, especially temperature, can be used in predicting the development of aphids, and air-parcel trajectories in estimating the source areas of migrants. These methods for forecasting aphid migrations, combined with radar observations, are useful for warning purposes and to intensify insect trapping. This would contribute to more efficient agricultural pest management.

Agriculture↗

Unexpected effects of chitinases on the peach-potato aphid (Myzus persicae Sulzer) when delivered via transgenic potato plants (Solanum tuberosum Linné) and in vitro.

With the aim of producing insect-resistant potato plants, internode explants of Solanum tuberosum L. cv. Désirée were transformed with an Agrobacterium strain C58pMP90 containing an insect (Phaedon cochleariae: Coleoptera, Chrysomelidae) chitinase gene and the neomycin phosphotransferase (nptII) gene as selectable marker, both under the control of the viral CaMV 35S promoter. Three transformed potato lines (CH3, CH5 and CH25) exhibiting the highest chitinolytic activities were selected for feeding experiments with the peach-potato aphid, Myzus persicae (Sulzer), under controlled photoperiod and temperature conditions. Aphids fed on transgenic potato plants showed a reduced pre-reproductive period and an enhanced daily fecundity. Transgenic potato lines did not affect nymphal mortality, but improved several biological parameters related to aphid population's growth. Artificial diets were used to provide active (1, 10, 100 and 500 microg ml(-1)) and inactive (500 microg ml(-1)) bacterial (Serratia marcescens) chitinase to M. persicae. These compounds increased nymph survival at all active chitinase doses when compared to the control diet, while inactive chitinase did not. Although the pre-reproductive period was slightly shortened and the daily fecundity slightly higher, active and inactive chitinase provided as food led a reduction from 1 to 1.5 day population's doubling time. Therefore chitinase activity was responsible for the probiotic effects on aphids. Our results question the relevance of a chitinase-based strategy in the context of potato culture protection.

Animals↗

Improved detection of barley yellow dwarf virus in single aphids using RT-PCR.

The detection of a British isolate of barley yellow dwarf virus (BYDV-G, PAV-like) from individual vector aphids, using a combined assay of reverse transcription and polymerase chain reaction (RT-PCR) is reported. The method makes use of a multiplex format, including internal control primers directed at conserved regions of insect actin. The actin primers serve as controls for each stage of the method and are suitable for use in a range of invertebrate species. Detection of BYDV in vector aphids for use in forecasting systems is at present carried out using an enzyme-amplified ELISA system. In direct comparisons with the amplified ELISA, RT-PCR shows an increase in sensitivity detecting 11 fg of purified virus. Detection of virus in ELISA-negative aphids by RT-PCR was also demonstrated, and its potential as a routine diagnostic tool for virus detection in aphids is discussed.

Animals↗

Simulation modelling of the population dynamics of cereal aphids.

A simulation model explaining the population dynamics of the grain aphid (Sitobion avenae), a serious pest of wheat in Western Europe, is described. The model includes the effects of crop development and some natural enemies on the biology of the aphid. It is concluded that although much of the population dynamics of the aphid, especially in relation to its host plant, is now well understood, many uncertainties still remain concerning natural enemies. As these organisms seem able to prevent cereal aphid outbreaks, in some years, these gaps are presently preventing the development of a reliable forecasting scheme.

Animals↗

Gut microbe of aphid closely related to its intracellular symbiont.

Pea aphid, Acyrthosiphon pisum harbors prokaryotic intracellular symbionts and, at least, two species of gut microbes. It has been shown that the aphid symbiont is more closely related to Escherichia coli than to any other free-living bacterium. Analysis of RFLP of their 16S rDNA and groE homologues suggested that one of the gut microbes is a close relative of E. coli and that the aphid symbiont is still closer to this microbe than to E. coli. In addition, the optimal temperature for the growth of this gut microbe was significantly lower than that of E. coli suggesting that this microbe has been adapted to the ambient temperature of the host insect and shares a very close ancestor with the aphid symbiont in common.

Animals↗

A microarray approach identifies ANT, OS-D and takeout-like genes as differentially regulated in alate and apterous morphs of the green peach aphid Myzus persicae (Sulzer).

Aphids exhibit a complex life cycle in which their phenotypes vary in response to environmental changes. Wing polyphenism is one of the phenotypes that have not been adequately studied. We developed a cDNA microarray from the green peach aphid Myzus persicae (Sulzer), and compared mRNA samples derived from alate and apterous adults. The microarray experiment resulted in 108 significantly changing clones that represented 31 unique ESTs. Among the highest and significantly expressed clones in alate adults, seven clones showed portions of a new adenine nucleotide translocase (ANT) gene from M. persicae and four clones contained sequences of the OS-D gene. Another clone showed significant homology with the takeout (TO) and TO-like proteins described from other insects. A full-length cDNA clone of M. persicae ANT (Mp ANT) was isolated and characterized. An reverse transcription-polymerase chain reaction (RT-PCR) analysis showed significant differences in Mp ANT, OS-D and Mp takeout-like genes (Mp TOL) expression in alate compared to apterous adults. A polyclonal antibody against the beef heart mitochondrial ANT reacted with a recombinant Mp ANT expressed in Escherichia coli, as it did with aphid extracts, showing higher amounts of the expected 33kDa band in alate adults. Spatial expression analysis showed higher Mp ANT expression in the thorax compared to head, abdomen, leg and antennae. OS-D and Mp TOL showed ubiquitous expression across the body, with OS-D being significantly higher in legs and antennae while Mp TOL being significantly higher in the head and abdomen. The possible causes for higher expression of ANT, OS-D and TOL in alates and the physiological effects on the aphid flight and dispersal are discussed.

Amino Acid Sequence↗

Density-dependent induction and suppression of soldier differentiation in an aphid social system.

We investigated the mechanism underlying the control of soldier production in colonies of a social aphid, Tuberaphis styraci, which has a sterile soldier caste in the second instar. High aphid density was shown to induce soldier production in T. styraci. Analysis of natural colonies revealed that the soldier proportion tended to increase with aphid density but reached a plateau. Artificial diet experiments identified a similar plateau of soldier proportion under high-density conditions. In order to gain insights into the controlling mechanism of soldier production, the effect of soldiers on reproducing adult aphids was examined using the artificial diet system. It was experimentally demonstrated that soldier production was suppressed by coexisting soldiers, whereas coexisting non-soldiers facilitated soldier production. These results suggested that caste ratio in the colony of T. styraci is controlled by positive and negative feedbacks consisting of density-dependent induction and suppression of soldier differentiation.

Aggression↗

Effect of low and high-saponin lines of alfalfa on pea aphid.

Pea aphid fed on a high-saponin line of alfalfa showed reduction of reproduction and survival, and disturbance in development of its population. This line negatively influenced aphid probing behaviour, particularly prolonging the non-probing period and probing of the peripheral tissues (epidermis and mesophyll) and shortening the period of phloem sap ingestion. The high-saponin line of alfalfa differed from the low-saponin one by the presence of zanhic acid tridesmoside and a higher level of 3-GlcA,28-AraRhaXyl medicagenic acid glycoside. The saponins incorporated into sucrose-agarose gels significantly reduced number of the aphid probes into the gels and extended their duration in comparison to the control gels (without tested compounds). Role of zanhic acid tridesmoside and 3-GlcA,28-AraRhaXyl medicagenic acid glycoside as potential factors for partial resistance of alfalfa towards the pea aphid is discussed.

Animals↗

Response of the entomopathogenic fungus Pandora neoaphidis to aphid-induced plant volatiles.

We used a model plant-aphid system to investigate whether the aphid-specific entomopathogenic fungus Pandora neoaphidis responds to aphid-induced defence by the broad-bean plant, Vicia faba. Laboratory experiments indicated that neither in vivo sporulation, conidia size nor the in vitro growth of P. neoaphidis was affected by Acyrthosiphon pisum-induced V. faba volatiles. The proportion of conidia germinating on A. pisum feeding on previously damaged plants was significantly greater than on aphids feeding on undamaged plants, suggesting a direct functional effect of the plant volatiles on the fungus. However, there were no significant differences in the infectivity of P. neoaphidis towards A. pisum feeding on either undamaged V. faba plants or plants previously infested with A. pisum. Therefore, these results provide no evidence to suggest that P. neoaphidis contributes to plant indirect defence strategies.

Animals↗

A hemiterpene glucoside as a probing deterrent of the bean aphid, Megoura crassicauda, from a non-host vetch, Vicia hirsuta.

The bean aphid, Megoura crassicauda Mordvilko, feeds selectively on plants belonging to the genus Vicia (Fabaceae). However, it never infests the tiny vetch, V. hirsuta (L.) Gray. The aphid appeared to discriminate between host and non-host plants by tasting specific chemicals during penetration of its stylet into the plant tissues. The aphid, after being stimulated by specific probing stimulants, deposited characteristic proteinous stylet sheaths through a parafilm membrane, which has one side in contact with an extract solution of Vicia angustifolia. However, an addition of a V. hirsuta extract to the medium strongly inhibited the salivary sheath formation. A specific probing deterrent was isolated from a V. hirsuta extract by monitoring the inhibitory effect, and identified as (E)-2-methyl-2-butene-1,4-diol 4-O-beta-d-glucopyranoside. A mixture of the glycoside and the stimulatory V. angustifolia fraction in the same equivalency found in plants significantly decreased the probing activity in M. crassicauda. Since the stylet insertion process is a crucial step for the aphid's settlement on a plant, the glycoside seems to act as an effective chemical barrier for V. hirsuta.

Animals↗