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Ant-aphid association: role of aphid alarm pheromone.

When attacked by predators, aphids secrete alarm pheromones that cause nearby aphids to disperse. Ant-associated (myrmecophilous) aphid species disperse less readily than nonmyrmecophilous species. The ant Formica subsericea responds to aphid alarm pheromone in a way that is beneficial to the aphid. These findings support our hypothesis that myrmecophilous aphids depend more on ants for protection from predators than on their own dispersive powers.

Animals

Evidence for the establishment of aphid-eubacterium endosymbiosis in an ancestor of four aphid families.

Aphids (superfamily Aphidoidea) contain eubacterial endosymbionts localized within specialized cells (mycetocytes). The endosymbionts are essential for the survival of the aphid hosts. Sequence analyses of the 16S rRNAs from endosymbionts of 11 aphid species from seven tribes and four families have indicated that the endosymbionts are monophyletic. Furthermore, phylogenetic relationships within the symbiont clade parallel the relationships of the corresponding aphid hosts. Our findings suggest that this endocytobiotic association was established in a common ancestor of the four aphid families with subsequent diversification into the present species of aphids and their endosymbionts.

Animals

Adaptation to Plant Defence in an Agricultural Insect Pest: Integrating Genome Scans and Gene Expression in the Soybean Aphid Reveals Multi-Genic Pathways.

In agroecosystems, intense selection pressures cause species to adapt and spread, often leading to the evolution and persistence of pests. Understanding how pests rapidly adapt can help develop sustainable strategies for their management and improve agroecosystem health. Pest adaptation involves stable variations in DNA sequence, as well as dynamic shifts in gene expression, often mediated by non-coding regulatory elements. We examined adaptation to plant defences in the soybean aphid, Aphis glycines, in which virulent aphids have overcome plant defences and avirulent aphids have not. Previous data with laboratory colonies suggested that virulent aphids have higher overall gene expression, including transposable elements, some of which influence gene regulation. However, we lack information on how genetic variation in natural populations impacts adaptation and potentially gene regulation. We integrated population genome scans of field-collected, soybean aphid populations with gene expression profiles of virulent and avirulent laboratory colonies to uncover connections between genetic differentiation and gene regulation for virulence. Genome scan methods found 2144 single nucleotide polymorphisms (SNPs) with significant genetic differentiation (i.e., outliers) in field-collected populations. These SNPs were near 1004 genes, representing 5.16% of the effective number of genes. Based on previous RNA-Seq data with laboratory colonies, we found 3160 genes and 147 long non-coding RNAs (lncRNAs) with differential expression among virulent and avirulent biotypes. By integrating both data sets, we identified 16 genes and 5 long non-coding RNAs with differential expression and that were associated with an outlier SNP (within 10 kbp). We validated SNPs with additional field collected aphids and found an aphid clone with stronger virulence than our laboratory virulent colony, surviving on 2 different aphid-resistant soybean varieties. This new virulent clone had fixed allele differences at 9 SNPs compared to our avirulent and other virulent colony. Field collected soybean aphids matching the phenotype of this new virulent clone had significant genetic differentiation with 3 outlier SNPs near genes related to zinc transport and lachesin compared to field collected avirulent aphids. Our entire data reinforced the importance of a potential multi-genetic response to overcome plant defence and generates new insights into complex genetic and regulatory mechanisms involved in insect-plant interactions.

Animals

Viruses of the Potyviridae with non-aphid vectors.

The large majority of members of the family Potyviridae are aphid-transmitted. However, 17 viruses whose vectors are unknown have been classified as members of the genus Potyvirus. Loss of aphid transmissibility has been observed in some strains of several potyviruses. There are currently 11 members of the Potyviridae whose vectors are not aphids. These viruses with non-aphid vectors exhibit most of the characteristics of the family. Viruses of the Potyviridae induce cytoplasmic cylindrical inclusions in their hosts whether their vectors are aphids, non-aphids, or are unknown. The virus genome produces the inclusion protein and thus the viruses have related inclusion body gene sequences. Non-aphid-transmitted viruses of the Potyviridae also are serologically related to aphid-transmitted potyviruses.

Animals

Aphid symbiotic virus engineered for in vivo expression of insecticidal effectors.

Microbial pesticides are eco-friendly alternatives to chemical pesticides. However, few viral pesticides have been developed. Insects harbor diverse symbiotic viruses, which have the potential to be engineered for translational applications in pest control. Here, we engineered Acyrthosiphon pisum virus (APV), a symbiotic virus of the pea aphid, to deliver anti-aphid effectors using reverse genetics technology. A cytomegalovirus (CMV) promoter-driven APV infectious clone was successfully rescued in pea aphids with the assistance of nanocarrier star polymer (SPc). Based on this infectious clone, the protein coding sequence of chymotrypsin inhibitor variant 8 (Chy8) and the double-stranded RNA sequence targeting the aphid clip-domain serine protease (SPLP) were separately assembled into the APV genome to generate APV-Chy8 and APV-dsSPLP infectious clones, respectively. The recombinant APV clones reduced aphid relative survival rates by 34% and 17% by microinjection, respectively. To enhance the transcriptional efficiency, the APV-Chy8 and APV-dsSPLP clones were transcribed in vitro using the T7 promoter. The in vitro-synthesized APV-Chy8 and APV-dsSPLP clones reduced aphid relative survival rates by 48% and 45% by microinjection, respectively. These results demonstrate that engineered APV can deliver cargos and reduce aphid survival under injection-based experimental conditions, highlighting the potential of symbiotic virus-based vectors for delivering insecticidal effectors.

Animals

The longest 18S ribosomal RNA ever known. Nucleotide sequence and presumed secondary structure of the 18S rRNA of the pea aphid, Acyrthosiphon pisum.

An EMBL4 recombinant phage which encodes one of the full length of the aphid ribosomal DNA has been isolated from the aphid genomic library. Determination of the complete nucleotide sequence of the aphid 18S rRNA gene revealed that it is 2469 bp with a G + C content of 59%. The aphid 18S rRNA gene studied here is the longest and has the highest G + C content among the 18S rRNA genes examined so far. Evidence provided by the S1 nuclease assay suggests that the aphid 18S rRNA gene examined in this study is not a pseudogene containing an insertion sequence. Based on the nucleotide sequence of the 18S rRNA gene, we constructed a presumed secondary-structure model of the aphid 18S rRNA. In the aphid 18S rRNA, the eucaryote-specific E21 and 41 region are supposed to be longer and more complex than the counterparts of other 18S rRNA.

Animals

[An example of research on biological control: Entomophthora fungi pathogenic for aphids].

The results obtained in 15 years of research on the Entomophthorales pathogen of aphids showed the importance of the action of these fungi in the regulation of natural aphid populations and their possible use in agriculture as a biological control agent. Recent ecological studies on natural populations of aphids established the seasonal variation of the different fungal species and the diverse degrees of specificity between the species or groups of species of aphid and the various species of Entomophthora. The study of populations dynamics of an aphid species on a cultivated plant permitted the determination of the way a certain number of biotic and abiotic factors, such as temperature, humidity, thresholds of the insect population and of the infecting fungus lead to an epizootic development. If the air propagation of the disease by conidia is understood for a long time, the role of the soil as a reservoir for the infecting fungus has been demonstrated recently. Under favourable climatic conditions, the use of industrially produced resistant resting spores would allow the regulation of aphid populations in nature.

Animals

Functional analysis of the role of a wound-induced leucine aminopeptidase gene homologue isolated from Rorippa indica in aphid herbivory.

Leucine aminopeptidases (LAPs) are multifunctional enzymes with roles in both defence and development. In plants, they are reported to be induced by wound-inflicting Lepidopteran insects and regulate wound response pathways leading to an effective defence response. Infestation by Hemipteran mustard aphid, Lipaphis erysimi (L.) Kaltenbach has been reported to induce wound response as well as a wound-responsive Arabidopsis thaliana Lap1 homologue (RI01; GenBank Accession: JK034053) in Rorippa indica (L.) Hiern. This is interesting as Hemipteran insects like aphids are assumed to inflict minimal wounding. In the present study, starting with the RI01 sequence information, we isolated the full length (1566 bp) sequence of a novel R. indica Lap (RiLap) gene, performed in silico analyses and developed transgenic R. indica plants with suppressed RiLAP activity by expressing a 565 bp antisense fragment of RiLap cDNA. We found that the isolated RiLAP is an acidic LAP of M17 family and suppressing it causes a significant increase in aphid herbivory but reduction in total chlorophyll content and possibly photosynthetic capacity in aphid infested transgenic plants of the T1 generation. These findings though preliminary suggest that RiLap could have a role in deterring aphids by acting as a regulatory protein simultaneously balancing defence response and photosynthetic capacity or plant growth. Noting the dearth of research in this area, this pilot study will be useful for designing future in depth analyses in understanding the role of Laps in defence response against Hemipteran insects. The study has implications in the development of sustainable pest management avenues.

Leucyl Aminopeptidase

What causes the aphid 28S rRNA to lack the hidden break?

In order to determine why the aphid 28S rRNA lacks the hidden break otherwise found in insects, the structure of the region of the aphid ribosomal DNA (rDNA) corresponding to the gap region, which in other insect rDNA transcripts is excised posttranscriptionally, was studied. Sequence comparison suggested that, in contradistinction to what is found in rDNA transcripts of other insects, a stem-loop structure formed in this region of the aphid rDNA transcript is not AU-rich. Nor did the loop of the aphid molecule contain the UAAU tract that can be a signal for the introduction of the hidden break, suggesting that in this particular region the aphid 28S rRNA resembles 28S rRNAs of deuterostomes, which do not contain the hidden break.

Animals

Polyamino acid induced aphid transmission of plant viruses.

Aphids transmitted poly-L-ornithine (PLO)-treated tobacco mosaic virus (TMV) when given acquistion and inoculation access periods as brief as 30 s and 2 min, respectively; the ability to transmit was lost within 90 min. Aphids without claws were able to transmit the virus. Transmission thus seems similar to that of nonpersistent viruses. The ratio of virus to polyamino acid, as well as the KCl concentration, markedly affected transmission. Transmission was best from mixtures which contained 250 mug/ml TMV, 2-5 MUG/ML PLO (mol. wt. 120000) and 0-6 M-KCl. A similar mixture favoured transmission when poly-L-lysine (mol. wt. 85000) was substituted for PLO, but with poly-L-lysine (mol. wt. 30 000) it was necessary to decrease the KCl to 0-3 M to obtain transmission. Less KCl (0-08 to 0-24 M) also favoured aphid transmission of PLO-treated potato virus X and tobacco rattle virus. PLO-treated TMV ultracentrifuged in the presence of, and resuspended in, 0-6 M-KCl remained aphid transmissible while PLO-treated virus in 2 M-DCl, which favours greater dissociation of the virus-PLO complex, was transmissible neither before nor after sedimentation by ultracentrifuging, and resuspension in 0-6 M-KCl. these results show that transmissibility is not due to a permanent alteration of the virus by PLO and indicate that the formation of a TMV-PLO complex is required for transmission. Sequential acquisition experiments suggest that PLO may act by binding TMV to receptor sites in aphids. However, the possibility that PLO affects the infection process was not ruled out.

Animals

Chimeric tomato plants show that aphid resistance and triacylglucose production are epidermal autonomous characters.

Graft chimeras were generated using Lycopersicon pennellii and L. esculentum to determine the contribution of the three meristem layers (L1, L2, and L3) to trichome density, sugar ester production, and aphid resistance. Sugar esters, in the form of triacylglucoses, have been implicated in the aphid resistance of pennellii. One chimera possessed the epidermal layer (L1) of pennellii and the internal tissues (L2 and L3) of the aphid-susceptible esculentum. The second chimera had both the L1 and L2 of pennellii and the L3 of esculentum. Type IV trichome densities did not differ significantly among the chimeras and pennellii. Both chimeras accumulated sugar esters with similar sugar and fatty acid composition as pennellii. The concentration of epicuticular sugar ester on the chimeras was also comparable with that of pennellii. Leaf cage and feeding studies demonstrated that both chimeras are as resistant to aphids as is pennellii. The resistance could be reduced similarly on all three types of plants by removal of the type IV trichome exudate. These results indicate that the presence and density of the type IV trichomes and the amount and type of sugar esters produced are features determined by the genotype of the epidermis. These epidermal features are sufficient to account for the aphid resistance observed in pennellii.

Animals

Honeydew of aphids as a source of sugar for Phlebotomus ariasi.

Wild-caught Phlebotomus ariasi Tonnoir, starved until their sugar meals had been digested, were caged for 24 h with plants or plants infested with aphids and then tested for fructose, a constituent of plant sap. No evidence was found that the flies took sap directly from nine types of plants present in their habitat. About two-thirds of flies caged with oak (Quercus ilex L.) infested with an aphid [Lachnus roboris (L.)] and about a fifth kept with leaves of the french bean (Phaseolus vulgaris L.) infested with an aphid took honeydew. Experiments with three other aphid species gave negative results. Of sandflies caught in an oak tree infested with aphids, half of the females and three-quarters of the males contained fructose. Tests on females caught in a house and grouped according to state of engorgement showed that, having taken blood, the females stop taking sugar until the bloodmeal is completely digested. It is predicted that honeydews are important in the development of Leishmania infantum Nicolle in the gut of P. ariasi.

Animals

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

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

The properties of a carboxylesterase from the peach-potato aphid, Myzus persicae (Sulz.), and its role in conferring insecticide resistance.

Carboxylesterases from different strains of Myzus persicae were examined to try to understand their contribution to insecticide resistance. Preliminary evidence that they are involved comes from the good correlation between the degree of resistance and the carboxylesterase and paraoxon-degrading activity in aphid homogenates. Furthermore the carboxylesterase associated with resistance could not be separated from the insecticide-degrading enzyme by electrophoresis or ion-exchange chromatography. Homogenates of resistant aphids hydrolysed paraoxon 60 times faster than did those of susceptible aphids, yet the purified enzymes from both sources had identical catalytic-centre activities towards this substrate and also towards naphth-1-yl acetate, the latter being hydrolysed by both 2x10(6) times faster than paraoxon. These observations provide evidence that the enzyme from both sources is identical, and that one enzyme hydrolyses both substrates. This was confirmed by relating the rate of paraoxon hydrolysis to the rate at which paraoxon-inhibited carboxylesterase re-activated. Both had the same first-order rate constant (0.01min(-1)), showing clearly that the hydrolysis of both substrates is brought about by the same enzyme. Its K(m) for naphth-1-yl acetate was 0.131mm, and for paraoxon 75pm. The latter very small value could not be measured directly, but was calculated from substrate-competition studies coupled with measurements of re-activation of the diethyl phosphorylated enzyme. Since the purified enzymes from resistant and susceptible aphids had the same catalytic-centre activity, the 60-fold difference between strains must be caused by different amounts of the same enzyme resulting from mutations of the regulator gene(s) rather than of the structural gene.

Animals