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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

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

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

[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

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

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

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

Reproductive strategy of winged and wingless morphs of the aphids Sitobion avenae and Metopolophium dirhodum.

The reproduction of apterous and alate morphs of the aphids Sitobion avenae and Metopolophium dirhodum is compared on the basis of fecundity in 5- and 10-day periods of adult life. Apterae of both species are consistently more fecund than alatae of comparable weight, producing about three more nymphs on average in any 5-day period. The reproductive differences are related to the number and quality of embryos at eclosion and to ovulation rates, both of which in turn appear to be linked to wing-muscle maintenance. These relationships between weight, embryos and reproduction may be used to predict a newly moulted adult aphid's fecundity, a method which may facilitate the assessment of resistance to aphids in new cereal varieties, by obviating lengthy recording of reproduction. The strategies by which alatae of these and other aphid species minimize the difference between their fecundity and that of apterae are discussed.

Age Factors

[A comparative study of the substrate and inhibitor specificity of the glutathione transferase from the spring grain aphid (Schizaphis gramina Rond.) and from rat liver].

Kinetic parameters of 9 substrates interaction with glutathione transferase (GST) from spring grain aphid and rat were studied. The most significant difference in Vmax values was noticed for 4-nitropyridine-N-oxide (6 times higher for aphid) and ethacrynic acid (7 times higher for rat). Km values were practically in all cases higher for aphid GST as compared to rat GST. New class of effectors of GST suggested by us, that is azimines (2 series), was used for the inhibitor analysis. GST interaction with these inhibitors was appreciated by three types of activity: nucleophilic replacement, thiolysis and N-deoxygenation. It has been shown that the degree of GST inhibition depended considerably both on the GST source and the substrate used. New high-effective inhibitors of GST were found among azimines and their higher specificity to rat GST as compared to aphid GST was demonstrated especially in thiolysis reaction.

Animals

Analysis of Duplication and Potential Functional Divergence of Wing Gene Network Components in Pea Aphids.

A fundamental focus of evolutionary developmental biology is uncovering the genetic mechanisms responsible for the gain and loss of characters. One approach to this question is to investigate changes in the coordinated expression of a group of genes important for the development of a character of interest (a gene regulatory network). Here we consider the possibility that modifications to the wing gene regulatory network (wGRN), as defined by work primarily done in Drosophila melanogaster, were involved in the evolution of wing dimorphisms of the pea aphid (Acyrthosiphon pisum). We hypothesize that this may have occurred via changes in expression levels or by duplication followed by divergence of wGRN components. To test this, we annotated members of the wGRN in the pea aphid genome and assessed their expression levels in first and third nymphal instars of winged and wingless morphs of males and asexual females. We find that only 2 of the 32 assessed genes exhibit morph-biased expression. We also find that three wing genes (apterous (ap), warts (wts), and decapentaplegic (dpp)) have undergone gene duplication. In each case, the resulting paralogs show signs consistent with functional divergence, exhibiting either sex-, morph-, or stage-specific expression. Two gene duplicates, wts2 and dpp3, are of particular interest with respect to wing dimorphism, as they exhibit male morph-specific isoforms and wingless male-biased expression, respectively. These gene expression results provide an important first step toward identifying members of the pea aphid wGRN that may play a causative role in differentiating winged from wingless morphs. These findings supplement our understanding of trends in developmental gene network evolution, such as side-stepping pleiotropic constraint via duplication and sub-functionalization, underlying the emergence of novel phenotypes.

Animals

In vivo sterol biosynthesis by pea aphid symbiotes as determined by digitonin and electron microscopic autoradiography.

Pea aphid primary symbiotes have previously been shown to synthesize cholesterol in vitro. Two electron microscopic techniques were used here to determine whether the symbiotes also synthesize cholesterol in vivo and whether this cholesterol is made available to the aphid. We also inquired into a possible role of secondary symbiotes in chosesterol biosynthesis. Treatment of aphids with digitonin resulted in significant alteration of ultrastructural sites in primary and secondary symbiote membranes. We concluded that these sites are areas of high cholesterol concentration in the symbiotes. Electron microscopic autoradiography with 3H-mevalonate precursor indicated that both primary and secondary symbiotes synthesize cholesterol; in both cases, the majority of grains were associated with the symbiote membranes. While the frequency of grains on the symbiotes remained constant, irrespective of incubation time in labelled media, the frequency of grains over surrounding tissues increased exponentially as the time of incubation was increased from 30 min to 8 h, indicating that symbiote cholesterol is transported to other tissues. High voltage electron microscopic autoradiography permitted thick section autoradiography, reducing the time of emulsion exposure from 54 days (thin section) to 12 days (0.5 mum sections).

Animals

Molecular and transcriptional regulation of plant defense responses to aphid infestation.

Aphids are one of the important agricultural pests causing substantial yield losses in crops grown across the globe. Aphids are known to cause direct feeding damages and indirect losses due to sooty mold development and plant virus transmission. Plants respond to these attacks by mounting a complex defense response at the infested sites and systemic levels. This multilayered defense response involves a highly coordinated network of phytohormones and other signalling components like Ca2+, mitogen activated protein kinases and reactive oxygen species. Key to these complex responses is a well-regulated gene expression involving several transcription factors. A wide range of transcription factors are structurally and functionally characterized across some model plants and in a few agronomically important crops. These transcription factors play diverse roles such as defense gene expression modulation, regulation of hormone signaling, secondary metabolism, oxidative stress response, cell wall modifications, and phloem-based defense. Understanding the integration of signaling pathways, hormone crosstalk, and transcription factor mediated regulation provides a framework for practical applications, including breeding, genome editing, and elicitor-based strategies. This review highlights how plant defense signaling and transcriptional regulation against aphids can be harnessed to develop sustainable and novel pest management solutions.

Aphid

Rapidly evolving aphid gall effector proteins exhibit saposin-like folds.

Many insects manipulate plants by injecting effector proteins. In one extreme example of this molecular "hijacking," Hormaphis cornu aphids inject bicycle proteins into Hamamelis virginiana, contributing to the development of novel organs called galls. Bicycle proteins share no amino acid sequence similarity with proteins of known function. Here, we report the crystal structures of two divergent bicycle proteins. Both proteins contain saposin-like folds: one with multiple disulfide bonds exhibits a swapped domain topology; the other has no disulfide bonds and possesses two distinct, tandem domains. To explore the structural evolution of bicycle proteins, we attempted to predict bicycle protein structures with Alphafold2 (AF2) and other deep learning programs. While AF2 did not recover the two experimental structures using existing databases, it succeeded when provided with multiple sequence alignments (MSAs) of protein sequences from newly sequenced closely related species. Using this approach, we generated 2,400 high-confidence bicycle protein predictions from seven aphid species. While all aphid bicycle proteins contain predicted saposin-like folds, they display a vast diversity of structural and physicochemical properties. While this diversity thwarts prediction of conserved functions encoded in structure, it suggests that bicycle proteins have evolved to target diverse plant processes and/or to evade plant immune surveillance. Our extension of AF2 with custom MSAs of proteins from closely related species provides a generalizable, powerful approach for predicting structures of rapidly evolving protein families.

Animals

Rapidly evolving aphid gall effector proteins exhibit saposin-like folds.

Many insects manipulate plants by injecting effector proteins. In one extreme example of this molecular "hijacking", Hormaphis cornu aphids inject bicycle proteins into Hamamelis virginiana (Witch Hazel), contributing to the development of novel organs called galls. Bicycle proteins share no amino acid sequence similarity with proteins of known function. Here, we report the crystal structures of two divergent bicycle proteins. Both proteins contain saposin-like folds: one with multiple disulfide bonds exhibits a helix swap; the other has no disulfide bonds and possesses two tandem domains. To explore the structural evolution of bicycle proteins, we predicted bicycle protein structures with Alphafold2 (AF2). While AF2 did not recover the two experimental structures using existing databases, it succeeded after we provided multiple sequence alignments (MSAs) containing protein sequences encoded in new genome sequences from closely related aphid species. Using this customized approach at scale, we generated 2400 high-confidence predictions for bicycle proteins from seven aphid species. This dataset revealed that bicycle proteins without cysteines are outliers in fold space and appear to have evolved from ancestral proteins with disulfide-bonded saposin-like folds. While all bicycle proteins contain predicted saposin-like folds, they display a vast diversity of structural and physicochemical properties. While this diversity thwarts prediction of conserved functions encoded in structure, it suggests that bicycle proteins have evolved to target diverse plant processes and/or to evade plant immune surveillance.

AlphaFold predictions

Ultrastructure of the antennal sensilla of aphids. I. Coeloconic and placoid sensilla.

An electron microscopical study was made of the coeloconic and placoid sensilla on the antennae of the aphids Aphis pomi, Macrosiphum euphorbiae, Nasonovia ribis-nigri, and Pemphigus bursarius. Scanning electron microscopy revealed some variation in morphology which may be functionally important but is more likely to reflect the evolution of these species. The placoid sensilla were shown by tranwo or three neurons is surrounded by two ensheathing cells. The ciliary regions of the dendrites pass through a vacuole into a cavity between an outer and an inner cuticle where they may be connected to the dendritic branches although such connections were no seen. Small pores (8 nm diameter) partially penetrate the cuticle implying that these sensilla have an olfactory function. They are suggested to be important in host selection by alate aphids. The coeloconic sensilla are poreless pegs with nonsensory cuticular projections at their tips. The distal portions of their dendrites contain densely packed microtubules and the cellular arrangement of the sensila is similar to that of the placoid sensilla. It is suggest that they may function as thermoreceptors.

Animals

Preliminary mapping of wheat (Triticum aestivum L.) tolerance genes to the English grain aphid (Sitobion avenae Fabricius) by genome-wide association study.

Six Sitobion avenae-tolerant wheat accessions, mapped 110 associated SNPs and six candidate genes were identified, providing valuable genetic resources for breeding wheat with tolerance to S. avenae. Wheat tolerance to the English grain aphid (Sitobion avenae) is rarely incorporated into integrated pest management strategies for wheat fields. The scarcity of tolerant accession and insufficient mapping of tolerance-related gene are key limiting factors. To address these gaps, 640 wheat accessions were evaluated for S. avenae tolerance, combined with genome-wide association study (GWAS) and qPCR validation. Six wheat accessions with stable tolerance were identified: Lerma Rojo 64, AC Vista, Hanxuan 10, Zimai, Ningnuomai 1, Louguding. A total of 110 single nucleotide polymorphism (SNP) loci associated with tolerance to S. avenae were mapped, and six candidate genes (TraesCS2D03G0041800, TraesCS2Dnew048215, TraesCS2D03G0046300, TraesCS6B03G0655800, TraesCS2Dnew048223, TraesCS2D03G0040800) were examined for transcriptional responses following aphid infestation via qRT-PCR. These genes are involved in cellular redox homeostasis, ADP-binding-mediated defense, and photosystem II (PSII) functionality. This study provides valuable genetic resources for breeding wheat with tolerance to S. avenae and lays a foundation for subsequent functional validation of these tolerance genes and its molecular mechanism exploration.

Animals

Arthropod ribosomes. Integrity of ribosomal ribonucleic acids from aphids and water fleas.

The 28 S aphid rRNA differs from those of the other insects in two points: (1) it lacks the primary nick; (2) it is larger by 0.2 - 106 daltons. The 28 S rRNA from water fleas also, like that from crayfish, is larger than those from comon insects by 0.2 - 106 daltons. These crustacean 28-S rRNAs were shown to contain the primary nick, which is probably not located in the central point. The 18-S rRNAs from aphids and water fleas had molecular weights significantly larger than 0.7 - 106, the common vaalue for the eukaryotic 18 S rRNA. It was was suggested that the 3'-terminal base sequences of these RNAs are different from the common sequences of these RNAs are different from the common sequence proposed for the 18 S rRNAA of eukaryotes. These exceptional characteristics of the rRNAs from the parthenogenetic animals may provide a probe for general functions of the rRNA in the eukaryotic ribosomes.

Animals