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The small size and extensive polymorphisms of aphids make it difficult to identify larvae and adults solely based on their morphology. Here, we present an identification tool for the developmental stages of Acyrthosiphon pisum (Hemiptera: Aphididae) based on deep learning as a proof of concept. You Only Look Once (YOLO) algorithm is one of the most effective deep learning techniques for object detection. Although several studies have been conducted using deep learning technology for the detection and counting of tiny pests, the type of light source and size of the images were the limiting factors, as training was highly focused on uniform datasets and small insects. One way to overcome this problem is to introduce many types of datasets obtained from various light sources and microscopic magnifications. This strategy minimizes errors and omissions in aphid detection across all developmental stages in aphid individuals to the greatest extent possible. The experimental results showed that our modified YOLOv8 model could obtain over 95.9% and 99% accuracy for mean average precision (mAP) and recall, respectively, under various light sources, such as yellow, white, and natural light, and stereomicroscope magnifications. This study showed an improved accuracy of aphid recognition at all developmental stages. The study presents a novel deep learning model utilizing the YOLO algorithm to identify developmental stages of A. pisum. This model achieves high accuracy across various light sources and magnifications, thereby enhancing aphid biology studies. [ABSTRACT FROM AUTHOR]

The alfalfa virome has been understudied. Existing research dealing with viral incidences within the crop primarily concentrate on local factors, rather than considering a broader perspective. In this comprehensive 2‐year study, we define the alfalfa virome and the main local and landscape factors affecting the incidence and annual increase of the Alfalfa mosaic virus (AMV), the most prevalent virus in alfalfa. The study was conducted in commercial alfalfa fields located along the highly productive northeast region of the Iberian Peninsula. For the first time in Europe, next‐generation sequencing revealed the presence of 14 different viruses representing the genera Cytorhabdovirus, Alphapartitivirus, Amalgavirus, Alfamovirus, Luteovirus, Enamovirus and Flavivirus. AMV was the most prevalent species, accounting for 89% of the identified viral contigs. Enzyme‐linked immunosorbent assays showed that the incidence of AMV varied between fields, with the average incidence doubling from 34% in 2019 to 65% in 2020. To assess the effect of local and landscape characteristics on the incidence of AMV and the observed annual increase, we selected different local variables and recorded landscape structure at three different buffer radii (250, 500 and 1000 m) from the centre of each field. Both the incidence and annual increase in AMV were driven mainly by local characteristics. The incidence of AMV showed a significant relationship with crop age and field area, whereas the annual increase was mainly influenced by crop age and the cumulative number of alate morphs of the aphid Acyrthosiphon pisum. Only one landscape composition variable, the percentage of alfalfa, showed a significant relationship with AMV incidence at the 250‐m scale. These results confirm the effect of local variables on the population structure of generalist viruses. [ABSTRACT FROM AUTHOR]

The host immunosuppression by parasitic wasps is an important component of the host regulation strategy. The venom injected at the oviposition is one of the key‐factors involved in this host alteration and, in some parasitoids, its immunosuppressive role is complemented by wasp's symbionts. Most studies in this research area are related to hosts belonging to Lepidoptera and Diptera, for which a strong immune response is observed, whereas little is known for hemimetabolous host species, characterized by apparently much weaker defense barriers. To fill this research gap, here we focus on the host–parasitoid system Acyrthosiphon pisum (Harris) (Hemiptera: Aphididae) – Aphidius ervi Haliday (Hymenoptera: Braconidae). We functionally characterized a serine protease homolog (AeSPH) protein in vivo, identified in the venom of the aphid endoparasitoid A. ervi, generating AeSPH‐depleted female wasps by RNA interference and evaluating their capacity to successfully parasitize the host. Parasitism success rate was negatively affected by AeSPH knockdown and associated with an increased phenoloxidase (PO) cascade activation in aphids, scored by measuring PO enzymatic activity and the expression of phenoloxidase activating factor 2, a proPO‐activating gene upregulated in response to A. ervi parasitism. Our results indicate that AeSPH contributes to parasitism success by inhibiting the melanization response of the host, which is therefore an important component of the defense barriers involved in the parasitoid egg suppression. The ongoing studies on other virulence factors in A. ervi venom will allow to further characterize the immunosuppression strategy and its possible broader role in the host regulation through its action on aphid symbiont development. [ABSTRACT FROM AUTHOR]

Simple Summary: This study explores the mechanisms affecting offspring sex ratios in the pea aphid, Acyrthosiphon pisum (Harris), a key issue in evolutionary ecology. It tests the impacts of the presence of both the same and different clones, as well as juvenile hormone III (JH III) levels, on offspring sex allocation. Using red and green clones and the agar method, this study set up three initial treatments using sexuparae and tracked daily offspring number and sex. In a mixed-clone treatment, 1 sexupara of the green clone and 20 oviparous females of either red (1G + 20Rov) or green (1G + 20Gov) clone were transferred onto agar leaves, with a control (1G) established for comparison. After the production of sexuparae, JH III doses were applied to them, and hormone titers and sex allocation in offspring were meticulously measured. The results highlighted marked variations in sex allocation, especially an increase in ovipara number in mixed-clone treatment groups. JH III application reversed this effect, indicating that mixed-clone treatment leads to lowered JH titers, which in turn affects sex allocation. This study concludes that A. pisum sexuparae can modify offspring sex allocation in response to adjacent clones, showing diverse mechanisms of sex allocation, where JH III plays a critical role. The mechanism controlling sex allocation in the pea aphid, Acyrthosiphon pisum (Harris), remains a crucial yet unresolved issue in the field of evolutionary ecology. This study aims to assess the influence of the presence of both self and non-self clones, along with juvenile hormone III (JH III) titer, on the sex allocation of aphid offspring. To this end, red and green clones were utilized as experimental subjects, and the agar method was employed. Initially, three distinct experimental treatments were established using sexuparae, and the daily offspring count and sex allocation in each treatment zone were recorded. Subsequently, an additional experimental condition involving mixed-clone treatments was introduced. This procedure entailed the transfer of a single sexupara and 20 oviparous females from either the red (1G + 20Rov) or green clone (1G + 20Gov) onto a leaf on agar medium. Simultaneously, a control setup with a new sexupara (1G) was established. Three days following sexupara production, a dose of 0, 25, or 50 ng of JH III was applied to the aphids' abdomens. Subsequently, the titers of JH III in the sexuparae across each treatment group were quantified, and the extent of sex allocation was tallied. The findings demonstrated pronounced disparities in sex allocation among the various treatments and, notably, a substantial increase in the total offspring and oviparous number in the mixed-clone treatment group. The effects of mixed-clone treatment on the sex allocation patterns of the sexupara progeny could be determined by the application of exogenous JH III, indicating that JH may mediate the effects of mixed-clone treatment on sex allocation. Consequently, it can be concluded that A. pisum sexuparae possess the capability to modulate their sex allocation in response to the nature of adjacent competitor clones, thereby demonstrating a variety of sex allocation patterns. Throughout this process, JH III plays a pivotal role. [ABSTRACT FROM AUTHOR]

One of the major insect pests in Pisum sativum L. (is Acyrthosiphon pisum Harris (Hemiptera: pests in Pisum sativum L. (Hemiptera: Aphididae) is Acyrthosiphon pisum Harris (Hemiptera: Aphididae). An effective strategy for aphid control is the resistant host plant use. The current study aimed to identify resistance mechanisms and assess biochemical and morphological markers of pea aphid resistance in pea accessions. Meteorological variables affected the pea aphid density, which positively correlated with temperature, while precipitation amount and humidity negatively impacted. The aphid number was significantly and positively associated with the leaf area and the nitrogen content but negatively correlated with calcium and phosphorus levels. The pea aphid-resistant cultivars L 123-7-11, L 128-1and L 125-5 had small leaf areas, and high phosphorus and calcium content but a low nitrogen level. In the mutual influence of the plant indicators, phosphorus concentration had the highest negative impact on pea aphid density, followed by calcium. The plant marker inclusion in the pea breeding process is an efficient tool for a substantial selection program improvement for aphid resistance. Therefore, resistant host plants are essential tools promoting considerable selection program improvement for aphid resistance in the P. sativum breeding process and helping develop sustainable and environmentally friendly agriculture. [ABSTRACT FROM AUTHOR]

BACKGROUND: In recent years, the use of the RNA interference technology (RNAi) has emerged as one of the new strategies for species‐specific control of insect pests. Its specificity depends on the distinctiveness of the target gene sequence for a given species. In this work, we assessed in the pea aphid Acyrthosiphon pisum (A. pisum) the use of a double‐stranded RNA (dsRNA) that targets the β2 divergent nicotinic acetylcholine receptor (nAChR) subunit (dsRNA‐β2), which shares low sequence identity with other subunits, to control populations of this pest at different developmental stages. Because nAChRs are targeted by neonicotinoid insecticides such as imidacloprid, we also assessed the effect of dsRNA‐β2 coupled to this insecticide on aphid survival. Finally, because the effect of a control agent on beneficial insect must be considered before any use of new pest management strategies, the acute toxicity of dsRNA‐β2 combined with imidacloprid was evaluated on honeybee Apis mellifera. RESULTS: In this work, we demonstrated that dsRNA‐β2 alone has an insecticidal effect on aphid larvae and adults. Moreover, dsRNA‐β2 and imidacloprid effects on aphid larvae and adults were additive, meaning that dsRNA‐β2 did not alter the efficacy of imidacloprid on these two developmental stages. Also, no obvious acute toxicity on Apis mellifera was reported. CONCLUSION: Using RNAi that targets β2 divergent nAChR subunit is effective alone or combined with imidacloprid to control A. pisum at larval and adult stages. Because no obvious Apis mellifera mortality has been reported, this RNAi‐based pest management strategy should be considered to control insect pest. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Simple Summary: Pea aphids (Acyrthosiphon pisum) serve as a model insect for ecological research and have recently emerged as a significant pest of alfalfa. Amino acid uptake and balance in insects typically rely on amino acid transporters. In this investigation, RT-qPCR was employed to examine the distinct gene expression patterns of seven amino acid transporters subsequent to pea aphid feeding on resistant and susceptible alfalfa varieties. RNA interference targeting the pea aphid ACYPI000536 and ACYPI004320 genes was carried out via a plant-mediated approach, elucidating the preliminary functions of these genes. The study revealed that down-regulating the ACYPI000536 gene led to increased histidine and lysine levels in pea aphids, subsequently elevating mortality post-feeding on the susceptible alfalfa variety "Lie Renhe". Conversely, down-regulating the ACYPI004320 gene resulted in elevated phenylalanine levels in pea aphids, leading to reduced mortality following feeding on the highly resistant alfalfa variety "Gannong 5". In recent years, pea aphids have become major pests of alfalfa. Our previous study found that "Gannong 5" is a highly aphid-resistant alfalfa variety and that "Lie Renhe" is a susceptible one. The average field susceptibility index of "Gannong 5" was 31.31, and the average field susceptibility index of "Lie Renhe" was 80.34. The uptake and balance of amino acids in insects are usually dependent on amino acid transporters. RT-qPCR was used to detect the relative expression levels of seven amino acid transporter differential genes in the different instar pea aphids fed on resistant and susceptible alfalfa varieties after 24 h, and two key genes were selected. When pea aphids fed on "Gannong 5", the expression of ACYPI004320 was significantly higher than that in pea aphids fed on "Lie Renhe"; however, the expression of ACYPI000536 was significantly lower than that in pea aphids fed on "Lie Renhe". Afterward, the RNA interference with pea aphid ACYPI000536 and ACYPI004320 genes was performed using a plant-mediated method, and gene function was verified via liquid chromatography–mass spectrometry and pea aphid sensitivity to aphid-resistant and susceptible alfalfa varieties. The results showed that the down-regulation of the ACYPI000536 gene expression led to an increase in the histidine and lysine contents in pea aphids, which, in turn, led to an increase in mortality when pea aphids fed on the susceptible variety "Lie Renhe". The down-regulation of the ACYPI004320 gene expression led to an increase in phenylalanine content in pea aphids, which, in turn, led to a decrease in mortality when pea aphids fed on the resistant variety "Gannong 5". [ABSTRACT FROM AUTHOR]

Introduction: The pea aphid, Acyrthosiphon pisum, is a typical sap-feeding insect and an important worldwide pest. There is a primary symbiont-Buchnera aphidicola, which can synthesize and provide some essential nutrients for its host. At the same time, the hosts also can actively adjust the density of bacterial symbiosis to cope with the changes in environmental and physiological factors. However, it is still unclear how symbionts mediate the interaction between herbivorous insects' nutrient metabolism and host plants. Methods: The current study has studied the effects of different host plants on the biological characteristics, Buchnera titer, and nutritional metabolismof pea aphids. This study investigated the influence of different host plants on biological characteristics, Buchnera titer, and nutritional metabolism of pea aphids. Results and discussion: The titer of Buchnera was significantly higher on T. Pretense and M. officinalis, and the relative expression levels were 1.966±0.104 and 1.621±0.167, respectively. The content of soluble sugar (53.46±1.97µg/mg), glycogen (1.12±0.07µg/mg) and total energy (1341.51±39.37µg/mg) of the pea aphid on V. faba were significantly higher and showed high fecundity (143.86 ±11.31) and weight (10.46±0.77µg/mg). The content of total lipids was higher on P. sativum and T. pretense, which were 2.82±0.03µg/mg and 2.92±0.07µg/mg, respectively. Correlation analysis found that the difference in Buchnera titer was positively correlated with the protein content in M. officinalis and the content of total energy in T. pratense (P < 0.05). This study confirmed that host plants not only affected the biological characteristics and nutritional metabolism of pea aphids but also regulated the symbiotic density, thus interfering with the nutritional function of Buchnera. The results can provide a theoretical basis for further studies on the influence of different host plants on the development of pea aphids and other insects. [ABSTRACT FROM AUTHOR]

Aphidius smithi (Aphidiinae: Hymenoptera) is an important endoparasitoid of pea aphid (Acyrthosiphon pisum) which has been utilized to determine the effectiveness of this agent in reducing pest damage. Biology of the A. smithi reared on A. pisum in the laboratory at 23± 1°C has been studied. The development cycle of A. smithi from larvae to adult was completed in about 11 days. The pre-mating period of males (n=10) varied between 5 and 8 min (mean: 4 min). Copulation time (n = 10 pairs) was between 33 and 55 sec (mean: 45.2 sec). Oviposition time (n = 10 females) was between 1 and 2 sec (mean: 1.5 sec). Female parasitoids lived longer (5.75 days) than male parasitoids (4 days) when offered honey and water as a food diet. Lifespan of adult male and females was shorter i.e. 2.25 and 2.75 days respectively when fed upon dissected aphid. When adult parasitoids were released on pea plant provided with pea aphid, mean period of life for male and female was 4.75 and 6 days respectively. Sex ratios of field collected mummies were female biased (60%). Two species of hyperparasitoids viz. Asaphes suspensus (68%) and Pachyneuron aphidis (54%) were involved. Examination for phenotypic polymorphism showed that field population of A. smithi contained both dark and light pigmentation pattern of abdominal segments while laboratory reared samples were only dark pigmented in both sexes. The findings of this study can help in defining strategies for the rearing to release this parasitoid in biological control programs against A. pisum in Pakistan. [ABSTRACT FROM AUTHOR]

The changing environmental conditions can affect insect biology over multiple generations and phenotypic plasticity is important for coping with these changes. Transgenerational plasticity occurs when the environment in which the parents developed influences the plastic response of the offspring phenotype. In the present study, the plastic effects of resource limitation on important life history traits such as body size, fecundity, survival, and resistance to starvation of the pea aphid Acyrthosiphon pisum were investigated over two generations. This study focused on understanding how resource limitation can determine an adaptive expression of maternal effects and transgenerational plasticity in fitness-related traits. Aphids showed phenotypic plasticity for the life history traits investigated, as they performed better when grown in an optimal environment than in a resource-poor one. Also, aphids had a poorer performance if their mothers were raised in a resource-poor environment. The effects of transgenerational plasticity were observed only in response to resistance to starvation, through increased survival in the offspring of the mother reared in a resource-poor environment, suggesting an evolutionary bet-hedging strategy. The results of this study showed that the effects of adaptive transgenerational plasticity may be partially masked in stressful environments, where developmental problems instead predominate. More information on the transgenerational response to resource limitation across generations can contribute to a better understanding of aphid biology.

Alfalfa (Medicago sativa L.) hosts several species of aphid, Acyrthosiphon pisum (Harris), Aphis craccivora Koch and Therioaphis trifolii (Monell). The preference of the aphids of alfalfa plants for dense assemblies or individual plants, as well as for healthy or infested plants, was investigated in the field as in the laboratory. Years of field research have revealed the specific preferences of all three species of aphid. A. pisum and T. trifolii are most commonly found in alfalfa crops, while A. craccivora is mostly found on alfalfa weeds. Also, a single species of aphid alone is usually present on a plant. In order to determine the reason for this clear preference and to establish whether at the very beginning, i.e. at the stage of choosing a host, aphid species distance themselves from each other, we tested the effect of the volatiles of healthy and infested plants on their attractiveness to aphids. A. craccivora is repelled by the volatiles of dense crops and plants previously infested with one of the other two species. A. pisum and T. trifolii choose a dense assembly of plants, repelled by the volatiles of plants previously infested with A. craccivora. A. pisum displays the weakest competitive traits, and A. craccivora the strongest. This research showed that competition between aphid species does not occur only when they find themselves on the same plant at the same time, fighting for resources, but also in the choice of plant, in order to avoid later competition. [ABSTRACT FROM AUTHOR]

Cordyceps javanica (Frieder. & Bally) (Hypocreales: Cordycipitaceae) is an important entomopathogenic fungus that is used as a mycoinsecticide to control pests such as aphids and whiteflies. However, few studies have been done on the pathogenicity of C. javanica to aphids. Using a virulence assay, we studied the entomopathogenic fungus C. javanica IJ-tg19, which has high virulence to the aphid Acyrthosiphon pisum (Harris) (Hemiptera: Aphididae). We compared conidial germination and activity of cuticle-degrading enzyme (subtilisin-like serine protease (Pr1) and chitinase) of C. javanica IJ-tg19 with the entomopathogenic strains Cordyceps fumosorosea (Wize) (Hypocreales: Cordycipitaceae) IF-1106 and Beauveria bassiana (Balsamo) (Hypocreales: Cordycipitaceae) BB-1339. The infection process of C. javanica IJ-tg19 strain to A. pisum was observed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The results showed that germination time of conidia of C. javanica IJ-tg19 and C. fumosorosea IF-1106 was shorter than that of B. bassiana BB-1339, and the germination rate was significantly higher than the BB-1339 strain. The germ tube of IJ-tg19 was accompanied by mucilage secretion during the body surface germination of A. pisum. The activity of total Pr1 and chitinase of IJ-tg19 reached their highest values at 48 h post-inoculation (PI), and the activities were significantly higher than those of IF-1106 and BB-1339. SEM and TEM observations showed that the infection peak was at 24 h PI, and mycelia had entered the hemocoel cavity of A. pisum at 48 h. These results indicate that C. javanica IJ-tg19 has several desirable attributes to act as an effective agent for the biological control of aphids. [ABSTRACT FROM AUTHOR]

Summary: Plant disease occurs simultaneously with insect attack. Arbuscular mycorrhizal fungi (AMF) modify plant biotic stress response. Arbuscular mycorrhizal fungi and pathogens may modify plant volatile organic compound (VOC) production and insect behavior. Nevertheless, such effects are rarely studied, particularly for mesocosms where component organisms interact with each other.Plant‐mediated effects of leaf pathogen (Phoma medicaginis) infection on aphid (Acyrthosiphon pisum) infestation, and role of AMF (Rhizophagus intraradices) in modifying these interactions were elucidated in a glasshouse experiment. We evaluated alfalfa disease occurrence, photosynthesis, phytohormones, trypsin inhibitor (TI) and total phenol response to pathogen and aphid attack, with or without AMF, and aphid behavior towards VOCs from AMF inoculated and non‐mycorrhizal alfalfa, with or without pathogen infection.AM fungus enhanced alfalfa resistance to pathogen and aphid infestation. Plant biomass, root : shoot ratio, net photosynthetic rate, transpiration rate, stomatal conductance, salicylic acid, and TI were significantly increased in AM‐inoculated alfalfa. Arbuscular mycorrhizal fungi and pathogen significantly changed alfalfa VOCs. Aphids preferred VOCs of AM‐inoculated and nonpathogen‐infected to nonmycorrhizal and pathogen‐infected alfalfa.We propose that AMF alter plant response to multiple biotic stresses in ways both beneficial and harmful to the plant host, providing a basis for strategies to manage pathogens and herbivore pests. [ABSTRACT FROM AUTHOR]

The mechanism controlling sex allocation in the pea aphid, Acyrthosiphon pisum (Harris), remains a crucial yet unresolved issue in the field of evolutionary ecology. This study aims to assess the influence of the presence of both self and non-self clones, along with juvenile hormone III (JH III) titer, on the sex allocation of aphid offspring. To this end, red and green clones were utilized as experimental subjects, and the agar method was employed. Initially, three distinct experimental treatments were established using sexuparae, and the daily offspring count and sex allocation in each treatment zone were recorded. Subsequently, an additional experimental condition involving mixed-clone treatments was introduced. This procedure entailed the transfer of a single sexupara and 20 oviparous females from either the red (1G + 20Rov) or green clone (1G + 20Gov) onto a leaf on agar medium. Simultaneously, a control setup with a new sexupara (1G) was established. Three days following sexupara production, a dose of 0, 25, or 50 ng of JH III was applied to the aphids’ abdomens. Subsequently, the titers of JH III in the sexuparae across each treatment group were quantified, and the extent of sex allocation was tallied. The findings demonstrated pronounced disparities in sex allocation among the various treatments and, notably, a substantial increase in the total offspring and oviparous number in the mixed-clone treatment group. The effects of mixed-clone treatment on the sex allocation patterns of the sexupara progeny could be determined by the application of exogenous JH III, indicating that JH may mediate the effects of mixed-clone treatment on sex allocation. Consequently, it can be concluded that A. pisum sexuparae possess the capability to modulate their sex allocation in response to the nature of adjacent competitor clones, thereby demonstrating a variety of sex allocation patterns. Throughout this process, JH III plays a pivotal role.

The pea aphid (PA), Acyrthosiphon pisum (Harris) (Hemiptera: Aphididae), can cause significant damage to dry and green peas, Pisum sativum L. (Fabaceae). Identifying pea lines with resistance to PA is important to advance available control measures. This research identified lines and putative genes with resistance to PA in the USDA Pisum sativum Collection located in Pullman, WA, USA, consisting of 301 lines collected worldwide. Eight commercial pea cultivars were also evaluated. A cluster analysis based on the mean number of adults, nymphs, and total aphids that developed on lines 10 days after the start of infestation by a single adult aphid per plant identified six clusters. Cluster 2, consisting of 48 lines, had the lowest aphid fecundity of all clusters with 2.1 adults, 7.3 nymphs, and 9.4 total aphids produced on average. 'Lifter' was the most resistant line based on low aphid fecundity. Lifter is a white‐flowered green dry pea cultivar that can advance PA resistance in food‐type peas. Genotyping by sequencing based on single‐nucleotide polymorphisms (SNPs) were used in genome‐wide association mapping to locate genes or quantitative trait loci (QTL) linked to PA resistance in which three, nine, and six SNPs were associated with resistance to adult, nymph, and total number of PAs that developed on lines, respectively. Seventeen candidate genes were identified in response to PA resistance. The results can effectively be used in breeding programs to target specific genomic regions with PA resistance and in traditional breeding using resistant lines as parents. [ABSTRACT FROM AUTHOR]

Wing polyphenism is a common phenomenon that plays key roles in environmental adaptation of insects. Insulin/insulin‐like growth factor signaling (IIS) pathway is a highly conserved pathway in regulation of metabolism, development, and growth in metazoans. It has been reported that IIS is required for switching of wing morph in brown planthopper via regulating the development of the wing pad. However, it remains elusive whether and how IIS pathway regulates transgenerational wing dimorphism in aphid. In this study, we found that pairing and solitary treatments can induce pea aphids to produce high and low percentage winged offspring, respectively. The expression level of ILP5 (insulin‐like peptide 5) in maternal head was significantly higher upon solitary treatment in comparison with pairing, while silencing of ILP5 caused no obvious change in the winged offspring ratio. RNA interference‐mediated knockdown of FoxO (Forkhead transcription factor subgroup O) in stage 20 embryos significantly increased the winged offspring ratio. The results of pharmacological and quantitative polymerase chain reaction experiments showed that the embryonic insulin receptors may not be involved in wing polyphenism. Additionally, ILP4 and ILP11 exhibited higher expression levels in 1st wingless offspring than in winged offspring. We demonstrate that FoxO negatively regulates the wing morph development in embryos. ILPs may regulate aphid wing polyphenism in a developmental stage‐specific manner. However, the regulation may be not mediated by the canonical IIS pathway. The findings advance our understanding of IIS pathway in insect transgenerational wing polyphenism. [ABSTRACT FROM AUTHOR]

Host plant shifts are central to diversification in insect herbivores. Many mechanisms can cause host shifts in insects, but one relatively unexplored mechanism is whole‐genome duplication (WGD) in the host plant. WGD, or polyploidy, is common in plants and causes spontaneous changes in physiology, morphology, and palatability that could impact the ability of herbivores to feed and develop on newly formed polyploids (neopolyploids).Here the authors tested if WGD affected the preference and performance of the specialist aphid, Acyrthosiphon pisum (pea aphids). Pea aphids seasonally form specialised lineages or 'host forms' on many host plant species including alfalfa and red clover. Aphid host forms on alfalfa and red clover naturally exist on different cytotypes of their respective hosts, with red clover aphids feeding on diploid clover and alfalfa aphids feeding on tetraploid alfalfa. Therefore, the authors predicted that these host forms would have a higher preference for and performance on their respective natal host cytotype.Neither host form exhibited a preference for a particular cytotype, but there were modest changes in aphid performance based on host cytotype. Specifically, aphids specialised to red clover had higher fecundity on diploid red clover than on neotetraploid red clover. Together, these results showed that both host forms were able to recognise and accept different cytotypes of the two host species, but only one host form experienced trade‐offs in performance when feeding on neotetraploids. These results suggest that WGD may act as a mechanism of host expansion in pea aphids as plants speciate via WGD. [ABSTRACT FROM AUTHOR]

Plants are a difficult food resource to use, and herbivorous insects have evolved a variety of mechanisms that allow them to fully exploit this poor nutritional resource. One such mechanism is the maintenance of bacterial symbionts that aid in host plant feeding and development. The majority of these intracellular symbionts have highly eroded genomes that lack many key regulatory genes; consequently, it is unclear if these symbionts can respond to changes in the insect's diet to facilitate host plant use. There is emerging evidence that symbionts with highly eroded genomes express small RNAs (sRNAs), some of which potentially regulate gene expression. In this study, we sought to determine if the reduced genome of the nutritional symbiont (Buchnera) in the pea aphid responds to changes in the aphid's host plant diet. Using transcriptome sequencing (RNA-seq), Buchnera sRNA expression profiles were characterized within two Buchnera life stages when pea aphids fed on either alfalfa or fava bean. Overall, this study demonstrates that Buchnera sRNA expression changes not only with life stage but also with changes in aphid host plant diet. Of the 321 sRNAs characterized in this study, 47% were previously identified and 22% showed evidence of conservation in two or more Buchnera taxa. Functionally, 13 differentially expressed sRNAs were predicted to target genes related to pathways involved in essential amino acid biosynthesis. Overall, results from this study reveal that host plant diet influences the expression of conserved and lineage-specific sRNAs in Buchnera and that these sRNAs display distinct host plant-specific expression profiles among biological replicates.IMPORTANCE In general, the genomes of intracellular bacterial symbionts are reduced compared to those of free-living relatives and lack many key regulatory genes. Many of these reduced genomes belong to obligate mutualists of insects that feed on a diet that is deficient in essential nutrients, such as essential amino acids. It is unclear if these symbionts respond with their host to changes in insect diet, because of their reduced regulatory capacity. Emerging evidence suggests that these symbionts express small RNAs (sRNAs) that regulate gene expression at the posttranscriptional level. Therefore, in this study, we sought to determine if the reduced genome of the nutritional symbiont Buchnera in the pea aphid responds to changes in the aphid's host plant diet. This study demonstrates for the first time that Buchnera sRNAs, some conserved in two or more Buchnera lineages, are differentially expressed when aphids feed on different plant species and potentially target genes within essential amino acid biosynthesis pathways.

Abstract Background Acyrthosiphon pisum or pea aphid is an insect of the Aphididae family, which attacks various species of legumes such as beans and peas. This pest causes economically heavy crop losses around the world. The use of conventional chemical insecticides is the only way to control its development. However, the harmful consequences of these chemicals are well known. They pollute various compartments of the environment, thus constituting a major risk for human and environmental health. The search for a more ecological alternative, respectful of the environment is, therefore, a necessity. Actinomycetes represent a source of biologically active secondary metabolites, such as antibiotics and biopesticidal agents. In this study, 150 secondary metabolites of actinomycetes have made the objective of an in silico research by molecular docking, by screening their potential inhibitors against the enzyme acetylcholinesterase (AChE) of A. pisum. Results The 3D structure of AChE, unavailable in the PDB database, was first modeled using the Modeller program, then the stereochemical quality of the model was validated. The molecular docking performed by the Autodock Vina algorithm allowed the selection of two metabolites giving binding energy equal to or lower than that of the co-crystallized inhibitor tetrahydro-acridine (−10.3Kcal/mol). The top-two metabolites are diazepinomicine (−10.9 Kcal/mol), and hygromycin (−10.3 Kcal/mol). These components have shown numerous interactions with the key residues of the catalytic site of the AChE enzyme, indicating their potential to inhibit its biological activity. The environmental and health safety of these components, as well as their bioavailability, were also studied by the verification of several pharmacokinetic and ADMET criteria. Diazepinomicine has shown excellent results verifying most of the criteria studied. A 50-ns MD simulation was also performed in order to test the stability of the complexes formed. Conclusions In addition to its favorable pharmacokinetic properties, the special chemical structure of diazepinomicin allows this molecule to interact intensely with AChE notably through the involvement of its two groups farnesyl diphosphate and dibenzodiazepinone which ensure several hydrogen and hydrophobic interactions, that offers very high stability to the complex AChE diazepinomicin. In conclusion, diazepinomicin can be suggested as a potential bioinsecticidal agent against the pest A. pisum.

Facultative bacterial endosymbionts are prevalent in aphid pest species and key organisms for the rapid adaptation to changing environment conditions under agricultural management. Facultative endosymbiont interactions allow beneficial phenotypes of insect hosts, providing resistance against natural enemies, fungal infections as well as enhanced resistance to thermal conditions, among others. These interactions have been highlighted as important modulators in the evolution of aphid defenses, being a key aspect for the invasive potential of aphids as pests. In this study, we have examined the bacterial community diversity associated to the pea aphid Acyrthosiphon pisum on alfalfa (Medicago sativa L.) crops in Central Chile by determining the presence and abundance of the seven most common facultative endosymbionts reported for aphid species, including: Hamiltonella defensa, Regiella insecticola, Serratia symbiotica, Spiroplasma sp., Rickettsia sp., Rickettsiella viridis and Fukatsuia symbiotica (or pea aphid X-type symbiont' PAXS), using a PCR-based approach across species-specific primers, in 16 alfalfa fields during the Chilean spring in the Maule Region. We reported six of the seven endosymbiont species studied, observing a high frequency of infected aphids being the endosymbiont H. defensa, the most predominant among the aphid populations in the different alfalfa fields studied. As H. defensa has been extensively studied in its defensive role to parasitoids, we discuss implications for biological control of this aphid pests. We also discuss the absence of a relevant defensive endosymbiont such as F. symbiotica in the Chilean pea aphid populations and potential implications for the success of the pea aphid biological control program. [ABSTRACT FROM AUTHOR]

Background: Acyrthosiphon pisum Harris is the most destructive pest worldwide because of its ability to feed on plants directly and transmit plant viruses as a vector. This study aims to identify triterpenoid saponins from Oxytropis hirta Bunge as biopesticides to control aphids. Results: Three new azukisapogenol triterpenoid saponins (1–3), a new pinoresinol lignan glycoside (8), and four known saponins (4–7) were identified from the root of O. hirta. Compounds 4–7 displayed significant aphicidal activities against A. pisum with oral toxicities (LC50 = 51.10–147.43 μg/mL, 72 h), deterrent effects (deterrence index = 1.00, 100–200 μg/mL, 24 h), and aphid reproduction inhibitory effects (inhibition rates = 75.91–86.73%, 400 μg/mL, 24 h), respectively. The carboxyl groups at C‐3 GlcA and C‐30 were functional groups for their aphicidal activities. The toxic symptoms caused by the optimal 5 involved insect body‐color changes from light green to dark or gray‐green, and then brown until death. The intestinal cavity, apical microvilli, nuclei, mitochondria, and electron dense granules in the midgut tissues of A. pisum were the target sites showing aphicidal activity. The suppression of pepsin and α‐amylase, and the activation of lipase and trypsin could be the signs of organelle damage in the midgut tissues. Conclusion: Azukisapogenol triterpenoid saponins from O. hirta could be used as biopesticides to control aphids for their multiple efficacies, including oral toxicity, deterrent activity, and reproduction inhibitory activity. The toxic symptoms involved insect body‐color changes. Midgut tissues and their related enzymes were the targets for saponins showing aphicidal activities. © 2022 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

In recent years, pea aphids have become major pests of alfalfa. Our previous study found that “Gannong 5” is a highly aphid-resistant alfalfa variety and that “Lie Renhe” is a susceptible one. The average field susceptibility index of “Gannong 5” was 31.31, and the average field susceptibility index of “Lie Renhe” was 80.34. The uptake and balance of amino acids in insects are usually dependent on amino acid transporters. RT-qPCR was used to detect the relative expression levels of seven amino acid transporter differential genes in the different instar pea aphids fed on resistant and susceptible alfalfa varieties after 24 h, and two key genes were selected. When pea aphids fed on “Gannong 5”, the expression of ACYPI004320 was significantly higher than that in pea aphids fed on “Lie Renhe”; however, the expression of ACYPI000536 was significantly lower than that in pea aphids fed on “Lie Renhe”. Afterward, the RNA interference with pea aphid ACYPI000536 and ACYPI004320 genes was performed using a plant-mediated method, and gene function was verified via liquid chromatography–mass spectrometry and pea aphid sensitivity to aphid-resistant and susceptible alfalfa varieties. The results showed that the down-regulation of the ACYPI000536 gene expression led to an increase in the histidine and lysine contents in pea aphids, which, in turn, led to an increase in mortality when pea aphids fed on the susceptible variety “Lie Renhe”. The down-regulation of the ACYPI004320 gene expression led to an increase in phenylalanine content in pea aphids, which, in turn, led to a decrease in mortality when pea aphids fed on the resistant variety “Gannong 5”.

Despite the fact that sap-feeding hemipterans are major agricultural pests, little is known about the pea aphid's (Acyrthosiphon pisum) nymphal development, compared to other insect models. Given our limited understanding of A. pisum nymphal development and variability in the naming/timing of its developmental events between different environmental conditions and studies, here, we address developmental knowledge gaps by elucidating how diet impacts A. pisum nymphal development for the LSR1 strain when it develops on its universal host plant (Vicia faba), isolated leaves, and artificial diet. Moreover, we test how plant age and transgenerational stressors, such as overcrowding and low plant vigor, can affect nymphal development. We also validate a morphological method to quickly confirm the life stage of each nymphal instar within a mixed population. Overall, we found extremely high variation in the timing of developmental events and a significant delay in nymphal (~5-25-h/instar) and pre-reproductive adult (~40-h) development when reared on isolated leaves and artificial diets, compared to intact host plants. Also, delays in development were observed when reared on older host plants (~9-17-h/event, post 2nd instar) or when previous generations were exposed to overcrowding on host plants (~20-h delay in nymph laying) compared to controls.

Insect chitinases play a crucial part to digest chitin in the exoskeleton during the molting process. However, research on insect chitinase related to the environmental stress response is very limited. This study was the first conducted to expression analysis of chitinase- related genes in A. pisum under abiotic stresses. Here, we identified five chitinase-like proteins (ApIDGF, ApCht3, ApCht7, ApCht10 and ApENGase), and clustered them into five groups (group II, III, V, X, and ENGase). Developmental expression analysis revealed that the five A. pisumchitinase-related genes were expressed at whole developmental stages with different relative expression patterns. When aphids were exposed to various abiotic stresses including temperature, insecticide and the stress 20-hydroxyecdysone (20E), all five chitinase genes were differentially expressed in A. pisum. The results showed that insecticide such as imidacloprid down-regulated the expression of these five Cht-related genes. Analysis of temperature stress of A. pisum chitinase suggested that ApCht7 expression was high at 10°C, which demonstrates its important role in pea aphids under low temperature. Conversely, ApCht10 was more active under high temperature stress, as it was significantly up-regulated at 30°C. Besides, 20E enhanced ApCht3 and ApCht10 expression in A. pisum, but reduced ApCht7 expression. These findings provide basic information and insights for the study of the role of these genes under abiotic stress, which advances our knowledge in the management of pea aphids under multiple stresses. [ABSTRACT FROM AUTHOR]

Background: Acyrthosiphon pisum or pea aphid is an insect of the Aphididae family, which attacks various species of legumes such as beans and peas. This pest causes economically heavy crop losses around the world. The use of conventional chemical insecticides is the only way to control its development. However, the harmful consequences of these chemicals are well known. They pollute various compartments of the environment, thus constituting a major risk for human and environmental health. The search for a more ecological alternative, respectful of the environment is, therefore, a necessity. Actinomycetes represent a source of biologically active secondary metabolites, such as antibiotics and biopesticidal agents. In this study, 150 secondary metabolites of actinomycetes have made the objective of an in silico research by molecular docking, by screening their potential inhibitors against the enzyme acetylcholinesterase (AChE) of A. pisum. Results: The 3D structure of AChE, unavailable in the PDB database, was first modeled using the Modeller program, then the stereochemical quality of the model was validated. The molecular docking performed by the Autodock Vina algorithm allowed the selection of two metabolites giving binding energy equal to or lower than that of the co-crystallized inhibitor tetrahydro-acridine (−10.3Kcal/mol). The top-two metabolites are diazepinomicine (−10.9 Kcal/mol), and hygromycin (−10.3 Kcal/mol). These components have shown numerous interactions with the key residues of the catalytic site of the AChE enzyme, indicating their potential to inhibit its biological activity. The environmental and health safety of these components, as well as their bioavailability, were also studied by the verification of several pharmacokinetic and ADMET criteria. Diazepinomicine has shown excellent results verifying most of the criteria studied. A 50-ns MD simulation was also performed in order to test the stability of the complexes formed. Conclusions: In addition to its favorable pharmacokinetic properties, the special chemical structure of diazepinomicin allows this molecule to interact intensely with AChE notably through the involvement of its two groups farnesyl diphosphate and dibenzodiazepinone which ensure several hydrogen and hydrophobic interactions, that offers very high stability to the complex AChE diazepinomicin. In conclusion, diazepinomicin can be suggested as a potential bioinsecticidal agent against the pest A. pisum. [ABSTRACT FROM AUTHOR]

An important contributing factor to the evolutionary success of insects is nutritional association with microbial symbionts, which provide the host insects with nutrients lacking in their unbalanced diets. These symbionts are often compartmentalized in specialized cells of the host, the bacteriocytes. Even though bacteriocytes were first described more than a century ago, few studies have explored their dynamics throughout the insect life cycle and in response to environmental stressors. Here, we use the Buchnera aphidicola/ pea aphid symbiotic system to study how bacteriocytes are regulated in response to nutritional stress throughout aphid development. Using artificial diets, we analyzed the effects of depletion or excess of phenylalanine or leucine, two amino acids essential for aphid growth and whose biosynthetic pathways are shared between the host and the symbiont. Bacteriocytes responded dynamically to those treatments, while other tissues showed no obvious morphological change. Amino acid depletion resulted in an increase in bacteriocyte numbers, with the extent of the increase depending on the amino acid, while excess either caused a decrease (for leucine) or an increase (for phenylalanine). Only a limited impact on survival and fecundity was observed, suggesting that the adjustment in bacteriocyte (and symbiont) numbers is sufficient to withstand these nutritional challenges. We also studied the impact of more extreme conditions by exposing aphids to a 24 h starvation period at the beginning of nymphal development. This led to a dramatic drop in aphid survival and fecundity and a significant developmental delay. Again, bacteriocytes responded dynamically, with a considerable decrease in number and size, correlated with a decrease in the number of symbionts, which were prematurely degraded by the lysosomal system. This study shows how bacteriocyte dynamics is integrated in the physiology of insects and highlights the high plasticity of these cells. [ABSTRACT FROM AUTHOR]

Botanical products have an important role in the development of sustainable and ecologically friendly agriculture. Therefore, this study is focused on assessing the insecticidal activity of botanical products (Pyrethro Natura and Rapax) applied individually and in combination with an organic fertilizer (Fitasio) against Acyrthosiphon pisum. Further the change in the plastid pigments concentrations, productivity as well as product selectivity on Coccinella septempunctata larvae was explored. The experiment was conducted at the fields of the Institute of Forage crops, Pleven, Bulgaria from 2019 to 2021 year in spring forage peas. An alternate method of assessing the insecticide efficacy was used based on insect days and cumulative insect days. It was found that the mixture of Pyrethro Natura and Phytasio was associated with the highest overall efficacy against A. pisum and a reduction of cumulative insect days by 64.4% through a span of 9 days with an additive interaction between compounds. The botanical products used had a beneficial effect on the vegetative development of the plants. The Pyrethro Natura in combination with Fitasio provided the highest productivity of 30.7%, content of plastid pigments, and the best plant physiological condition. Botanicals were harmless with toxicity not exceeding 25% against ladybug larvae and displayed an important perspective due to their multiple benefits. [ABSTRACT FROM AUTHOR]

Although tannins are widely distributed in broad beans and alfalfa, the pea aphid (Acyrthosiphon pisum) can still destroy them. The ATP binding cassette (ABC) transporters participate in the metabolism of plant secondary metabolites and pesticides in insects. However, whether ABC transporter genes play a role in the metabolism of tannins in the pea aphid is unclear. Here, we found that verapamil (an ABC transporter inhibitor) significantly increased the mortality of tannic acid to pea aphid, which indicated that ABC transporter gene was related to the metabolism of tannic acid by pea aphid. Then, we identified 54 putative ABC transporter genes from the genome database of A. pisum. These genes were divided into eight subfamilies, ApABCA to ApABCH , of which subfamily G has the largest number of genes with 19, followed by the subfamily C with 14. RT-qPCR results show that the expression levels of ApABCA2 , ApABCC7 , ApABCG2 , and ApABCG3 were highly expressed in the first instar, while those of ApABCA3 , ApABCG6 , ApABCG7 , ApABCH3 , and ApABCH4 were highly expressed in adults. Furthermore, transcription levels of many ABC transporter genes were induced by tannic acid. Especially, ApABCG17 and ApABCH2 were obviously induced after being exposed to tannic acid. Meanwhile, knockdown of ApABCG17 by RNA interference resulted in increased sensitivity of pea aphid to tannic acid. These results suggest that ApABCG17 may be involved in tannic acid metabolism in pea aphid. This study will help us to understand the mechanism of tannic acid metabolism in pea aphid, and provides a basis for further research on the physiological function of ABC transporter genes in pea aphid. • The verapamil (an ABC transporter inhibitor) significantly increased the mortality rate of tannic acid to pea aphids. • The domain structures and classification of ABC transporter genes in A. pisum was analyzed. • The expression profiles of ABC transporter genes at different stages and after tannic acid treatment were analyzed. • ApABCG17 and ApABCH2 were significantly induced after exposure to tannic acid • Knockdown of ApABCG17 by RNA interference resulted in increased sensitivity of pea aphid to tannic acid. [ABSTRACT FROM AUTHOR]

Alfalfa (Medicago sativa) is the most important forage legume and one of the most widely grown crops in Serbia. Aphids (Hemiptera: Aphididae) are one of the most significant groups of pests of that crop. Three aphid species, Acyrthosiphon pisum, Aphis craccivora and Therioaphis trifolii, are considered as important pests of alfalfa in Serbia. The most abundant of them, T. trifolii, is more adapted to warmer temperatures and it is the predominant aphid species during summer months, while A. pisum is prevalent during the spring period. In warmer climates, an increasing abundance of T. trifolii and decreasing populations of A. pisum have been observed in alfalfa in Serbia. On the other hand, A. craccivora is a minor pest of alfalfa, which rarely occurs in high numbers. The present paper synthesizes the existing knowledge about these three aphid pests of alfalfa, including their morphology, biology, seasonal dynamics, pest status and effects of temperature on their development.

An important contributing factor to the evolutionary success of insects is nutritional association with microbial symbionts, which provide the host insects with nutrients lacking in their unbalanced diets. These symbionts are often compartmentalized in specialized cells of the host, the bacteriocytes. Even though bacteriocytes were first described more than a century ago, few studies have explored their dynamics throughout the insect life cycle and in response to environmental stressors. Here, we use the Buchnera aphidicola/pea aphid symbiotic system to study how bacteriocytes are regulated in response to nutritional stress throughout aphid development. Using artificial diets, we analyzed the effects of depletion or excess of phenylalanine or leucine, two amino acids essential for aphid growth and whose biosynthetic pathways are shared between the host and the symbiont. Bacteriocytes responded dynamically to those treatments, while other tissues showed no obvious morphological change. Amino acid depletion resulted in an increase in bacteriocyte numbers, with the extent of the increase depending on the amino acid, while excess either caused a decrease (for leucine) or an increase (for phenylalanine). Only a limited impact on survival and fecundity was observed, suggesting that the adjustment in bacteriocyte (and symbiont) numbers is sufficient to withstand these nutritional challenges. We also studied the impact of more extreme conditions by exposing aphids to a 24 h starvation period at the beginning of nymphal development. This led to a dramatic drop in aphid survival and fecundity and a significant developmental delay. Again, bacteriocytes responded dynamically, with a considerable decrease in number and size, correlated with a decrease in the number of symbionts, which were prematurely degraded by the lysosomal system. This study shows how bacteriocyte dynamics is integrated in the physiology of insects and highlights the high plasticity of these cells.

In insects, the loss of flight typically involves a dispersal-reproduction transition, but the underlying molecular mechanisms remain poorly understood. In the parthenogenetic pea aphid Acyrthosiphon pisum, winged females undergo flight-muscle degeneration after flight and feeding on new host plants. Similarly, topical application of a juvenile hormone (JH) mimic to starved aphids also induces flight-muscle degeneration. We found that feeding preferentially upregulated the expression of the JH receptor gene Met and a JH-inducible gene, Kr-h1, in the flight muscles, and, thus, enhanced tissue-specific JH sensitivity and signaling. RNAi-mediated knockdown of Kr-h1 prevented flight-muscle degeneration. Likewise, blocking nutritional signals by pharmacological inhibition of the target of rapamycin complex 1 (TORC1) impaired JH sensitivity of the flight muscles in feeding aphids and subsequently delayed muscle degeneration. RNA-sequencing analysis revealed that enhanced JH signaling inhibited the transcription of genes involved in the tricarboxylic acid cycle, likely resulting in reduction of the energy supply, mitochondrial dysfunction and muscle-fiber breakdown. This study shows that nutrient-dependent hormone sensitivity regulates developmental plasticity in a tissue-specific manner, emphasizing a relatively underappreciated mechanism of hormone sensitivity in modulating hormone signaling. [ABSTRACT FROM AUTHOR]

Nicotinamide adenine dinucleotide phosphate (NADPH)‐cytochrome P450 reductase (CPR) is involved in the metabolism of endogenous and exogenous substances, and detoxification of insecticides. RNA interference (RNAi) of CPR in certain insects causes developmental defects and enhanced susceptibility to insecticides. However, the CPR of Acyrthosiphon pisum has not been characterized, and its function is still not understood. In this study, we investigated the biochemical functions of A. pisum CPR (ApCPR). ApCPR was found to be transcribed in all developmental stages and was abundant in the embryo stage, and in the gut, head, and abdominal cuticle. After optimizing the dose and silencing duration of RNAi for downregulating ApCPR, we found that ApCPR suppression resulted in a significant decrease in the production of cuticular and internal hydrocarbon contents, and of cuticular waxy coatings. Deficiency in cuticular hydrocarbons (CHCs) decreased the survival rate of A. pisum under desiccation stress and increased its susceptibility to contact insecticides. Moreover, desiccation stress induced a significant increase in ApCPR mRNA levels. We further confirmed that ApCPR participates in CHC production. These results indicate that ApCPR modulates CHC production, desiccation tolerance, and insecticide susceptibility in A. pisum, and presents a novel target for pest control. [ABSTRACT FROM AUTHOR]

Prey mothers at risk of predation sometimes change the morphology and/or antipredator behaviour of their offspring to reduce predation risk. When maternal exposure to predation risk changes the morphology of some offspring, it is unclear whether and how the other offspring, which have normal morphology, exhibit antipredator behaviour. We aimed to clarify these behaviours using pea aphids, Acyrthosiphon pisum Harris (Hemiptera: Aphididae), and aphidophagous Asian ladybird beetles, Harmonia axyridis Pallas (Coleoptera: Coccinellidae), which induce aphids to release the alarm pheromone (E)-β-farnesene (EBF) and to exhibit antipredator responses. Pea aphids exposed to EBF reduced reproduction through changes in the number of wingless offspring, producing more winged offspring than unexposed conspecifics. Wingless aphids whose mothers had been exposed to EBF showed higher dispersal from host plants with predators than the offspring of unexposed mothers. These results suggest that pea aphids at risk of predation increase their offspring survival by increasing the number of winged offspring and antipredator dispersal of wingless offspring. [ABSTRACT FROM AUTHOR]

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The endosymbiotic bacterium Buchnera aphidicola allows its host Acyrthosiphon pisum to utilise a nutritionally limited phloem sap diet without significant mortality by providing essential amino acids (EAAs), which it biosynthesises de novo via complex pathways consisting of multiple enzymes. Previous studies have reported how non-essential amino acids (NEAAs) provided by the host are utilised by B. aphidicola, along with how genes within the biosynthetic pathways respond to amino acid deficiency. Although the effect on B. aphidicola gene expression upon the removal of a single EAA and multiple NEAAs from the A. pisum diet has been reported, little is known about the effects of the complete simultaneous removal of multiple EAAs, especially branched-chain amino acids (BCAAs). To investigate this, A. pisum was provided with amino acid deficient diets ilv- (lacking isoleucine, leucine, valine) or thra- (lacking threonine, methionine, lysine). Due to their involvement in the production of several amino acids, the expression of genes ilvC, ilvD (both involved in isoleucine, leucine and valine biosynthesis) and thrA (involved in threonine, methionine and lysine biosynthesis) was analysed and the expression of trpC (involved in tryptophan biosynthesis) was used as a control. Survival was reduced significantly when A. pisum was reared on ilv- or thra- (P < 0.001 and P = 0.000 respectively) compared to optimal artificial diet and was significantly lower on ilv- (P < 0.001) than thra-. This is likely attributed to the EAAs absent from ilv- being required at higher concentrations for aphid growth, than those EAAs absent from thra-. Expression of ilvC and ilvD were upregulated 2.49- and 2.08-fold (respectively) and thrA expression increased 2.35- and 2.12-fold when A. pisum was reared on ilv- and thra- (respectively). The surprisingly large upregulation of thrA when reared on ilv- is likely due to threonine being an intermediate in isoleucine biosynthesis. Expression of trpC was not affected by rearing on either of the two amino acid deficient diets. To our knowledge this study has shown, for the first time, how genes within the biosynthetic pathways of an endosymbiont respond to the simultaneous complete omission of multiple EAAs as well as all three BCAAs (leucine, isoleucine, valine), from the host diet., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The pea aphid Acyrthosiphon pisum is a worldwide pest that causes serious losses in agricultural production. Drunken horse grass (Achnatherum inebrians) is a widely distributed perennial poisonous grass of grasslands of Northern and Northwestern China. Changes in adult pea aphid transcripts after exposure to aqueous extracts of endophyte‐infected (E+) and endophyte‐free (E−) plants and distilled water controls (CK) were compared to identify candidate genes and pathways of the pea aphid. The resulting 31,858 unigenes annotations provide useful data for future molecular studies of pea aphid. A total of 147 mRNA transcripts were differentially expressed genes (DEGs): 30 genes were up‐regulated and 6 were down‐regulated in the E+ versus E− treated aphids; 45 up‐regulated and 40 down‐regulated genes were identified in E+ versus CK; 23 up‐regulated and 3 down‐regulated genes were identified in the E− versus CK. KEGG analysis mapped 16, 24 and 42 differentially expressed genes to 13, 20 and 22 substantially enriched pathways for E+ versus CK, E+ versus CK and E− versus CK, respectively. A number of fecundity factors were annotated from 266 unigenes, including fatty acid synthase, aminopeptidase N‐like, and cuticular protein genes that were highly expressed in aphids treated with extracts of both E+ and E− plants. Many of these genes were up‐regulated at least twofold in E− compared with the E+ treated aphids. The resultant transcriptomic database was authenticated by the transcript levels of five selected fecundity‐related genes quantified from different treatment material in a real‐time quantitative polymerase chain reaction (PCR). Interestingly, the genes related to lysosome that the most abundant in the E+ and E− treated aphids. Our studies provided the first comprehensive insights into varying treatments and effects on toxicity related genes in aphids. This work will facilitate efforts to explore new biocontrol strategies against pea aphid. [ABSTRACT FROM AUTHOR]

Natural enemies that can use multiple habitats are thought to better withstand disturbances in agricultural systems than natural enemies that are habitat specialists. This is because habitat generalists have populations in multiple habitats that can serve as sources of immigrants into an agricultural crop following a disturbance. In contrast, the dynamics of habitat specialists are tightly coupled with those of one agricultural crop. Nonetheless, some habitat specialists are successful in highly disturbed environments. To test how the magnitude of within‐field disturbance affects biological control agents, we conducted a large‐scale field manipulation in alfalfa fields and monitored the response of pea aphids, habitat‐generalist predators, a habitat‐specialist parasitoid (Aphidius ervi), and hyperparasitoids of A. ervi. The manipulation involved three treatments: harvesting normally (intermediate disturbance); spraying insecticide immediately after harvesting (high disturbance); and harvesting in strips (low disturbance). As a group, the habitat‐generalist predator species showed a range of responses to disturbances, from no response to decreases in abundance in the high‐disturbance treatment, indicating differences in their response to the density of pea aphids following disturbances. Surprisingly, percentage parasitism by the habitat‐specialist parasitoid was little affected by experimental disturbance manipulations. Furthermore, two of the four hyperparasitoids of A. ervi were negatively affected by the magnitude of disturbance, suggesting that disturbance could have an indirect positive effect on A. ervi. These results suggest that a habitat specialist can overcome the detrimental effects of disturbances without using alternative habitats. In addition, disturbance can sometimes benefit biological control agents by disproportionally negatively affecting their enemies from the fourth trophic level. [ABSTRACT FROM AUTHOR]

Abstract Background Aphids can be positioned as robust pest insects in farming and as ones of the model organisms for arthropods in molecular biology. Carotenoids are pigments that protect organisms from photooxidative damage caused by excessive light. Aphids were shown to possess genes of fungal origin for carotenoid biosynthesis, whereas a little knowledge was available about the functions of the genes and the biosynthetic pathway. Even carotenoid species contained in aphids were not enough understood. Main purpose of this study is to clarify these insufficient findings. Results The whole carotenoid biosynthetic pathway of the pea aphid (Acyrthosiphon pisum) was elucidated at the gene level, through comprehensive functional analysis of its carotenogenic genes, using Escherichia coli that synthesized carotenoid substrates, along with structural and quantitative analysis of carotenoids from various aphid species. Four genes were needed to synthesize all carotenoids accumulated in aphids from geranylgeranyl diphosphate. The tor gene mediated desaturation reaction from phytoene to 3,4-didehydrolycopene. It was revealed that a gene designated ApCrtYB3, which was considered to have functionally evolved in aphids, can convert lycopene into uncommon carotenoids with the γ-ring such as (6′S)-β,γ-carotene and γ,γ-carotene. We further demonstrated that the atypical carotenoids work as ecological indicators for estimating the food chain from aphids to predatory arthropods, and showed that aphids contributed with significant levels to the food chain from insect herbivores to several predatory arthropods, i.e., the red dragonfly (Sympetrum frequens; adults), seven-spotted ladybird (Coccinella septempunctata), and two spiders, Oxyopes sertatus and Nephila clavata. Gut microflora of the dragonfly (mature adults) was also found to include endosymbiotic bacteria such as Serratia symbiotica specific to the black bean aphid (Aphis fabae). Conclusions We revealed the whole carotenoid biosynthetic pathway of aphids, including functional identification of the corresponding genes. Subsequently, we showed that arthropodal food chain can be estimated using the uncommon carotenoids of aphids as ecological indicators. This result indicated that aphids made significant contributions to the food chain of several predatory arthropods including the red-dragonfly adults. Aphids are likely to be positioned as an important “phytochemicals” source for some predatory insects and arachnids, which are often active under bright sunlight.

Abstract Natural enemies that can use multiple habitats are thought to better withstand disturbances in agricultural systems than natural enemies that are habitat specialists. This is because habitat generalists have populations in multiple habitats that can serve as sources of immigrants into an agricultural crop following a disturbance. In contrast, the dynamics of habitat specialists are tightly coupled with those of one agricultural crop. Nonetheless, some habitat specialists are successful in highly disturbed environments. To test how the magnitude of within‐field disturbance affects biological control agents, we conducted a large‐scale field manipulation in alfalfa fields and monitored the response of pea aphids, habitat‐generalist predators, a habitat‐specialist parasitoid (Aphidius ervi), and hyperparasitoids of A. ervi. The manipulation involved three treatments: harvesting normally (intermediate disturbance); spraying insecticide immediately after harvesting (high disturbance); and harvesting in strips (low disturbance). As a group, the habitat‐generalist predator species showed a range of responses to disturbances, from no response to decreases in abundance in the high‐disturbance treatment, indicating differences in their response to the density of pea aphids following disturbances. Surprisingly, percentage parasitism by the habitat‐specialist parasitoid was little affected by experimental disturbance manipulations. Furthermore, two of the four hyperparasitoids of A. ervi were negatively affected by the magnitude of disturbance, suggesting that disturbance could have an indirect positive effect on A. ervi. These results suggest that a habitat specialist can overcome the detrimental effects of disturbances without using alternative habitats. In addition, disturbance can sometimes benefit biological control agents by disproportionally negatively affecting their enemies from the fourth trophic level.

The aim of this study was to evaluate the antioxidant effect of selenium in Pisum sativum L. plants pre-treated with sodium selenite or sodium selenate at a concentration of 10 and 20 μM, and then colonized by pea aphid Acyrthosiphon pisum (Harris). It has been hypothesized that selenium at low concentrations alleviates oxidative stress caused by aphid feeding on pea leaves. The study focused on the generation of reactive oxygen species (superoxide anion, hydrogen peroxide and hydroxyl radical), the activities of the antioxidant enzymes (superoxide dismutase and ascorbate peroxidase) scavenging the reactive oxygen species levels, as well as on total antioxidant activity in pea leaves. Selenium in pea leaves exposed to aphid feeding affected changes in the levels of reactive oxygen species, the activity of studied antioxidant enzymes, and the total antioxidant capacity. Effects depended on the form and concentration of selenium, as well as on the time after the colonization of pea plants by aphids. Obtained results showed beneficial effects of selenium in alleviating oxidative stress in pea leaves caused by aphid feeding.

Biofortification of plants with selenium (Se) is one of the current trends in agriculture. This treatment enriches plants with this essential element for humans and animals, and also increases plant resistance to stress factors. The current knowledge of the interactions between Se and plants is well documented but between the plants biofortificated with Se and phytophages is insufficient and this problem should be further investigated. The research has focused on the effect of the pre-treatment of pea with Se on the pea aphid feeding behaviour and demographic parameters of the aphid population, and also has shown the activity of antioxidant (superoxide dismutase and catalase), detoxifying (S-glutathione transferase and β-glucosidase) and redox (peroxidase and polyphenol oxidase) enzymes in tissues of Acyrthosiphon pisum. The analysis of feeding models recorded using the Electronic Penetration Graph technique has shown that the pre-treatment of peas with Se affects the activity of aphids during the feeding. The use of 10 and 20 µM sodium selenite and sodium selenate caused changes in A. pisum probing and feeding behavior. What is more, Se reduced the activity of aphids associated with the phloem phase and caused an extension of non-probing time and non-phloem penetration. The effect of Se on the demographic population parameters of the aphid was also observed in the significant reduction of female fertility and the reproductive potential of the insects. Selenium enrichment of plants colonized by aphids increased the activity of antioxidant, detoxifying and redox enzymes in the tissues of A. pisum. It was also found that both Se taken up by plants, as well as, the feeding of aphids resulted in greater synthesis of flavonols and carotenoids in pea leaves.

The Janus kinase/signal transducers and activators of transcription (JAK/STAT) signaling is activated by infections of bacteria, fungi, viruses and parasites and mediated cellular and humoral immune responses. In the pea aphid Acyrthosiphon pisum little is known about the function of JAK/STAT signaling in its immune system. In this study, we first showed that expression of genes in the JAK/STAT signaling, including the receptors Domeless1/2 , Janus kinase (JAK) and transcriptional factor Stat92E , is up-regulated upon bacteria Escherichia coli and Staphylococcus aureus and fungus Beauveria bassiana infections. After knockdown of expression of these genes by means of dsRNA injection, the aphids harbored more bacteria and suffered more death after infected with E. coli and S. aureus , but showed no significant change after B. bassiana infection. Our study suggests the JAK/STAT signaling contributes to the defense against bacterial infection in the pea aphid. [Display omitted] • Pea aphid JAK/STAT signaling was up-regulated by bacterial and fungal infections. • Knockdown of expression of JAK/STAT signaling caused higher bacterial loads in pea aphids. • Knockdown of expression of JAK/STAT signaling resulted in higher mortality of aphids after bacterial infection. [ABSTRACT FROM AUTHOR]

Insect herbivores, such as aphids, are common on plants, yet how they interact with plant microbiomes remains largely unknown. For instance, for the widespread bacterial epiphyte and potential aphid pathogen Pseudomonas syringae , aphids could impact bacterial populations by serving as secondary hosts or by altering the epiphytic habitat through feeding and/or waste secretion. Here, we examined whether the pea aphid, Acyrthosiphon pisum , could influence epiphytic populations of P. syringae. First, we quantified epiphytic growth ability without aphids and virulence to aphids across 21 diverse P. syringae strains. For eight strains that varied in these traits we then assessed the influence of aphid presence on epiphytic bacterial growth. In some cases P. syringae benefited significantly from the presence of aphids, with up to 3.8 times more cell doublings. This benefit was not correlated with strain traits but rather with initial population densities; smaller bacterial populations received relatively more benefit from aphids, and larger populations received less benefit. Honeydew, the sugary waste product of aphids, in the absence of aphids was sufficient to increase P. syringae density on leaves. We conclude that aphid honeydew can sometimes increase P. syringae epiphytic growth but that the bacteria may not benefit from using aphids as hosts. [ABSTRACT FROM AUTHOR]

Insects are known to be negatively impacted by noise from anthropogenic sources. Recently, pest control methods have been developed using specific airborne (sound) and substrate-borne vibratory signals. However, there is limited information available on the effects of non-specific vibrations on pest insects, especially those not known to use sound or vibration for communication. Here, we tested whether non-specific substrate-borne or airborne vibrations (white noise) cause reductions in the number of aphids of three species on single plants in short-term (24 h) laboratory trials. Substrate-borne vibrations transmitted through the host plant reduced total numbers of all three species (45 to 62% reductions in total aphid numbers), while airborne vibrations played through speakers had no detectable effect. The effect of substrate-borne vibrations on aphid numbers appeared to be mostly due to inducing aphids to leave the plant, rather than reducing the reproductive output of remaining aphids. To our knowledge, our results provide the first proof-of-concept indicating that non-specific substrate-borne vibrations can affect pests not known to use vibrations to communicate and should be explored further as a physical pest control method. Future research in this area is needed to investigate longer-term effects of non-specific vibrations on pests, their host plants, and their natural enemies. [ABSTRACT FROM AUTHOR]

Simple Summary: We know that neuropeptides and G protein-coupled receptors regulate the physiology and behavior of animals and that the pea aphid (Acyrthosiphon pisum) is a serious agricultural pest and model insect. In this study, we investigated the short neuropeptide F and its receptor in pea aphid. Feeding analysis showed that the probing time and total phloem duration significantly decreased in response to sNPF and predicted sNPFR gene silencing in RNAi assays. The silencing of sNPF significantly reduced the aphid's reproduction but not survival. Our findings will help in the design of control strategies by using the molecular biological approach. Insect short neuropeptide F (sNPF), an ortholog of prolactin-releasing peptide of invertebrates, regulates diverse biological processes, including feeding, olfaction, locomotion, and sleep homeostasis in insects. However, its function is still unclear in an important model insect and agricultural pest, the pea aphid (Acyrthosiphon pisum). Here, we investigated short neuropeptide F (ApsNPF) and its receptor (ApsNPFR) in A. pisum. The sNPF gene contains three exons and two long introns. In addition, the genome contains a single sNPF receptor with seven transmembrane domains. Stage- and tissue-specific transcript profiling by qRT-PCR revealed that ApsNPF and ApsNPFR were mainly expressed in the central nervous system. The receptor was also detected in antennae, midgut, and integument. The highest expression levels were found in first instar nymphs compared to other developmental stages. Besides, the starvation-induced pattern indicated that the sNPF network depends on the nutritional state of the insect. An electrical penetration graph showed that probing time and phloem duration of A. pisum on broad bean plants decreased in response to dssNPF and dssNPFR in RNAi assays. sNPF silencing reduced the number of nymphs per female but not aphid survival. We believe that our results advance in-depth knowledge of the sNPF/sNPFR signaling cascade and its place in regulating feeding behavior in insects. In turn, it may contribute to the potential design of new strategies to control aphids, with a focus on the sNPF system. [ABSTRACT FROM AUTHOR]

Simple Summary: Resistance of a Pisum fulvum and five Pisum sativum genotypes to Acyrthosiphon pisum pea and alfalfa-adapted biotypes was evaluated by measuring aphid body mass, confirming the variable resistance level of these genotypes. The feeding behavior of the aphids on the Pisum genotypes was then examined by electropenetrography (EPG). The EPG results suggested that the resistance of Pisum genotypes to non-adapted A. pisum resides in mesophyll and phloem tissues while the resistance variation of P. sativum to pea adapted aphids may be influenced by the quality of phloem sap. Resistant genotypes of crops have emerged as an alternative and sustainable solution to pesticide use against pest insects. The resistance depends on the genetic diversity of the host plant and the pest species and can cause an alteration of the insect behavior. The aim of this work was to characterize the resistance level of different Pisum genotypes (one P. fulvum and five P. sativum genotypes) to two biotypes of the aphid Acyrthosiphon pisum, respectively adapted to pea and alfalfa, by measuring the individual aphid weight and analyzing aphid feeding behavior by electropenetrography (EPG). Aphid body mass was influenced by Pisum genotypes reflecting variation in their resistance level. P. fulvum was the most resistant to the A. pisum pea biotype (ArPo28 clone) and showed intermediate resistance to the A. pisum alfalfa biotype (LSR1 clone). The resistance levels of the five P. sativum genotypes to the two aphid biotypes were variable and more pronounced for the alfalfa biotype. EPG data showed that ArPo28 on P. fulvum and LSR1 on all the Pisum genotypes spent shorter time phloem feeding compared to ArPo28 on P. sativum genotypes, indicating that the resistance of Pisum genotypes to non-adapted A. pisum resides in mesophyll and phloem cells. In the meantime, ArPo28 on P. sativum genotypes with a different level of resistance spent a similar length of time phloem feeding, indicating that the quality of phloem sap of the resistance genotypes may not be optimal for the aphid. The study indicated that the resistance of Pisum genotypes to the two A. pisum biotypes involves different genetic factors and mechanisms that affect the aphid differently. [ABSTRACT FROM AUTHOR]

Although most aphid species living on leaves have a green body color, little is known regarding the biosynthetic pathways of green pigments. We found that a clone of the pea aphid, Acyrthosiphon pisum (Harris) produced both green‐ and yellow‐colored males. The females of this clone were green in color, while 8.4% of the males produced were yellow. To date, yellow body color has been reported only in a single mutant clone in A. pisum. To explore the genetic pattern of yellow body color, green or yellow males were mated with green females of the same clone. The hatchability of the eggs sired by yellow males (26.2%) was less than half that of the eggs sired by green males (79.0%). The hatched foundresses of both groups were all green, with no yellow foundresses. Because aphids have an XX‐XO sex determination system, color polymorphism in males suggests that male body color may be governed by an X‐linked locus. If females possess heterozygosity at the putative locus, they can produce alternative phenotypes in males. The small proportion of yellow males and absence of yellow foundresses imply that the allele responsible for yellow body color has a deleterious effect. The present study suggests that this clone could be used to elucidate the biosynthetic pathways and underlying genetics of green pigments in aphids. [ABSTRACT FROM AUTHOR]

Many insect herbivores engage in apparent competition whereby two species interact through shared natural enemies. Upon insect attack, plants release volatile blends that attract natural enemies, but whether these volatiles mediate apparent competition between herbivores is not yet known. We investigate the role of volatiles that are emitted by bean plants upon infestation by Acyrthosiphon pisum aphids on the population dynamics and fitness of Sitobion avenae aphids, and on wheat phloem sap metabolites. In a field experiment, the dynamics of S. avenae aphids on wheat were studied by crossing two treatments: exposure of aphid colonies to A. pisum-induced bean volatiles and exclusion of natural enemies. Glasshouse experiments and analyses of primary metabolites in wheat phloem exudates were performed to better understand the results from the field experiment. In the field, bean volatiles did not affect S. avenae dynamics or survival when aphids were exposed to natural enemies. When protected from them, however, volatiles led to larger aphid colonies. In agreement with this observation, in glasshouse experiments, aphid-induced bean volatiles increased the survival of S. avenae aphids on wheat plants, but not on an artificial diet. This suggests that volatiles may benefit S. avenae colonies via metabolic changes in wheat plants, although we did not find any effect on wheat phloem exudate composition. We report a potential case of associational susceptibility whereby plant volatiles weaken the defences of receiving plants, thus leading to increased herbivore performance. [ABSTRACT FROM AUTHOR]