Electrical Penetration Graph Research Articles

Identification and Characterization of Resistance of Three Aphid Species on Contrasting Alfalfa Cultivars.

Simple SummaryAphis craccivora Koch (cowpea aphid, CPA), Acyrthosiphon pisum Harris (pea aphid, PA) and Therioaphis trifolii Buckton (spotted alfalfa aphid, SAA) are the three species of devastating pests on alfalfa in China. A study was conducted in the laboratory for identification and characterization of resistance to these three aphids among 16 of the main alfalfa cultivars planted in China. Resistance was indicated by antibiosis, antixenosis, and measuring feeding behavior using EPG (electrical penetration graph). The results indicated that different alfalfa cultivars have significantly different resistance levels to a particular species of aphid, and the same alfalfa variety also has different resistance to the three aphid species. Specifically, we evaluated the resistance of different alfalfa cultivars to CPA, which can help us for further study on the defense mechanism against CPA and for better management of this pest.Aphids on alfalfa (Medicago sativa) including Aphis craccivora Koch (cowpea aphid, CPA), Acyrthosiphon pisum Harris (pea aphid, PA) and Therioaphis trifolii Buckton (spotted alfalfa aphid, SAA) cause significant yield losses worldwide. In this experiment, the development of these three species of aphids on 16 alfalfa cultivars was compared. The results showed that the plant cultivar had a significant influence on the development of aphids as there are significant differences in the body weight of aphids reared on different alfalfa cultivars. In addition, antibiosis between the alfalfa cultivars Pegasis and Gannong NO.9 and the three species of aphids was evaluated by measuring aphid body weight and fecundity. Antixenosis was measured using choice tests, and feeding behavior was quantified using electrical penetration graphs (EPG). The Pegasis cultivar was observed to have both antibiosis and antixenosis effects with CPA, but was susceptible to PA and SAA compared with the Gannong NO.9 cultivar. CPA had less mean body weight, less fecundity, and shorter feeding time on the Pegasis cultivar, and preferred to settle on Gannong NO.9 cultivar. In contrast, Gannong NO.9 exhibits antibiosis and antixenosis to PA and SAA compared with Pegasis, as shown by lower body weight, lower fecundity and chose to settle less often, but EPG data showed that PA and SAA showed no different significance in feeding behavior between Pegasis and Gannong NO.9.

Evaluation of resistance to black bean aphid (Aphis fabae) in selected varieties and mutant genotypes of common bean (Phaseolus vulgaris)

AbstractCommon bean (Phaseolus vulgaris) is an important food crop across sub‐Saharan Africa. In Zambia, actual common bean yields are typically lower than potential yields because of the impact of invertebrate pests and plant diseases. Black bean aphids (Aphis fabae) (Hemiptera: Aphididae) negatively impact bean productivity directly by ingesting plant assimilates and indirectly by vectoring diseases such as bean common mosaic virus. Current breeding programmes aim to develop bean cultivars with improved yield and tolerance to pests. The objective of this study was to screen five common bean varieties (Rozi Koko, Mwezi Moja, Majesty, KK25 and AO‐1012‐29‐3A) and four mutation‐derived genotypes (CA 3, CA 15, CA 24 and CA 38) for resistance to black bean aphid. Commercial bean cultivars such as Kabulangeti and Carioca (variety from which all mutants were derived) were used as controls for the selected varieties and mutant genotypes, respectively. Several parameters of aphid resistance traits were assessed. Deterrence to aphid was assessed by settling preference while physical barriers to aphid feeding were evaluated by nymph survival. Reduction in palatability of phloem sap was evaluated by nymph development and mean relative growth rate. Electrical penetration graph recordings of feeding behaviour were performed in order to localise aphid‐resistant factors. Nymph development was significantly longer on AO‐1012‐29‐3A compared to Kabulangeti despite the fact that there were significantly fewer glandular trichomes on this line. The variety AO‐1012‐29‐3A can be used in genetic improvement of common bean for aphid resistance.

Probing behavior of the corn leafhopper Dalbulus maidis on susceptible and resistant maize hybrids.

The corn leafhopper Dalbulus maidis is the main vector of the pathogens that cause corn stunt, a major disease of maize in the Americas. In line with plant resistance being an efficient tool to control diseases, the findings of a previous work showed that some corn hybrids are resistant to D. maidis. In this work, we assessed the probing behavior of D. maidis on susceptible and resistant corn hybrids using EPG (Electrical Penetration Graph) technology. Feeding of fifteen-day-old, non-inoculative females was recorded for 20 hours, with access to hybrids DK390, DK670, DK79-10, and DK72-10. Compared to the susceptible hybrid DK670, the other hybrids shifted D. maidis probing behavior in a way consistent with plant resistance to insects. This shift consisted of a higher number of probes of short duration, difficulties in attaining phloem ingestion and increase in xylem ingestion. In addition to this common shift in probing behavior, a phloem-located resistance factor was inferred in DK72-10 based on the longer time spent in phloem conditioning to attain phloem ingestion. In contrast, DK390 expressed the highest level of mesophyll and phloem-based resistance, in both cases seen with repeated attempts of short duration, a behavior typically associated with failed attempts to ingest. These findings support and are consistent with previous research, providing useful information to characterize maize hybrids resistant to D. maidis, and consequently to corn stunt.

Feeding Behavior of Riptortus pedestris (Fabricius) on Soybean: Electrical Penetration Graph Analysis and Histological Investigations.

Simple SummaryRiptortus pedestris (Fabricius) (Hemiptera: Alydidae) is a major soybean pest with the peak population occurrence during the seed maturity stage from pod filling to harvest. Soybean pods and/or seeds are required for R. pedestris development. However, the feeding strategies employed by this stink bug to feed on soybean are still not clear. In the present study, we recorded the feeding behaviors of R. pedestris on soybean using electropenetrography (EPG). The biological meaning of each waveform was confirmed by histological examination of plant tissues containing stylets or salivary sheath. In total, five phases of waveforms were identified: non-probing, pathway (Rp1), xylem sap ingestion (Rp2), salivation and ingestion (Rp3), and interruption (Rp4). Xylem ingestion (Rp2) was observed during R. pedestris feeding on soybean leaflets, stems, and pods, demonstrating that the stink bug was ingesting xylem sap from vascular tissue. Cell rupture (salivation/ingestion, Rp3) was only recorded during R. pedestris feeding on cotyledon and pods. Histological images showed that stylet tips and the salivary sheath were positioned in the tissues of cotyledon and pods. Taken together, our results demonstrate that R. pedestris uses a cell-rupture strategy to acquire nutrients from soybean pods and/or seeds while utilizing salivary sheath tactics to obtain water from xylem sap. These findings provide insightful information to understand the interactions between R. pedestris and the soybean plant. Riptortus pedestris (Fabricius) is a major agricultural pest feeding on soybean pods and seeds. The large populations occur during seed maturity stages from pod filling to harvest. Its infestation results in shriveled and dimpled seeds while vegetative structures (leaflet and stem) remain green, known as “Stay Green” syndrome. Additional evidence also demonstrates that soybean pods and seeds are required for Riptortus pedestris development. However, the feeding behavior strategies employed by this stink bug to feed on soybean plants are still not clear. In the present study, the feeding behaviors of R. pedestris on soybean plants were recorded by electropenetrography (EPG), and a waveform library was created for this species. A total of five phases of waveforms—nonprobing, pathway (Rp1), xylem sap ingestion (Rp2), salivation and ingestion (Rp3), and interruption (Rp4)—were identified. Non-probing waveforms Z and NP and pathway (Rp1) were found in all tested plant structures (leaflet, stem, cotyledon, and pods). Waveform Rp2 (xylem sap ingestion, xylem ingestion) was primarily recorded during R. pedestris feeding on leaflets and stems, while Rp3 (salivation/ingestion) was only observed during feeding on cotyledon and pods. Histological examinations confirmed that correlation between Rp2 and stylet tip positioning in the xylem vessel in leaflets and stems. Stylet tips end in the tissues of cotyledon and pods when Rp3 is recorded. Taken together, our results demonstrate that R. pedestris ingests xylem sap from vegetative tissues of soybean (leaflet and stem) via a salivary sheath strategy to obtain water. It mainly acquires nutrients from soybean pods and/or seeds using cell-rupture tactics. This study provided insightful information to understand the field occurrence patterns of “Stay Green” syndrome, which may have important implications for pest control.

Real-Time Feeding Behavior Monitoring by Electrical Penetration Graph Rapidly Reveals Host Plant Susceptibility to Crapemyrtle Bark Scale (Hemiptera: Eriococcidae).

Simple SummaryCrapemyrtle bark scale (CMBS; Acanthococcus lagerstroemiae), an invasive polyphagous sap feeder in the United States, has spread across 16 U.S. states in less than two decades, posing potential risks to the Green Industry. Confirming the host range is crucial for effective pest management of invasive insects. However, host range confirmation relying on greenhouse or field trials is often inefficient and time-consuming. In this study, we used the electrical penetration graph (EPG) to monitor the stylet penetration of CMBS in plant tissue in real-time. An R programming-based application was developed to better characterize the insect EPG waveforms recorded by EPG. By analyzing EPG-based EPG parameters, we demonstrated that CMBS has difficulty accessing the phloem tissue (salivation and ingestion) of a resistant plant compared to a susceptible plant. Importantly, we hereby present CMBS typical feeding behaviors on susceptible and non-susceptible plants comparatively, which provides direct evidence for revealing unknown hosts rapidly.Host range confirmation of invasive hemipterans relies on the evaluation of plant susceptibility though greenhouse or field trials, which are inefficient and time-consuming. When the green industry faces the fast-spreading threat of invasive pests such as crapemyrtle bark scale (Acanthococcus lagerstroemiae), it is imperative to timely identify potential host plants and evaluate plant resistance/susceptibility to pest infestation. In this study, we developed an alternative technology to complement the conventional host confirmation methods. We used electrical penetration graph (EPG) based technology to monitor the A. lagerstroemiae stylet-tip position when it was probing in different plant tissues in real-time. The frequency and relative amplitude of insect EPG waveforms were extracted by an R programming-based software written to generate eleven EPG parameters for comparative analysis between plant species. The results demonstrated that the occurrences of phloem phase and xylem phase offered conclusive evidence for host plant evaluation. Furthermore, parameters including the percentage of insects capable of accessing phloem tissue, time duration spent on initiating phloem phase and ingesting phloem sap, provided insight into why host plant susceptibility differs among similar plant species. In summary, this study developed a novel real-time diagnostic tool for quick A. lagerstroemiae host confirmation, which laid the essential foundation for effective pest management.

Seed Coating with Triflumezopyrim Induces the Rice Plant’s Defense and Inhibits the Brown Planthopper’s Feeding Behavior

Triflumezopyrim (TFM), a novel mesoionic insecticide, has been effective in controlling sucking insect pests such as the brown planthopper (BPH). However, the mechanism of TFM as a seed coating agent in paddy fields remains unclear. Here, we investigate the seed germination rates (>80%) and seedling emergence rates (>85%) of rice treated with TFM at 0, 22.5, 45.0, 67.5, and 90.0 g a.i. ha−1 with no significant effect on germination rates. In addition, the low TFM residue concentration (0.04 mg. kg−1) is maintained in the rice stem. Meanwhile, the TFM seeds’ treatments lead to increased oxalic acid, flavonoids, total phenol, callose contents, and elevated C/N ratio in rice plants at 60 and 90 days after sowing (DAS). The electrical penetration graph (EPG) results indicate that TFM as a seed coating treatment prolongs the non-probing period and inhibits phloem sap ingestion at 90 DAS. Furthermore, the mechanically transplanted rice treated with TFM provides long-term prevention against the BPH infestation. This study demonstrates that seeds treated with TFM play a vital role in controlling the BPH population up to >90%. These results provide a novel valuable control strategy for BPH in the rice fields.

Entomopathogenic fungus disrupts the phloem-probing behavior of Diaphorina citri and may be an important biological control tool in citrus

Citrus is among the most important fruit crops worldwide; however, numerous pests and diseases affect the orchards, increasing production costs. The psyllid Diaphorina citri, is a vector of the phloem-limited bacteria ‘Candidatus Liberibacter spp.’, the causal agent of Huanglongbing (HLB) disease. The lack of a cure for HLB requires management of the vector, mainly by intensive use of chemical insecticides, leading to the selection of resistant populations. Our study determined the effects of the entomopathogenic fungus Cordyceps fumosorosea on the probing behavior of D. citri at different time points after the fungus was applied by spraying. The electrical penetration graph technique was used to monitor the stylet activities of D. citri after application of the microbiological product. The effects were more pronounced between 30 and 96 h after the insects were sprayed, with significant disruption of the stylet activities related to the phloem and directly associated with the transmission of HLB. Our study indicated that the microbiological product Challenger®, with the active ingredient C. fumosorosea fungus, can significantly change the probing behavior of D. citri, may be helpful in more-sustainable management of the vector, and can be used to reduce the spread of HLB.

Reprogramming of sorghum proteome in response to sugarcane aphid infestation

Sugarcane aphid (SCA; Melanaphis sacchari Zehntner) is a key piercing-sucking pest of sorghum (Sorghum bicolor) that cause significant yield losses. While feeding on host plants, complex signaling networks are invoked from recognition of insect attack to induction of plant defenses. Consequently, these signaling networks lead to the production of insecticidal compounds or limited access of nutrients to insects. Previously, several studies were published on the transcriptomics analysis of sorghum in response to SCA infestation, but no information is available on the physiological changes of sorghum at the proteome level. We used the SCA resistant sorghum genotype SC265 for the global proteomics analysis after 1 and 7 days of SCA infestation using the TMT-plex technique. Peptides matching a total of 4211 proteins were identified and 158 proteins were differentially expressed at day 1 and 7. Overall, proteome profiling of SC265 after SCA infestation at days 1 and 7 revealed the suppression of plant defense-related proteins and upregulation of plant defense and signaling-related proteins, respectively. The plant defense responses based on proteome data were validated using electrical penetration graph (EPG) technique to observe changes in aphid feeding. Feeding behavior analyses revealed that SCA spent significantly longer time in phloem phase on SCA infested plants for day 1 and lesser time in day 7 SCA infested sorghum plants, compared to their respective control plants. Overall, our study provides insights into underlying mechanisms that contribute to sorghum resistance to SCA.

Screening and Evaluation for Antixenosis Resistance in Wheat Accessions and Varieties to Grain Aphid, Sitobion miscanthi (Takahashi) (Hemiptera: Aphididae).

The grain aphid, Sitobion miscanthi causes serious damage by removing nutritional content from wheat plants and transmitting viral diseases. The use of resistant wheat cultivars is an effective method of aphid management. To identify S. miscanthi resistant cultivars, preliminary antixenosis resistance screening was conducted on 112 Ethiopian and 21 Chinese wheat accessions and varieties along with bioassay to test for further antixenosis resistance, identification of aphid feeding behavior using electrical penetration graph (EPG), and imaging of leaf trichome densities using a 3D microscope. According to antixenosis resistance screening, one highly-resistant, 25 moderately-resistant, and 38 slightly-resistant wheat cultivars to S. miscanthi were identified. Aphid choice tests showed that Luxuan266, 243726, and 213312 were the least preferred after 12, 24, 48, and 72 h of S. miscanthi release. Longer duration of Np, longer time to first probe, and shorter duration of E2 waveforms were recorded in Lunxuan266, 243726, and 213312 than in Beijing 837. The trichome density on adaxial and abaxial leaf surfaces of Lunxuan266, 243726 and 213312 was significantly higher than on those of Beijing 837. We concluded that Lunxuan266, 243726, and 213312 were antixenosis resistant to S. miscanthi based on the choice test, EPG results, and leaf trichome densities.

Armet from whitefly saliva acts as an effector to suppress plant defences by targeting tobacco cystatin.

Arginine rich, mutated in early stage of tumours (Armet), is a well-characterized bifunctional protein as an unfolded protein response component intracellularly and a neurotrophic factor extracellularly in mammals. Recently, a new role of Armet as an effector protein mediating insect-plant interactions has been reported; however, its molecular mechanisms underlying the regulation of plant defences remain unclear. We investigated the molecular mechanisms underlying whitefly-secreted Armet-mediated regulation of insect-plant interaction by agrobacterium-mediated transient expression, RNA interference, electrical penetration graph, protein-protein interaction studies, virus-induced gene silencing assay, phytohormone analysis and whitefly bioassays. Armet, secreted by Bemisia tabaci whitefly, is highly expressed in the primary salivary gland and is delivered into tobacco plants during feeding. Overexpression of the BtArmet gene in tobacco enhanced whitefly performance, while silencing the BtArmet gene in whitefly interrupted whitefly feeding and suppressed whitefly performance on tobacco plants. BtArmet was shown to interact with NtCYS6, a cystatin protein essential for tobacco anti-whitefly resistance, and counteract the negative effects of NtCYS6 on whitefly. These results indicate that BtArmet is a salivary effector and acts to promote whitefly performance on tobacco plants through binding to the tobacco cystatin NtCYS6. Our findings provide novel insight into whitefly-plant interactions.

Helicoverpa armigera herbivory negatively impacts Aphis gossypii populations via inducible metabolic changes.

Helicoverpa armigera and Aphis gossypii are two important insect species that feed on cotton plants. These insects have distinct abilities to induce plant resistance and tolerate plant toxins, which results in interspecific competition imbalance that may be fatal to the low-tolerance A. gossypii and force these insects to develop avoidance behaviors and subsequently separate from their niche. We implemented ecological experiments to test the effects of H. armigera-induced plant resistance and behavioral avoidance in A. gossypii, and employed transcriptomics and metabolomics analyses to reveal changes in resistance genes and metabolites in plants. Our results demonstrate that cotton plants induced by H. armigera cause significant inhibitory and avoidance effects on A. gossypii insect populations. Electrical penetration graph (EPG) analysis showed changes in plant resistance induced by H. armigera leading to a decreased feeding efficiency of A. gossypii. In addition, genes associated with jasmonic acid and ethylene signaling pathways were significantly up-regulated in cotton plants after H. armigera induction, which led to a significant up-regulation of metabolites inducing plant resistance. These observations were corroborated by bioactivity analysis on metabolites, which showed that jasmonic acid, gossypol and tannins have significant inhibitory effects on A. gossypii populations. In contrast, methylparaben is associated with avoidance behaviors on A. gossypii populations. Our study suggests that the differences in the ability to induce plant resistance and tolerance between two non-predatory insects were lethal to low-tolerance A. gossypii insects, which might be a major factor determining their niche differentiation. This was further demonstrated by screening anti-insect and bio-hormonal metabolites. Our study provides a reference for investigating the evolutionary relationship between non-predatory insects and insights to implement effective insect biocontrol. © 2022 Society of Chemical Industry.

Short Neuropeptide F and Its Receptor Regulate Feeding Behavior in Pea Aphid (Acyrthosiphon pisum).

Simple SummaryWe 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.

Virus Infection and Host Plant Suitability Affect Feeding Behaviors of Cannabis Aphid (Hemiptera: Aphididae), a Newly Described Vector of Potato Virus Y.

Aphids are the most prolific vectors of plant viruses resulting in significant yield losses to crops worldwide. Potato virus Y (PVY) is transmitted in a non-persistent manner by 65 species of aphids. With the increasing acreage of hemp (Cannabis sativa L.) (Rosales: Cannabaceae) in the United States, we were interested to know if the cannabis aphid (Phorodon cannabis Passerini) (Hemiptera: Aphididae) is a potential vector of PVY. Here, we conduct transmission assays and utilize the electrical penetration graph (EPG) technique to determine whether cannabis aphids can transmit PVY to hemp (host) and potato (non-host) (Solanum tuberosum L.) (Solanales: Solanaceace). We show for the first time that the cannabis aphid is an efficient vector of PVY to both hemp (96% transmission rate) and potato (91%) using cohorts of aphids. In contrast, individual aphids transmitted the virus more efficiently to hemp (63%) compared to potato (19%). During the initial 15 min of EPG recordings, aphids demonstrated lower number and time spent performing intracellular punctures on potato compared to hemp, which may in part explain low virus transmission to potato using individual aphids. During the entire 8-hour recording, viruliferous aphids spent less time ingesting phloem compared to non-viruliferous aphids on hemp. This reduced host acceptance could potentially cause viruliferous aphids to disperse thereby increasing virus transmission. Overall, our study shows that cannabis aphid is an efficient vector of PVY, and that virus infection and host plant suitability affect feeding behaviors of the cannabis aphid in ways which may increase virus transmission.

Did Myzus persicae (Sulzer) from potato reared on a novel host for 15 years retain its host-related properties?

Myzus persicae (Sulzer) is an important agricultural pest worldwide causing major economic losses due to its ability to transmit over 100 viruses including Potato virus Y (PVY). Myzus persicae shows considerable variation with respect to performance on its host plants. The objective of this study was to use a survival experiment, behavioural observations, including observations of probing and feeding behaviour obtained using the electrical penetration graph (EPG) technique, and a PVY acquisition experiment to determine whether or not potato was still the more suitable host for M. persicae originating on potato and reared on a novel host, table beet, for over 15 years. In a survival experiment, the pre-reproductive period was significantly longer while adult survival and whole longevity were significantly lower for M. persicae reared on beet fed beet leaves compared to M. persicae reared on potato fed potato leaves. The number of progenies produced and fecundity were both significantly reduced (90 and 85%, respectively) for M. persicae reared on beet fed beet leaves. Ethological observations and EPG assessment of M. persicae behaviour reared on beet placed on beet leaves showed significantly impaired behavioural responses compared to M. persicae reared on potato placed on potato leaves. The rate of PVY acquisition was the same for M. persicae reared on beet and on potato. These results indicate that after 15 years on table beet, M. persicae still performs better on its original host, potato, and appears to be a specialized potato-adapted genotype.

Biocontrol Potential of Novel Emerging Multiacting Bacterium Bacillus amyloliquefaciens NC6 against Brevicoryne brassicae in Brassica rapa ssp. Pekinensis

The emerging elicitor protein PeBA1, extracted from Bacillus amyloliquefaciens NC6, was tested against the cabbage aphid (Brevicoryne brassicae) for its biocontrol potential. Its effects on the survival, lifespan, immature development, adult reproductive performance, and pest–pathogen interaction were assessed using electrical penetration graphs (EPGs) against B. brassicae. Furthermore, the direct effects of PeBA1 with temperature and climate change in Brassica rapa ssp. Pekinensis plants were investigated by the characterization of active compounds in B. amyloliquefaciens with multi-acting entomopathogenic effects. Compared with controls, PeBA1 treatments decreased (second- and third-generation) B. brassicae population growth rates. In a host selection test, control plants were colonized faster by B. brassicae than PeBA1-treated B. rapa plants. The B. brassicae nymphal development was extended by PeBA1 concentrations. Likewise, fecundity was reduced in PeBA1-treated seedlings compared with control, with fewer offspring produced. The trichomes and wax production on PeBA1-treated leaves resulted in a hostile environment for B. brassicae. PeBA1 altered the surface structure of the leaves, reducing B. brassicae reproduction and preventing colonization. Systemic defensive processes also included the activation of pathways (JA, SA, and ET). Based on these findings against B. brassicae, integrated pest management and bio control with PeBA1 in the agroecosystem appears to be suitable.

Local and systemic effect of azadirachtin on host choice and feeding activity of Macrosiphum rosae on rose plants

Many studies in the last decades addressed the antifeedant and repellent effects of neem (Azadirachta indica)-based pesticides on arthropod herbivores with azadirachtin as the leading active ingredient. However, results for aphids regarding repellence and feeding were often contradictory and mainly based on laboratory studies with self-made neem extracts in high concentrations. Moreover, no data are available on the presence of azadirachtin in the phloem of treated plants—the tissue that most aphids feed on. We investigated how the application of the commercial neem product NeemAzal-T/S onto only one plant half (upper or lower) affects nymphs of the aphid Macrosiphum rosae in clip cages on the untreated plant half. Results for aphid mortality indicate that active ingredients of NeemAzal-T/S are translocated both upwards and downwards in rose plants and that active ingredients are transported in the phloem. Furthermore, we investigated whether NeemAzal-T/S has a repellent or antifeedant effect on M. rosae. A choice test in the greenhouse as well as the quantification of honeydew excretion and electrical penetration graph analysis do not show differences in settling or feeding on untreated and NeemAzal-T/S-treated rose plants. Collectively, our data show that the effect of approved concentrations of NeemAzal-T/S on M. rosae is based on the toxicity after feeding on treated plants, not on starvation.

Leafhopper feeding behaviour on three grapevine cultivars with different susceptibilities to Flavescence dorée

Scaphoideus titanus (Ball) is a grapevine-feeder leafhopper, and the most important vector of Flavescence dorée of grapevine (FD), a disease associated with phytoplasmas belonging to ribosomal subgroups 16Sr-V–C and –D. FD is a major constraint to viticulture in several European countries and, so far, its control has relied on roguing of infected plants and insecticide applications against the vector. Detailed knowledge on different levels of the multifaceted phytoplasma-plant-vector relationship is required to envisage and explore more sustainable ways to control the disease spread. In the present work, S. titanus feeding behaviour was described on three grapevine cultivars: Barbera (susceptible to FD), Brachetto, and Moscato (tolerant to FD) using the Electrical Penetration Graph (EPG) technique. Interestingly, no differences were highlighted in the non-phloem feeding phases, thus suggesting that the tested cultivars have no major differences in the biochemical composition or structure of the leaf cuticle, epidermis or mesophyll, that can affect the first feeding activities. On the contrary, the results showed significant differences in leafhopper feeding behaviour in terms of the duration of the phloem feeding phase, longer on Barbera and shorter on Brachetto and Moscato, and of the frequency of interruption-salivation events inside the phloem, higher on Brachetto and Moscato. These findings indicate a possible preference for the Barbera cultivar, a better host for the leafhopper. Scaphoideus titanus feeding behaviour on Barbera correlates with an enhanced FDp transmission efficiency, thus explaining, at least in part, the higher susceptibility of this cultivar to FD. The mechanisms for the possible non-preference for Brachetto and Moscato are discussed, and an antixenosis is hypothesized. We propose that breeding for resistance against FD should take into account both plant traits associated with the response to the phytoplasmas and to the vector.

Host Acceptance and Plant Resistance: A Comparative Behavioral Study of Myzus persicae and Acyrthosiphon pisum.

Simple SummaryAphids are one of the most destructive insect pests worldwide. The green peach aphid (Myzus persicae) feeds on a broad range of plants, whereas the pea aphid (Acyrthosiphon pisum) only feeds on legumes. In this study, these two aphid species were used to investigate host acceptance and plant resistance to aphid feeding. Experiments on host plant preference and aphid performance (with regard to survival, development, and fecundity) confirmed that rape (Brassica rapa) is a suitable host and that faba bean (Vicia faba) is a poor host for the green peach aphid; for the pea aphid, faba bean is a suitable host, whereas rape is a nonhost. The probing and feeding behavior of these two aphid species on rape and faba bean was examined, and the results demonstrated the feeding preferences of these two aphid species. The green peach aphid had difficulty ingesting the phloem sap of faba bean. For the nonhost, the pea aphid spent relatively little time on mesophyll probing and did not achieve phloem sap ingestion. Furthermore, the effects of the probing and feeding behavior of specialist and generalist aphids on the spread of plant diseases caused by viruses were discussed.Aphids are prominent phloem-feeding insect pests. Myzus persicae and Acyrthosiphon pisum are generalist and specialist species, respectively. In this study, these two aphid species were used to investigate host acceptance and plant resistance to aphid feeding. M. persicae survived and reproduced on rape (Brassica rapa), but few individuals (9%) survived on faba bean (Vicia faba). A. pisum survived and reproduced on faba bean, but no A. pisum survived on rape. The probing and feeding behavior of M. persicae and A. pisum on rape and faba bean was examined using an electrical penetration graph (EPG) technique. The results demonstrated the feeding preferences of these two aphid species. The EPG results suggest that the resistance of faba bean to M. persicae and that of rape to A. pisum are likely residing in the phloem and mesophyll tissues, respectively. Due to the distinct probing and feeding behaviors, specialist and generalist aphids would have different impacts on the epidemiology of plant viral diseases. The findings can be applied to the management of viral diseases transmitted by specialist or generalist aphids in crop production.

Plant pathogen-mediated rapid acclimation of a host-specialized aphid to a non-host plant.

Polyphagous aphids often consist of host‐specialized lineages, which have greater fitness on their native hosts than on others. The underlying causes are important for understanding of the evolution of diet breadth and host shift of aphids. The cotton‐melon aphid Aphis gossypii Glover is extremely polyphagous with many strict host‐specialized lineages. Whether and how the lineage specialized on the primary host hibiscus shifts to the secondary host cucumber remains elusive. We found that the hibiscus‐specialized lineage suffered high mortality and gave birth to very few nymphs developing into yellow dwarfs on fresh cucumber leaves, and did not inflict any damage symptoms on cucumber plants. The poor performance did not improve with prolonged exposure to cucumber; however, it did significantly improve when the cucumber leaves were pre‐infected with a biotrophic phytopathogen Pseudoperonospora cubensis. More importantly, the hibiscus‐specialized lineage with two‐generation feeding experience on pre‐infected cucumber leaves performed as well as the cucumber‐specialized lineage did on fresh cucumber leaves, and inflicted typical damage symptoms on intact cucumber plants. Electrical penetration graph (EPG) indicated that the hibiscus‐specialized lineage did not ingest phloem sap from fresh cucumber leaves but succeeded in ingesting phloem sap from pre‐infected cucumber leaves, which explained the performance improvement of the hibiscus‐specialized lineage on pre‐infected cucumber leaves. This study revealed a new pathway for the hibiscus‐specialized lineage to quickly acclimate to cucumber under the assistance of the phytopathogen. We considered that the short feeding experience on pre‐infected cucumber may activate expression of effector genes that are related to specific host utilization. We suggest to identify host‐specific effectors by comparing proteomes or/and transcriptomes of the hibiscus‐specialized lineage before and after acclimating to cucumber.

Probing behavior of Neophilaenus campestris on various plant species

AbstractThe recent history of Xylella fastidiosa Wells et al. introduction in Europe illustrates how the lack of knowledge about the bacterium‐vector‐host interactions hinders the application of effective containment strategies, with the bacterial spread that currently appears relentless. Vector behavior is a key component in plant pathogen transmission; therefore, detailed knowledge of vector probing behavior on various host plants would furnish useful data for containing the bacterium. Aiming at this goal, we carried out electrical penetration graph (EPG)‐assisted probing behavior observations of the spittlebug, Neophilaenus campestris (Fallen) (Hemiptera: Aphrophoridae), a candidate vector widespread in all the areas where X. fastidiosa is currently established. Spittlebug probing was first characterized on one of its preferred host plants, Bromus madritensis L. (Poaceae), over short (6 h) and long (16 h) time spans. Thereafter, we performed comparative observations of the spittlebug probing on B. madritensis, olive, and grapevine. Overall, the probing behavior of N. campestris, that is, the main characteristic waveforms, did not differ substantially from that of Philaenus spumarius (L.) (Hemiptera: Aphrophoridae); however, here we report and describe a new kind of xylem activity interruption, provisionally termed N2, not previously observed in P. spumarius. Considering the time spent in xylem ingestion as a percentage of the total probing time (host‐suitability indicator used for spittlebugs), grapevine and B. madritensis are the preferred hosts for N. campestris, with olive as the least suitable among the plants tested. Successful probes, that is, probes during which xylem ingestion occurred, on grapevine were 2.5× greater than olive. Therefore, our data on N. campestris probing behavior suggest that the spittlebug role in X. fastidiosa epidemiology in European vineyards should be given more attention. In the present manuscript, we also discuss the current difficulties in interpreting and analyzing the outcomes of EPG studies on spittlebugs.