Electropenetrography Research Articles

Electropenetrography Study of Euschistus heros (F.) (Heteroptera: Pentatomidae) Nymphs Feeding on Vegetative Soybean Structures.

The study aimed to understand the feeding behavior of Neotropical brown stink bug nymphs Euschistus heros (F.) on soybean plants during vegetative stage through electropenetrography (EPG) technique. Three distinct phases were identified: non-feeding, pathway, and ingestion. Waveforms representing these phases were consistent across nymphal instars and plant structures, and named Np, Eh1, and Eh2, respectively. Biological interpretations of the waveforms were proposed by integrating visual observations, comparisons with adult waveforms, and histological studies. The waveforms Np, Eh1, and Eh2 were associated with resting/walking, initial stylet-plant contact, and xylem sap ingestion, respectively. Notably, nymphs showed a higher number of Eh1 events and longer durations when feeding on petioles compared to leaves, particularly in younger instars. However, differences between instars diminished in older nymphs. Fifth instars consistently exhibited the highest Eh1 values, and displayed longer xylem ingestion durations compared to other instars. Second instars demonstrated increased xylem ingestion events on petioles compared to leaves. Across plant structures, on petioles, nymphs generally showed longer xylem ingestion durations than on leaf surfaces. Fifth instar consistently had the longest ingestion durations overall. Additionally, statistical differences in xylem ingestion duration were observed between instars within each plant structure, with fifth instars displaying the longest durations. These findings offer valuable insights into the feeding behavior of E. heros nymphs, which could inform the development of more effective pest management strategies for soybean crops.

Effects of host plants on aphid feeding behavior, fitness, and Buchnera aphidicola titer.

Aphids are sap-feeding plant pests that depend on their symbiotic relationships with the primary endosymbiont Buchnera aphidicola to adapt to impoverished diets. However, how the host plant affects the aphid primary symbiont and aphid adaptation to host plant transfer are poorly known. In this study, aphid symbiont screening and genotype identification were used to establish 2 aphid strains (Rhopalosiphum maidis [Rm] and Rhopalosiphum padi [Rp] strains) containing only Buchnera without any secondary symbionts for both wheat aphid species (R. maidis and R. padi). Aphid fitness and Buchnera titers were unstable on some of these host plants after transferring to novel host plants (G1-G5), which were influenced by host plant species and generations; however, they stabilized after prolonged feeding on the same plants for 10 generations. The electropenetrography (EPG) records showed that the allocation of aphid feeding time was significantly distinct in the 6 host plants; aphids had more intracellular punctures and spent more nonprobing time on green bristlegrass which was not conducive to its growth compared with other plants. The content of soluble sugar, soluble protein, and amino acid in the leaves of the 6 host plants were also clearly separated. The correlation coefficient analysis showed that the nutrient contents of host plants had significant correlations with aphid feeding behaviors, fitness, and Buchnera titers. In the meantime, aphid fitness, and Buchnera titers were also affected by aphid feeding behaviors. Also, Buchnera titers of aphid natural populations on 6 host plants showed a visible difference. Our study deepened our understanding of the interaction among aphids, endosymbionts, and host plants, indicating that the host plant nutrient content is a predominant factor affecting aphid adaptation to their diet, initially affecting aphid feeding behaviors, and further affecting aphid fitness and Buchnera titers, which would further contribute to exploiting new available strategies for aphid control.

Adaptation of feeding behaviors on two Brassica species by colonizing and noncolonizing Bemisia tabaci (Hemiptera: Aleyrodidae) NW whiteflies.

Bemisia tabaci New World (NW)(Gennadius) (Hemiptera: Aleyrodidae), a whitefly in the B. tabaci species complex, is polyphagous on many plant species. Yet, it has been displaced, albeit not entirely, by other whitefly species. Potential causes could include issues with adaptation, feeding, and the colonization of new-hosts; however, insights that would help clarify these possibilities are lacking. Here, we sought to address these gaps by performing electropenetrography (EPG) recordings of NW whiteflies, designated "Napus" and "Rapa," reared on 2 colony hosts, Brassica napus and B. rapa, respectively. Analysis of 17 probing and pathway (pw) phase-related EPG variables revealed that the whiteflies exhibited unique probing behaviors on their respective colony hosts, with some deterrence being encountered on B. rapa. Upon switching to B. rapa and B. napus, the probing patterns of Napus and Rapa whiteflies, respectively, adapted quickly to these new-hosts to resemble that of whiteflies feeding on their colony hosts. Results for 3 of the EPG variables suggested that B. rapa's deterrence against Napus whitefly was significant prior to the phloem phase. This also suggested that adaptation by Rapa whitefly improved its pw probing on B. rapa. Based on analysis of 24 phloem phase-related EPG variables, Napus and Rapa whiteflies performed equally well once they entered phloem phase and exhibited comparable phloem acceptability on both the colony- and new-hosts. These findings demonstrate that NW whiteflies reared on a colony host are highly adaptable to feeding on a new host despite encountering some deterrence during the nonphloem phases in B. rapa plant.

Evaluation of Cotton Fleahopper (Pseudatomoscelis seriatus (Reuter)) Feeding on Mpp51Aa2-Traited Cotton Utilizing Electrical Penetration Graph (EPG) Waveforms.

Prior to the recent implementation of the Mpp51Aa2 pesticidal protein (ThryvOn), transgenic cotton cultivars have historically offered no control of the cotton fleahopper (Pseudatomocelis seriatus (Reuter)). To evaluate the feeding behavior of cotton fleahoppers on ThryvOn cotton, electropenetrography (EPG) using a Giga-8 DC instrument was used to monitor the probing activity of fourth- and fifth-instar cotton fleahopper nymphs on both ThryvOn and non-ThryvOn cotton squares. Nymphs were individually placed on an excised cotton square for 8 h of EPG recording, after which resulting waveforms were classified as non-probing, cell rupturing, or ingestion. Although there were significantly more cell rupturing events per insect on ThryvOn (mean ± SEM, 14.8 ± 1.7) than on non-ThryvOn squares (mean ± SEM, 10.3 ± 1.6), there was no difference attributable to ThryvOn in the average number of ingestion events per insect. However, the average duration of ingestion events was significantly shorter on squares with ThryvOn (mean ± SEM, 509 ± 148 s) than on squares without (mean ± SEM, 914 ± 135 s). This suggests that cotton fleahoppers continued to probe despite their inability to sustain ingestion. These results provide conclusive evidence that the Mpp51Aa2 pesticidal protein affects the feeding behavior of cotton fleahopper nymphs.

Volatiles from essential oils of three Lamiaceae plants repel the winged cotton aphid, disturb its feeding behavior and reduce its fecundity.

Insects use odor detection to sense their surroundings. Use of volatile compounds, such as essential oils (EOs) of plants, to repel pests and disrupt their olfaction-driven behaviors has great practical potential for use in integrated pest management. Despite the available information on the repellent effects of EOs on herbivorous insects, the olfaction-based mechanisms remain unknown. Y-tube olfactometer tests showed that the EOs of three Lamiaceae plants - Mentha arvensis L., Mentha piperita L. and Lavandula angustifolia Mill. - were significantly repellent to winged cotton aphid, Aphis gossypii Glover. Electrical penetration graph (EPG) tests indicated the EOs reduced phloem feeding and increased the level of non-productive probing by the aphids. The EOs also reduced the fecundity of winged Aphis gossypii. Electrophysiological bioassays and gas chromatography-mass spectrometry (GC-MS) identified five physiologically active volatiles, that is menthone, isomenthone, neomenthol and menthol from Mentha piperita; menthone and menthol from Mentha arvensis; and linalool from L. angustifolia. Behavioral tests confirmed that all five compounds repelled winged Aphis gossypii. Under field conditions, the growth rate of aphid populations after 7 days was significantly lower in fields treated with these compounds than in the control fields. Our findings demonstrated that three EOs not only repelled winged Aphis gossypii but also interfered with the aphid's feeding behavior and reduced its fecundity. These EOs and their active constituents have great potential as eco-friendly control products for use against Aphis gossypii. The effects of these EOs also exceed other repellents that only keep pests away from host plants. © 2024 Society of Chemical Industry.

Aphids May Facilitate the Spread of Sclerotinia Stem Rot in Oilseed Rape by Carrying and Depositing Ascospores.

Aphids and Sclerotinia stem rot in oilseed rape are often studied in isolation, and their relationship is rarely explored. Our field studies have revealed a significant positive correlation between the number of aphids and the incidence of Sclerotinia stem rot. Hence, starting with the colonizing stages of the two pests, Breveroryne brassicae was assessed for its potential to acquire, transmit, and inoculate Sclerotinia sclerotiorum by being sprayed with an ascospore suspension. Moreover, distinctions in aphid feeding behavior were examined between aphids on inoculated/uninoculated winter and spring oilseed rape plants or aphids, both with and without S. sclerotiorum ascospores, using electropenetrography (EPG). The results showed that aphid feeding followed by dropping ascospore suspension significantly increased the incidence of S. sclerotiorum. Ascospores were able to adhere to aphids and were carried by aphids to healthy plants, causing disease. The results of the EPG analysis indicated that aphid feeding behavior was significantly altered in all leaf tissue levels following infection with S. sclerotiorum. Specifically, aphids initiated their first puncture significantly sooner, began probing mesophyll cells earlier, had a significantly shorter pathway duration, and secreted saliva more frequently but reduced salivation prior to feeding and ingestion compared to aphids feeding on uninfected oilseed rape. Additionally, the feeding behavior of aphids carrying ascospores was markedly different from that of aphids not carrying ascospores, implying that ascospores directly influence aphid feeding behavior but that this influence appeared to be beneficial only for S. sclerotiorum infection. Aphids carrying ascospores started to puncture cells more quickly, with a significant increase in the frequency and duration of short probes and cell punctures, shortened pathway durations, and reduced salivation before feeding compared to aphids not carrying ascospores. It is clear that there is an interaction between aphids and S. sclerotiorum. The impact of S. sclerotiorum on aphid feeding behavior is directional, favoring the spread of the fungus.

Characterization and correlation of the probing behaviors of Macrosteles quadrilineatus (Hemiptera: Cicadellidae) with electropenetrography (EPG) waveforms.

Aster leafhopper (Hemiptera: Cicadellidae: Macrosteles quadrilineatus Forbes) is a polyphagous insect species that migrates into the upper Midwest of the United States and the Western Canadian Prairies. Populations of this insect are associated with the transmission of a plant pathogen (Candidatus Phytoplasma asteris, 16SrI) to several annual crops and perennial plant species. Previous studies suggest that aster leafhoppers can sometimes prefer less suitable hosts for their development and survival, yet it is unclear if this lower performance on certain plant species is associated with reduced or impaired probing behaviors due to characteristics of the plants. To characterize the probing behaviors of aster leafhoppers, direct current electropenetrography recordings of male and female adults on barley (Polaes: Poaceae: Hordeum vulgare L.) were combined with plant histology, allowing the identification of nine waveforms and their proposed biological meanings. For each waveform, the number of waveform events per insect (NWEI), the waveform duration per insect (WDI), the waveform duration per event per insect (WDEI), and the percentage of recording time were calculated and statistically compared between sexes. Male and female aster leafhoppers exhibited similar behavioral responses for most of these variables, except for the NWEI for waveforms associated with nonprobing activities and the pathway phase. In these cases, male aster leafhoppers exhibited a higher number of events than females. Comparison of the proposed waveforms in this study with previous work on other hemipteran species provided additional support to the interpretation of the biological activities associated with each waveform.

AC-DC Electropenetrography as a Tool to Quantify Probing and Ingestion Behaviors of the Yellow Fever Mosquito (Aedes aegypti) on Mice in Biocontainment.

A detailed understanding of mosquito probing and ingestion behaviors is crucial in developing novel interventions to interrupt the transmission of important human and veterinary pathogens, but these behaviors are difficult to observe as the mouthparts are inserted into the skin of the host. Electropenetrography (EPG) allows indirect observation, recording, and quantification of probing and ingestion behaviors of arthropods by visualizing the electrical waveform associated with these behaviors. The study of mosquito probing and ingestion behaviors has been limited to the use of human hands as host, which is not suitable for pathogen transmission studies. Mouse models of mosquito-borne diseases are a widely used and indispensable tool in this research, but previous attempts to use direct current EPG to study mosquito probing behaviors on mice have been unsuccessful. Accordingly, the present study used alternating or direct current (AC-DC) EPG to observe the ingestion behaviors of adult Aedes aegypti mosquitoes on a mouse host in real time under BSL-2 containment conditions with enhanced BSL-3 practices. Our results show that waveform families previously identified during Ae. aegypti probing and ingestion on human hands were observed using 100 mV of AC at an input resistance (Ri) of 107 Ohms (Ω) on CD1 mice. This work is a proof of concept for using mouse models for studying mosquito probing and ingestion behaviors with AC-DC EPG. In addition, these data show that the experimental setup used in these experiments is sufficient for conducting studies on mosquito probing and ingestion behaviors under BSL-2 containment conditions enhanced with BSL-3 practices. This work will serve as a foundation for using EPG to investigate the effects of pathogen infection on mosquito probing behaviors and to understand the real-time dynamics of pathogen transmission.

Bt Soybean Cry1Ac Does Not Affect Development, Reproduction, or Feeding Behavior of Red-Banded Stink Bug Piezodorus guildinii (Hemiptera: Pentatomidae).

Bt soybean cultivation is increasing worldwide. The Cry1Ac protein expressed in Bt soybean efficiently controls several lepidopteran pests. The stink bug, Piezodorus guildinii (Westwood), a major pest for soybean in the Americas, is not controlled by Bt crops, although possible sub-lethal effects may occur. Even if there were no negative effects for sting bug, ingesting toxins could affect its bio-controllers. We tested through ELISA detection if P. guildinii ingests Cry1Ac from Bt soybean and possible effects on its development, reproduction, survival, and feeding behavior. Biological traits were evaluated under controlled conditions of nymphs and adults feeding on pods of near-isogenic cultivars DM5958iPRO (Bt) and DM59i (non-Bt). Feeding behavior was recorded using an AC-DC electropenetrography (EPG) device. Results indicated that P. guildinii ingested the Cry1Ac protein; however, nymphal period and accumulated survival percentage did not differ between cultivars. Feeding on Bt soybean pods did not affect fecundity (i.e., number of egg masses and eggs/female) nor egg viability. Different feeding behaviors were only detected on the pathway phase (stylet penetration into plant tissue), which was more pronounced in the Bt cultivar. However, the total duration of the feeding activities on seeds was numerically higher (ca. 2X) on Bt plants compared to non-Bt. This is the first study to demonstrate that P. guildinii does ingest the Cry1Ac protein and excrete it without being absorbed, probably explaining the lack of direct adverse effects on its biological parameters. EPG could indicate that Bt soybean plants might be less palatable than non-Bt to red-banded stink bug.

Probing behavior of the leafhopper analyzed through DC electropenetrography and microscopy

Yamatotettix flavovittatus Matsumara is a new leafhopper species vector of sugarcane white leaf (SCWL) phytoplasma that causes sugarcane chlorosis symptoms. The effects of probing behavior of Y. flavovittatus on sugarcane and its implication for SCWL phytoplasma transmission are yet to be studied. In this research, we used DC electropenetrography (EPG) to define waveforms produced by adult and fifth-instar nymphal Y. flavovittatus on sugarcane and correlated them with salivary sheath termini (likely stylet tip locations) via light and scanning electron microscopy. The following six waveforms and associated activities are described: (NP) non-probing, (Yf1) stylet probing into epidermal cells, (Yf2) stylet probing through mesophyll/parenchyma, (Yf3) stylet contact with phloem and likely watery salivation, (Yf4) active ingestion of sap from phloem, probably sieve elements, and (Yf5) unknown stylet activity in multiple cell types. Study findings reveal that the Y. flavovittatus vector ingests sieve tube element more frequently and for longer durations than any other cell type, supporting that Y. flavovittatus is primarily a phloem feeder. Adult Y. flavovittatus show a longer total probing duration and produces a high density of puncture holes on sugarcane leaves. Moreover, probing behaviors revealed that adults typically ingest phloem sap more frequently and for longer durations than fifth-instar nymphs, enhancing sap ingestion. Furthermore, we propose that adults are more likely to acquire (during Yf4) and inoculate (during Yf3) higher amounts of phytoplasma than fifth-instar nymphs. This information on the penetration behavior of leafhopper Y. flavovittatus serves as a basis for advanced studies on the transmission mechanism of SCWL phytoplasma.

An electropenetrography waveform library for the probing and ingestion behaviors of Culex tarsalis on human hands.

Culex tarsalis Coquillett (Diptera: Culicidae) mosquitoes are capable of vectoring numerous pathogens affecting public and animal health. Unfortunately, the probing behaviors of mosquitoes are poorly understood because they occur in opaque tissues. Electropenetrography (EPG) has the potential to elucidate these behaviors by recording the electrical signals generated during probing. We used an AC-DC EPG with variable input resistors (Ri levels) to construct a waveform library for Cx. tarsalis feeding on human hands. Biological events associated with mosquito probing were used to characterize waveforms at four Ri levels and with two electrical current types. The optimal settings for EPG recordings of Cx. tarsalis probing on human hands was an Ri level of 107 Ohms using an applied signal of 150 millivolts alternating current. Waveforms for Cx. tarsalis included those previously observed and associated with probing behaviors in Aedes aegypti L. (Diptera: Culicidae): waveform families J (surface salivation), K (stylet penetration through the skin), L (types 1 and 2, search for a blood vessel/ingestion site), M (types 1 and 2, ingestion), N (type 1, an unknown behavior which may be a resting and digestion phase), and W (withdrawal). However, we also observed variations in the waveforms not described in Ae. aegypti, which we named types L3, M3, M4, and N2. This investigation enhances our understanding of mosquito probing behaviors. It also provides a new tool for the automated calculation of peak frequency. This work will facilitate future pathogen acquisition and transmission studies and help identify new pest and disease management targets.

Electropenetrography study of Euschistus heros (F.) (Heteroptera: Pentatomidae) nymphs feeding at different sites of the soybean pod.

This study aimed to characterize and correlate the stylet penetration behaviors of nymphs of the Neotropical brown stink bug, Euschistus heros (F.), on immature soybean pods. Waveforms were obtained using electropenetrography (EPG). The findings revealed that the nymphs exploited the xylem vessels and the seed tegument or endosperm. Primarily 4 phases were characterized: nonfeeding, pathway, salivation, and ingestion. The waveforms of each phase were similar in appearance across instars. The biological meanings of waveforms were based on visual observations, comparison with waveforms of adults, and histological studies. Np represents the insect resting or walking on soybean pod surface. Eh1 represents the first contact between the mouthparts (stylets) and plant tissue. Eh2 represents xylem sap ingestion, and Eh3 represents seed activities (including tegument and endosperm). The number of waveform events did not differ among instars for all waveforms. However, for Eh3, fifth instars performed more activities than other instars. The second instars had the smallest value, and third and fourth instars had intermediate values. For total duration, all waveforms differed among instars. Np duration was shorter for third compared with second and fourth instars and intermediate for fifth instar. For Eh1, second and third instars had the longest duration (1.5× to 2× greater) compared with fourth and fifth instars. For Eh2 and Eh3, the second-instar showed the longest (~2× greater) and shortest durations, respectively. Overall, this study provides important insights into the feeding behavior of E. heros nymphs so that effective pest management programs can be developed to contain this pest.

Aphids on Aphid-Susceptible Cultivars Have Easy Access to Turnip Mosaic Virus, and Effective Inoculation on Aphid-Resistant Cultivars of Oilseed Rape (Brassica napus).

Plant viruses improve transmission efficiency by directly and indirectly influencing vector behavior, but the impact of plant cultivars on these modifications is rarely studied. Using electropenetrography (EPG) technology, a comparative study of the effects of turnip mosaic virus (TuMV) infection on quantitative probing behaviors of the cabbage aphid (Brevicoryne brassicae) was conducted on two oilseed rape cultivars ('Deleyou6' and 'Zhongshuang11'). Compared to mock-inoculated plants, cabbage aphids on infected plants increased the frequency of brief probing, cell penetration, and salivation. Additionally, aphids on infected 'Deleyou6' prolonged cell penetration time and decreased ingestion, but not on infected 'Zhongshuang11', suggesting that aphids were more likely to acquire and vector TuMV on the aphid-susceptible cultivar 'Deleyou6' than on resistant cultivars. TuMV also affected aphid probing behavior directly. Viruliferous aphids reduced the pathway duration, secreted more saliva, and ingested less sap than non-viruliferous aphids. In comparison with non-viruliferous aphids, viruliferous aphids started the first probe earlier and increased brief probing and cell penetration frequencies on the aphid-resistant cultivar 'Zhongshuang11'. Based on these observations, viruliferous aphids can be inoculated with TuMV more efficiently on 'Zhongshuang11' than on 'Deleyou6'. Although aphid resistance and TuMV infection may influence aphid probing behavior, oilseed rape resistance to aphids does not impede TuMV transmission effectively.

Investigation of an expanded, lumped-element model of electropenetrography (EPG) and the accuracy of the traditional R and emf model

Electropenetrography (EPG) allows researchers to measure feeding behaviors of piercing-sucking pests such as aphids and mosquitoes. The presently accepted electrical circuit model of an EPG monitor shows excitation voltage (Vin) applied to a feeding substrate (such as a plant), then interacting with a variable resistor (Ra) representing the feeding insect, plus biopotentials generated in the plant-insect interface (Vemf). Together, Ra and Vemf then interact with the input resistor (Ri) of the first amplifier in the monitor. After signal processing, variable voltage output (Vout) results in waveforms that represent insect feeding. The present work performed a lumped-element circuit analysis to determine whether the above simple model was correct by today’s electrical engineering standards. This work showed that the above model was too simple to explain EPG signals. The newly developed model adds valuable information on parasitic effects such as capacitance in the amplifier (Ci) and from poor insect tethering (Cai), as well as effects of filtering due to induction (Lp) and capacitance (Cp) in the plant. This work recommends specific steps to avoid these problems. In addition, it was found that the insect is a complex electrical system, now symbolized by Za for impedance. Yet, despite the findings of both parasitic and complex, new elements in the EPG model, this work fundamentally supports the foundation of EPG science. That is, it confirms that the emf and still-named R components exist and can provide the basis for interpretation of EPG waveforms. Nonetheless, designers and researchers must be alert to potential problems with parasitic elements in the EPG circuit and take active steps to reduce them. These findings provide important benchmark data for future studies of the nature of Za, especially its importance in interpreting waveforms in the context of the crucial emf and R responsiveness curves. Together with the present study, such analysis will enable improvements in future EPG monitor designs.

Feeding Behavior Comparison of Bean Bugs, Riptortus pedestris and Halyomorpha halys on Different Soybean Cultivars

Riptortus pedestris (Fabricius) and Halyomorpha halys (Stål) are the major pests that feed on soybean pods, seeds, and fruits. Higher populations and damage occur during the soybean maturity stages (podding to harvest). To compare the feeding behavior of R. pedestris and H. halys, we used the six most cultivated cultivars (Daepung-2ho, Daechan, Pungsannamul, Daewon, Seonpung, and Seoritae) in Korea using the electropenetrography (EPG) technique. Both R. pedestris and H. halys, the NP (non-penetration), a non-probing waveform, was the shortest in the Pungsannamul (298 and 268 min) and the longest in the Daepung-2ho (334 and 339 min), respectively. The feeding waveforms Pb (phloem feeding: E1-Salivation and E2-Sap feeding) and G (xylem feeding) were the longest in Pungsannamul and the shortest in Daepung-2ho. In addition, as a result of investigating the damage rate by planting six cultivars of beans in the field, as expected, the proportions of damage types B and C were highest in Pungsannamul and lowest in Daepung-2ho. These results reveal that both bug species ingest xylem sap from leaflets and stems using a salivary sheath strategy to acquire water and nutrients from soybean pods/seeds with cell-rupture tactics. This study provides perceptive information to understand the feeding behavior, field occurrence, and damage patterns of R. pedestris and H. halys, which may have key implications for the management of hemipteran pests by determining the specificity and susceptibility of host plants.

Turnip Mosaic Virus Infection Differentially Modifies Cabbage Aphid Probing Behavior in Spring and Winter Oilseed Rape (Brassica napus).

Simple SummaryWinter oilseed rape accounts for about 90% of total oilseed rape planting area in China, with the majority of it concentrated in the Yangtze River basin. The remaining 10% is mostly found in the provinces of China’s northwest plateau. Winter oilseed rape areas in China have gradually expanded to the north in the last decade, resulting in cabbage aphids and turnip mosaic virus (TuMV). The aphid is a vector for TuMV and is gradually increasing on winter and spring oilseed rape. Quantifying the probing behaviors of the aphids on spring oilseed rape and winter oilseed rape helps us to understand TuMV regulation of the aphids. We found that compared to mock-inoculated plants, cabbage aphids on infected plants increased brief probing frequency, cell penetration frequency, intracellular probing time, decreased time to first probe and pathway duration, potentially promoting viral acquisition and minimizing viral loss and plant damage. Viruliferous aphids had reduced pathway duration, increased cell penetration frequency, increased intracellular probing time, increased salivation frequency, and ingested less sap compared with non-viruliferous aphids, primed for viral infection. TuMV infection also differentially modified aphid feeding behavior on winter and spring oilseed rape cultivars, primarily on uninfected plants.Direct and indirect effects of plant virus infection on vector behavior have been discovered to improve virus transmission efficiency, but the impact of plant cultivars in virus–vector–plant interactions has received little attention. Electropenetrography (EPG) allows real-time tracking and quantification of stylet penetration behaviors, pathogen transmission, and plant resistance mechanisms. Quantitative probing behaviors on a spring oilseed rape cultivar, ‘Xinyou17’, and a winter oilseed rape cultivar, ‘Zheping4’, were investigated using EPG to compare turnip mosaic virus (TuMV) regulation of cabbage aphid probing behavior. Results for indirect effects showed that compared to mock-inoculated plants, cabbage aphids on infected plants increased brief probing frequency, cell penetration frequency, intracellular probing time, and decreased time to first probe and pathway time, potentially promoting viral acquisition. TuMV also directly influences aphid probing behavior. Viruliferous aphids had reduced pathway time, increased cell penetration frequency, increased intracellular probing time, increased salivation frequency, and ingested less sap than non-viruliferous aphids, primed for viral infection. Although oilseed rape cultivars can also influence aphid behavior, the main effect of cultivars was not significant on TuMV-infected plants.

Behavioral disruption of two orchard hemipteran pests by garlic essential oil

AbstractEssential oils (EOs) are promising sources of effective bioactive compounds for insect pest control, especially in heavily treated agrosystems like orchards. Plants belonging to the Amaryllidaceae family – onion, garlic (Allium sativum L.), and others – present an interesting potential in apple (Malus domestica Borkh.) and pear (Pyrus communis L., both Rosaceae) tree integrated pest management due to the repellent and deterrent properties of their chemical compounds. This study explores the potential disruption by garlic EOs of the behavior of two hemipteran pests towards their host trees: rosy apple aphid, Dysaphis plantaginea (Passerini) (Aphididae), on cultivated apple, and pear psyllid, Cacopsylla pyri (L.) (Psyllidae), on pear. Effects on orientation and feeding behavior were evaluated using choice tests and the electropenetrography (EPG) technique. The results revealed that garlic EOs delivered through rubber dispensers were intrinsically repellent, but when combined with the attractive host plant, the repellents and attractants canceled each other out for the two studied hemipterans. Garlic EOs caused disruptions in the feeding behavior of the two insects, particularly the psyllids, which showed a decrease in phloem sap intake. This study highlights the mechanisms disrupting the process of host‐plant colonization, in particular the possible masking of host‐locating cues or the involvement of confusing chemical stimuli due to EO volatiles.

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.

Monarda didyma Hydrolate Affects the Survival and the Behaviour of Drosophila suzukii.

Simple SummaryDuring the steam distillation of aromatic plants, two main fractions are usually obtained: the hydrophobic essential oils and the hydrophilic fraction commonly known as hydrolate (HY). The essential oils are largely used in several industry fields, including the agricultural industry as biopesticides. Residual HYs, instead, are often discarded as by-products of little or no value. Our research pointed out that also HYs have biological activity, suggesting their potential use in plant-based strategy for the pest control. In more detail, we investigated the insecticidal properties of the hydrolate from Monarda didyma, scarlet beebalm, towards Drosophila suzukii. Using specific molecular and behavioural assays, we showed that M. didyma hydrolate affected the fitness and behaviour of D. suzukii, providing new insights in the D. suzukii control strategies through M. didyma hydrolate.Drosophila suzukii (Matsumara) is an herbivorous pest whose control in the field with conventional chemical is particularly difficult and has important drawbacks. Here, we investigated the insecticidal properties of hydrolate from Monarda didyma, scarlet beebalm, an aromatic herb in the Lamiaceae family. The identification of volatile organic compounds (VOCs) by CG–MS systems revealed that thymol (38%) and carvacrol (59%) were the most abundant VOCs in the hydrolate. M. didyma hydrolate did not show fumigant toxicity. Conversely, in contact assays, M. didyma hydrolate showed a LC50 of 5.03 µL mL−1, 48 h after the application on D. suzukii adults. Expression of detoxification genes increased in flies that survived the LC50 application. Furthermore, toxicity persisted for 7 days after the treatment in the survival evaluation. Artificial diet assays with 100 and 1000 µL mL−1 of M. didyma hydrolate resulted in a significant decrease in total food intake in both male and female D. suzukii adults. In addition, electropenetrography (EPG) showed that the D. suzukii females’ feeding behaviour was altered in hydrolate-treated diets. The hydrolate also caused a significant reduction in the number of eggs laid in two different oviposition assays. Overall, our findings provide a new perspective for the improvement of D. suzukii control strategies through M. didyma hydrolate.

Seed Damage by the Neotropical Brown Stink Bug, Euschistus heros (F.) to Resistant Soybean Cultivars with the Block Technology Versus a Susceptible Cultivar.

Laboratory and greenhouse studies were conducted with the Neotropical brown stink bug, Euschistus heros (F.), to evaluate and compare adult damage at two infestation levels (2 and 4 adults/plant) and feeding behavior on seeds of three resistant soybean cultivars bearing the Block technology ('BRS 1003 IPRO', 'BRS 543 RR', and 'BRS 391') compared to a susceptible cultivar ('BRS 5601 RR'). No difference in number or weight of damaged seeds (as percentages) was found among the cultivars at either infestation level. Differences were only observed between infestation levels within cultivar; higher values were reported with 4 adults/plant, except for 'BRS 543 RR'. At 2 adults/plant, total seed area damaged (mm2) and percentage of seed area damaged were significantly lower only on 'BRS 1003 IPRO'; significant differences among cultivars were found in damage to internal but not external seed surfaces. At 4 adults/plant, all Block cultivars differed from 'BRS 5601 RR' in overall seed damage, and greater percent damage occurred on both seed surfaces on 'BRS 5601 RR'. Electropenetrography (EPG) demonstrated that adults reached and fed in the seeds of all soybean cultivars. However, the feeding event duration in seeds of Block cultivars was much shorter than on 'BRS 5601 RR', which likely explains differences between internal and external seed damage. Furthermore, the total duration of feeding activities on seeds of Block cultivars was ca. 4-6 times shorter than on the susceptible cultivar; these two EPG feeding variables account for the lower seed damage observed for the Block cultivars. These cultivars represent an important new strategy for pest control on soybean.