Leaffooted Bugs: Insect Pest Species of Growing Concern for Agriculture.

Leptoglossus (Hemiptera: Coreidae) are a diverse genus of phytophagous insects. Literature regarding Leptoglossus has increased as species are identified as emerging agricultural pests or reported outside of their native range. Within Leptoglossus, five species dominate the literature and are known pests and vectors of plant pathogenic microbes in several major crops. Despite the increasing profile of Leptoglossus, current monitoring and management methods rely primarily on visual inspection, and semiochemical tools have yet to be developed. This Perspective identifies and discusses gaps in the Leptoglossus literature as well as areas of research needed for the development of effective tools for monitoring insect populations and enabling informed pest-management practices.

Maize terpene synthase 1 impacts insect behavior via the production of monoterpene volatiles ß-myrcene and linalool.

Plant-derived volatiles are important mediators of plant-insect interactions as they can provide cues for host location and quality, or act as direct or indirect defense molecules. The volatiles produced by Zea mays (maize) include a range of terpenes, likely produced by several of the terpene synthases (TPS) present in maize. Determining the roles of specific terpene volatiles and individual TPSs in maize-insect interactions is challenging due to the promiscuous nature of TPSs in vitro and their potential for functional redundancy. In this study, we used metabolite GWAS of a sweetcorn diversity panel infested with Spodoptera frugiperda (fall armyworm) to identify genetic correlations between TPSs and individual volatiles. This analysis revealed a correlation between maize terpene synthase 1 (ZmTPS1) and emission of the monoterpene volatiles linalool and ß-myrcene. Electroantennogram assays showed gravid S. frugiperda could detect both linalool and ß-myrcene. Quantification of headspace volatiles in a maize tps1 loss-of-function mutant confirmed that ZmTPS1 is an important contributor to linalool and ß-myrcene emission in maize. Furthermore, pairwise choice assays between tps1 mutant and wild-type plants showed that ZmTPS1, and by extension its volatile products, aid host location in the chewing insect S. frugiperda, yet repel the sap-sucking pest, Rhopalosiphum maidis (corn leaf aphid). On the other hand, ZmTPS1 had no impact on indirect defense via the recruitment of the parasitoid Cotesia marginiventris. ZmTPS1 is therefore an important mediator of the interactions between maize and its insect pests.

Blueberries infected with the fungal pathogen Colletotrichum fioriniae release odors that repel Drosophila suzukii.

BACKGROUND: Spotted-wing drosophila, Drosophila suzukii, is a serious pest of thin-skinned fruits. Alternative methods to control this pest are needed to reduce insecticide use, including new repellents. Previous research demonstrated that D. suzukii adults use odor cues to avoid blueberries infected with the fungal pathogen Colletotrichum fioriniae, which causes the disease anthracnose. To identify novel D. suzukii repellents, we investigated the volatile emission from experimentally-infected fruit, which were inoculated with C. fioriniae isolates in the laboratory, and from field-collected fruit, which were naturally infected and harvested from a field. We then tested the pathogen-induced volatiles on D. suzukii adult behavior. RESULTS: Volatile emission was similar between all five C. fioriniae strains, with good agreement between experimentally-infected and field-collected berries. In total, 14 volatiles were found to be more abundant in infected versus uninfected fruit headspace. In multiple-choice bioassays, nine of the 14 volatiles elicited repellency responses from adult D. suzukii. These nine volatiles were further evaluated in dual choice assays, where all nine reduced fly capture by 43-96% compared to the control. The most repellent compounds tested were the esters ethyl butanoate and ethyl (E)-but-2-enoate, which were more or equally repellent to the known D. suzukii repellents 1-octen-3-ol, geosmin, and 2-pentylfuran. Dose-response assays identified concentration-dependent effects on D. suzukii repellency and oviposition when applied individually and consistent aversion observed across doses of a 1:1 blend. CONCLUSION: We report two repellents from C. fioriniae-infected blueberries that could be useful semiochemicals for the behavioral manipulation of D. suzukii in the field. © 2023 Society of Chemical Industry. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Field evaluation of glutathione and glutamic acid as host marking pheromones for control of Tephritid fruit flies in a mango orchard in Kenya.

BACKGROUND: Tephritid fruit flies are a major constraint to the global production of horticultural crops. In Africa, an array of native and alien invasive fruit fly species represents a key challenge to the horticultural industry. In an effort to develop a safer management tool for these pests, we previously identified glutathione (GSH) and glutamic acid (GA), as the host marking pheromones (HMPs) of Ceratitis cosyra and Ceratitis rosa, respectively. Here, we report on the effectiveness of these compounds (GSH and GA) in reducing natural fruit fly population infestations in mango orchards. Mango trees at two different agroecological zones in Kenya were sprayed with HMPs, and the fruits sampled periodically and assessed for fruit fly emergence. Fruit fly emergence data were compared to two controls, the positive control using spot spraying of food bait (SS), and the negative control using water (W). RESULTS: The two HMPs and SS substantially decreased fruit fly emergence from the sampled mangoes. GSH and GA treated mangoes showed reduced C. cosyra and C. rosa infestation by ~70-75% relative to control (W), and with variation noted in the pheromone applied and the recovered fruit fly species. The adult emergence in pheromone-treated plants compared favorably with the positive control SS. However, the HMPs had little impact on reducing Bactrocera dorsalis infestation of mangoes. CONCLUSION: The decrease in fruit fly emergence in sampled mango fruits from HMP treated trees corroborate previous laboratory results and support the prospect of using HMPs in the management of African fruit fly species. © 2022 Society of Chemical Industry.

Early infestation volatile biomarkers of fruit fly Bactrocera dorsalis (Hendel) ovipositional activity in mango (Mangifera indica L.).

Infestation of agricultural commodities by insect pests results in significant economic, import and export, food safety, and invasive insect introduction issues for growers, consumers, and inspectors. The Oriental fruit fly (Bactrocera dorsalis) is considered a highly invasive insect pest with populations reported in more than 60 countries, with prevalent distributions in Asia and Africa. B. dorsalis is phytophagous with a host range encompassing hundreds of fruits and vegetables. Damage to the fruit or vegetable is inflicted through oviposition and subsequent larval feeding resulting in spoilage. Early detection of insect pest infestations is a critical component for ensuring food safety as well as controlling introduction and spread of invasive insects. However, detection of ovipositional activity and early larval development is visually difficult, thus rapid and non-destructive detection often relies on odors associated with infestation. We investigated the odors of mangoes (Mangifera indica L.) infested with B. dorsalis and compared the volatile profiles of infested mangoes to non-infested and mechanically damaged mangoes 24 h post-infestation. GC-MS and multivariate analyses provided the identification of eleven compounds unique to infested mangoes compared to mechanically damaged or non-infested fruit. Results indicated compositional and quantitative differentiation of volatile profiles among treatments for detection of infested fruit at quality checks or points of commerce.

Chemo-Ecological Insights into the Use of the Non-Host Plant Vegetable Black-Jack to Protect Two Susceptible Solanaceous Crops from Root-Knot Nematode Parasitism.

Plant parasitic nematodes (PPNs) develop through three major stages in their life cycle: hatching, infection, and reproduction. Interruption of any of these stages can affect their growth and survival. We used screenhouse pot experiments, laboratory in vitro hatching and mortality assays, and chemical analysis to test the hypothesis that the non-host Asteraceae plant vegetable black-jack (Bidens pilosa) suppresses infection of the PPN Meloidogyne incognita in two susceptible Solanaceae host plants, tomato (Solanum lycopersicum) and black nightshade (S. nigrum). In intercrop and drip pot experiments, B. pilosa significantly reduced the number of galls and egg masses in root-knot nematode (RKN)-susceptible host plants by 3-9-fold compared to controls. Chemical analysis of the most bioactive fraction from the root exudates of B. pilosa identified several classes of compounds, including vitamins, a dicarboxylic acid, amino acids, aromatic acids, and a flavonoid. In in vitro assays, the vitamins and aromatic acids elicited the highest inhibition in egg hatching, whereas ascorbic acid (vitamin) and 2-hydroxybenzoic acid (aromatic acid) elicited strong nematicidal activity against M. incognita, with LC50/48 h values of 12 and 300 ng/µL, respectively. Our results provide insights into how certain non-host plants can be used as companion crops to disrupt PPN infestation.

Identification of Repellents from Four Non-Host Asteraceae Plants for the Root Knot Nematode, Meloidogyne incognita.

Olfactory cues guide plant parasitic nematodes (PPNs) to their host plants. We tested the hypothesis that non-host plant root volatiles repel PPNs. To achieve this, we compared the olfactory responses of infective juveniles (J2s) of the PPN Meloidogyne incognita to four non-host Asteraceae plants, namely, black-jack (Bidens pilosa), pyrethrum (Chrysanthemum cinerariifolium), marigold (Tagetes minuta), and sweet wormwood (Artemisia annua), traditionally used in sub-Saharan Africa for the management of PPNs. Chemical analysis by coupled gas chromatography-mass spectrometry (GC/MS) combined with random forest analysis, followed by behavioral assays, identified the repellents in the root volatiles of B. pilosa, T. minuta, and A. annua as (E)-ß-farnesene and 1,8-cineole, whereas camphor was attractive. In contrast, random forest analysis predicted repellents for C. cinerariifolium and A. annua as ß-patchoulene and isopropyl hexadecanoate. Our results suggested that terpenoids generally account for the repellency of non-host Asteraceae plants used in PPN management.

Sampling of Volatiles in Closed Systems: A Controlled Comparison of Three Solventless Volatile Collection Methods.

Complex inter-organismal communication among plants, insects, and microbes in natural and agricultural ecological systems is typically governed by emitted and perceived semiochemicals. To understand and ultimately utilize the role of volatile semiochemicals in these interactions, headspace volatiles are routinely collected and analyzed. Numerous collection systems are available (e.g., static or dynamic; adsorption or absorption) where the choice of technique should be dependent upon the plant, insect, or microbial ecological system studied, the information sought, and the limitations of each method. Within these constraints, it remains necessary that each method detects and provides the accurate in situ, or in vitro, volatile profile of the studied system. Herein, we analyzed and compared the pros and cons of three solventless, thermal desorption systems (SPME, Tenax/cold trap, SPDE) using a synthetic standard blend of compounds mimicking a simple natural blend (benzaldehyde, b-caryophyllene, (Z)-3-hexenol, 6-methyl-5-hepten-2-one, and limonene). Direct splitless injection and Super Q collections of the standard blend were used as controls. The results indicated that related qualitative, as well as quantitative differences, could be correlated with adsorbent sampling capacity and structural bias. The results for Tenax/cold trap and SPDE also were affected by sampled headspace volumes. All solventless techniques exhibited high analytical reproducibility, with SPME and SPDE providing ease of use, low cost, and minimal instrument modifications. The more complex Tenax/cold trap technique provided higher collection efficiency. Using these results, we provide guidance for technique selection for chemical communication applications.

Zea mays Volatiles that Influence Oviposition and Feeding Behaviors of Spodoptera frugiperda.

Fall armyworm (Spodoptera frugiperda) is a major global pest of many crops, including maize (Zea mays). This insect is known to use host plant-derived volatile organic compounds to locate suitable hosts during both its adult and larval stages, yet the function of individual compounds remains mostly enigmatic. In this study, we use a combination of volatile profiling, electrophysiological assays, pair-wise choice behavioral assays, and chemical supplementation treatments to identify and assess specific compounds from maize that influence S. frugiperda host location. Our findings reveal that methyl salicylate and (E)-alpha-bergamotene are oviposition attractants for adult moths but do not impact larval behavior. While geranyl acetate can act as an oviposition attractant or repellent depending on the host volatile context and (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT) is an oviposition deterrent. These compounds can also be attractive to the larvae when applied to specific maize inbreds. These data show that S. frugiperda uses different plant volatile cues for host location in its adult and larval stage and that the background volatile context that specific volatiles are perceived in, alters their impact as behavioral cues.

Pollen and yeast change nectar aroma and nutritional content alone and together, but honey bee foraging reflects only the avoidance of yeast.

Floral nectar often contains pollen and microorganisms, which may change nectar's chemical composition, and in turn impact pollinator affinity. However, their individual and combined effects remain understudied. Here, we examined the impacts of the nectar specialist yeast, Metschnikowia reukaufii, and the addition of sunflower (Hellianthus annus) pollen. Pollen grains remained intact, yet still increased yeast growth and amino acid concentrations in nectar, whereas yeast depleted amino acids. Pollen, but not yeast, changed nectar sugar concentrations by converting sucrose to its monomers. Both pollen and yeast contributed emissions from nectar, though yeast volatiles were more abundant than pollen volatiles. Yeast volatile emission was positively correlated with pollen concentration and cell density, and yeast depleted a subset of pollen-derived volatiles. Honey bees avoided foraging on yeast-inoculated nectar and foraged equally among uninoculated nectars regardless of pollen content, underscoring the importance of microbial metabolites in mediating pollinator foraging.

Volatile microbial semiochemicals and insect perception at flowers.

Many plant-associated microbial communities produce volatile signals that influence insect responses, yet the impact of floral microorganisms has received less attention than other plant microbiomes. Floral microorganisms alter plant and floral odors by adding their own emissions or modifying plant volatiles. These contextual and microbe species-specific changes in floral signaling are detectable by insects and can modify their behavior. Opportunities for future work in floral systems include identifying specific microbial semiochemicals that underlie insect behavioral responses and examining if insect species vary in their responses to microbial volatiles. Examining if documented patterns are consistent across diverse plant-microbe-insect interactions and in realistic plant-based studies will improve our understanding of how microbes mediate pollination interactions in complex system.

Compounds Associated with Infection by the Root-Knot Nematode, Meloidogyne javanica, Influence the Ability of Infective Juveniles to Recognize Host Plants.

Plant root chemistry is altered by the parasitism of plant-parasitic nematodes (PPN). Here, we investigated the influence of the infective stage juveniles (J2) of Meloidogyne javanica in inducing tomato (Solanum lycopersicum) root volatiles and chemotactic effect on conspecifics. In olfactometer assays, J2 avoided the roots of 2-day infected plants but preferred 7-day-infected tomato compared to healthy plants. Chemical analysis showed a 2-7-fold increase in the amounts of monoterpenes emitted from tomato roots infected with M. javanica relative to healthy roots. In further bioassays, the monoterpenes ß-pinene, (+)-(2)-carene, α-phellandrene, and ß-phellandrene differentially attracted (51-87%) J2 relative to control. Concurrent reduction and increase in the levels of methyl salicylate and (Z)-methyl dihydrojasmonate, respectively, in the root volatiles reduced J2 responses. These results demonstrate that the host plant can alter its root volatile composition to inhibit PPN attack. The observed plant-produced inhibition of J2 warrants further investigation as a potential management tool for growers.

Microbial Co-Occurrence in Floral Nectar Affects Metabolites and Attractiveness to a Generalist Pollinator.

Microbial metabolism can shape cues important for animal attraction in service-resource mutualisms. Resources are frequently colonized by microbial communities, but experimental assessment of animal-microbial interactions often focus on microbial monocultures. Such an approach likely fails to predict effects of microbial assemblages, as microbe-microbe interactions may affect in a non-additive manner microbial metabolism and resulting chemosensory cues. Here, we compared effects of microbial mono- and cocultures on growth of constituent microbes, volatile metabolite production, sugar catabolism, and effects on pollinator foraging across two nectar environments that differed in sugar concentration. Growth in co-culture decreased the abundance of the yeast Metschnikowia reukaufii, but not the bacterium Asaia astilbes. Volatile emissions differed significantly between microbial treatments and with nectar concentration, while sugar concentration was relatively similar among mono- and cocultures. Coculture volatile emission closely resembled an additive combination of monoculture volatiles. Despite differences in microbial growth and chemosensory cues, honey bee feeding did not differ between microbial monocultures and assemblages. Taken together, our results suggest that in some cases, chemical and ecological effects of microbial assemblages are largely predictable from those of component species, but caution that more work is necessary to predict under what circumstances non-additive effects are important.

Traps and Attractants for Monitoring Navel Orangeworm (Lepidoptera: Pyralidae) in the Presence of Mating Disruption.

The use of aerosol mating disruption for management of the navel orangeworm Amyelois transitella (Walker) in California tree nuts has increased markedly. This treatment suppresses pheromone monitoring traps in neighboring orchards as well as in the orchard under treatment. The current study, therefore, addresses the interrelated topics of which attractant is most effective, how the number of adults captured is affected by trap design, and what sex is captured. Under most circumstances, wing traps baited with phenyl propionate (PPO) captured more adults than those baited with a 5-compound kairomone blend. Adding a pheromone lure along with the dispenser for the experimental attractant increased the number of adults captured with PPO both in and near-mating disruption. In contrast, addition of a pheromone lure along with the kairomone blend only increased the number of adults captured in orchards near but not under mating disruption. Delta traps, which are preferred by the industry, captured fewer adults than wing traps. Improvements in the number of adults captured with PPO in delta traps from addition of a pheromone lure and from design modifications were additive. Both sexes were captured by all attractants and trap designs tested, and the sex ratio was highly variable. Open traps like the wing trap captured a slightly higher proportion of males than closed traps. These findings are discussed in the context of management of the navel orangeworm.

Microbial metabolites elicit distinct olfactory and gustatory preferences in bumblebees.

Animals such as bumblebees use chemosensory cues to both locate and evaluate essential resources. Increasingly, it is recognized that microbes can alter the quality of foraged resources and produce metabolites that may act as foraging cues. The distinct nature of these chemosensory cues however and their use in animal foraging remain poorly understood. Here, we test the hypothesis that species of nectar-inhabiting microbes differentially influence pollinator attraction and feeding via microbial metabolites produced in nectar. We first examined the electrophysiological potential for bumblebee (Bombus impatiens) antennal olfactory neurons to respond to microbial volatile organic compounds (mVOCs), followed by an olfactory preference test. We also assessed gustatory preferences for microbial-altered nectar through both no-choice and choice feeding assays. Antennal olfactory neurons responded to some mVOCs, and bees preferred nectar solutions inoculated with the bacterium Asaia astilbes over the yeast Metschnikowia reukaufii based on volatiles alone. However, B. impatiens foragers consumed significantly more Metschnikowia-inoculated nectar, suggesting distinct roles for mVOCs and non-volatile metabolites in mediating both attraction and feeding decisions. Collectively, our results suggest that microbial metabolites have significant potential to shape interspecific, plant-pollinator signalling, with consequences for forager learning, economics and floral host reproduction.

Elicitation of Differential Responses in the Root-Knot Nematode Meloidogyne incognita to Tomato Root Exudate Cytokinin, Flavonoids, and Alkaloids.

Root exudates of plants mediate interactions with a variety of organisms in the rhizosphere, including root-knot nematodes (RKNs, Meloidogyne spp.) We investigated the responses of the motile stage second-stage juveniles (J2s) of Meloidogyne incognita to non-volatile components identified in the root exudate of tomato. Using stylet thrusting, chemotaxis assays, and chemical analysis, we identified specific metabolites in the root exudate that attract and repel J2s. Liquid chromatography quadrupole time-of-flight mass spectrometry analysis of bioactive fractions obtained from the root exudate revealed a high diversity of compounds, of which five were identified as the phytohormone zeatin (cytokinin), the flavonoids quercetin and luteolin, and alkaloids solasodine and tomatidine. In stylet thrusting and chemotaxis assays, the five compounds elicited concentration-dependent responses in J2s relative to 2% dimethyl sulfoxide (negative control) and methyl salicylate (positive control). These results indicate that J2 herbivory is influenced by root exudate chemistry and concentrations of specific compounds, which may have potential applications in RKN management.

Identification of Glutamic Acid as a Host Marking Pheromone of the African Fruit Fly Species Ceratitis rosa (Diptera: Tephritidae).

Host marking pheromones (HMPs) deposited by female fruit flies deter other females from overexploiting the same fruit for egg laying. Using a bioassay-guided approach, we identified the HMP of the Natal fruit fly species Ceratitis rosa as glutamic acid, 1, from the aqueous fecal matter extract of ovipositing females by liquid chromatography-quadrupole time-of-flight-mass spectrometry (LC-QTOF-MS). Dual choice oviposition assays showed that both the fecal matter extract and 1 significantly reduced oviposition responses in conspecific females of C. rosa. Glutamic acid levels were 10-20 times higher in fecal matter than in the ovipositor or hemolymph extracts of females. Identification of 1 as a host marking pheromone in females of C. rosa improves our understanding of fruit fly chemical ecology and provides evidence that it could be used as a potential component in the integrated management of this fruit fly species.

Cucumber and Tomato Volatiles: Influence on Attraction in the Melon Fly Zeugodacus cucurbitate (Diptera: Tephritidae).

The main hosts of the melon fly Zeugodacus cucurbitate are cultivated and wild cucurbitaceous plants. In eastern Africa, the melon fly is a major pest of the Solanaceae plant Solanum lycopersicum (tomato). We hypothesized that shared species-specific volatiles may play a role in host attraction. We tested this hypothesis by comparing the olfactory responses of the melon fly to Cucumis sativus (cucumber) (Cucurbitaceae) and tomato plant odors in behavioral and electrophysiological assays, followed by chemical analysis to identify the key compounds mediating the interactions. Our results identified 13 shared components between cucumber and tomato plant odors. A synthetic blend of seven of the shared components dominated by monoterpenes at concentrations mimicking the volatile bouquet of cucumber and tomato attracted both sexes of the melon fly. Our results suggest that the presence and quantity of specific compounds in host odors are the main predictors for host recognition in Z. cucurbitate.

Identification of Key Root Volatiles Signaling Preference of Tomato over Spinach by the Root Knot Nematode Meloidogyne incognita.

The root knot nematode, Meloidogyne incognita (Kofoid and White) Chitwood, is a serious pest of tomato ( Solanum lycopersicum) and spinach ( Spinacea oleracea) in sub-Saharan Africa. In East Africa these two crops are economically important and are commonly intercropped by smallholder farmers. The role of host plant volatiles in M. incognita interactions with these two commodities is currently unknown. Here, we investigate the olfactory basis of attraction of tomato and spinach roots by the infective second stage juveniles (J2s) of M. incognita. In olfactometer assays, J2s were attracted to root volatiles from both crops over moist sand (control), but in choice tests using the two host plants, volatiles of tomato roots were more attractive than those released by spinach. Root volatiles sampled by solid phase microextraction (SPME) fiber and analyzed by gas chromatography/mass spectrometry (GC/MS) identified a total of eight components, of which five (2-isopropyl-3-methoxypyrazine, 2-(methoxy)-3-(1-methylpropyl)pyrazine, tridecane, and α- and ß-cedrene) occurred in the root-emitted volatiles of both plants, with three (δ-3-carene, sabinene, and methyl salicylate) being specific to tomato root volatiles. In a series of bioassays, methyl salicylate contributed strongly to the attractiveness of tomato, whereas 2-isopropyl-3-methoxypyrazine and tridecane contributed to the attractiveness of spinach. M. incognita J2s were also more attracted to natural spinach root volatiles when methyl salicylate was combined than to spinach volatiles alone, indicating that the presence of methyl salicylate in tomato volatiles strongly contributes to its preference over spinach. Our results indicate that since both tomato and spinach roots are attractive to M. incognita, identifying cultivars of these two plant species that are chemically less attractive can be helpful in the management of root knot nematodes.

Interactions Among Plants, Insects, and Microbes: Elucidation of Inter-Organismal Chemical Communications in Agricultural Ecology.

The last 2 decades have witnessed a sustained increase in the study of plant-emitted volatiles and their role in plant-insect, plant-microbe, and plant-plant interactions. While each of these binary systems involves complex chemical and biochemical processes between two organisms, the progression of increasing complexity of a ternary system (i.e., plant-insect-microbe), and the study of a ternary system requires nontrivial planning. This planning can include an experimental design that factors in potential overarching ecological interactions regarding the binary or ternary system, correctly identifying and understanding unexpected observations that may occur during the experiment and thorough interpretation of the resultant data. This challenge of planning, performing, and interpreting a plant's defensive response to multiple biotic stressors will be even greater when abiotic stressors (i.e., temperature or water) are factored into the system. To fully understand the system, we need to not only continue to investigate and understand the volatile profiles but also include and understand the biochemistry of the plant's response to these stressors. In this review, we provide examples and discuss interaction considerations with respect to how readers and future authors of the Journal of Agricultural and Food Chemistry can contribute their expertise toward the extraction and interpretation of chemical information exchanged between agricultural commodities and their associated pests. This holistic, multidisciplinary, and thoughtful approach to interactions of plants, insects, and microbes, and the resultant response of the plants can lead to a better understanding of agricultural ecology, in turn leading to practical and viable solutions to agricultural problems.