Soil moisture conditions alter behavior of entomopathogenic nematodes.

BACKGROUND: A variety of environmental factors can disrupt biotic interactions between plants, insects and soil microorganisms with consequences for agricultural management and production. Many of these belowground interactions are mediated by volatile organic compounds (VOCs) which can be used for communication under appropriate environmental conditions. Behavioral responses to these compounds may likewise be dependent on varying soil conditions which are influenced by a changing climate. To determine how changing environmental conditions may affect VOC-mediated biotic interactions, we used a belowground system where entomopathogenic nematodes (EPNs) - tiny roundworm parasitoids of soil-borne insects - respond to VOCs by moving through the soil pore matrix. Specifically, we used two genera of EPNs - Heterorhabditis and Steinernema - that are known to respond to four specific terpenes - α-pinene, linalool, d-limonene and pregeijerene - released by the roots of plants in the presence of herbivores. We assessed the response of these nematodes to these terpenes under three moisture regimes to determine whether drier conditions or inundated conditions may influence the response behavior of these nematodes. RESULTS: Our results illustrate that the recovery rate of EPNs is positively associated with soil moisture concentration. As soil moisture concentration increases from 6% to 18%, substantially more nematodes are recovered from bioassays. In addition, we find that soil moisture influences EPN preference for VOCs, as illustrated in the variable response rates. Certain compounds shifted from acting as a repellent to acting as an attractant and vice versa depending on the soil moisture concentration. CONCLUSION: On a broad scale, we demonstrate that soil moisture has a significant effect on EPN host-seeking behavior. EPN efficacy as biological control agents could be affected by climate change projections that predict varying soil moisture concentrations. We recommend that maintaining nematodes as biological control agents is essential for sustainable agriculture development, as they significantly contribute not only to soil health but also to efficient pest management. © 2024 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

The Ecology of Salicylic Acid Signaling: Primary, Secondary and Tertiary Effects with Applications in Agriculture.

The salicylic acid pathway is one of the primary plant defense pathways, is ubiquitous in vascular plants, and plays a role in rapid adaptions to dynamic abiotic and biotic stress. Its prominence and ubiquity make it uniquely suited for understanding how biochemistry within plants can mediate ecological consequences. Induction of the salicylic acid pathway has primary effects on the plant in which it is induced resulting in genetic, metabolomic, and physiologic changes as the plant adapts to challenges. These primary effects can in turn have secondary consequences for herbivores and pathogens attacking the plant. These secondary effects can both directly influence plant attackers and mediate indirect interactions between herbivores and pathogens. Additionally, stimulation of salicylic acid related defenses can affect natural enemies, predators and parasitoids, which can recruit to plant signals with consequences for herbivore populations and plant herbivory aboveground and belowground. These primary, secondary, and tertiary ecological consequences of salicylic acid signaling hold great promise for application in agricultural systems in developing sustainable high-yielding management practices that adapt to changing abiotic and biotic environments.

Machine Learning for Characterization of Insect Vector Feeding.

Insects that feed by ingesting plant and animal fluids cause devastating damage to humans, livestock, and agriculture worldwide, primarily by transmitting pathogens of plants and animals. The feeding processes required for successful pathogen transmission by sucking insects can be recorded by monitoring voltage changes across an insect-food source feeding circuit. The output from such monitoring has traditionally been examined manually, a slow and onerous process. We taught a computer program to automatically classify previously described insect feeding patterns involved in transmission of the pathogen causing citrus greening disease. We also show how such analysis contributes to discovery of previously unrecognized feeding states and can be used to characterize plant resistance mechanisms. This advance greatly reduces the time and effort required to analyze insect feeding, and should facilitate developing, screening, and testing of novel intervention strategies to disrupt pathogen transmission affecting agriculture, livestock and human health.

Disruption of Vector Host Preference with Plant Volatiles May Reduce Spread of Insect-Transmitted Plant Pathogens.

Plant pathogens can manipulate the odor of their host; the odor of an infected plant is often attractive to the plant pathogen vector. It has been suggested that this odor-mediated manipulation attracts vectors and may contribute to spread of disease; however, this requires further broad demonstration among vector-pathogen systems. In addition, disruption of this indirect chemical communication between the pathogen and the vector has not been attempted. We present a model that demonstrates how a phytophathogen (Candidatus Liberibacter asiaticus) can increase its spread by indirectly manipulating the behavior of its vector (Asian citrus psyllid, Diaphorina citri Kuwayama). The model indicates that when vectors are attracted to pathogen-infected hosts, the proportion of infected vectors increases, as well as, the proportion of infected hosts. Additionally, the peak of infected host populations occurs earlier as compared with controls. These changes in disease dynamics were more important during scenarios with higher vector mortality. Subsequently, we conducted a series of experiments to disrupt the behavior of the Asian citrus psyllid. To do so, we exposed the vector to methyl salicylate, the major compound released following host infection with the pathogen. We observed that during exposure or after pre-exposure to methyl salicylate, the host preference can be altered; indeed, the Asian citrus psyllids were unable to select infected hosts over uninfected counterparts. We suggest mechanisms to explain these interactions and potential applications of disrupting herbivore host preference with plant volatiles for sustainable management of insect vectors.

Entomopathogenic Nematodes for Field Control of Onion Maggot (Delia antiqua) and Compatibility with Seed Treatments.

Onion maggot (Delia antiqua) is a prominent pest of allium crops in temperate zones worldwide. Management of this pest relies on prophylactic insecticide applications at planting that target the first generation. Because effective options are limited, growers are interested in novel tactics such as deployment of entomopathogenic nematodes. We surveyed muck soils where onions are typically grown to determine if entomopathogenic nematode species were present, and then evaluated the compatibility of entomopathogenic nematode species with the insecticides commonly used to manage D. antiqua. We also evaluated the efficacy of these entomopathogenic nematodes for reducing D. antiqua infestations in the field. No endemic entomopathogenic nematodes were detected in surveys of muck fields in New York. Compatibility assays indicated that, although insecticides such as spinosad and, to some extent, cyromazine did cause mortality of entomopathogenic nematodes, these insecticides did not affect infectivity of the entomopathogenic nematodes. Field trials indicated that applications of entomopathogenic nematodes can reduce the percentage of onion plants killed by D. antiqua from 6% to 30%. Entomopathogenic nematodes reduced D. antiqua damage and increased end of season yield over two field seasons. Applications of entomopathogenic nematodes may be a viable option for reducing D. antiqua populations in conventional and organic systems. Together with other management tactics, like insecticide seed treatments, applications of entomopathogenic nematodes can provide a yield boost and a commercially acceptable level of D. antiqua control.

NOAA Open Data Dissemination: Petabyte-scale Earth system data in the cloud.

NOAA Open Data Dissemination (NODD) makes NOAA environmental data publicly and freely available on Amazon Web Services (AWS), Microsoft Azure (Azure), and Google Cloud Platform (GCP). These data can be accessed by anyone with an internet connection and span key datasets across the Earth system including satellite imagery, radar, weather models and observations, ocean databases, and climate data records. Since its inception, NODD has grown to provide public access to more than 24 PB of NOAA data and can support billions of requests and petabytes of access daily. Stakeholders routinely access more than 5 PB of NODD data every month. NODD continues to grow to support open petabyte-scale Earth system data science in the cloud by onboarding additional NOAA data and exploring performant data formats. Here, we document how this program works with a focus on provenance, key datasets, and use. We also highlight how to access these data with the goal of accelerating use of NOAA resources in the cloud.

The Smart Soil Organism Detector: An instrument and machine learning pipeline for soil species identification.

Understanding the diversity of soil organisms is complicated by both scale and substrate. Every footprint we leave in the soil covers hundreds to millions of organisms yet we cannot see them without extremely laborious extraction and microsopy endeavors. Studying them is also challenging. Keeping them alive so that we can understand their lifecycles and ecological roles ranges from difficult to impossible. Functional and taxonomic identification of soil organisms, while possible, is also challenging. Here we present the Smart Soil Organism Detector, an instrument and machine learning pipeline that combines high-resolution imaging, multi-spectral sensing, large-bore flow cytometry, and machine learning to extract, isolate, count, identify, and separate soil organisms in a high-throughput, high-resolution, non-destructive, and reproducible manner. This system is not only capable of separating alive nematodes, dead nematodes, and nematode cuticles from soil with 100% out-of-sample accuracy, but also capable of identifying nematode strains (sub-species) with 95.5% out-of-sample accuracy and 99.4% specificity. Soil micro-arthropods were identified to class with 96.1% out-of-sample accuracy. Broadly applicable across soil taxa, the Smart SOD system is a tool for understanding global soil biodiversity.

Phenology and Monitoring of the Lesser Chestnut Weevil (Curculio sayi).

With the introduction in recent years of high-yield blight-resistant chestnut varieties, the commercial chestnut industry in the United States is expanding. Accompanying this expansion is a resurgence in a primary pest of chestnut: C. sayi, the lesser chestnut weevil. This weevil damages the nut crop and infestations can surge from 0 to close to 100% in as little as two years. Understanding the dynamics of this pest has been challenging. Most work was conducted in the 1900s and only recently has this weevil garnered renewed interest. Recent work on C. sayi phenology has been completed in Missouri but conflicted with anecdotal reports from northern growers. From 2019 to 2020, we used a combination of trapping and microcosm studies to understand both C. sayi phenology and the means of monitoring this pest. C. sayi populations were univoltine and peaked in mid-October. Pyramid traps were the most effective at capturing adult C. sayi. C. sayi larvae, pupae, eclosed adults, and emerging adults were recovered from microcosm experiments. These results suggest that C. sayi emerges later in the northern US with the potential for a single generation to emerge over multiple subsequent years. Understanding C. sayi phenology along with the means of monitoring forms the basis for effective management and control in commercial chestnut orchards.

The Lesser Chestnut Weevil (Curculio sayi): Damage and Management with Biological Control Using Entomopathogenic Fungi and Entomopathogenic Nematodes.

The lesser chestnut weevil, Curculio sayi (Gyllenhal), can cause irreparable damage to chestnuts through direct consumption and/or introduction of secondary pathogens. With the resurgence of blight resistant American Chestnut plantings both for commercial production and for habitat restoration, C. sayi has become a similarly resurgence pest. Here, we investigated the nature and extent of C. sayi larval damage on individual nuts and collected harvests with an eye toward the quantifying impacts. Next, we explored management options using biological control including entomopathogenic fungi and entomopathogenic nematodes. Nut damage from C. sayi can be extensive with individual nuts hosting several larvae, larvae emerging from nuts several weeks post harvest, and nut weight loss even after C. sayi have emerged from the nut. Applications of entomopathogenic fungi reduced chances of chestnut infestation, while certain strains of entomopathogenic nematodes increased the probability of C. sayi larval mortality. Understanding C. sayi damage and exploring biological control management options could be a useful tool in the effective management of this resurgent pest.

Non-lethal effects of entomopathogenic nematode infection.

Entomopathogenic nematodes are typically considered lethal parasites of insect hosts. Indeed they are employed as such for biological control of insect pests. The effects of exposure to entomopathogenic nematodes are not strictly limited to mortality, however. Here we explore non-lethal effects of exposure to entomopathogenic nematodes by introducing the relatively non-susceptible pupal stage of Delia antiqua to thirteen different strains. We specifically chose to inoculate the pupal stage because it tends to be more resistant to infection, yet resides in the soil where it could come into contact with EPN biological control agents. We find that there is no significant mortality at the pupal stage, but that there are a host of strain-dependent non-lethal effects during and after the transition to adulthood including altered developmental times and changes in risk of death compared to controls. We also find that exposure to specific strains can reduce risk of mortality. These results emphasize the strain-dependent nature of entomopathogenic nematode infection and highlight the positive and negative ramifications for non-lethal effects for biological control of insect pests. Our work emphasizes the need for strain-specific screening of biological control agents before wide-spread adoption.

Reduced Infestation by Xylosandrus germanus (Coleoptera: Curculionidae: Scolytinae) in Apple Trees Treated with Host Plant Defense Compounds.

The ambrosia beetle Xylosandrus germanus (Blandford) is an invasive pest that has caused tree decline and death in numerous NY dwarf apple orchards during the past ten years, despite efforts to control them using trunk sprays of chlorpyrifos or pyrethroids, either alone or combined with the repellent verbenone. From 2017 to 2019, we tested trunk applications of different repellents and plant defense compounds for protection against X. germanus in potted apple trees adjacent to infested orchards. Treatments included topical formulations of verbenone and methyl salicylate (MeSa), alone and in combination, at different rates and timings. Additional treatments evaluated included the systemic acquired resistance activators acibenzolar-S-methyl, Reynoutria sachalinensis extract, and salicylic acid. The combination verbenone+MeSa treatments had the lowest incidences of attack sites and galleries containing adults or brood, although results varied among years. In a separate trial, we found no significant difference in numbers of adults caught in ethanol-baited traps placed 5-20 m from an apple bolt treated with the verbenone+MeSa repellent, suggesting that the repellent's effect did not extend to those distances from the treated target. Cross-sectional discs of trunk tissue sampled in August were analyzed for levels of phytohormones. Quantities of ergosterol, abscissic acid, salicylic acid, jasmonic acid, methyl salicylate, methyl jasmonate, trans-cinnamic acid, and indole-3-cinnamic acid did not significantly vary across treatments; however, trees with greater beetle damage contained higher levels of jasmonic and salicylic acid, which are key molecules in plant defense pathways.

Attract and kill: spinosad containing spheres to control onion maggot (Delia antiqua).

BACKGROUND: Onion maggot (Delia antiqua) is a pest of onions worldwide. Current means of managing this pest rely heavily on prophylatic insecticide treatments at planting. These options may not be viable in organic production systems or situations where insecticide-resistant populations occur. Here we explore the efficacy of an attract and kill strategy for control of D. antiqua evaluating the ability of attractive, spinosad containing spheres to kill adult D. antiqua and reduce crop losses. RESULTS: Spinosad containing spheres were able to consistently kill D. antiqua adults over the course of the field season (mortality range: between 49% and 59% on average). Pairing spinosad spheres with Delia Lure increased efficacy by 72% compared with the spheres alone. Performance of this attract and kill strategy also can reduce damage by D. antiqua larvae in the field, but it did not achieve a level of control comparable to the level provided by a conventional insecticide treatment. CONCLUSION: Implementation of this attract and kill strategy could be a valuable tool in situations where conventional pesticides are either not available or desired, where additional control techniques are needed, or to provide a season-long option for control of D. antiqua populations. © 2020 Society of Chemical Industry.

Sting nematodes modify metabolomic profiles of host plants.

Plant-parasitic nematodes are devastating pathogens of many important agricultural crops. They have been successful in large part due to their ability to modify host plant metabolomes to their benefit. Both root-knot and cyst nematodes are endoparasites that have co-evolved to modify host plants to create sophisticated feeding cells and suppress plant defenses. In contrast, the ability of migratory ectoparasitic nematodes to modify host plants is unknown. Based on global metabolomic profiling of sting nematodes in African bermudagrass, ectoparasites can modify the global metabolome of host plants. Specifically, sting nematodes suppress amino acids in susceptible cultivars. Upregulation of compounds linked to plant defense have negative impacts on sting nematode population densities. Pipecolic acid, linked to systemic acquired resistance induction, seems to play a large role in protecting tolerant cultivars from sting nematode feeding and could be targeted in breeding programs.

A multi-omics approach to solving problems in plant disease ecology.

The swift rise of omics-approaches allows for investigating microbial diversity and plant-microbe interactions across diverse ecological communities and spatio-temporal scales. The environment, however, is rapidly changing. The introduction of invasive species and the effects of climate change have particular impact on emerging plant diseases and managing current epidemics. It is critical, therefore, to take a holistic approach to understand how and why pathogenesis occurs in order to effectively manage for diseases given the synergies of changing environmental conditions. A multi-omics approach allows for a detailed picture of plant-microbial interactions and can ultimately allow us to build predictive models for how microbes and plants will respond to stress under environmental change. This article is designed as a primer for those interested in integrating -omic approaches into their plant disease research. We review -omics technologies salient to pathology including metabolomics, genomics, metagenomics, volatilomics, and spectranomics, and present cases where multi-omics have been successfully used for plant disease ecology. We then discuss additional limitations and pitfalls to be wary of prior to conducting an integrated research project as well as provide information about promising future directions.

Risk taking of educated nematodes.

Nematode parasites rely on successful host infection to perpetuate their species. Infection by individual nematode parasites can be risky, however; any one individual could be killed by the host's immune response. Here we use a model system to show that environmental cues and parasite past experience can be used by entomopathogenic nematodes to reduce individual risk of infection. Past parasite experience can more than double the infective virulence (number of host invaders) of a given cohort of entomopathogenic nematode parasites. This plasticity in individual parasite risk-taking and associated infection can be used to manage infection of parasitic nematodes: enhancing biological control with entomopathogenic nematodes and developing behavioral and chemical strategies to reduce infection by vertebrate and plant parasitic nematodes.

Behavioral Evidence for Host Transitions in Plant, Plant Parasite, and Insect Interactions.

Specialized herbivorous insects have the ability to transition between host plant taxa, and considering the co-evolutionary history between plants and the organisms utilizing them is important to understanding plant insect interactions. We investigated the role of a pine tree parasite, dwarf mistletoe (Arceuthobium spp.) M. Bieb. Santalales: Viscaceae, in mediating interactions between Neophasia (Lepidoptera: Pieridae) butterflies and pine trees, the butterflies' larval hosts. Mistletoe is considered the butterflies' ancestral host, and the evolutionary transition to pine may have occurred recently. In Arizona, United States, we studied six sites in pine forest habitats: three in Neophasia menapia (Felder and R. Felder, 1859) habitat and three in Neophasia terlooii Behr, 1869 habitat. Each site contained six stands of trees that varied in mistletoe infection severity. Butterfly behavior was observed and ranked at each stand. Volatile compounds were collected from trees at each site and analyzed using gas chromatography-mass spectroscopy. Female butterflies landed on or patrolled around pine trees (i.e., interacted) more than males, and N. terlooii interacted more with pine trees than N. menapia. Both butterfly species interacted more with tree stands harboring greater mistletoe infection, and N. terlooii interacted more with heavily infected tree stands than did N. menapia. The influence of mistletoe on Neophasia behavior may be mediated by differences in tree volatiles resulting from mistletoe infection. Volatile profiles significantly differed between infected and uninfected pine trees. The role of mistletoe in mediating butterfly interactions with pines has implications for conservation biology and forest management, and highlights the importance of understanding an organism's niche in an evolutionary context.

Multitrophic Effects of Belowground Parasitoid Learning.

The ability to learn allows organisms to take advantage of dynamic and ephemeral opportunities in their environment. Here we show that learning in belowground entomopathogenic nematodes has cascading multitrophic effects on their hosts, other nematodes, and nematophagous fungal predators. In addition to quantifying these effects, we show that social behavioral plasticity in these belowground parasitoids can amplify signaling by plant defense pathways and results in an almost doubling of insect herbivore infection by entomopathogenic nematodes. Cumulatively, these effects point to the critical role of plant signaling in regulating community structure while suggesting an equally important role for behavioral plasticity in shaping community dynamics.

Silo-Stored Pistachios at Varying Humidity Levels Produce Distinct Volatile Biomarkers.

Fungal-contaminated tissues are known to produce volatile profiles that are different from uncontaminated tissues. Fungi require certain water activity levels before growth can occur. For nonxerophilic fungi, a water activity of 0.85 is typical for growth, and for extreme xerophilic fungi, the water activity can be as low as 0.64. Recent investigations with stored pistachios (kernels in shell, no hull tissue) at varying relative humidities showed differences among the collected volatile profiles at the tested humidities (ambient, 63, 75, and 84%). Water activities of the kernel and shell were also measured. Results showed significant changes in volatile profiles as a function of water activity of the corresponding pistachio tissue with measured water activity levels at or below that of what is considered extreme xerophilic activities. Because fungal growth, including mycotoxigenic fungi, is dependent upon water activity, the detected volatile profiles could be used for early detection of fungal presence. Multivariate analysis of the volatile data demonstrated significant differences among the volatile profiles at the tested relative humidity levels, and several volatiles were identified as biomarkers of increased humidity and likely fungal development.

Concurrence in the ability for lipid synthesis between life stages in insects.

The ability to synthesize lipids is critical for an organism's fitness; hence, metabolic pathways, underlying lipid synthesis, tend to be highly conserved. Surprisingly, the majority of parasitoids deviate from this general metabolic model by lacking the ability to convert sugars and other carbohydrates into lipids. These insects spend the first part of their life feeding and developing in or on an arthropod host, during which they can carry over a substantial amount of lipid reserves. While many parasitoid species have been tested for lipogenic ability at the adult life stage, it has remained unclear whether parasitoid larvae can synthesize lipids. Here we investigate whether or not several insects can synthesize lipids during the larval stage using three ectoparasitic wasps (developing on the outside of the host) and the vinegar fly Drosophila melanogaster that differ in lipogenic ability in the adult life stage. Using feeding experiments and stable isotope tracing with gas chromatography/mass spectrometry, we first confirm lipogenic abilities in the adult life stage. Using topical application of stable isotopes in developing larvae, we then provide clear evidence of concurrence in lipogenic ability between larval and adult life stages in all species tested.

Eliciting maize defense pathways aboveground attracts belowground biocontrol agents.

Plant defense pathways mediate multitrophic interactions above and belowground. Understanding the effects of these pathways on pests and natural enemies above and belowground holds great potential for designing effective control strategies. Here we investigate the effects of aboveground stimulation of plant defense pathways on the interactions between corn, the aboveground herbivore adult Diabrotica speciosa, the belowground herbivore larval D. speciosa, and the subterranean ento-mopathogenic nematode natural enemy Heterorhabditis amazonensis. We show that adult D. speciosa recruit to aboveground herbivory and methyl salicylate treatment, that larval D. speciosa are relatively indiscriminate, and that H. amazonensis en-tomopathogenic nematodes recruit to corn fed upon by adult D. speciosa. These results suggest that entomopathogenicnematodes belowground can be highly attuned to changes in the aboveground parts of plants and that biological control can be enhanced with induced plant defense in this and similar systems.