Blueberry floral probiotics: nectar microbes inhibit the growth of Colletotrichum pathogens.

AIMS: To identify whether microorganisms isolated from blueberry flowers can inhibit the growth of Colletotrichum, an opportunistic plant pathogen that infects flowers and threatens yields, and to assess the impacts of floral microbes and Colletotrichum pathogens on artificial nectar sugars and honey bee consumption. METHODS AND RESULTS: The growth inhibition of Colletotrichum (Colletotrichum acutatum, Colletotrichum fioriniae, and Colletotrichum gloeosporioides) was screened using both artificial nectar co-culture and dual culture plate assays. All candidate nectar microbes were screened for antagonism against a single C. acutatum isolate. Then, the top four candidate nectar microbes showing the strongest inhibition of C. acutatum (Neokomagataea thailandica, Neokomagataea tanensis, Metschnikowia rancensis, and Symmetrospora symmetrica) were evaluated for antagonism against three additional C. acutatum isolates, and single isolates of both C. fioriniae and C. gloeosporioides. In artificial nectar assays, single and three-species cultures inhibited the growth of two of four C. acutatum isolates by ca. 60%, but growth of other Colletotrichum species was not affected. In dual culture plate assays, inhibition was observed for all Colletotrichum species for at least three of four selected microbial antagonists (13%‒53%). Neither honey bee consumption of nectar nor nectar sugar concentrations were affected by any microbe or pathogen tested. CONCLUSIONS: Selected floral microbes inhibited growth of all Colletotrichum species in vitro, although the degree of inhibition was specific to the assay and pathogen examined. In all microbial treatments, nectar sugars were preserved, and honey bee preference was not affected.

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.

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.

Fighting on two fronts: Elevated insect resistance in flooded maize.

To grow and thrive plants must be able to adapt to both adverse environmental conditions and attack by a variety of pests. Elucidating the sophisticated mechanisms plants have developed to achieve this has been the focus of many studies. What is less well understood is how plants respond when faced with multiple stressors simultaneously. In this study, we assess the response of Zea mays (maize) to the combinatorial stress of flooding and infestation with the insect pest Spodoptera frugiperda (fall armyworm). This combined stress leads to elevated production of the defence hormone salicylic acid, which does not occur in the individual stresses, and the resultant salicylic acid-dependent increase in S. frugiperda resistance. Remodelling of phenylpropanoid pathways also occurs in response to this combinatorial stress leading to increased production of the anti-insect C-glycosyl flavones (maysins) and the herbivore-induced volatile phenolics, benzyl acetate, and phenethyl acetate. Furthermore, changes in cellular redox status also occur, as indicated by reductions in peroxidase and polyphenol oxidase activity. These data suggest that metabolite changes important for flooding tolerance and anti-insect defence may act both additively and synergistically to provide extra protection to the plant.

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.

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.

Contrasting insect attraction and herbivore-induced plant volatile production in maize.

MAIN CONCLUSION: The maize inbred line W22 has lower herbivore-induced volatile production than B73 but both fall armyworm larvae and the wasps that parasitize them prefer W22 over B73. Maize inbred line W22 is an important resource for genetic studies due to the availability of the UniformMu mutant population and a complete genome sequence. In this study, we assessed the suitability of W22 as a model for tritrophic interactions between maize, Spodoptera frugiperda (fall armyworm) and the parasitoid wasp Cotesia marginiventris. W22 was found to be a good model for studying the interaction as S. frugiperda prefers W22 over B73 and a higher parasitism rate by C. marginiventris was observed on W22 compared to the inbred line B73. W22 also produced lower amounts of many herbivore-induced volatile terpenes and indole emission upon treatment with S. frugiperda oral secretions. We propose that some of the major herbivore-induced terpene volatiles are perhaps impeding S. frugiperda and C. marginiventris preference and that as yet unidentified compounds are produced at low abundance may be positively impacting these interactions.

Nectar-inhabiting microorganisms influence nectar volatile composition and attractiveness to a generalist pollinator.

The plant microbiome can influence plant phenotype in diverse ways, yet microbial contribution to plant volatile phenotype remains poorly understood. We examine the presence of fungi and bacteria in the nectar of a coflowering plant community, characterize the volatiles produced by common nectar microbes and examine their influence on pollinator preference. Nectar was sampled for the presence of nectar-inhabiting microbes. We characterized the headspace of four common fungi and bacteria in a nectar analog. We examined electrophysiological and behavioral responses of honey bees to microbial volatiles. Floral headspace samples collected in the field were surveyed for the presence of microbial volatiles. Microbes commonly inhabit floral nectar and the common species differ in volatile profiles. Honey bees detected most microbial volatiles tested and distinguished among solutions based on volatiles only. Floral headspace samples contained microbial-associated volatiles, with 2-ethyl-1-hexanol and 2-nonanone - both detected by bees - more often detected when fungi were abundant. Nectar-inhabiting microorganisms produce volatile compounds, which can differentially affect honey bee preference. The yeast Metschnikowia reukaufii produced distinctive compounds and was the most attractive of all microbes compared. The variable presence of microbes may provide volatile cues that influence plant-pollinator interactions.

A quantitative survey of the blueberry (Vaccinium spp.) culturable nectar microbiome: variation between cultivars, locations, and farm management approaches.

Microbes in floral nectar can impact both their host plants and floral visitors, yet little is known about the nectar microbiome of most pollinator-dependent crops. In this study, we examined the abundance and composition of the fungi and bacteria inhabiting Vaccinium spp. nectar, as well as nectar volume and sugar concentrations. We compared wild V. myrsinites with two field-grown V. corymbosum cultivars collected from two organic and two conventional farms. Differences in nectar traits and microbiomes were identified between V. corymbosum cultivars but not Vaccinium species. The microbiome of cultivated plants also varied greatly between farms, whereas management regime had only subtle effects, with higher fungal populations detected under organic management. Nectars were hexose-dominant, and high cell densities were correlated with reduced nectar sugar concentrations. Bacteria were more common than fungi in blueberry nectar, although both were frequently detected and co-occurred more often than would be predicted by chance. "Cosmopolitan" blueberry nectar microbes that were isolated in all plants, including Rosenbergiella sp. and Symmetrospora symmetrica, were identified. This study provides the first systematic report of the blueberry nectar microbiome, which may have important implications for pollinator and crop health.

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.

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.

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.

Setaria viridis as a model for translational genetic studies of jasmonic acid-related insect defenses in Zea mays.

Little is known regarding insect defense pathways in Setaria viridis (setaria), a model system for panicoid grasses, including Zea mays (maize). It is thus of interest to compare insect herbivory responses of setaria and maize. Here we use metabolic, phylogenetic, and gene expression analyses to measure a subset of jasmonic acid (JA)-related defense responses to leaf-chewing caterpillars. Phylogenetic comparisons of known defense-related maize genes were used to identify putative orthologs in setaria, and candidates were tested by quantitative PCR to determine transcriptional responses to insect challenge. Our findings show that while much of the core JA-related metabolic and genetic responses appear conserved between setaria and maize, production of downstream secondary metabolites such as benzoxazinoids and herbivore-induced plant volatiles are dissimilar. This diversity of chemical defenses and gene families involved in secondary metabolism among grasses presents new opportunities for cross species engineering. The high degree of genetic similarity and ease of orthologous gene identification between setaria and maize make setaria an excellent species for translational genetic studies, but the species specificity of downstream insect defense chemistry makes some pathways unamenable to cross-species comparisons.

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.

Comparison of Direct and Indirect Photolysis in Imazosulfuron Photodegradation.

Imazosulfuron, a sulfonylurea herbicide used in rice cultivation, has been shown to undergo photodegradation in water, but neither the photochemical mechanism nor the role of indirect photolysis is known. The purpose of this study was to investigate the underlying processes that operate on imazosulfuron during aqueous photodegradation. Our data indicate that in the presence of oxygen, most photochemical degradation proceeds through a direct singlet-excited state pathway, whereas triplet-excited state imazosulfuron enhanced decay rates under low dissolved oxygen conditions. Oxidation by hydroxyl radical and singlet oxygen were not significant. At dissolved organic matter (DOM) concentrations representative of rice field conditions, fulvic acid solutions exhibited faster degradation than rice field water containing both humic and fulvic acid fractions. Both enhancement, via reaction with triplet-state DOM, and inhibition, via competition for photons, of degradation was observed in DOM solutions.

Photochemical degradation of imazosulfuron under simulated California rice field conditions.

BACKGROUND: The photodegradation of imazosulfuron (IMZ), a potent broad-spectrum herbicide, was investigated under simulated rice field conditions. Previous reports have indicated that it is photolabile, but have failed to report radiation intensity or determine a quantum yield, precluding extrapolation to environmental rates. Therefore, the objective of this investigation was to determine the photolytic rate of IMZ under simulated rice field conditions and how it is influenced by environmental factors such as turbidity, salinity and temperature. RESULTS: IMZ was efficiently photolyzed in all solutions and fitted pseudo-first-order kinetics. Degradation was faster in HPLC-grade water than in field water. Field-relevant variances in temperature, turbidity and salinity did not significantly influence degradation. The experimentally derived quantum yield for direct photolysis (2.94 × 10(3) ) was used to predict the half-life of IMZ in a California rice field (3.6 days). CONCLUSIONS: Aqueous photolysis is predicted to be an important process in the overall degradation of IMZ in the environment, regardless of variances in salinity, organic matter and temperature. Based on the predicted half-life of IMZ in a California rice field (3.6 days), state-mandated holding periods for field water post-IMZ application (30 days) are expected to allow for sufficient clearance of the herbicide (>98%), preventing significant contamination of the environment upon release of tailwater. © 2015 Society of Chemical Industry.