Plant-pathogen System Research Articles

On the large time behaviour of the solutions of an evolutionary-epidemic system with spatial dispersal.

This paper investigates some properties of the large time behaviour of the solutions of a spatially distributed system of equations modelling the evolutionary epidemiology of a plant-pathogen system. The model takes into account the phenotypic trait and the mutation of the pathogen, which is described by a non-local operator. We roughly speaking prove that the solutions separate the phenotype trait from the spatio-temporal evolution in the large time asymptotic. This feature is obtained by investigating the positive and bounded entire solutions of the problem, that are shown to exhibit such a separation of the variables property, by reformulating them as the positive solutions of suitable integral equations in some ordered Banach space. In addition, some numerical simulations are performed to support our theoretical results.

Tailor-made green composts with suppressive properties against tracheofusariosis of wild rocket (Diplotaxis tenuifolia): Useful option for sustainable circular horticulture

Intensive cultivation systems of wild rocket (Diplotaxis tenuifolia) are threatened by the outbreak of tracheofusariosis caused by the soil-borne fungal pathogen Fusarium oxysporum f. sp. raphani. The disease is difficult to eradicate by curative methods due to the endophytic progression of the pathogen. Suppressive composts can help to prevent this adversity by providing organic matter carrying chemical and biological components that can directly and/or indirectly interfere with the plant-pathogen system. In this study, a collection of seven promising composts from different selected green feedstocks were investigated for their suppressive properties towards wild rocket Fusarium wilting. In planta assays identified two composts, obtained from olive tree pruning + plant and tomato residues, and from olive waste + straw + wool residues, with the highest levels of suppressiveness, influenced by both application dose and plant development stage. These active composts showed overlapping 13C NMR profiles as in vivo suppressiveness, being positively correlated with the organic aliphatic alkyl, O-Aryl and carboxyl C fractions and subsequently with the hydrophobic index and alkyl ratio, and negatively with the lignin ratio. On the contrary, although biotic compost components showed antifungal effects, biological properties did not correlate with in planta wilting suppression. Consistently, reflectance spectroscopy in the Vis-NIR-SWIR highlighted the clustering of samples depending on their feedstock and chemical composition. The results allow the hypothesis of a plant-mediated suppression mechanism. The composting of crop residues improves the circularity of horticulture and makes it possible to produce, from selected matrices, tailored compost towards the sustainable cultivation of wild rocket.

Within-plant genetic drift to control virus adaptation to host resistance genes.

Manipulating evolutionary forces imposed by hosts on pathogens like genetic drift and selection could avoid the emergence of virulent pathogens. For instance, increasing genetic drift could decrease the risk of pathogen adaptation through the random fixation of deleterious mutations or the elimination of favorable ones in the pathogen population. However, no experimental proof of this approach is available for a plant-pathogen system. We studied the impact of pepper (Capsicum annuum) lines carrying the same major resistance gene but contrasted genetic backgrounds on the evolution of Potato virus Y (PVY). The pepper lines were chosen for the contrasted levels of genetic drift (inversely related to Ne, the effective population size) they exert on PVY populations, as well as for their contrasted resistance efficiency (inversely related to the initial replicative fitness, Wi, of PVY in these lines). Experimental evolution was performed by serially passaging 64 PVY populations every month on six contrasted pepper lines during seven months. These PVY populations exhibited highly divergent evolutionary trajectories, ranging from viral extinctions to replicative fitness gains. The sequencing of the PVY VPg cistron, where adaptive mutations are likely to occur, allowed linking these replicative fitness gains to parallel adaptive nonsynonymous mutations. Evolutionary trajectories were well explained by the genetic drift imposed by the host. More specifically, Ne, Wi and their synergistic interaction played a major role in the fate of PVY populations. When Ne was low (i.e. strong genetic drift), the final PVY replicative fitness remained close to the initial replicative fitness, whereas when Ne was high (i.e. low genetic drift), the final PVY replicative fitness was high independently of the replicative fitness of the initially inoculated virus. We show that combining a high resistance efficiency (low Wi) and a strong genetic drift (low Ne) is the best solution to increase resistance durability, that is, to avoid virus adaptation on the long term.

Investigating the Effects of Temperature on Pathogen Propagation in Arabidopsis.

Temperature is one of the most prominent environmental factors that influence plant immunity. Depending on the plant-pathogen system, increased temperature may inhibit or enhance disease resistance or immunity in plants. Measuring the effect of temperature on plant immunity is the first step toward revealing climate effects on plant-pathogen interactions and molecular regulators of temperature sensitivity of plant immunity. Quantification of plant disease resistance or susceptibility under different temperatures can be accomplished by assessing pathogen growth over time in infected plants or tissues. Here, we present a protocol for quantifying pathogen growth in the most studied system of Arabidopsis thaliana and Pseudomonas syringae pathovar tomato (Pst) DC3000. We discuss important factors to consider for assaying pathogen growth in plants under different temperatures. This protocol can be used to assess temperature sensitivity of resistance in different plant genotypes and to various pathovars.

Leaf functional traits and pathogens: Linking coffee leaf rust with intraspecific trait variation in diversified agroecosystems.

Research has demonstrated that intraspecific functional trait variation underpins plant responses to environmental variability. However, few studies have evaluated how trait variation shifts in response to plant pathogens, even though pathogens are a major driver of plant demography and diversity, and despite evidence of plants expressing distinct strategies in response to pathogen pressures. Understanding trait-pathogen relationships can provide a more realistic understanding of global patterns of functional trait variation. We examined leaf intraspecific trait variability (ITV) in response to foliar disease severity, using Coffea arabica cv. Caturra as a model species. We quantified coffee leaf rust (CLR) severity-a fungal disease prominent in coffee systems-and measured key coffee leaf functional traits under contrasting, but widespread, management conditions in an agroforestry system. We found that coffee plants express significant ITV, which is largely related to shade tree treatment and leaf position within coffee canopy strata. Yet within a single plant canopy stratum, CLR severity increased with increasing resource conserving trait values. However, coffee leaves with visible signs of disease expressed overall greater resource acquiring trait values, as compared to plants without visible signs of disease. We provide among the first evidence that leaf traits are correlated with foliar disease severity in coffee, and that functional trait relationships and syndromes shift in response to increased disease prevalence in this plant-pathogen system. In doing so, we address a vital gap in our understanding of global patterns of functional trait variation and highlight the need to further explore the potential role of pathogens within established global trait relationships and spectra.

SlS5H silencing reveals specific pathogen-triggered salicylic acid metabolism in tomato

BackgroundSalicylic acid (SA) is a major plant hormone that mediates the defence pathway against pathogens. SA accumulates in highly variable amounts depending on the plant-pathogen system, and several enzyme activities participate in the restoration of its levels. Gentisic acid (GA) is the product of the 5-hydroxylation of SA, which is catalysed by S5H, an enzyme activity regarded as a major player in SA homeostasis. GA accumulates at high levels in tomato plants infected by Citrus Exocortis Viroid (CEVd), and to a lesser extend upon Pseudomonas syringae DC3000 pv. tomato (Pst) infection.ResultsWe have studied the induction of tomato SlS5H gene by different pathogens, and its expression correlates with the accumulation of GA. Transient over-expression of SlS5H in Nicotiana benthamiana confirmed that SA is processed by SlS5H in vivo. SlS5H-silenced tomato plants were generated, displaying a smaller size and early senescence, together with hypersusceptibility to the necrotrophic fungus Botrytis cinerea. In contrast, these transgenic lines exhibited an increased defence response and resistance to both CEVd and Pst infections. Alternative SA processing appears to occur for each specific pathogenic interaction to cope with SA levels. In SlS5H-silenced plants infected with CEVd, glycosylated SA was the most discriminant metabolite found. Instead, in Pst-infected transgenic plants, SA appeared to be rerouted to other phenolics such as feruloyldopamine, feruloylquinic acid, feruloylgalactarate and 2-hydroxyglutarate.ConclusionUsing SlS5H-silenced plants as a tool to unbalance SA levels, we have studied the re-routing of SA upon CEVd and Pst infections and found that, despite the common origin and role for SA in plant pathogenesis, there appear to be different pathogen-specific, alternate homeostasis pathways.

Validation of reference genes as an internal control for studying Avena sativa–Puccinia coronata interaction by RT-qPCR

In this study we evaluated eleven candidate reference genes in Avena sativa during compatible and incompatible interactions with two different pathotypes of Puccinia coronata f. sp. avenae in six time points post-inoculation. The identification of genes with high expression stability was performed by four algorithms (geNorm, NormFinder, BestKeeper and ΔCt method). The results obtained confirmed that the combination of two genes would be sufficient for reliable normalization of the expression data. In general, the most stable in the tested plant-pathogen system were HNR (heterogeneous nuclear ribonucleoprotein 27C) and EF1A (elongation factor 1-alpha). ARF (ADP-ribosylation factor) and EIF4A (eukaryotic initiation factor 4A-3) could also be considered as exhibiting high expression stability. CYP (cyclophilin) was shown by all assessment methods to be the worst candidate for normalization in this dataset. To date, this is the first report of reference genes selection in A. sativa–P. coronata interaction system. Identified reference genes enable reliable and comprehensive RT-qPCR analysis of oat gene expression in response to crown rust infection. Understanding the molecular mechanisms involved in the host–pathogen interactions may expand knowledge of durable resistance strategies beneficial to modern oat breeding.

Reprogramming of phytopathogen transcriptome by a non-bactericidal pesticide residue alleviates its virulence in rice

Bacteria equipped with virulence systems based on highly bioactive small molecules can circumvent their host's defense mechanisms. Pathogens employing this strategy are currently threatening global rice production. In the present study, variations in the virulence of the highly destructive Burkholderia plantarii were observed in different rice-producing regions. The environment-linked variation was not attributable to any known host-related or external factors. Co-occurrence analyses indicated a connection between reduced virulence and 5-Amino-1,3,4-thiadiazole-2-thiol (ATT), a non-bactericidal organic compound. ATT, which accumulates in rice plants during metabolization of specific agrochemicals, was found to reduce virulence factor secretion by B. plantarii up to 88.8% and inhibit pathogen virulence by hijacking an upstream signaling cascade. Detailed assessment of the newly discovered virulence inhibitor resulted in mechanistic insights into positive effects of ATT accumulation in plant tissues. Mechanisms of virulence alleviation were deciphered by integrating high-throughput data, gene knockout mutants, and molecular interaction assays. TroK, a histidine protein kinase in a two-component system that regulates virulence factor secretion, is likely the molecular target antagonized by ATT. Our findings provide novel insights into virulence modulation in an important plant-pathogen system that relies on the host's metabolic activity and subsequent signaling interference.

Analysis of Virus Spread Around the Cell Death Zone at Spatiotemporal Resolution Using Confocal Microscopy.

The role of programmed cell death (PCD) in hypersensitive response (HR)-conferred resistance depends on the type of host-pathogen interaction and therefore has to be studied for each individual pathosystem. Here we present and explain the protocol for studying the role of PCD in HR-conferred resistance in potato plants in the interaction with the viral pathogen. As an experimental system, we use genotype Rywal, where the virus spread is restricted and HR PCD develops 3days post potato virus Y (PVY) inoculation. As a control of virus multiplication and spread, we include its transgenic counterpart impaired in salicylic acid (SA) accumulation (NahG-Rywal), in which the HR-PCD occurs but the spread of the virus is not restricted. To follow the occurrence of virus-infected cells and/or virus multiplication outside the cell death zone, we use GFP-tagged PVY (PVY-N605(123)-GFP) which can be monitored by confocal microscopy. Any other plant-pathogen system which results in PCD development could be studied using a modified version of this protocol.

Effects of Microelements on Downy Mildew (Peronospora belbahrii) of Sweet Basil.

We recently demonstrated that spraying or irrigating with Ca, Mg and K reduces the severity of sweet basil downy mildew (SBDM). Here, the effects of Mn, Zn, Cu and Fe on SBDM were tested in potted plants. The effects of Mn and Zn were also tested under semi-commercial and commercial-like field conditions. Spray applications of a mixture of EDTA-chelated microelements (i.e., Fe-EDTA, Mn-EDTA, Zn-EDTA, Cu-EDTA and Mo) reduces SBDM severity. The application of EDTA chelates of individual microelements (i.e., Fe-EDTA, Mn-EDTA and Zn-EDTA) significantly reduces SBDM in potted plants. Foliar applications of Mn-EDTA and Zn-EDTA are found to be effective under semi-commercial conditions and were, thus, further tested under commercial-like conditions. Under commercial-like conditions, foliar-applied Mn-EDTA and Zn-EDTA decreased SBDM severity by 46–71%. When applied through the irrigation solution, those two microelements reduce SBDM by more than 50%. Combining Mg with Mn-EDTA and Zn-EDTA in the irrigation solution does not provide any additional disease reduction. In the commercial-like field experiment, the microelement-mixture treatment, applied as a spray or via the irrigation solution, was combined with fungicides spray treatments. This combination provides synergistic disease control. The mode of action in this plant–pathogen system may involve features of altered host resistance.

Fungal-derived extracts induce resistance against Botrytis cinerea in Arabidopsis thaliana

Development of a sustainable agriculture is a current trend searching for alternative and more environmentally friendly strategies to control crop diseases. The necrotrophic fungus Botrytis cinerea, the causal agent of grey mould, affects crops pre- and post-harvest and cause huge economical losses. Microorganisms and microbially-derived elicitors are known to induce plant defences. Here, we study two fungal extracts derived from Colletotrichum acutatum (isolate M11). The crude extract, called CF (culture filtrate) and the semi-purified fraction ACF (axenic culture filtrate) consist in a blend of fungal-derived elicitors. Their activities were evaluated in the plant-pathogen system Arabidopsis thaliana-B. cinerea. Both CF and ACF induced local and systemic resistance, rendering A. thaliana resistant against grey mould disease. The extracts showed no antifungal activity since no inhibition of mycelium growth, changes in hyphal ultrastructure or damage in cell membrane permeability were observed. These findings would indicate that the protective effect was indirect and it was found that CF and ACF induced up-regulation of the defence-related genes PR1, WRKY70 and ERF6, as well as giving local callose deposition in treated leaves. Both fungal extracts have the potential to be used as a main component of a biologically based disease control product with activity against grey mould, with the advantage that the purification process of CF is simpler than for ACF.

Decomposition and organic amendments chemistry explain contrasting effects on plant growth promotion and suppression of Rhizoctonia solani damping off.

Organic Amendments (OAs) has been used in agroecosystems to promote plant growth and control diseases caused by soilborne pathogens. However, the role of OAs chemistry and decomposition time on plant growth promotion and disease suppression is still poorly explored. In this work, we studied the effect of 14 OAs at four decomposition ages (3, 30, 100, and 300 days) on the plant—pathogen system Lactuca sativa–Rhizoctonia solani. OAs chemistry was characterized via 13C-CPMAS NMR spectroscopy as well as for standard chemical (i.e. N content, pH, EC) and biological parameters (i.e. phytotoxicity and R. solani proliferation bioassay). OAs have shown variable effects, ranging from inhibition to stimulation of Lactuca sativa and Lepidium sativum growth. We recorded that N rich OAs with high decomposability were conducive in the short-term, while converting suppressive in the long term (300 days). On the other hand, cellulose-rich OAs with high C/N ratio impaired L. sativa growth but were more consistent in providing protection from damping-off, although this property has significantly shifted during decomposition time. These results, for the first time, highlight a consistent trade-off between plant growth promotion and disease control capability of OAs. Finally, we found that OAs effects on growth promotion and disease protection can be hardly predictable based on the chemical characteristic, although N content and some 13C CPMAS NMR regions (alkyl C, methoxyl C, and carbonyl C) showed some significant correlations. Therefore, further investigations are needed to identify the mechanism(s) behind the observed suppressive and conducive effects and to identify OAs types and application timing that optimize plant productivity and disease suppression in different agro-ecosystems.

Electrome alterations in a plant-pathogen system: Toward early diagnosis

This work aimed to verify the existence of patterns on the electrophysiological systemic responses of tomato plants inoculated with a pathogenic fungus in an environment with controlled light and temperature. Electrical signalling was measured before and after inoculation in the same plants, and data were analysed with time series techniques and approximate multi-scale entropy (ApEn). Machine learning algorithms were utilised in order to classify data before and after infection throughout the five days of experiments. The obtained results have shown that it is possible to distinguish differences in the plant’s electrome activity before and after the fungus inoculation. In some cases, we have found scale invariance quantified by the power law decay in the distribution histogram. We also found a higher degree of internal organisation quantified by ApEn. The results of the classification algorithms achieved higher accuracy of infection detection at the initial stage of pathogen recognition by the plant. Besides, this study showed evidence that long-distance electrical signalling is likely involved in the plant-pathogen interaction, since signals were obtained in the stem and the inoculum applied on the plant leaves. This might be useful for the early detection of plant infections.

Functional Variation of Plant-Pathogen Interactions: New Concept and Methods for Virulence Data Analyses.

Classical virulence analysis is based on discovering virulence phenotypes of isolates with regard to a composition of resistance genes in a differential set of host genotypes. With such a vision, virulence phenotypes are usually treated in a genetic manner as one of two possible alleles, either virulence or avirulence in a binary locus. Therefore, population genetics metrics and methods have become prevailing tools for analyzing virulence data at multiple loci. However, a basis for resolving binary virulence phenotypes is infection type (IT) data of host-pathogen interaction that express functional traits of each specific isolate in a given situation (particular host, environmental conditions, cultivation practice, and so on). IT is determined by symptoms and signs observed (e.g., lesion type, lesion size, coverage of leaf or leaf segments by mycelium, spore production and so on), and assessed by IT scores at a generally accepted scale for each plant-pathogen system. Thus, multiple IT profiles of isolates are obtained and can be subjected to analysis of functional variation within and among operational units of a pathogen. Such an approach may allow better utilization of the information available in the raw data, and reveal a functional (e.g., environmental) component of pathogen variation in addition to the genetic one. New methods for measuring functional variation of plant-pathogen interaction with IT data were developed. The methods need an appropriate assessment scale and expert estimations of dissimilarity between IT scores for each plant-pathogen system (an example is presented). Analyses of a few data sets at different hierarchical levels demonstrated discrepancies in results obtained with IT phenotypes versus binary virulence phenotypes. The ability to measure functional IT-based variation offers promise as an effective tool in the study of epidemics caused by plant pathogens.

Probing of Phytofungal Proteins for Fungicidal Activity by Molecular Docking

Background: Plant fungal diseases are the primary causes of foliage and crop loss eventually affecting the overall economic outcome and yield quality. Hence, various chemical compounds are employed to eradicate the fungi in agriculture. Methods: Virtual screening and molecular docking strategies provide themselves as great alternatives to find lead compounds. Lead compounds for each fungal infection was docked to target protein sequence and assessed for the strongest interaction. Findings: Various molecules were taken under the study, for being the target ligands to bring about a fungicidal reaction in the plant pathogen system. The screening of molecules was done thoroughly to produce the results. Ligands identified through this study allow us to make plant host fight against the fungal pathogen and prevent the occurrence of the disease. The interactions have been thoroughly studied with various softwares like SPDBV and PyMol and through various online databases like STRING, GenePept, PDB, UniProt, PatchDock, Protein structure prediction server -2 and others for the overall evaluation of the drug molecule designed and to study its overall effects for the overall higher efficacy and to prevent the occurrence of the fungal disease and management of the fungal pathogens in agriculture against various economically valuable plants. The lead compounds revealed several hydrophobic and polar contacts were demonstrated by comparing interactions. Applications: The molecular affinity of the fungicidal compound has been tested against the target pathogen as well as the host system components to understand the interaction and to draw out the functioning and the analysis. The compatibility between the molecule and the protein has been studied to decipher the effectiveness of the molecule and its effects in the system. The present results let us establish lead compounds that can be used for the development of antifungal drugs although structural activity relationship studies have to be undertaken.

Modeling nutrient and disease dynamics in a plant-pathogen system.

Human activities alter elemental nutrient cycling, which can have profound impacts on agriculture, grasslands, lakes, and other systems. It is becoming increasingly clear that enhanced nitrogen and phosphorus levels can affect disease dynamics across a range of taxa. However, there are few mathematical models that explicitly incorporate nutrients into host-pathogen interactions. Using viral load and plant mass data from an experiment with cereal yellow dwarf virus and its host plant, Avena sativa, we propose and compare two models describing the overall infection dynamics. However, the first model considers nutrient-limited virus production while the other considers a nutrient-induced viral production delay. A virus reproduction number is derived for this nutrient model, which depends on environmental and physiological attributes. Results suggest that including nutrient mediated viral production mechanisms can give rise to robust models that can be used to untangle how nutrients impact pathogen dynamics.

The Use of Features from Fluorescence, Thermography, and NDVI Imaging to Detect Biotic Stress in Lettuce.

Fluorescence, normalized difference vegetation index, and thermal imaging are three frequently used nondestructive methods to detect biotic stress in plants. Due, in part, to the inconsistent results reported in the literature and the lack of measurements on the whole-plant scale, we tested the suitability of a wide variety of variables obtained using these three imaging methods to classify young plants into biotically stressed and nonstressed plants. To this end, we applied the model plant-pathogen system lettuce-Rhizoctonia solani. The relevant data from each image and plant (healthy and diseased) was extracted semiautomatically using sophisticated image processing algorithms. This method enabled us to identify the most appropriate variables via discriminant function and logistic regression analysis: photosystem II maximum quantum yield (Fv/Fm) and fluorescence decline ratio can be used to classify variables with an error ≤0.052. Lettuce seedlings with an Fv/Fm ratio > 0.73 were consistently healthy. In some cases, it was possible to detect infection prior to the appearance of symptoms. Possibilities to transfer the method to horticultural practice are discussed.

Statistical modeling of long-term grapevine response to \u2018Candidatus Phytoplasma solani\u2019 infection in the field

Bois noir (BN) is the most widespread European grapevine yellows disease caused by ‘Candidatus Phytoplasma solani’. Although our knowledge of the mechanisms of interactions of this pathogenic bacteria with host is largely unknown, the plant-pathogen system of BN is commonly used as a model system for studying grapevine yellows diseases. We applied here a conceptual model of general plant pathology – a disease triangle for describing interactions among the host plant, the pathogen and the environment. We generated a proof-of-concept statistical model for disease triangle using original experimental data and different statistical and data mining approaches for a selected system of ‘Ca. P. solani’ infection of cv. ‘Chardonnay’ grapevine plants. We monitored individual plants from a single vineyard over a period of six years. Phytoplasma content, the expression of 21 selected grapevine genes and environmental conditions were recorded and related to disease severity. Our model predicts that in described conditions BN is a function of the expression of grapevine gene VvDMR6, summer rainfall and abundance of ‘Ca. P. solani’. The greatest impact among elements of the disease triangle is attributed to the pathogen, and is independent of the pathogen titer. We showed that this first de facto representation of the disease triangle is useful for showing disease dynamics over several years and could be applied to other plant-pathogen systems. The overall results of this study will contribute to understanding of ‘Ca. P. solani’ biology and its interactions with grapevine host.

Transmission and temporal dynamics of anther‐smut disease (Microbotryum) on alpine carnation (Dianthus pavonius)

Summary Theory has shown that sterilizing diseases with frequency‐dependent transmission (characteristics shared by many sexually transmitted diseases) can drive host populations to extinction. Anther‐smut disease (caused by Microbotryum sp.) has become a model plant pathogen system for studying the dynamics of vector‐ and sexually transmitted diseases: infected individuals are sterilized, producing spores instead of pollen, and the disease is spread between reproductive individuals by insect pollinators. We investigated anther‐smut disease in a heavily infected population of Dianthus pavonius (alpine carnation) over an 8‐year period to determine disease impacts on host population dynamics. Over the 8 years, disease prevalence remained consistently high (>40%), while the host population numbers declined by over 50%. The observed rate of vector transmission to reproductive, adult hosts was inadequate to explain the high disease prevalence. Additional density‐dependent aerial transmission to highly susceptible juveniles, indicated from experimental field and greenhouse studies, is likely to play a key role in maintaining the high disease prevalence. Epidemiological models that accounted for the mixed transmission mode predicted an eventual decline in disease. Synthesis. Our results demonstrate that high prevalence of a sterilizing disease does not necessarily drive host populations towards extinction and also highlights the importance of demographic studies for establishing the presence of alternative transmission modes.

Floral Scent Mimicry and Vector-Pathogen Associations in a Pseudoflower-Inducing Plant Pathogen System.

Several fungal plant pathogens induce ‘pseudoflowers’ on their hosts to facilitate insect-mediated transmission of gametes and spores. When spores must be transmitted to host flowers to complete the fungal life cycle, we predict that pseudoflowers should evolve traits that mimic flowers and attract the most effective vectors in the flower-visiting community. We quantified insect visitation to flowers, healthy leaves and leaves infected with Monilinia vaccinii-corymbosi (Mvc), the causative agent of mummy berry disease of blueberry. We developed a nested PCR assay for detecting Mvc spores on bees, flies and other potential insect vectors. We also collected volatiles from blueberry flowers, healthy leaves and leaves infected with Mvc, and experimentally manipulated specific pathogen-induced volatiles to assess attractiveness to potential vectors. Bees and flies accounted for the majority of contacts with flowers, leaves infected with Mvc and healthy leaves. Flowers were contacted most often, while there was no difference between bee or fly contacts with healthy and infected leaves. While bees contacted flowers more often than flies, flies contacted infected leaves more often than bees. Bees were more likely to have Mvc spores on their bodies than flies, suggesting that bees may be more effective vectors than flies for transmitting Mvc spores to flowers. Leaves infected with Mvc had volatile profiles distinct from healthy leaves but similar to flowers. Two volatiles produced by flowers and infected leaves, cinnamyl alcohol and cinnamic aldehyde, were attractive to bees, while no volatiles manipulated were attractive to flies or any other insects. These results suggest that Mvc infection of leaves induces mimicry of floral volatiles, and that transmission occurs primarily via bees, which had the highest likelihood of carrying Mvc spores and visited flowers most frequently.