The cost of phage resistance in a plant pathogenic bacterium is context-dependent.

Parasites are ubiquitous features of living systems and many parasites severely reduce the fecundity or longevity of their hosts. This parasite-imposed selection on host populations should strongly favor the evolution of host resistance, but hosts typically face a trade-off between investment in reproductive fitness and investment in defense against parasites. The magnitude of such a trade-off is likely to be context-dependent, and accordingly costs that are key in shaping evolution in nature may not be easily observable in an artificial environment. We set out to assess the costs of phage resistance for a plant pathogenic bacterium in its natural plant host versus in a nutrient-rich, artificial medium. We demonstrate that mutants of Pseudomonas syringae that have evolved resistance via a single mutational step pay a substantial cost for this resistance when grown on their tomato plant hosts, but do not realize any measurable growth rate costs in nutrient-rich media. This work demonstrates that resistance to phage can significantly alter bacterial growth within plant hosts, and therefore that phage-mediated selection in nature is likely to be an important component of bacterial pathogenicity

Understanding bacteriophage specificity in natural microbial communities.

This is an open access article that is freely available in ORE or from the publisher's web site. Please cite the published version.Studying the coevolutionary dynamics between bacteria and the bacteriophage viruses that infect them is critical to understanding both microbial diversity and ecosystem functioning. Phages can play a key role in shaping bacterial population dynamics and can significantly alter both intra- and inter-specific competition among bacterial hosts. Predicting how phages might influence community stability and apparent competition, however, requires an understanding of how bacteria-phage interaction networks evolve as a function of host diversity and community dynamics. Here, we first review the progress that has been made in understanding phage specificity, including the use of experimental evolution, we then introduce a new dataset on natural bacteriophages collected from the phyllosphere of horse chestnut trees, and finally we highlight that bacterial sensitivity to phage is rarely a binary trait and that this variation should be taken into account and reported. We emphasize that there is currently insufficient evidence to make broad generalizations about phage host range in natural populations, the limits of phage adaptation to novel hosts, or the implications of phage specificity in shaping microbial communities. However, the combination of experimental and genomic approaches with the study of natural communities will allow new insight to the evolution and impact of phage specificity within complex bacterial communities

Exploring the risks of phage application in the environment

This is an open access article that is freely available in ORE or from the publisher's web site. Please cite the published version.Interest in using bacteriophages to control the growth and spread of bacterial pathogens is being revived in the wake of widespread antibiotic resistance. However, little is known about the ecological effects that high concentrations of phages in the environment might have on natural microbial communities. We review the current evidence suggesting phage-mediated environmental perturbation, with a focus on agricultural examples, and describe the potential implications for human health and agriculture. Specifically, we examine the known and potential consequences of phage application in certain agricultural practices, discuss the risks of evolved bacterial resistance to phages, and question whether the future of phage therapy will emulate that of antibiotic treatment in terms of widespread resistance. Finally, we propose some basic precautions that could preclude such phenomena and highlight existing methods for tracking bacterial resistance to phage therapeutic agents

The tri-­trophic interaction of plants, pathogenic bacteria and bacteriophages

The ecology and evolution of pathogens are key factors in predicting the severity and spread of disease, as well as treatment outcomes. However, the effects of multiple trophic levels that include host, microbial competitors and viruses are typically overlooked. In this thesis I develop our understanding of bacteria-­phage coevolution, microbial dispersal and the role of the microbiome in disease. The results of these experiments have direct implications for phage therapy: the use of bacteriophages to treat bacterial infections. Firstly, I explore the risks of phage application in the environment and draw parallels with the misuse of antibiotics in selecting for bacterial resistance. I then demonstrate that the evolution of resistance to phages in a plant pathogenic bacterium is context-­dependent. Notably, I find a fitness cost in plant infections that is absent when the bacteria are cultured solely in the laboratory. I then characterize four novel phages and use a simple laboratory based assay to predict their potential as phage therapy agents in an agricultural context. Next I show that reservoir species of plant hosts can affect the evolution of virulence, when bacteria are passaged on both a focal and distant host, but find no evidence of local adaptation. I also show that the evolution of such traits can occur in a parallel manner at the genetic level. I then determine a compositional shift in the microbiota associated with the symptoms of bleeding canker disease in Horse Chestnut trees across the length of the UK. Finally, I find an age-­related decline in bacterial species richness and evidence for niche-­assembly theories by investigating bacterial dispersal in UK Oak trees in a single woodland.University of Exete

Individual bacteria in structured environments rely on phenotypic resistance to phage

This is the final version. Available on open access from Public Library of Science via the DOI in this recordData Availability: All relevant data are within the paper and its Supporting Information files.Bacteriophages represent an avenue to overcome the current antibiotic resistance crisis, but evolution of genetic resistance to phages remains a concern. In vitro, bacteria evolve genetic resistance, preventing phage adsorption or degrading phage DNA. In natural environments, evolved resistance is lower possibly because the spatial heterogeneity within biofilms, microcolonies, or wall populations favours phenotypic survival to lytic phages. However, it is also possible that the persistence of genetically sensitive bacteria is due to less efficient phage amplification in natural environments, the existence of refuges where bacteria can hide, and a reduced spread of resistant genotypes. Here, we monitor the interactions between individual planktonic bacteria in isolation in ephemeral refuges and bacteriophage by tracking the survival of individual cells. We find that in these transient spatial refuges, phenotypic resistance due to reduced expression of the phage receptor is a key determinant of bacterial survival. This survival strategy is in contrast with the emergence of genetic resistance in the absence of ephemeral refuges in well-mixed environments. Predictions generated via a mathematical modelling framework to track bacterial response to phages reveal that the presence of spatial refuges leads to fundamentally different population dynamics that should be considered in order to predict and manipulate the evolutionary and ecological dynamics of bacteria-phage interactions in naturally structured environments.Medical Research Council (MRC)Engineering and Physical Sciences Research Council (EPSRC)Gordon and Betty and Gordon Moore FoundationEuropean Research Council (ERC)Biotechnology and Biological Sciences Research Council (BBSRC)Natural Environment Research Council (NERC)Marie Skłodowska-Curie ActionsDefence Science and Technology Laboratory (Dstl)Royal Societ

Resource quality determines the evolution of resistance and its genetic basis

This is the final version. Available on open access from Wiley via the DOI in this recordData Availability: All the experimental data to support the findings of this study including all virus assay and development data is available at DataDryad. https://doi.org/10.5061/dryad.k98sf7m4g. The complete sequencing data in CRAM format is available from the European Bioinformatics Institute (EBI), under accession number PRJEB27964.Parasites impose strong selection on their hosts, but the level of any evolved resistance may be constrained by the availability of resources. However, studies identifying the genomic basis of such resource‐mediated selection are rare, particularly in non‐model organisms. Here, we investigated the role of nutrition in the evolution of resistance to a DNA virus (PiGV), and any associated trade‐offs in a lepidopteran pest species (Plodia interpunctella). Through selection experiments and whole genome re‐sequencing we identify genetic markers of resistance that vary between the nutritional environments during selection. We do not find consistent evolution of resistance in the presence of virus but rather see substantial variation among replicate populations. Resistance in a low nutrition environment is negatively correlated with growth rate, consistent with an established trade‐off between immunity and development, but this relationship is highly context dependent. Whole genome resequencing of the host shows that resistance mechanisms are likely to be highly polygenic and although the underlying genetic architecture may differ between high and low nutrition environments, similar mechanisms are commonly used. As a whole, our results emphasise the importance of the resource environment on influencing the evolution of resistance.Natural Environment Research Council (NERC)National Institutes of Health (NIH

full_assay_dataset

Data from plant infections of Tomato and Arabidopsis with P.syringae lines from an experimental evolution experiment