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Review 2: "A Weaponized Phage Suppresses Competitors in Historical and Modern Metapopulations of Pathogenic Bacterial"

Overall, the reviews are very positive, offering constructive suggestions to further enhance the manuscript's impact and clarity.

Published onApr 12, 2024
Review 2: "A Weaponized Phage Suppresses Competitors in Historical and Modern Metapopulations of Pathogenic Bacterial"
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A weaponized phage suppresses competitors in historical and modern metapopulations of pathogenic bacteria
A weaponized phage suppresses competitors in historical and modern metapopulations of pathogenic bacteria
Description

Abstract Bacteriophages, the viruses of bacteria, are proposed to drive bacterial population dynamics, yet direct evidence of their impact on natural populations is limited. Here we identified viral sequences in a metapopulation of wild plant-associated Pseudomonas spp. genomes. We discovered that the most abundant viral cluster does not encode an intact phage but instead encodes a tailocin - a phage-derived element that bacteria use to kill competitors for interbacterial warfare. Each pathogenic Pseudomonas sp. strain carries one of a few distinct tailocin variants, which target variable polysaccharides in the outer membrane of co-occurring pathogenic strains. Analysis of historic herbarium samples from the last 170 years revealed that the same tailocin and receptor variants have persisted in the Pseudomonas populations for at least two centuries, suggesting the continued use of a defined set of tailocin haplotypes and receptors. These results indicate that tailocin genetic diversity can be mined to develop targeted “tailocin cocktails” for microbial control.One-Sentence Summary Bacterial pathogens in a host-associated metapopulation use a repurposed prophage to kill their competitors.

RR:C19 Evidence Scale rating by reviewer:

  • Strong. The main study claims are very well-justified by the data and analytic methods used. There is little room for doubt that the study produced has very similar results and conclusions as compared with the hypothetical ideal study. The study’s main claims should be considered conclusive and actionable without reservation.

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Review: The authors' main claims involve the identification of a highly conserved viral cluster (VC2) within the pathogenic clade of P. viridiflava through Pseudomonas spp. sequencing, alongside predictions and experimental validation of tailocin functionality. They emphasize tailocins' origin from phages, their potential as effective agricultural antimicrobials, and the significant relationship between diverse tailocin variants and their targeting of pathogenic Pseudomonas strains, inferring co-evolution with O-antigens.

The manuscript utilizes Pseudomonas spp. sequencing to demonstrate the presence of a highly conserved viral cluster (VC2) within the pathogenic clade of P. viridiflava. Further predictions regarding the functional aspects of tailocins are made and experimentally validated. Tailocins, which originate from phages, show promising potential as agricultural antimicrobials due to their efficient bactericidal properties and precise targeting abilities specific to particular strains. It also has been demonstrated in P. syringae. In addition to demonstrating the presence and functionality of VC2, the authors elucidate the significant relationship between highly diverse tailocin variants and the effective targeting of closely related pathogenic Pseudomonas strains. They also infer the co-evolution of tailocins and O-antigens. This aspect is crucial for understanding the application of phage therapy, as bacteriophage resistance often arises from competition with receptors. The logical coherence of the argumentation and the research methodology design adequately support the study's findings.

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