Multi-Omic Analysis of Bacteriophage-Insensitive Mutants Reveals a Putative Role for the Rcs Two-Component Phosphorelay System in Phage Resistance Development in Erwinia amylovora
Bibliographic record
Abstract
Phage therapy has garnered significant attention due to the rise of life-threatening multidrug-resistant pathogenic bacteria and the growing awareness of the transfer of resistance genes between pathogens. Considering this, phage therapy applications are being extended to target plant pathogenic bacteria, such as Erwinia amylovora, which causes fire blight in apple and pear orchards. Understanding the mechanisms involved in phage resistance is crucial for enhancing the effectiveness of phage therapy. Despite the challenges of naturally developing a bacteriophage-insensitive mutant (BIM) of E. amylovora (without traditional mutagenesis methods), this study successfully created a BIM against the podovirus ϕEa46-1-A1. The parent strain, E. amylovora D7, and the BIM B6-2 were extensively compared at genomic, transcriptomic, and phenotypic levels. The phenotypic comparison included the metabolic behavior, biofilm formation, and in planta evaluations of pathogenicity. The results revealed a mutation in strain B6-2 in the rcsB gene, which encodes a second regulator in the Rcs two-component phosphorelay system (TCS). This mutation resulted in significant changes in the B6-2 BIM, including downregulation of amylovoran gene expression (e.g., an average log2 fold change of −4.35 across amsA-L), visible alterations in biofilm formation, increased sensitivity to antibiotics (22.4% more sensitive to streptomycin), and a loss of pathogenicity as assessed in an apple seedling virulence model in comparison to the wildtype strain. The findings presented in this study highlight the critical role of the Rcs phosphorelay system in phage resistance in E. amylovora. Based on these findings, we have proposed a model that explains the effect of the B6-2 rcsB mutation on the Rcs phosphorelay system and its contribution to the development of phage resistance in E. amylovora.
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How this classification was reachedexpand
Full frame distilled prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.
Codex and Gemma teacher scores by category
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.001 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.001 | 0.000 |
| Bibliometrics | 0.000 | 0.001 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.000 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one teacher head, not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".