DNA Methylome Regulates Virulence and Metabolism inPseudomonas syringae

Bacterial pathogens employ epigenetic mechanisms, including DNA methylation, to adapt to environmental changes, and these mechanisms play important roles in various biological processes.Pseudomonas syringaeis a model phytopathogenic bacterium, but its methylome is less well known than that of other species. In this study, we conducted single-molecule real-time sequencing to profile the DNA methylation landscape in three model pathovars ofP. syringae. We identified one Type-I restriction-modification system (HsdMSR), including the conserved sequence motif associated with N⁶-methyladenine (6mA). About 25%–40% of the genes involved in DNA methylation were conserved in two or more of the strains, revealing the functional conservation of methylation inP. syringae. Subsequent transcriptomic analysis highlighted the involvement of HsdMSR in virulent and metabolic pathways, including the Type III secretion system, biofilm formation, and translational efficiency. The regulatory effect of HsdMSR on transcription was dependent on both strands being fully 6mA methylated. Overall, this work illustrated the methylation profile inP. syringaeand the critical involvement of DNA methylation in regulating virulence and metabolism. Thus, this work contributes to a deeper understanding of epigenetic transcriptional control inP. syringaeand related bacteria.