Single-cell profiling of complex plant responses to Pseudomonas syringae infection

SUMMARY Plant response to pathogen infection varies within a leaf, yet this heterogeneity is not well resolved. We exposed Arabidopsis to Pseudomonas syringae or mock treatment and profiled >11,000 individual cells using single-cell RNA sequencing. Integrative analysis of cell populations from both treatments identified distinct pathogen responsive cell clusters exhibiting transcriptional responses ranging from immunity to susceptibility. Pseudotime analyses through pathogen infection revealed a continuum of disease progression from an immune to susceptible state. Confocal imaging of promoter reporter lines for transcripts enriched in immune cell clusters expressed surrounding substomatal cavities colonized or in close proximity to bacterial colonies, suggesting cells within immune clusters represent sites of early pathogen invasion. Susceptibility clusters exhibited more general localization and were highly induced at later stages of infection. Overall, our work uncovers cellular heterogeneity within an infected leaf and provides unique insight into plant differential response to infection at a single-cell level.