Pseudomonas syringae subpopulations cooperate by coordinating flagellar and type III secretion spatiotemporal dynamics to facilitate plant infection

Abstract

Isogenic bacterial populations can display probabilistic cell-to-cell variation in response to challenges. This phenotypic heterogeneity can affect virulence in animals, but its impact on plant pathogens is unknown. Previously, we showed that expression of the type III secretion system (T3SS) of the plant pathogen Pseudomonas syringae displays phenotypic variation in planta. Here we use flow cytometry and microscopy to investigate single-cell flagellar expression in relation to T3SS expression, showing that both systems undergo phenotypic heterogeneity in vitro in apoplast-mimicking medium and within apoplastic microcolonies throughout colonization of Phaseolus vulgaris. Stochastic, spatial and time factors shape the dynamics of a phenotypically diverse pathogen population that displays division of labour during colonization: effectors produced by T3SS-expressing bacteria act as ‘common goods’ to suppress immunity, allowing motile flagella-expressing bacteria to increase and leave infected tissue before necrosis. These results showcase the mechanisms of bacterial specialization during plant colonization in an environmentally and agriculturally relevant system. Single-cell gene expression analysis reveals phenotypic heterogeneity to enable bacterial specialization over the course of plant colonization.