Motility-induced buckling and glassy dynamics regulate three-dimensional transitions of bacterial monolayers

Abstract

Many mature bacterial colonies and biofilms are complex three-dimensional (3D) structures. One key step in their developmental program is a transition from a two-dimensional (2D) monolayer into a 3D architecture. Despite the importance of controlling the growth of microbial colonies and biofilms in a variety of medical and industrial settings, the underlying physical mechanisms behind single-cell dynamics, collective behaviors of densely-packed cells, and 3D complex colony expansion remain largely unknown. In this work, we explore the mechanisms behind the 2D-to-3D transition of motile Pseudomonas aeruginosa colonies; we provide a new motility-induced, rate-dependent buckling mechanism for their out-of-plane growth. We find that swarming of motile bacterial colonies generate sustained in-plane flows. We show that the viscous shear stresses and dynamic pressures arising from these flows allow cells to overcome cell-substrate adhesion, leading to buckling of bacterial monolayers and growth into the third dimension. Modeling bacterial monolayers as 2D fluid films, we identify universal relationships that elucidate the competition between in-plane viscous stresses, pressure and cell-substrate adhesion. Furthermore, we show that bacterial monolayers can exhibit crossover from swarming to kinetically-arrested, glassy-like states above an onset density, resulting in distinct 2D-to-3D transition mechanisms. Combining experimental observations of P. aeruginosa colonies at single-cell resolution, molecular dynamics simulations of active systems, and theories of glassy dynamics and 2D fluid films, we develop a dynamical state diagram that predicts the state of the colony, and the mechanisms governing their 2D-to-3D transitions.