title: Vibrio cholerae biofilm growth program and architecture revealed by single-cell live imaging.
abstract: Biofilms are surface-associated bacterial communities that are crucial in nature and during infection. Despite extensive work to identify biofilm components and to discover how they are regulated, little is known about biofilm structure at the level of individual cells. Here, we use state-of-the-art microscopy techniques to enable live single-cell resolution imaging of a Vibrio cholerae biofilm as it develops from one single founder cell to a mature biofilm of 10,000 cells, and to discover the forces underpinning the architectural evolution. Mutagenesis, matrix labeling, and simulations demonstrate that surface adhesion-mediated compression causes V. cholerae biofilms to transition from a 2D branched morphology to a dense, ordered 3D cluster. We discover that directional proliferation of rod-shaped bacteria plays a dominant role in shaping the biofilm architecture in V. cholerae biofilms, and this growth pattern is controlled by a single gene, rbmA Competition analyses reveal that the dense growth mode has the advantage of providing the biofilm with superior mechanical properties. Our single-cell technology can broadly link genes to biofilm fine structure and provides a route to assessing cell-to-cell heterogeneity in response to external stimuli.
authors.affiliation: Department of Mechanical and Aerospace Engineering, Princeton University, Princeton, NJ 08544; Department of Molecular Biology, Princeton University, Princeton, NJ 08544;
authors.forename: Jing
authors.initials: J
authors.lastname: Yan
authors.affiliation: Department of Physics, Princeton University, Princeton, NJ 08544;
authors.forename: Andrew G
authors.initials: AG
authors.lastname: Sharo
authors.affiliation: Department of Mechanical and Aerospace Engineering, Princeton University, Princeton, NJ 08544;
authors.forename: Howard A
authors.initials: HA
authors.lastname: Stone
authors.affiliation: Department of Molecular Biology, Princeton University, Princeton, NJ 08544;
authors.forename: Ned S
authors.initials: NS
authors.lastname: Wingreen
authors.affiliation: Department of Molecular Biology, Princeton University, Princeton, NJ 08544; Howard Hughes Medical Institute, Chevy Chase, MD 20815 bbassler@princeton.edu.
authors.forename: Bonnie L
authors.initials: BL
authors.lastname: Bassler
mesh_terms: D001422:Bacterial Adhesion / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics*; D018441:Biofilms / Q000254:growth & development*; D049109:Cell Proliferation / Q000235:genetics; D006801:Humans; D059010:Single-Cell Analysis / Q000379:methods*; D014734:Vibrio cholerae / Q000235:genetics / Q000254:growth & development / Q000472:pathogenicity / Q000648:ultrastructure*
chemical_list: D001426:Bacterial Proteins
keywords: biofilm; biomechanics; community; self-organization; single cell