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The effects of coculture on cellular spatial heterogeneity in colony and plant-associated biofilms


EMSL Project ID
51105

Abstract

Bacteria live everywhere, from the soil and sea to the surface of every plant and animal on earth. Most bacteria exist within diverse, multispecies communities that exert profound impacts on their hosts as well as on ecosystem-level processes. In spite of the critical importance bacteria have on diverse ecosystems, how bacteria establish and interact within such communities remains largely unknown. The phylogenetic diversity contained within natural microbial communities is well established; in addition, even genetically identical bacterial cells exhibit phenotypic heterogeneity. For example, the environmentally relevant model microbe Bacillus subtilis differentiates into a range of transcriptionally distinct cell types, including those that are producing biofilm matrix, sporulating, or secreting extracellular enzymes for biomass degradation (among many others), activities that may have important environmental impacts. This cellular heterogeneity is frequently generated through the activity of secreted metabolites that act as cell-cell communication molecules. In ongoing work, we are defining how such secreted metabolites spatially correlate with the distribution of diverse cell types within mature B. subtilis biofilms. Here, we propose to expand on this work to examine how these patterns of cellular heterogeneity are perturbed by coculture, both during early biofilm formation and in colony and plant-associated biofilms. Specifically, we will examine B. subtilis' interactions with an environmental isolate of Pantoea agglomerans; we have shown that a coculture between these two strains results in a distinctive biofilm morphology that also alters cellular heterogeneity in B. subtilis. Pursuing these questions will require the ability to visualize both chemical and cellular heterogeneity within fixed and live, intact biofilm colonies. These capabilities are uniquely available at EMSL through their multispectral and super-resolution fluorescence confocal microscopes and their imaging mass spectrometry platforms. The scientific expertise of EMSL staff will be critical to handling the collection, integration, and analysis of these complex spatial datasets. We anticipate that the data from the work proposed here will reveal fundamental insights into how bacteria establish cellular and metabolic heterogeneity within biofilm communities and how interactions with other bacteria perturb the dynamics of these spatial distributions. Our results will yield a more mechanistic understanding of how chemical and cellular heterogeneity is generated within microbial consortia, and thus inform future studies focused on generating predictive models of microbial interactions or manipulating microbial behavior within multispecies microbial communities.

Project Details

Project type
Exploratory Research
Start Date
2019-11-26
End Date
2021-09-30
Status
Closed

Team

Principal Investigator

Elizabeth Shank
Institution
University of Massachusetts Medical School

Team Members

Gregory Vandergrift
Institution
Pacific Northwest National Laboratory

Eric Beaudoin
Institution
University of North Carolina at Chapel Hill

Sarah Yannarell
Institution
University of North Carolina at Chapel Hill