Extensive cellular multi-tasking within Bacillus subtilis biofilms
ABSTRACT Bacillus subtilis is a soil-dwelling bacterium that can form biofilms, or communities of cells surrounded by a self-produced extracellular matrix. In biofilms, genetically identical cells often exhibit heterogeneous transcriptional phenotypes, so that subpopulations of cells carry out essen...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
American Society for Microbiology
2023-08-01
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| Series: | mSystems |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/msystems.00891-22 |
| _version_ | 1797730339073818624 |
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| author | Sarah M. Yannarell Eric S. Beaudoin Hunter S. Talley Alexi A. Schoenborn Galya Orr Christopher R. Anderton William B. Chrisler Elizabeth A. Shank |
| author_facet | Sarah M. Yannarell Eric S. Beaudoin Hunter S. Talley Alexi A. Schoenborn Galya Orr Christopher R. Anderton William B. Chrisler Elizabeth A. Shank |
| author_sort | Sarah M. Yannarell |
| collection | DOAJ |
| description | ABSTRACT Bacillus subtilis is a soil-dwelling bacterium that can form biofilms, or communities of cells surrounded by a self-produced extracellular matrix. In biofilms, genetically identical cells often exhibit heterogeneous transcriptional phenotypes, so that subpopulations of cells carry out essential yet costly cellular processes that allow the entire population to thrive. Surprisingly, the extent of phenotypic heterogeneity and the relationships between subpopulations of cells within biofilms of even in well-studied bacterial systems like B. subtilis remains largely unknown. To determine relationships between these subpopulations of cells, we created 182 strains containing pairwise combinations of fluorescent transcriptional reporters for the expression state of 14 different genes associated with potential cellular subpopulations. We determined the spatial organization of the expression of these genes within biofilms using confocal microscopy, which revealed that many reporters localized to distinct areas of the biofilm, some of which were co-localized. We used flow cytometry to quantify reporter co-expression, which revealed that many cells “multi-task,” simultaneously expressing two reporters. These data indicate that prior models describing B. subtilis cells as differentiating into specific cell types, each with a specific task or function, were oversimplified. Only a few subpopulations of cells, including surfactin and plipastatin producers, as well as sporulating and competent cells, appear to have distinct roles based on the set of genes examined here. These data will provide us with a framework with which to further study and make predictions about the roles of diverse cellular phenotypes in B. subtilis biofilms. IMPORTANCE Many microbes differentiate, expressing diverse phenotypes to ensure their survival in various environments. However, studies on phenotypic differentiation have typically examined only a few phenotypes at one time, thus limiting our knowledge about the extent of differentiation and phenotypic overlap in the population. We investigated the spatial organization and gene expression relationships for genes important in B. subtilis biofilms. In doing so, we mapped spatial gene expression patterns and expanded the number of cell populations described in the B. subtilis literature. It is likely that other bacteria also display complex differentiation patterns within their biofilms. Studying the extent of cellular differentiation in other microbes may be important when designing therapies for disease-causing bacteria, where studying only a single phenotype may be masking underlying phenotypic differentiation relevant to infection outcomes. |
| first_indexed | 2024-03-12T11:42:51Z |
| format | Article |
| id | doaj.art-b4c992fa01f3441a82316256f850d6a7 |
| institution | Directory Open Access Journal |
| issn | 2379-5077 |
| language | English |
| last_indexed | 2024-03-12T11:42:51Z |
| publishDate | 2023-08-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mSystems |
| spelling | doaj.art-b4c992fa01f3441a82316256f850d6a72023-08-31T13:00:43ZengAmerican Society for MicrobiologymSystems2379-50772023-08-018410.1128/msystems.00891-22Extensive cellular multi-tasking within Bacillus subtilis biofilmsSarah M. Yannarell0Eric S. Beaudoin1Hunter S. Talley2Alexi A. Schoenborn3Galya Orr4Christopher R. Anderton5William B. Chrisler6Elizabeth A. Shank7Department of Microbiology and Immunology, University of North Carolina , Chapel Hill, North Carolina, USADepartment of Systems Biology, University of Massachusetts Chan Medical School , Worcester, Massachusetts, USADepartment of Biology, University of North Carolina , Chapel Hill, North Carolina, USADepartment of Microbiology and Immunology, University of North Carolina , Chapel Hill, North Carolina, USAEnvironmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory , Richland, Washington, USAEnvironmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory , Richland, Washington, USAEnvironmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory , Richland, Washington, USADepartment of Systems Biology, University of Massachusetts Chan Medical School , Worcester, Massachusetts, USAABSTRACT Bacillus subtilis is a soil-dwelling bacterium that can form biofilms, or communities of cells surrounded by a self-produced extracellular matrix. In biofilms, genetically identical cells often exhibit heterogeneous transcriptional phenotypes, so that subpopulations of cells carry out essential yet costly cellular processes that allow the entire population to thrive. Surprisingly, the extent of phenotypic heterogeneity and the relationships between subpopulations of cells within biofilms of even in well-studied bacterial systems like B. subtilis remains largely unknown. To determine relationships between these subpopulations of cells, we created 182 strains containing pairwise combinations of fluorescent transcriptional reporters for the expression state of 14 different genes associated with potential cellular subpopulations. We determined the spatial organization of the expression of these genes within biofilms using confocal microscopy, which revealed that many reporters localized to distinct areas of the biofilm, some of which were co-localized. We used flow cytometry to quantify reporter co-expression, which revealed that many cells “multi-task,” simultaneously expressing two reporters. These data indicate that prior models describing B. subtilis cells as differentiating into specific cell types, each with a specific task or function, were oversimplified. Only a few subpopulations of cells, including surfactin and plipastatin producers, as well as sporulating and competent cells, appear to have distinct roles based on the set of genes examined here. These data will provide us with a framework with which to further study and make predictions about the roles of diverse cellular phenotypes in B. subtilis biofilms. IMPORTANCE Many microbes differentiate, expressing diverse phenotypes to ensure their survival in various environments. However, studies on phenotypic differentiation have typically examined only a few phenotypes at one time, thus limiting our knowledge about the extent of differentiation and phenotypic overlap in the population. We investigated the spatial organization and gene expression relationships for genes important in B. subtilis biofilms. In doing so, we mapped spatial gene expression patterns and expanded the number of cell populations described in the B. subtilis literature. It is likely that other bacteria also display complex differentiation patterns within their biofilms. Studying the extent of cellular differentiation in other microbes may be important when designing therapies for disease-causing bacteria, where studying only a single phenotype may be masking underlying phenotypic differentiation relevant to infection outcomes.https://journals.asm.org/doi/10.1128/msystems.00891-22biofilmscell typesconfocal microscopyflow cytometryfluorescent reporters |
| spellingShingle | Sarah M. Yannarell Eric S. Beaudoin Hunter S. Talley Alexi A. Schoenborn Galya Orr Christopher R. Anderton William B. Chrisler Elizabeth A. Shank Extensive cellular multi-tasking within Bacillus subtilis biofilms mSystems biofilms cell types confocal microscopy flow cytometry fluorescent reporters |
| title | Extensive cellular multi-tasking within Bacillus subtilis biofilms |
| title_full | Extensive cellular multi-tasking within Bacillus subtilis biofilms |
| title_fullStr | Extensive cellular multi-tasking within Bacillus subtilis biofilms |
| title_full_unstemmed | Extensive cellular multi-tasking within Bacillus subtilis biofilms |
| title_short | Extensive cellular multi-tasking within Bacillus subtilis biofilms |
| title_sort | extensive cellular multi tasking within bacillus subtilis biofilms |
| topic | biofilms cell types confocal microscopy flow cytometry fluorescent reporters |
| url | https://journals.asm.org/doi/10.1128/msystems.00891-22 |
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