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Coculturing bacteria leads to reduced phenotypic heterogeneities

Jasmine Heyse (UGent) , Benjamin Buysschaert (UGent) , Ruben Props (UGent) , Peter Rubbens (UGent) , Andre Skirtach (UGent) , Willem Waegeman (UGent) and Nico Boon (UGent)
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Abstract
Isogenic bacterial populations are known to exhibit phenotypic heterogeneity at the single-cell level. Because of difficulties in assessing the phenotypic heterogeneity of a single taxon in a mixed community, the importance of this deeper level of organization remains relatively unknown for natural communities. In this study, we have used membrane-based microcosms that allow the probing of the phenotypic heterogeneity of a single taxon while interacting with a synthetic or natural community. Individual taxa were studied under axenic conditions, as members of a coculture with physical separation, and as a mixed culture. Phenotypic heterogeneity was assessed through both flow cytometry and Raman spectroscopy. Using this setup, we investigated the effect of microbial interactions on the individual phenotypic heterogeneities of two interacting drinking water isolates. Through flow cytometry we have demonstrated that interactions between these bacteria lead to a reduction of their individual phenotypic diversities and that this adjustment is conditional on the bacterial taxon. Single-cell Raman spectroscopy confirmed a taxon-dependent phenotypic shift due to the interaction. In conclusion, our data suggest that bacterial interactions may be a general driver of phenotypic heterogeneity in mixed microbial populations. IMPORTANCE: Laboratory studies have shown the impact of phenotypic heterogeneity on the survival and functionality of isogenic populations. Because phenotypic heterogeneity plays an important role in pathogenicity and virulence, antibiotic resistance, biotechnological applications, and ecosystem properties, it is crucial to understand its influencing factors. An unanswered question is whether bacteria in mixed communities influence the phenotypic heterogeneity of their community partners. We found that coculturing bacteria leads to a reduction in their individual phenotypic heterogeneities, which led us to the hypothesis that the individual phenotypic diversity of a taxon is dependent on the community composition.
Keywords
Raman spectroscopy, axenic culture, coculture, flow cytometry, microbial interactions, phenotypic heterogeneity, single cell, synthetic ecosystems, GENE-EXPRESSION, FLOW-CYTOMETRY, DRINKING-WATER, SINGLE CELLS, INDIVIDUALITY, NOISE, MICROBIOLOGY, POPULATIONS, DIVERSITY, SYSTEMS

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Chicago
Heyse, Jasmine, Benjamin Buysschaert, Ruben Props, Peter Rubbens, Andre Skirtach, Willem Waegeman, and Nico Boon. 2019. “Coculturing Bacteria Leads to Reduced Phenotypic Heterogeneities.” Applied and Environmental Microbiology 85 (8).
APA
Heyse, Jasmine, Buysschaert, B., Props, R., Rubbens, P., Skirtach, A., Waegeman, W., & Boon, N. (2019). Coculturing bacteria leads to reduced phenotypic heterogeneities. APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 85(8).
Vancouver
1.
Heyse J, Buysschaert B, Props R, Rubbens P, Skirtach A, Waegeman W, et al. Coculturing bacteria leads to reduced phenotypic heterogeneities. APPLIED AND ENVIRONMENTAL MICROBIOLOGY. 2019;85(8).
MLA
Heyse, Jasmine et al. “Coculturing Bacteria Leads to Reduced Phenotypic Heterogeneities.” APPLIED AND ENVIRONMENTAL MICROBIOLOGY 85.8 (2019): n. pag. Print.
@article{8614858,
  abstract     = {Isogenic bacterial populations are known to exhibit phenotypic heterogeneity at the single-cell level. Because of difficulties in assessing the phenotypic heterogeneity of a single taxon in a mixed community, the importance of this deeper level of organization remains relatively unknown for natural communities. In this study, we have used membrane-based microcosms that allow the probing of the phenotypic heterogeneity of a single taxon while interacting with a synthetic or natural community. Individual taxa were studied under axenic conditions, as members of a coculture with physical separation, and as a mixed culture. Phenotypic heterogeneity was assessed through both flow cytometry and Raman spectroscopy. Using this setup, we investigated the effect of microbial interactions on the individual phenotypic heterogeneities of two interacting drinking water isolates. Through flow cytometry we have demonstrated that interactions between these bacteria lead to a reduction of their individual phenotypic diversities and that this adjustment is conditional on the bacterial taxon. Single-cell Raman spectroscopy confirmed a taxon-dependent phenotypic shift due to the interaction. In conclusion, our data suggest that bacterial interactions may be a general driver of phenotypic heterogeneity in mixed microbial populations. 
IMPORTANCE: Laboratory studies have shown the impact of phenotypic heterogeneity on the survival and functionality of isogenic populations. Because phenotypic heterogeneity plays an important role in pathogenicity and virulence, antibiotic resistance, biotechnological applications, and ecosystem properties, it is crucial to understand its influencing factors. An unanswered question is whether bacteria in mixed communities influence the phenotypic heterogeneity of their community partners. We found that coculturing bacteria leads to a reduction in their individual phenotypic heterogeneities, which led us to the hypothesis that the individual phenotypic diversity of a taxon is dependent on the community composition.},
  articleno    = {e02814-18},
  author       = {Heyse, Jasmine and Buysschaert, Benjamin and Props, Ruben and Rubbens, Peter and Skirtach, Andre and Waegeman, Willem and Boon, Nico},
  issn         = {0099-2240},
  journal      = {APPLIED AND ENVIRONMENTAL MICROBIOLOGY},
  keywords     = {Raman spectroscopy,axenic culture,coculture,flow cytometry,microbial interactions,phenotypic heterogeneity,single cell,synthetic ecosystems,GENE-EXPRESSION,FLOW-CYTOMETRY,DRINKING-WATER,SINGLE CELLS,INDIVIDUALITY,NOISE,MICROBIOLOGY,POPULATIONS,DIVERSITY,SYSTEMS},
  language     = {eng},
  number       = {8},
  pages        = {13},
  title        = {Coculturing bacteria leads to reduced phenotypic heterogeneities},
  url          = {http://dx.doi.org/10.1128/AEM.02814-18},
  volume       = {85},
  year         = {2019},
}

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