Abstract
Ecological interactions between different species of yeasts have been observed and described extensively, but the mechanisms of interaction remain poorly understood. A hindrance to the characterization of multispecies yeast ecosystems is the lack of accurate methods for rapid real-time analysis of population dynamics in synthetic multispecies consortia. Here, we sought to accelerate and improve the sensitivity of ecological modelling and characterization of a synthetic yeast ecosystem by developing a flow cytometry–based method that tracks and sorts fluorescently tagged individual yeast species in real time during growth in model multispecies consortia. A protocol for integrative genetic modification of non-conventional yeasts was developed. The application of the method was demonstrated in a model four-species synthetic wine-yeast ecosystem that consisted of species commonly isolated from natural wine fermentations. The data show that this method allows for rapid generation of meaningful ecological data that contributes to our understanding of multispecies synthetic yeast ecosystems. Furthermore, interspecies interactions have been shown to impact the evolution of yeasts in natural ecosystems, and this platform will provide an ideal tool to better evaluate the impact of biotic selection pressures.
Key Points • Fluorescent labelling of yeast species in a consortium for multicolour flow cytometry • Method developed to track population dynamics of multispecies yeast consortia • Enables real-time visualization, manipulation and response analyses of population dynamics • Produces accurate, reproducible data with powerful visual analyses potential at a rapid rate |
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Acknowledgements
We acknowledge the flow cytometry technical assistance of Mrs. Lize Engelbrecht at the fluorescence microscopy unit of the central analytical facility of Stellenbosch University.
Funding
The following funders supported this research: The National Research Foundation of South Africa through Grant number 83471 (SA Research Chair in Integrated Wine Science) to FFB and the Royal Society (Future Leaders - Independent African Researchers (FLAIR)) to DR.
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DR and FFB initiated the research and conceived and conceptualised the project; CC designed and conducted the experimental work; CC, RNB, DR and FFB analysed and interpreted the data; CC wrote the draft version; and CC, DR, RNB and FFB edited the final manuscript.
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Conacher, C.G., Naidoo-Blassoples, R.K., Rossouw, D. et al. Real-time monitoring of population dynamics and physical interactions in a synthetic yeast ecosystem by use of multicolour flow cytometry. Appl Microbiol Biotechnol 104, 5547–5562 (2020). https://doi.org/10.1007/s00253-020-10607-x
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DOI: https://doi.org/10.1007/s00253-020-10607-x