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A rapid, high-throughput method for quantitative determination of ethanol tolerance in Saccharomyces cerevisiae

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Abstract

There are a variety of methods available for determining ethanol tolerance phenotypes in Saccharomyces cerevisiae. Many of these methods are limited in through-put and/or application. We describe here a microtiter-plate, growth-based, liquid-culture, rapid ethanol tolerance assay (RETA) that overcomes these limitations. Determinations of ethanol tolerance gave results that were comparable to shake-flask cultures, and there was a high level of intra- and inter-plate reproducibility. We report the successful application of this assay to determining the segregation pattern of ethanol tolerance in backcrosses of two ethanol-tolerant mutants (SM and CM) to a non-tolerant parent (W303-1A); RETA was used to determine ethanol tolerance in numerous progeny resulting from these crosses. This assay, or variations thereof, will prove be of great value for anyone attempting to develop quantitative, high-throughput growth assays for yeast or other microorganisms.

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Acknowledgments

This project was supported by Australia’s grape growers and winemakers through their investment body, the Grape and Wine Research and Development Corporation, with matching funds from the Australian Government. The Australian Wine Research Institute is a member of the Wine Innovation Cluster.

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Author notes

  1. Grant A. Stanley

    Present address: School of Medical and Applied Sciences, University of Central Queensland, Rockhampton, Queensland, 4702, Australia

Authors and Affiliations

  1. The Australian Wine Research Institute, PO Box 197, Glen Osmond, SA, 5064, Australia

    Tina M. T. Tran, Paul J. Chambers & Simon A. Schmidt

  2. School of Molecular Sciences, Victoria University of Technology, Werribee Campus (W008), P.O. Box 14428, Melbourne, Victoria, 8001, Australia

    Tina M. T. Tran & Grant A. Stanley

Authors
  1. Tina M. T. Tran
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  2. Grant A. Stanley
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  3. Paul J. Chambers
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  4. Simon A. Schmidt
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Correspondence to Simon A. Schmidt.

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Tran, T.M.T., Stanley, G.A., Chambers, P.J. et al. A rapid, high-throughput method for quantitative determination of ethanol tolerance in Saccharomyces cerevisiae . Ann Microbiol 63, 677–682 (2013). https://doi.org/10.1007/s13213-012-0518-4

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  • DOI: https://doi.org/10.1007/s13213-012-0518-4

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