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Determination of the Mating Efficiency of Haploids in Saccharomyces cerevisiae
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An efficient method for isolating mating-competent cells from bottom-fermenting yeast using mating

Taku Ota1, Keiko Kanai2, Hisami Nishimura1

  • 1Kirin Company Ltd, Research Laboratories for Alcoholic Beverage Technologies, 1-17-1 Namamugi, Tsurumi-ku, Yokohama, 230-8628, Japan.

Yeast (Chichester, England)
|October 28, 2017
PubMed
Summary

A new method efficiently isolates mating-competent cells (MCCs) from bottom-fermenting yeast without genetic modification. This technique simplifies crossbreeding for industrial yeast strains, improving brewing processes.

Keywords:
S. pastorianusbottom-fermenting yeastcrossbreedingmatingmating pheromone

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Area of Science:

  • Microbiology
  • Yeast Genetics
  • Industrial Biotechnology

Background:

  • Crossbreeding novel yeast strains is crucial for developing improved industrial characteristics.
  • Obtaining mating-competent cells (MCCs) from Saccharomyces pastorianus (bottom-fermenting yeast) is challenging due to inefficient existing methods.
  • Genetic modification is often required to facilitate yeast crossbreeding.

Purpose of the Study:

  • To develop a productive and non-genetic modification method for isolating MCCs from bottom-fermenting yeast.
  • To enable efficient crossbreeding of industrial yeast strains for practical applications.

Main Methods:

  • Utilized mating pheromone-supersensitive mutants (Δbar1 and Δsst2) that exhibit growth defects in the presence of mating pheromones.
  • Developed a plate assay where MCCs secreting α-factor and a-factor create a growth-defect halo on lawns of MATa Δbar1 and MATα Δsst2, respectively.
  • Isolated meiotic segregants by selecting cells causing growth defects on pheromone-supersensitive cells.

Main Results:

  • Successfully identified MCCs from bottom-fermenting yeast strains using the developed plate assay.
  • Isolated 40 α/α-type and six a/a-type meiotic segregants from bottom-fermenting yeast strains.
  • Verified the mating types of isolated segregants through crossing with tester strains.

Conclusions:

  • The described method efficiently isolates MCCs without artificial genetic modification.
  • This technique is applicable to various industrial yeast strains, including wine, sake, and distiller's yeasts.
  • The method offers a convenient and efficient tool for crossbreeding industrial yeast strains, benefiting practical brewing.