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Direct mating between diploid sake strains of Saccharomyces cerevisiae

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Abstract

Various auxotrophic mutants of diploid heterothallic Japanese sake strains of Saccharomyces cerevisiae were utilized for selecting mating-competent diploid isolates. The auxotrophic mutants were exposed to ultraviolet (UV) irradiation and crossed with laboratory haploid tester strains carrying complementary auxotrophic markers. Zygotes were then selected on minimal medium. Sake strains exhibiting a MATa or MATα mating type were easily obtained at high frequency without prior sporulation, suggesting that the UV irradiation induced homozygosity at the MAT locus. Flow cytometric analysis of a hybrid showed a twofold higher DNA content than the sake diploid parent, consistent with tetraploidy. By crossing strains of opposite mating type in all possible combinations, a number of hybrids were constructed. Hybrids formed in crosses between traditional sake strains and between a natural nonhaploid isolate and traditional sake strains displayed equivalent fermentation ability without any apparent defects and produced comparable or improved sake. Isolation of mating-competent auxotrophic mutants directly from industrial yeast strains allows crossbreeding to construct polyploids suitable for industrial use without dependence on sporulation.

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Acknowledgements

We would like to thank Dr. Harumi Suzuki in Yamaguchi University Medical School, Japan, for his help with flow cytometric analysis, and Mr. Takahiro Nagayama in Nagayama Honke Shuzo, Japan, for his help in making sake. This work was supported in part by the RSP program of Japan Science and Technology Corporation.

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Correspondence to Rinji Akada.

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Hashimoto, S., Aritomi, K., Minohara, T. et al. Direct mating between diploid sake strains of Saccharomyces cerevisiae . Appl Microbiol Biotechnol 69, 689–696 (2006). https://doi.org/10.1007/s00253-005-0039-1

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Keywords

  • Industrial Strain
  • Isoamyl Acetate
  • Auxotrophic Mutation
  • Yeast Peptone Dextrose
  • Opposite Mating Type