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. 2003 Jan;69(1):649-53.
doi: 10.1128/AEM.69.1.649-653.2003.

The spoilage yeast Zygosaccharomyces bailii forms mitotic spores: a screening method for haploidization

Fernando Rodrigues  1 , Paula Ludovico, Maria João Sousa, H Yde Steensma, Manuela Côrte-Real, Cecília Leão
Affiliations

The spoilage yeast Zygosaccharomyces bailii forms mitotic spores: a screening method for haploidization

Fernando Rodrigues et al. Appl Environ Microbiol. 2003 Jan.

Abstract

Zygosaccharomyces bailii ISA 1307 and the type strain of this spoilage yeast show a diploid DNA content. Together with a rather peculiar life cycle in which mitotic but no meiotic spores appear to be formed, the diploid DNA content explains the observed difficulties in obtaining auxotrophic mutants. Mitotic chromosome loss induced by benomyl and selection on canavanine media resulted in three haploid strains of Z. bailii. This new set of Z. bailii strains allows the easy isolation of recessive mutants and is suitable for further molecular genetic studies.

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Figures

FIG. 1.
FIG. 1.
Survival curves for Z. bailii ISA 1307 (filled circles), Z. bailii H15 (one of the isolated haploid mutants [open circles]), and S. cerevisiae W303 1A (filled squares) after irradiation with UV light.
FIG. 2.
FIG. 2.
Disruption of ZbACS2 (accession number AJ314837) by APT1. (A) Schematic representation of the APT1 expression cassette for disruption experiments. (B) Southern blot of the Z. bailii ISA 1307 wild-type strain (lanes 1 and 3) and mutant 14, which is resistant to Geneticin (Z. bailii ISA 1307-14) (lanes 2 and 4). The genomic DNA isolated was digested with the restriction enzyme HindIII (lanes 1 and 2) or PstI-XhoI (lanes 3 and 4).
FIG. 3.
FIG. 3.
Microscopy of stained Z. bailii cells. (A) Bright field; (B) blue fluorescence; (C) overlap of bright field and blue fluorescence. (Section I) Conjugated cells stained with calcofluor white; (section II) conjugated but not sporulated cells stained with DAPI; (section III) sporulated cells stained with DAPI. The fluorescence of calcofluor white and DAPI is blue.
FIG. 4.
FIG. 4.
Green fluorescence histograms of S. cerevisiae W303 1A (A), Z. bailii ISA 1307 (B), Z. bailii H15 (C), and a mixture of Z. bailii ISA 1307 and Z. bailii H15 (D) after SYBR Green I staining.
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