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Electrophoretic Karyotypes of Wine Strains of Saccharomyces cerevisiae

Contributed by Alan T. Bakalinsky Department of Food Science and Technology, Oregon State University, Corvallis, OR 97331-6602


Abstract
Chromosomal profiles of wine strains of Saccharomyces cerevisiae were compared to those of a standard laboratory karyotype. Qualitative analysis supported previous genetic findings that two of the wine strains are aneuploid and two diploid. Obstacles to direct and meaningful quantitation of the electrophoretic profiles are discussed.


Methods
Yeast Strains
The homothallic wine yeasts UCD Enology 505 (California Champagne), UCD Enology 595 (Pasteur Champagne), UCD Enology 522 (Montrachet), and 522X14 have been described.1 2407-1a (a haploid isogenic to X2180-1A), 2407 (diploid), and 2794 (triploid) are wildtype heterothallic laboratory strains.

Pulsed-Field Gel Electrophoresis
The electrophoretic apparatus (CHEF-DR II cell, Bio-Rad Laboratories) has been described.6 Standard conditions were a 30- hour run (23 hours with a 60 second switching interval followed by 7 hours with 100 second intervals) at 200 volts, 1.8 liters of 0.5X TBE buffer, 65 ml of 1% agarose (Ultra Pure DNA Grade, Bio-Rad Laboratories) cast in a 5 x 5.5 mold, and a buffer temperature maintained at 12C by constant recycling through a cooling bath. Samples of cell-containing agarose beads, 35 l, were loaded into wells which were subsequently sealed by addition of a few drops of molten 1% Low Melt Preparative Grade Agarose (Bio- Rad Laboratories). Gels were stained in used buffer containing 0.5 g/ml ethidium bromide for 30 minutes and destained overnight.
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