Isolation of Meiotic Segregants from a Bottom Fermenting Yeast

Abstract

Viable meiotic segregants were isolated from a bottom fermenting brewing yeast transformed by IME1 using a high copy number plasmid. These transformants sporulated under sporulation conditions and viable meiotic segregants were obtained from the spore asci. Flow cytometry showed that the meiotic segregants had a lower DNA content per cell than the parental strains, indicating meiosis had proceeded normally. Chromosomal observation by pulsed-field gel electrophoresis and array comparative genomic hybridization (array-CGH) showed that meiotic segregants, like the parent strains, contained two types of chromosome: the S. cerevisiae type and the S. bayanus type. In certain meiotic segregants, some chromosomes were missing, and the chromosome copy number changed. Isolation of meiotic segregants resulted in a ploidy reduction, which can be applied in the breeding of yeast strains.