Genomic stability of Saccharomyces cerevisiae baker's yeasts.
The objective of this study has been to gather data on genomic stability of baker's yeast strains during long-term mitotic growth under restrictive conditions so that comparisons could be made to other studies indicating genomic instability during meiosis. The work describes the analysis of mitotic stability of the nuclear and mitochondrial genomes in the baker's yeast strain V1 during incubation in continuous culture for 190 generations (300 days). The cells were cultured in complete medium containing 2% glucose and 8 to 12% ethanol, as a mutagenic agent specific for mtDNA. The high concentration of ethanol severely limited the growth rate of the cells. DNA samples were monitored for chromosomal pattern, polymorphisms in selected nuclear genes (SUC2, MALIT, ADH1) and mobile genetic elements (Ty1 and Y'), and for RFLPs in mtDNA. The results show that both the nuclear and mitochondrial genomes of grande cells were very stable. However, the frequency of petite mutants in the population varied dramatically during the course of the experiment, reaching as high as 87% petite during the first 27 days of the experiment and declining to 5.8% petite at the end. This decline can be attributed to selection against petite mutants in media containing high concentrations of ethanol. Moreover, when samples and the parental strain were compared at the end of the experiment, no change could be observed in parameters such as their growth rate in different media, capacity to leave doughs, viability in ethanol or frequency of petite mutants. Results therefore indicated that the majority of the cells in the population were very similar to the parental throughout the experiments, with no apparent molecular or phenotypical changes.
Gasent-Ramírez
J M
JM
Departamento de Genética, Facultad de Biología, Universidad de Sevilla, Spain.
Castrejón
F
F
Querol
A
A
Ramón
D
D
Benítez
T
T
D015139:Blotting, Southern; D002467:Cell Nucleus / Q000235:genetics; D003470:Culture Media / Q000737:chemistry; D004271:DNA, Fungal / Q000187:drug effects / Q000235:genetics; D004272:DNA, Mitochondrial / Q000032:analysis / Q000187:drug effects / Q000235:genetics; D000431:Ethanol / Q000633:toxicity; D016681:Genome, Fungal; D005947:Glucose; D006026:Glycoside Hydrolases / Q000032:analysis; D008938:Mitosis / Q000235:genetics*; D009153:Mutagens / Q000633:toxicity; D009154:Mutation; D010641:Phenotype; D012150:Polymorphism, Restriction Fragment Length; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development; D043324:beta-Fructofuranosidase
D003470:Culture Media; D004271:DNA, Fungal; D004272:DNA, Mitochondrial; D009153:Mutagens; D000431:Ethanol; D006026:Glycoside Hydrolases; D043324:beta-Fructofuranosidase; D005947:Glucose