Abstract
ABSTRACTIn order to facilitate the mitochondrial DNA-RFLP analysis, a simplified procedure for isolation of mtDNA was developed. It reduces the time for analysis and could be applied for specific detection of genetic changes in the genome of industrially important S. cerevisiae strains. Furthermore, genetic stability control of brewing S. carlsbergensis strains was performed. Upon applying of PCR fingerprinting, chromosomal karyotyping, and mtDNA-RFLP techniques, only two very similar characteristic DNA profiles were observed in yeast strains isolated from different sources. These data confirmed that mtDNA-RFLP analysis is as reliable as other conventionally used molecular methods, giving same high discriminative results when genome rearrangements were analyzed. Contrary to the data, obtained from the analysis of brewing Saccharomyces strains, a difference between the PCR-profile and mtDNA restriction patterns was observed when distillery strains were studied. Although mtDNA-RFLP profiles of the ethanol producing strains were highly similar, slight divergence between them was observed, which was probably due to the damaging effect of ethanol and acetaldehyde on the mtDNA. Application of this technique, contrary to whole genome typing methods, could detect the presence even of small divergence in the tested strains and could be used as highly specific tool for genetic control of industrial fermentations.
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