To evaluate the influence of the genomic properties of yeasts on the formation of wine flavour, genotypic diversity among natural Saccharomyces cerevisiae strains originating from grapes collected in four localities of three Austrian vine-growing areas (Thermenregion: locations Perchtoldsdorf and Pfaffstätten, Neusiedlersee-Hügelland: location Eisenstadt, Neusiedlersee: location Halbturn) was investigated and the aroma compounds produced during fermentation of the grape must of 'Grüner Veltliner' were identified. Amplified fragment length polymorphism analysis (AFLP) showed that the yeast strains cluster in four groups corresponding to their geographical origin. The genotypic analysis and sequencing of the D1/D2 domain of 26S rRNA encoding gene and ITS1/ITS2 regions indicated that the Perchtoldsdorf strains were putative interspecies hybrids between S. cerevisiae and Saccharomyces kudriavzevii. Analysis of the aroma compounds by GS/MS indicated a region-specific influence of the yeasts on the chemical composition of the wines. The aroma compound profiles generated by the Perchtoldsdorf strains were more related to those produced by the Pfaffstätten strains than by the Eisenstadt and Halbturn strains. Similar to the Pfaffstätten yeasts, the putative hybrid strains were good ester producers, suggesting that they may influence the wine quality favourably.

New double-and triple-hybrid Saccharomyces yeasts were characterized using PCRrestriction fragment length polymorphism of 35 nuclear genes, located on different chromosome arms, and the sequencing of one nuclear and one mitochondrial gene. Most of these new hybrids were originally isolated from fermentations; however, two of them correspond to clinical and dietary supplement isolates. This is the first time that the presence of double-hybrid S. cerevisiae  S. kudriavzevii in non-fermentative substrates has been reported and investigated. Phylogenetic analysis of the MET6 nuclear gene confirmed the double or triple parental origin of the new hybrids. Restriction analysis of gene regions in these hybrids revealed a high diversity of genome types. From these molecular characterizations, a reduction of the S. kudriavzevii fraction of the hybrid genomes is observed in most hybrids. Mitochondrial inheritance in hybrids was deduced from the analysis of mitochondrial COX2 gene sequences, which showed that most hybrids received the mitochondrial genome from the S. kudriavzevii parent. However, two strains inherited a S. cerevisiae COX2, being the first report of S. cerevisiae  S. kudriavzevii hybrids with S. cerevisiae mitochondrial genomes. These two strains are those showing a higher S. kudriavzevii nuclear genome reduction, especially in the wine hybrid AMH. This may be due to the release of selective pressures acting on the other hybrids to maintain kudriavzevii mitochondriainteracting genes.

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The yeasts of the Saccharomyces genus exhibit a low pre-zygotic barrier and readily form interspecies hybrids. Following the hybridization event, the parental genomes undergo gross chromosomal rearrangements and genome modifications that may markedly influence the metabolic activity of descendants. In the present study, two artificially constructed hybrid yeasts (Saccharomyces cerevisiae x Saccharomyces uvarum and S. cerevisiae x Saccharomyces kudriavzevii) were used in order to evaluate the influence of high-sugar wine fermentation on the evolution of their genotypic and phenotypic properties. It was demonstrated that the extent of genomic modifications differs among the hybrids and their progeny, but that stress should not always be a generator of large genomic disturbances. The major genome changes were observed after meiosis in the F1 segregants in the form of the loss of different non-S. cerevisiae chromosomes. Under fermentation condition, each spore clone from a tetrad developed a mixed population characterized by different genotypic and phenotypic properties. The S. cerevisiae x S. uvarum spore clones revealed large modifications at the sequence level of the S. cerevisiae sub-genome, and some of the clones lost a few additional S. cerevisiae and S. uvarum chromosomes. The S. cerevisiae x S. kudriavzevii segregants were subjected to consecutive loss of the S. kudriavzevii markers and chromosomes. Both the hybrid types showed increased ethanol and glycerol production as well as better sugar consumption than their parental strains. The hybrid segregants responded differently to stress and a correlation was found between the observed genotypes and fermentation performances.

The aim of the present study was to evaluate the autochthonous yeast population during spontaneous fermentations of grape musts in Austrian wine-producing areas. Investigation of genomic and genetic variations among wine yeasts was a first step towards a long-term goal of selecting strains with valuable enological properties typical for this geographical region. An approach, combining sequences of the D1/D2 domain of the 26S rRNA gene and random amplified polymorphic DNA fingerprinting, was used to characterize yeasts at the species level, whereas the differentiation of Saccharomyces strains was accomplished by amplified fragment length polymorphism fingerprinting. At the beginning of fermentation, representatives of nine genera were identified, with Hanseniaspora and Metschnikowia species characterized most frequently. Saccharomyces cerevisiae and Saccharomyces bayanus var. uvarum strains, which were identified throughout the entire fermentation process, showed a high level of genetic diversity. A number of S. cerevisiae strains were common at multiple wineries, but a wide range of strains with characteristic profiles were characterized at individual locations. This biodiversity survey represents a contribution to the investigation and preservation of genetic diversity of biotechnologically relevant yeasts in Austrian wine-making areas.