Indigenous yeast population from Georgian aged wines produced by traditional "Kakhetian" method.

The yeast microbiota present in wines produced by the ancient "Kakhetian" method in Georgia (EU) was studied. This technique involves the use of terracotta vessels (amphoras), during spontaneous fermentation, maceration phase and wine ageing. The analysed yeasts were collected from wines after maturation for one year in ten amphoras from a Georgian winery. The 260 isolates were all identified as Saccharomyces cerevisiae, and the majority were classified as flor yeasts by restriction analysis of ITS region. A first technological and molecular screening was used to select 70 strains for further characterization. Both genetic and metabolic characterization discriminated flor from non-flor strains. The combined results obtained by analysis of interdelta region and mtDNA-RFLP yielded 23 different biotypes; no biotype was common to flor and non-flor strains. The wines produced by flor yeasts showed a high content in acetaldehyde, acetic acid, acetoin, whereas the level of other compounds was similar to wines obtained by non-flor strains. This study represents the first report on the composition of yeast microbiota involved in the maturation of this traditional wine. These flor strains represent an interesting yeast population, in possession of peculiar characteristics allowing them to survive during wine ageing, becoming the dominant flora in the final wine.

Capece A ，Siesto G ，Poeta C ，Pietrafesa R ，Romano P ... -  《-》

French Jura flor yeasts: genotype and technological diversity.

Fifty-four Saccharomyces cerevisiae strains were isolated from Jura "Vin Jaune" velum and characterized by conventional physiological and molecular tests including ITS RFLP and sequence analysis, karyotyping and inter delta typing. ITS RFLP and sequence revealed a specific group of related strains different from the specific profile of Sherry flor yeast caused by a 24 bp deletion in the ITS1 region described by Esteve-Zarzoso et al. (Antonie Van Leeuwenhoek 85:151-158, 2004). Interdelta typing, the most discriminative method, revealed a high diversity of Jura flor yeast strains and gathered strains in clusters unequally shared between the northern and southern part of the Jura vineyard. The assessment of phenotypic diversity among the isolated strains was investigated for three wine metabolites (ethanal, acetic acid, and sotolon) from micro scale velum tests. Except at an early stage of ageing, the production of these metabolites was not correlated to the five genetic groups obtained by interdelta typing, but correlated to the cellar where strains had been isolated. The different strains isolated in a cellar produced mostly one type of velum (thin or thick, grey or white); but thin and grey velums, recognized as responsible for high quality wines, were obtained more frequently for one of the five groups of delta genotypes.

Charpentier C ，Colin A ，Alais A ，Legras JL ... -  《-》

Yeast population dynamics during the fermentation and biological aging of sherry wines.

Molecular and physiological analyses were used to study the evolution of the yeast population, from alcoholic fermentation to biological aging in the process of "fino" sherry wine making. The four races of "flor" Saccharomyces cerevisiae (beticus, cheresiensis, montuliensis, and rouxii) exhibited identical restriction patterns for the region spanning the internal transcribed spacers 1 and 2 (ITS-1 and ITS-2) and the 5.8S rRNA gene, but this pattern was different, from those exhibited by non-flor S. cerevisiae strains. This flor-specific pattern was detected only after wines were fortified, never during alcoholic fermentation, and all the strains isolated from the velum exhibited the typical flor yeast pattern. By restriction fragment length polymorphism of mitochondrial DNA and karyotyping, we showed that (i) the native strain is better adapted to fermentation conditions than commercial strains; (ii) two different populations of S. cerevisiae strains are involved in the process of elaboration, of fino sherry wine, one of which is responsible for must fermentation and the other, for wine aging; and (iii) one strain was dominant in the flor population integrating the velum from sherry wines produced in González Byass wineries, although other authors have described a succession of races of flor S. cerevisiae during wine aging. Analyzing all these results together, we conclude that yeast population dynamics during biological aging is a complex phenomenon and differences between yeast populations from different wineries can be observed.

Esteve-Zarzoso B ，Peris-Torán MJ ，García-Maiquez E ，Uruburu F ，Querol A ... -  《-》

Acetaldehyde and ethanol are responsible for mitochondrial DNA (mtDNA) restriction fragment length polymorphism (RFLP) in flor yeasts.

Flor yeasts grow and survive in fino sherry wine where the frequency of respiratory-deficient (petite) mutants is very low. Mitochondria from flor yeasts are highly acetaldehyde- and ethanol-tolerant, and resistant to oxidative stress. However, restriction fragment length polymorphism (RFLP) of mtDNA from flor yeast populations is very high and reflects variability induced by the high concentrations of acetaldehyde and ethanol of sherry wine on mtDNA. mtDNA RFLP increases as the concentration of these compounds also increases, but is followed by a total loss of mtDNA in petite cells. Yeasts with functional mitochondria (grande) are target of continuous variability, so that flor yeast mtDNA can evolve extremely rapidly and may serve as a reservoir of genetic diversity, whereas petite mutants are eventually eliminated because metabolism in sherry wine is oxidative.

Castrejón F ，Codón AC ，Cubero B ，Benítez T ... -  《SYSTEMATIC AND APPLIED MICROBIOLOGY》

Genetic characterization of strains of Saccharomycescerevisiae responsible for 'refermentation' in Botrytis-affected wines.

Saccharomyces cerevisiae is responsible for alcoholic fermentation of wines. However, some strains can also spoil sweet Botrytis-affected wines. Three 'refermentation' strains were isolated during maturation. Characterization of those strains in regards to their fingerprint, rDNA sequence and resistance to SO2, which constituted the main source of stress in Botrytis-affected wines, was carried out. Refermentation strains could be clearly discriminated by interdelta fingerprinting. However, they exhibited close relationships by karyotyping. A part of RDN1 locus sequence was examined by using PCR-RFLP and PCR-DGGE. The resistance of refermentation strains to SO2 was performed by using real time quantitative PCR focusing on SSU1 gene. Results suggested that refermentation strains were heterozygote in 26S rDNA and their ITS1-5.8S rDNA-ITS2 region sequence revealed relationships with 'flor' strains. As described in the literature for flor strain, two out of three refermentation strains constitutively developed a higher level of SSU1 expression than the reference strains, improving their putative tolerance to SO2. Therefore, refermentation strains of S. cerevisiae had developed many strategies to survive during maturing sweet wines. Singularities in rDNA sequence and SSU1 overexpression revealed a natural adaptation. Moreover, genomic relationship between flor and refermentation strains suggested that stress sources could induced selection of survivor strains.

Divol B ，Miot-Sertier C ，Lonvaud-Funel A 《JOURNAL OF APPLIED MICROBIOLOGY》