engleski

Use of Different Molecular Methods in the Identification of Brettanomyces Spp. from Croatian Grapes and Wines

A wide variety of yeast species have been implicated in wine spoilage. Of the spoilage yeasts, species of Brettanomyces (imperfect state Dekkera) are probably the most serious and controversial. They are most frequently found in red wine, 6 to 10 months after barrelling but this yeast have been isolated from many other fermented beverages, such as beer and cider. These potential spoilage yeasts have been identified in almost every wine-producing area of the world. In this investigation we reported the first systematic approach of study Brettanomyces spp. on grapes and in wines from Croatia. We applied classical and different molecular methods (DGGE, RE-PCR and PCR-HMA) for this research. Also, we have investigated three different total DNA extraction methods from wine and identification of viable cells B.bruxellensis in wine. The best evaluated method was heteroduplex mobility assay and these are the first report of using this method in Brettanomyces spp. identification. The use of PCR-DGGE needs a sequencing step of the PCR product. The Brett primer set used for PCR screenings gave us untrustworthy results. Bands expected size for Brett were also obtained for a commercial yeast strain S.cerevisiae. Using HMA we were able to identify four positive Brett wines of 80 analysed. Only in two case the identify population was viable. Also, in one case (of 20 analyzed) was able to identify B.bruxellensis in barriques barrels. The identification, monitoring of that population and the origin are very important. We need to investigate the ecology of Brettanomyces spp. to monitor the spread of this yeast.

Brettanomyces spp.; wine

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