Selection and identification of autochthonous yeasts in Slovakian wine samples using a rapid and reliable three-step approach


Domenico Pangallo, Institute of Molecular Biology, Slovak Academy of Sciences, Dúbravská cesta 21, 845 51 Bratislava, Slovakia. E-mail:


Aims:  The investigation of yeast microflora during the must fermentation of two wine varieties (Frankovka modra – Blaufränkisch and Veltlinske zelene – Grüner Veltliner) from two consecutive vintages was performed using a three-step approach.

Methods and Results:  The investigation strategy consisted of the combination of yeast cultivation, selection of the isolated yeasts based on the amplification of internal transcribed spacer 2 using a fluorescence-labelled primer (f-ITS-PCR) and a final identification step based on amplification and sequencing of the ITS1-5.8S rDNA-ITS2 region of the selected yeasts. By this three-step approach, it was possible to screen 433 yeasts isolates that belonged to 13 different species.

Conclusions:  The f-ITS-PCR allowed the unambiguous differentiation of all isolated yeast species that produced their typical f-ITS-PCR profile.

Significance and Impact of the Study:  This is one of few reports that treat the yeast diversity in Slovakian wines and in two varieties largely cultivated in Central Europe. The three-step approach permitted the rapid and reliable identification of isolated yeasts. The f-ITS-PCR with its good discrimination power can represent a suitable molecular tool for the selection of yeast members recovered from food or other environments.