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

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.