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Identification of Saccharomyces paradoxus and Saccharomyces cerevisiae isolated from Zagreb wine region

Vineyard and wine production process represent an environment shaped by man trough centuries. The most important members of that enviroment are yeasts, primarily Saccharomyces sensu stricto (S. cerevisiae, S. bayanus, S. pastorianus, S. paradoxus). S. paradoxus is considered as a indigenous type possible to find only in natural conditions without antropological influence. The main goal of investigation was isolation of indigenous Saccharomyces sensu stricto strains and differentiation on the species level. Vineyard yeast isolation was carried out from berries and must during the fementation in laboritry conditions. Identification on the genus level was carried out by using Lysine medium and WL nutrient agar. Strain identification was done trough physiological tests (37 C growth and manitol assimilation) and amplification of the 18S rDNA+ITS1 regions followed by restriction enzymes digestion (HaeIII and MspI). Total of 20 strais were tested ; 15 of them were indigenous isolates of Saccharomyces sensu strico from vineyard Jazbina of Zagreb wine region ; one comercial S. bayanus strain, and four type strains for Saccharomyces sensu stricto group. Physiological mehods revealed that three indigenous isolates belong to the S. cerevisiae species, while others belong to the species S. paradoxus. Molecular identification method gave somewhat different results. rDNA-RFLP method determined that two isolates, previously identified as a S. paradoxus belong to the S. cervisiae species. Phisioogical test identification must be supported by genetic identificaton. Saccharomyces paradoxus isolation from vineyard, where an influence of man is considerably present, emerges numerous questions regarding this species ecology as well as interactions with other members of that group.

Saccharomyces paradoxus; Saccharomyces cerevisiae; rDNA-RFLP

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