Diversity of Candida zemplinina isolates inferred from RAPD, micro/minisatellite and physiological analysis

Among non-Saccharomyces wine yeasts, Candida zemlpinina is one of the frequently isolated and oenologically important species. It is mostly known from European winemaking areas and it has become one of the key species of non-Saccharomyces wine yeasts to study. Investigating the diversity of C. zemplinina isolates is important for a deeper understanding of the non-Saccharomyces wine yeasts and for the yeast starter industry, as numerous researches have pointed to the potential use of this species in winemaking. For assessing the biodiversity of a larger number of strains, RAPD and micro/minisatellite PCR is often the method of choice, however, this technique is often unstandardized. Whereas some laboratories use these methods for species identifications, others apply RAPD primers for determining intraspecies diversity. In this study, we have tested 5 different RAPD and micro/minisatellite primers on strains of C. zemplinina isolated from different locations. We show that after a rigorous PCR-optimization aimed at reproducibility and comparability of band patterns with these PCR-reactions, diversity of different strains from a wide range of geographic locations is relatively low. The analysis of several oenologically important physiological traits of the strains showed a relatively low level of diversity as well. We also demonstrate that the intraspecific diversity of C. zemplinina observable with different techniques (RAPD, micro/minisatellite or physiological analysis) may be fairly different and not necessarily comparable.