Inter- and Intra-Specific Differentiation of Natural Wine Strains of Hanseniaspora (Kloeckera) by Physiological and Molecular Methods
Gellért Bujdosó1*, Christoph M. Egli2 and Thomas Henick-Kling2
1Department of Microbiology and Biotechnology, Faculty of Food Science, Szent István University, Somlói út 14–16, Budapest 1118, Hungary
2Department of Food Science and Technology, New York State Agricultural Experiment Station, Cornell University, Geneva 14456–0462, NY, USA
Article history:
Received June 13, 2000
Accepted October 26, 2000
Key words:
wine yeast, Hanseniaspora (Kloeckera), RAPD-PCR, RFLP (restriction fragment length polymorphism), yeast identification
Summary:
Six different species and five different strains within one species of the Hanseniaspora (anamorph Kloeckera) were obtained from CBS Culture Collection, Delft, Netherlands, to analyze and compare with unidentified Hanseniaspora strains isolated from juice and fermenting wine. Identification and differentiation were done using physiological and molecular methods. When defining the species of the genera Hanseniaspora (Kloeckera) by phenotypic characteristics, misidentification occurred for growth at 37 °C, for the assimilation of sucrose and 2-keto D-gluconate. For specific and reliable genus, species, and strain identification we evaluated both amplification of ITS1–5.8S-ITS4 rDNA, cut with various restriction enzymes, and the application of random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) using microsatellite and oligonucleotide (10-mers) primers. All the different primers (microsatellite, RAPD) worked properly and identically at both species and strain discrimination. The procedures were repeated several times and the techniques were found to be accurate and dependable.
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