Use of RAPD Analysis for Differentiation among Six Enological Saccharomyces spp. Strains

Angela Xufre, Fernanda Simões, Francisco Gírio*, Alda Clemente and M. Teresa Amaral-Collaço

INETI, Departamento de Biotecnologia, Estrada do Paço do Lumiar, 22, 1649-038 Lisboa, Portugal

Article history:
Received November 10, 1999
Accepted January 18, 2000

Key words:
DNA fingerprinting, RAPD, ARDRA, wine yeast identification, Saccharomyces cerevisiae 

Summary:
Two molecular typing techniques, amplified ribosomal DNA restriction analysis and random amplified polymorphic DNA assay, were evaluated for discrimination among six strains previously identified as synonymous of Saccharomyces cerevisiae by classical physiological characteristics and enological criteria. The first method that used restriction patterns originated by the amplification of internal transcribed spacers (ITS1 and ITS2) and the ribosomal gene 5.8S did not allow the differentiation among the six strains. On the contrary, reproducible amplicon fingerprints for the Saccharomyces spp. strains were obtained by random amplified polymorphic DNA assay using twenty one different primers. Among them, only four primers allowed discrimination among the six Saccharomyces spp. tested. The percentage of similarity between all species tested ranged from 40 to 80 %. The potential of the random amplified polymorphic DNA assay analysing the diversity of wine yeast species and developing polymerase chain reaction primers for wine yeast characterisation is discussed.  

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