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Differentiation and species identification of yeasts using PCR

¹The Australian Wine Research Institute, PO Box 197, Glen Osmond, SA 5064, Australia
²Department of Plant Science, Waite Agricultural Research Institute, The University of Adelaide, SA 5064, Australia
⁴Department of Horticulture, Viticulture and Oenology, Waite Agricultural Research Institute, The University of Adelaide, SA 5064, Australia
³Cooperative Research Centre for Viticulture, Plant Research Centre, Hartley Grove, Urrbrae, SA 5064, Australia
⁵Petaluma Pty Ltd, Spring Gully Road, Piccadilly, SA 5151, Australia

Author for correspondence: Miguel de Barros Lopes. Tel: +618 8303 7331. Fax: +61 8 8303 7102. e-mail: mlopes{at}waite.adelaide.edu.au

ABSTRACT

A PCR-based method has been developed that permits both intraspecies differentiation and species identification of yeast isolates. Oligonucleotide primers that are complementary to intron splice sites were used to produce PCR fingerprints that display polymorphisms between different species of indigenous wine yeasts. Although polymorphisms existed between isolates of the same species, the banding patterns shared several amplification products that allowed species identification. Importantly, the method was able to distinguish between species of the closely related Saccharomyces sensu stricto yeasts. In two cases where isolates could not be positively identified there was discrepancy between the phenetic and phylogenetic species concept. The method has applications in yeast ecological studies, enabling the rapid grouping of isolates with related genomes and the investigation of population dynamics of strains of the same species.

Key Words: PCR • identification • yeasts

The GenBank accession numbers for the 26S rRNA sequences determined in this study are AF020437 (CBS 187^T), AF020438 (CBS 107^T), AF020435 (AWRI 1271) and AF020436 (AWRI 1272).

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