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Applied and Environmental Microbiology, October 2002, p. 4884-4893, Vol. 68, No. 10
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.10.4884-4893.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Yeast Diversity and Persistence in Botrytis-Affected Wine Fermentations

David A. Mills,* Eric A. Johannsen, and Luca Cocolin{dagger}

Department of Viticulture and Enology, University of California, Davis, California 95616-8749

Received 11 February 2002/ Accepted 25 June 2002

Culture-dependent and -independent methods were used to examine the yeast diversity present in botrytis-affected ("botrytized") wine fermentations carried out at high (~30°C) and ambient (~20°C) temperatures. Fermentations at both temperatures possessed similar populations of Saccharomyces, Hanseniaspora, Pichia, Metschnikowia, Kluyveromyces, and Candida species. However, higher populations of non-Saccharomyces yeasts persisted in ambient-temperature fermentations, with Candida and, to a lesser extent, Kluyveromyces species remaining long after the fermentation was dominated by Saccharomyces. In general, denaturing gradient gel electrophoresis profiles of yeast ribosomal DNA or rRNA amplified from the fermentation samples correlated well with the plating data. The direct molecular methods also revealed a Hanseniaspora osmophila population not identified in the plating analysis. rRNA analysis also indicated a large population (>106 cells per ml) of a nonculturable Candida strain in the high-temperature fermentation. Monoculture analysis of the Candida isolate indicated an extreme fructophilic phenotype and correlated with an increased glucose/fructose ratio in fermentations containing higher populations of Candida. Analysis of wine fermentation microbial ecology by using both culture-dependent and -independent methods reveals the complexity of yeast interactions enriched during spontaneous fermentations.

* Corresponding author. Mailing address: Department of Viticulture and Enology, University of California, Davis, One Shields Ave., Davis, CA 95616-8749. Phone: (530) 754-7821. Fax: (530) 752-0382. E-mail: damills{at}ucdavis.edu.

{dagger} Present address: Department of Food Science, University of Udine, 33100 Udine, Italy.

Applied and Environmental Microbiology, October 2002, p. 4884-4893, Vol. 68, No. 10
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.10.4884-4893.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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