Antonie van Leeuwenhoek

, Volume 83, Issue 2, pp 149–154 | Cite as

Yeasts present during spontaneous fermentation of Lake Erie Chardonnay, Pinot Gris and Riesling

  • Harry van Keulen
  • Donald G. Lindmark
  • Kathleen E. Zeman
  • Wes Gerlosky


The composition of wine yeast populations, present during spontaneous fermentation of Chardonnay, Pinot Gris and Riesling from the Lake Erie Region was studied. A combination of biochemical and molecular techniques was used to identify non-Saccharomyces and Saccharomyces yeast isolates. The biochemical techniques included analysis of yeast isolates by sugar fermentation and carbon and nitrogen assimilation. Molecular techniques involved ribotyping of a highly variable segment in the 26S rRNA gene using DNA sequence analysis and restriction fragment length polymorphism of amplified DNA. The results show that of the non-Saccharomyces yeasts, several related species of Hanseniaspora, were the most abundant yeasts present during early stages of fermentation. Later in fermentation S. cerevisiae dominated, and based on biochemical analyses consisted of a heterogeneous group of genotypes. There were no major differences in yeast populations among the three types of juice analyzed.

Indigenous yeast rDNA RFLP Ribotyping Spontaneous fermentation 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Araujo S., Ferrer A., Sulbáran de Ferrer B., Nava C., Ojeda de Rodríguez G. and Nava R.A. 1998. Yeasts isolated from fermenting juice extracted from white-wine grape varieties in Zulia state, Venezuela. Rev. Fac. Agron. 15: 249–255.Google Scholar
  2. Boulton R.B., Singleton V.L., Bisson L.F. and Kunkee R.E. 1996. Principles and practices of winemaking. Aspen Publishers, Gaithersburg, MD.Google Scholar
  3. Ciani M. and Maccarelli F. 1998. Oenological properties of non-Saccharomyces yeasts associated with wine-making. World J. Microbiol. Biotechnol. 14: 199–203.CrossRefGoogle Scholar
  4. Ciani M. and Picciotti G. 1995. The growth kinetics and fermentation behavior of some non-Saccharomyces yeasts associated with wine making. Biotech. Lett. 17: 1247–1250.CrossRefGoogle Scholar
  5. Degré R., Thomas D.Y., Ash J., Mailhiot K., Morin A. and Dubord C. 1998. Wine yeasts strain identification. Am. J. Enol.Vitic. 40: 309–319.Google Scholar
  6. Fernández M.T., Ubeda J.F. and Briones A.I. 1999. Comparative study of non-Saccharomyces microflora of musts in fermentation, by physiological and molecular methods. FEMS Microbiol. Lett. 173: 223–229.CrossRefGoogle Scholar
  7. Fleet G.H. and Heard G.M. 1993. Yeast growth during fermentation. In: Fleet G.H. (ed.), Wine Microbiology and Biotechnology. Harwood Academic Press, Switzerland, pp. 27–75.Google Scholar
  8. Fugelsang K.C. 1997. Wine Microbiology. Chapman & Hall, New York.Google Scholar
  9. Guillamón J.M., Sabate J., Barrio E., Cano J. and Querol A. 1998. Rapid identification of wine yeast species based on RFLP analysis of the ribosomal internal transcribed spacer (ITS) region. Arch. Microbiol. 169: 387–392.PubMedCrossRefGoogle Scholar
  10. Heard G.M. and Fleet G.H. 1985. Growth of natural yeast flora during the fermentation of inoculated wines. Appl. Environ. Microbiol. 50: 727–728.PubMedGoogle Scholar
  11. Kurtzman C.P. and Robnett C.J. 1998. Identification and phylogeny of ascomycetous yeasts from analysis of nuclear large subunit (26S) ribosomal DNA partial sequences. Antonie van Leeuwenhoek 73: 331–371.PubMedCrossRefGoogle Scholar
  12. Longo E., Cansado D., Agrelo D. and Villa T.G. 1991. Effect of climatic condition on yeast diversity in grape musts from Northwest Spain. Am. J. Enol. Vitic. 42: 141–144.Google Scholar
  13. Martini A., Ciani M. and Scorzetti G. 1996. Direct enumeration and isolation of wine yeasts from grape surfaces. Am. J. Enol. Vitic. 47: 435–440.Google Scholar
  14. Michot B. and Bachellerie J.-P. 1987. Comparison of large subunit rRNAs reveal some eukaryote-specific elements of secondary structure. Biochimie 69: 11–23.PubMedCrossRefGoogle Scholar
  15. Mortimer R.K., Romano P., Suzzi G. and Polsinelli M. 1994. Genome renewal: a new phenomenon revealed from a genetic study of 43 strains of Saccharomyces cerevisiae derived from natural fermentation of grape musts. Yeast 10: 1543–1552.PubMedCrossRefGoogle Scholar
  16. Pardo I., Garcia M.J., Zungina M. and Uruburu F. 1989. Dynamics of microbial populations during fermentation of wines from the Utiel-Requena region of Spain. Appl. Environ. Microbiol. 55: 539–541.PubMedGoogle Scholar
  17. Pramateftaki P.V., Landaris P. and Typas M.A. 2000. Molecular identification of wine yeasts at species or strain level: a case study with strains from two wine growing areas of Greece. J. Appl. Microbiol. 89: 236–248.PubMedCrossRefGoogle Scholar
  18. Rankine B.C. 1967. Formation of higher alcohols by wine yeasts and relationship to taste threshold. J. Sci. Food Agric. 18: 583–589.Google Scholar
  19. Ribéreau-Gayon P. 1985. New developments in wine microbiology. Am. J. Enol. Vitic. 36: 1–10.Google Scholar
  20. Sambrook J., Fritsch E.F. and Maniatis T. 1989. Molecular Cloning. A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.Google Scholar
  21. Schütz M. and Gafner J. 1993. Analysis of yeast diversity during spontaneous and induced alcoholic fermentations. J. Appl. Bacteriol. 75: 551–557.Google Scholar
  22. Sifritt S.K. 1976. The Ohio wine and wine grape industries, PhD, Kent State University.Google Scholar
  23. Thompson J.D., Higgins D.G. and Gibson T.J. 1994. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, positions-specific gap penalties and weight matrix choice. Nucl. Acids Res. 22: 4673–4680.PubMedGoogle Scholar
  24. Torija M.J., Rozès N., Poblet M., Guillamón J.M. and Mas A. 2001. Yeast population dynamics in spontaneous fermentation: Comparison between two different wine-producing areas over a period of three years. Antonie van Leeuwenhoek 79: 345–352.PubMedCrossRefGoogle Scholar
  25. Vaughan-Martini A. and Martini A. 1995. Facts, myths and legends on the prime industrial microorganism. J. Ind. Microbiol. 14: 514–522.PubMedCrossRefGoogle Scholar
  26. Weiss J.B., van Keulen H. and Nash T.E. 1992. Classification of subgroups of Giardia lamblia based upon ribosomal RNA gene sequence using the polymerase chain reaction. Mol. Biochem. Parasitol. 54: 73–86.PubMedCrossRefGoogle Scholar

Copyright information

© Kluwer Academic Publishers 2003

Authors and Affiliations

  • Harry van Keulen
    • 1
  • Donald G. Lindmark
    • 1
  • Kathleen E. Zeman
    • 1
  • Wes Gerlosky
    • 2
  1. 1.Department of Biological, Geological and Environmental SciencesCleveland State UniversityClevelandUSA
  2. 2.Harpersfield VineyardGenevaUSA

Personalised recommendations