Skip to main content
SpringerLink
Log in

Detection of non-Saccharomyces yeast strains in alcoholic fermentations by direct PCR and/or plating methods

  • Original Paper
  • Published:
European Food Research and Technology Aims and scope Submit manuscript

Abstract

Since the population of indigenous non-Saccharomyces yeasts is displaced from an alcoholic fermentation within a few hours, the use of a selective enrichment broth would be a valid approach for their detection. To accomplish this, yeast carbon base (YCB)–lysine broth without or with different concentrations of cycloheximide was tested. Lysine is an amino acid that cannot serve as the only nitrogen source for Saccharomyces spp., which is in contrast to non-Saccharomyces yeast, while cycloheximide is an antibiotic that is more or less effective against Saccharomyces spp. strains. Metagenomics, the study of genetic material from mixed microbial community as a grape must fermentation has the potential for operating in a culture-independent environment. Direct identification by PCR (dPCR) without previous isolation and a classical approach to analyse the yeast population in a wine inoculated with a mixed population of Saccharomyces cerevisiae and non-Saccharomyces yeasts were compared. Results showed that pre-incubation on YCB–lysine and dPCR combined with classic approaches based on the use of solid media is a suitable strategy for detecting yeast species involved in vinifications.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  1. Karasu-Yalcin S, Senses-Ergul S, Ozbas ZY (2013) Molecular identification of the yeast strains associated with spontaneous wine fermentation of Kalecik Karasi and Emir grapes. J Pure Appl Microbiol 7(4):2653–2672

    CAS  Google Scholar 

  2. Li EH, Liu AG, Xue B, Liu YL (2011) Yeast species associated with spontaneous wine fermentation of Cabernet Sauvignon from Ningxia, China. World J Microbiol Biotechnol 27(10):2475–2482

    Article  CAS  Google Scholar 

  3. Baffi MA, Bezerra CD, Arevalo-Villena M, Briones-Perez AI, Gomes E, Da Silva R (2011) Isolation and molecular identification of wine yeasts from a Brazilian vineyard. Ann Microbiol 61(1):75–78

    Article  CAS  Google Scholar 

  4. Zott K, Miot-Sertier C, Claisse O, Lonvaud-Funel A, Masneuf-Pomarede I (2008) Dynamics and diversity of non-Saccharomyces yeasts during the early stages in winemaking. Int J Food Microbiol 125(2):197–203

    Article  CAS  Google Scholar 

  5. Sannino C, Francesca N, Corona O, Settanni L, Cruciata M, Moschetti G (2013) Effect of the natural winemaking process applied at industrial level on the microbiological and chemical characteristics of wine. J Biosci Bioeng 116(3):347–356

    Article  CAS  Google Scholar 

  6. Beltran G, Torija MJ, Novo M, Ferrer N, Poblet M, Guillamon JM, Rozes N, Mas A (2002) Analysis of yeast populations during alcoholic fermentation: a six year follow-up study. Syst Appl Microbiol 25(2):287–293

    Article  CAS  Google Scholar 

  7. Ocon E, Gutierrez AR, Garijo P, Tenorio C, Lopez I, Lopez R, Santamaria P (2010) Quantitative and qualitative analysis of non-Saccharomyces yeasts in spontaneous alcoholic fermentations. Eur Food Res Technol 230(6):885–891

    Article  CAS  Google Scholar 

  8. Capece A, Salzano G, Romano P (2003) Molecular typing techniques as a tool to differentiate non-Saccharomyces wine species. Int J Food Microbiol 84(1):33–39

    Article  CAS  Google Scholar 

  9. Sun Y, Guo JJ, Liu FB, Liu YL (2014) Identification of indigenous yeast flora isolated from the five wine grape varieties harvested in Xiangning, China. Antonie Van Leeuwenhoek Int J Gen Molecular Microbiol 105(3):533–540

    Article  CAS  Google Scholar 

  10. Echeverrigaray S, Randon M, da Silva K, Zacaria J, Delamare APL (2013) Identification and characterization of non-Saccharomyces spoilage yeasts isolated from Brazilian wines. World J Microbiol Biotechnol 29(6):1019–1027

    Article  CAS  Google Scholar 

  11. Cocolin L, Campolongo S, Alessandria V, Dolci P, Rantsiou K (2011) Culture independent analyses and wine fermentation: an overview of achievements 10 years after first application. Ann Microbiol 61(1):17–23

    Article  Google Scholar 

  12. Fernandez M, Ubeda JF, Briones AI (2000) Typing of non-Saccharomyces yeasts with enzymatic activities of interest in wine-making. Int J Food Microbiol 59(1–2):29–36

    Article  CAS  Google Scholar 

  13. David V, Terrat S, Herzine K, Claisse O, Rousseaux S, Tourdot-Marechal R, Masneuf-Pomarede I, Ranjard L, Alexandre H (2014) High-throughput sequencing of amplicons for monitoring yeast biodiversity in must and during alcoholic fermentation. J Ind Microbiol Biotechnol 41(5):811–821

    Article  CAS  Google Scholar 

  14. Pfliegler WP, Horvath E, Kallai Z, Sipiczki M (2014) Diversity of Candida zemplinina isolates inferred from RAPD, micro/minisatellite and physiological analysis. Microbiol Res 169(5–6):402–410

    Article  CAS  Google Scholar 

  15. Medina K, Boido E, Farina L, Gioia O, Gomez ME, Barquet M, Gaggero C, Dellacassa E, Carrau F (2013) Increased flavour diversity of Chardonnay wines by spontaneous fermentation and co-fermentation with Hanseniaspora vineae. Food Chem 141(3):2513–2521

    Article  CAS  Google Scholar 

  16. Di Maio S, Polizzotto G, Planeta D, Oliva D (2011) A method to discriminate between the Candida stellata and Saccharomyces cerevisiae in mixed fermentation on WLD and Lysine agar media. S Afr J Enol Vitic 32(1):35–41

    Google Scholar 

  17. White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds) PCR Protocols, a guide to methods and applications. Academic Press, San Diego, pp 315–322

    Google Scholar 

Download references

Acknowledgments

This research was supported by the Regional Project Program (JCCM-PEII-2014-007-A).

Author information

Authors and Affiliations

  1. IRICA, Instituto Regional de Investigación Científica Aplicada, Universidad de Castilla La Mancha, Avda. Camilo J. Cela s/n, 13071, Ciudad Real, Spain

    Juan Ubeda-Iranzo, Patricia Díaz-Hellín, María Chacón-Ocaña & Ana Briones

Authors
  1. Juan Ubeda-Iranzo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Patricia Díaz-Hellín
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. María Chacón-Ocaña
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ana Briones
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Juan Ubeda-Iranzo.

Ethics declarations

Conflict of interest

None.

Compliance of Ethics Requirements

This article does not contain any studies with human or animal subjects.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ubeda-Iranzo, J., Díaz-Hellín, P., Chacón-Ocaña, M. et al. Detection of non-Saccharomyces yeast strains in alcoholic fermentations by direct PCR and/or plating methods. Eur Food Res Technol 241, 843–850 (2015). https://doi.org/10.1007/s00217-015-2509-8

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00217-015-2509-8

Keywords

Search

Navigation