Skip to main content
SpringerLink
Log in

Influence of organic viticulture on non-Saccharomyces wine yeast populations

  • Original paper
  • Published:
Annals of Microbiology Aims and scope Submit manuscript

Abstract

This study evaluated the population dynamics of non-Saccharomyces biota during spontaneous fermentation of organic musts. Non-Saccharomyces yeasts were found to be present at high levels during all fermentations. A total of 543 yeast colonies were isolated, 190 from Lysine-Medium (LM) agar, 254 from Wallerstein Laboratory Nutrient (WLN) agar and 99 from YPD agar. To estimate yeast population dynamics during spontaneous fermentation a genotypic approach was applied. PCR-RFLP of the ITS1-5.8S rDNA-ITS2 region and sequence determination of the D1/D2 region of the 26S rRNA gene enabled identification of the yeast isolates at the species level. Hanseniaspora uvarum, Metschnikowia fructicola and Candida zemplinina predominated, while Issatchenkia terricola, Issatchenkia orientalis and Pichia sp. were identified with a lower frequency. Hanseniaspora uvarum, M. fructicola and C. zemplinina represented 43%, 31% and 11% of the total non-Saccharomyces population isolated, respectively. Some yeast isolates were shown to be closely related to Hanseniaspora spp. and Candida spp. on the basis of the D1/D2 sequences. Based on those results, the coexistence of different Hanseniaspora and Pichia species in grape musts was supposed, and their complete identification was achieved using additional molecular markers. Moreover, strain typing and differentiation was carried out by RAPD-PCR. High strain polymorphisms were observed in the different species. For some strains, appreciable properties were demonstrated both in vitro by the API-ZYM test and in must. In must microvinification some strains showed good fermentation performances, low production of acetic acid and a partial capability to degrade malic acid.

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

  • Cavazza A, Grando MS, Zini C (1992) Rilevazione della flora microbica di mosti e vini. Vignevini 9:17–20

    Google Scholar 

  • Charoenchai C, Fleet GH, Henschke PA, Tood BEN (1997) Screening of non-Saccharomyces wine yeasts for the presence of extracellular hydrolytic enzymes. Austr J Grape Wine Res 3:2–8

    Article  CAS  Google Scholar 

  • Ciani M, Beco L, Comitini F (2006) Fermentation behaviour and metabolic interactions of multistarter wine yeast fermentations. Int J Food Microbiol 108:239–245

    Article  CAS  PubMed  Google Scholar 

  • Ciani M, Comitini F, Mannazzu I, Domizio P (2010) Controlled mixed culture fermentation: a new perspective on the use of non-Saccharomyces yeasts in wine making. FEMS Yeast Res 10:123–133

    Google Scholar 

  • Di Maro E, Ercolini D, Coppola S (2007) Yeasts dynamics during spontaneous wine fermentation of the Catalanesca grape. Int J Food Microbiol 117:201–210

    Article  PubMed  Google Scholar 

  • Esteve-Zarzoso B, Belloch C, Uruburu F, Querol A (1999) Identification of yeasts by RFLP analysis of the 5.8 S rRNA gene and the two ribosomal internal transcribed spacers. Int J Syst Bacteriol 49:329–337

    Article  CAS  PubMed  Google Scholar 

  • Fell JW, Boekhout T, Fonseca A, Scorzetti G, Statzell-Tallman A (2000) Biodiversity and systematic of basidiomycetous yeasts as a determined by large-subunit rDNA D1/D2 domain sequence analysis. Int J Syst Evol Microbiol 20:1351–1371

    Google Scholar 

  • Fernández MT, Ubeda JF, Briones AI (2000) Typing of non-Saccharomyces yeasts with enzymatic activities of interest in winemaking. Int J Food Microbiol 59:29–36

    Article  PubMed  Google Scholar 

  • Fleet GH, Lafon-Lafourcade S, Ribéreau-Gayon P (1984) Evolution of yeasts and lactic acid bacteria during fermentation and storage of Bordeaux Wines. Appl Environ Microbiol 48:1034–1038

    CAS  PubMed  Google Scholar 

  • Gafner J, Schultz M (1996) Impact of glucose-fructose ratio on stuck fermentations: practical experiences to restart stuck fermentation. Vitic Enol Sci 51:214–218

    CAS  Google Scholar 

  • Gil JV, Mateo JJ, Jiménez M, Pastor A, Huerta T (1996) Aroma compounds in wine as influenced by apiculate yeasts. J Food Sci 61:1247–1266

    Article  CAS  Google Scholar 

  • González SS, Barrio E, Gafner J, Querol A (2006) Natural hybrids from Saccharomyces cerevisiae, Saccharomyces bayanus and Saccharomyces kudriavzevii in wine fermentations. FEMS Yeast Res 6:1221–1234

    Article  PubMed  Google Scholar 

  • González SS, Barrio E, Querol A (2007) Molecular identification and characterization of wine yeast isolated from Tenerife (Canary Island, Spain). J Appl Microbiol 102:1018–1025

    PubMed  Google Scholar 

  • Huey B, Hall J (1989) Hypervariable DNA fingerprinting in Escherichia coli. Minisatellite probe from bacteriophage M13. J Bacteriol 171:2528–2532

    CAS  PubMed  Google Scholar 

  • IFOAM (2005) IFOAM Basic standards for organic production and processing. Bonn-Germany http://www.ifoam.org

  • Jemec KP, Cadez N, Zagorc T, Bubic V, Zupec A, Raspor P (2001) Yeast population dynamics in five spontaneous fermentations of Malvasia must. Food Microbiol 18:247–259

    Article  Google Scholar 

  • Kurtzman CP, Robnett CJ (1998) Identification and phylogeny of ascomycetous yeasts from analysis of nuclear large-subunit (26 S) ribosomal DNA partial sequences. Antonie van Leeuwenhoek 73:331–371

    Article  CAS  PubMed  Google Scholar 

  • Kurtzman CP, Droby S (2001) Metschnikowia fructicola, a new ascosporic yeast with potential for biocontrol of postharvest fruit rots. System Appl Microbiol 24:395–399

    Article  CAS  Google Scholar 

  • Kurtzman CP, Robnett C (2003) Phylogenetic relationships among yeasts of the Saccharomyces complex determined from multigene sequence analysis. FEMS Yeast Res 3:417–432

    Article  CAS  PubMed  Google Scholar 

  • Lema C, Garcia-Jares C, Orriols I, Angulo L (1996) Contribution of Saccharomyces and non-Saccharomyces populations to the production of some components of Albarino wine aroma. Am J Enol Vitic 47:206–216

    CAS  Google Scholar 

  • Longo E, Cansado J, Agrelo D, Villa TG (1991) Effect of climatic conditions on yeast diversity in grape musts from Northwest Spain. Am J Enol Vitic 42:141–144

    Google Scholar 

  • Lopandic K, Tiefenbrunner W, Gangl H, Mandl K, Berger S, Leitner G, Abd-Ellah GA, Querol A, Gardner RC, Sterflinger K, Prillinger H (2008) Molecular profiling of yeasts isolated during spontaneous fermentations of Austrian wines. FEMS Yeast Res 8:1063–1075

    Article  CAS  PubMed  Google Scholar 

  • Martín B, Garriga M, Hugas M, Bover-Cid S, Veciana-Nogués MT, Aymerich T (2006) Molecular, technological and safety characterization of Gram-positive catalase-positive cocci from slightly fermented sausages. Int J Food Microbiol 107:148–158

    Article  PubMed  Google Scholar 

  • Mills DA, Johannsen EA, Cocolin L (2002) Yeast diversity and persistence in Botrytis-affected wine fermentations. Appl Environ Microbiol 68:4884–4893

    Article  CAS  PubMed  Google Scholar 

  • Morris EO, Eddy MA (1957) Method for measurement of wild yeast infection in pitching yeast. J Inst Brew 63:34–43

    Google Scholar 

  • Nisiotou A, Nychas GJE (2007) Yeast populations residing on healthy or Botrytis-infected grapes from a vineyard in Attica, Greece. Appl Environ Microbiol 73:2765–2768

    Article  CAS  PubMed  Google Scholar 

  • Pallmann CL, Brown JA, Olineka TL, Cocolin L, Mills DA, Bisson L (2001) Use of WL medium to profile native flora fermentations. Am J Enol Vitic 52:198–203

    CAS  Google Scholar 

  • Phister TG, Mills DA (2003) Real-Time PCR assay for detection and enumeration of Dekkera bruxellensis in wine. Appl Environ Microbiol 69:7430–7434

    Article  CAS  PubMed  Google Scholar 

  • Prakitchaiwattana CJ, Fleet GH, Heard GM (2004) Application and evaluation of denaturing gradient gel electrophoresis to analyse the yeast ecology of wine grapes. FEMS Yeast Res 4:865–877

    Article  CAS  PubMed  Google Scholar 

  • Pretorius IS, van der Westhuizen TJ, Augustyn OPH (1999) Yeast biodiversity in vineyards and wineries and its importance to the South African wine industry. S Afr J Enol Vitic 20:61–74

    Google Scholar 

  • Querol A, Barrio E, Huerta T, Ramón D (1992) Molecular monitoring of wine fermentations conduced by active dry yeast system. Appl Environ Microbiol 58:2948–2953

    CAS  PubMed  Google Scholar 

  • Raspor P, Milek DM, Polanc J, Mozina SS, Cadez N (2006) Yeasts isolated from three varieties of grapes cultivated in different locations of the Dolenjska vine-growing region, Slovenia. Int J Food Microbiol 109:97–102

    Article  CAS  PubMed  Google Scholar 

  • Romano P, Paraggio M, Turbanti L (1998) Stability in by-product formation as a strain selection tool of Saccharomyces cerevisiae wine yeasts. J Appl Microbiol 84:336–341

    Article  CAS  PubMed  Google Scholar 

  • Sipiczki M (2003) Candida zemplinina sp. Nov., an osmotolerant and psychrotolerant yeast that ferments sweet botrytized wines. Int J Syst Evol Microbiol 53:2079–2083

    Article  CAS  PubMed  Google Scholar 

  • Suzzi G, Tofalo R, Chaves- López C, Ramazzotti S, Stagnari F, Di Fabio F, Barca E, Castrignanò A, Pisante M (2008) Isolation and selection of Saccharomyces cerevisiae from Montepulciano d’Abruzzo “Colline Teramane” areas with different microclimatic characteristics. In: Proceedings of OIV 2008—31st World Congress of Vine and Wine. 15–20 June 2008, Verona, Italy

  • Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599

    Article  CAS  PubMed  Google Scholar 

  • Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The CLUSTAL-X windows interface: flexible strategies for multiple sequence alignment aided by quantity analysis tools. Nucleic Acids Res 25:4876–4882

    Article  CAS  PubMed  Google Scholar 

  • Tofalo R, Chaves-López C, Di Fabio F, Schirone M, Felis GE, Torriani S, Paparella A, Suzzi G (2009) Molecular identification and osmotolerant profile of wine yeasts that ferment a high sugar grape must. Int J Food Microbiol 130:179–187

    Article  CAS  PubMed  Google Scholar 

  • Torija MJ, Rozès N, Poblet M, Guillamòn JM, Mas A (2001) Yeast population dynamics in spontaneous fermentations: comparison between two different wine-producing areas over a period of three years. Antonie van Leeuwenhoek 79:345–352

    Article  CAS  PubMed  Google Scholar 

  • Toro ME, Vazquez F (2002) Fermentation behaviour of controlled mixed and sequential cultures of Candida cantarellii and Saccharomyces cerevisiae wine yeasts. World J Microbiol Biotechnol 18:347–354

    Article  CAS  Google Scholar 

  • Trioli G, Hofmann U (2009) ORWINE: code of good organic viticulture and wine-making. ECOVIN-Federal Association of Organic Wine-Producer. Oppenheim

  • Viana F, Gil JV, Genovés S, Vallés S, Manzanares P (2008) Rational selection of non-Saccharomyces wine yeasts for mixed starters based on ester formation and enological traits. Food Microbiol 25:778–785

    Article  CAS  PubMed  Google Scholar 

  • Zironi R, Romano P, Suzzi G, Battistuta F, Corni G (1993) Volatile metabolites produced in wine by mixed and sequential cultures of Hanseniaspora guilliermondii or Kloeckera apiculata and Saccharomyces cerevisiae. Biotechnol Lett 15:235–238

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors are grateful to “Azienda BioVitivinicola Pepe” for providing samples and for kind cooperation. The valuable technical assistance of L. Di Giuseppe, F. Tittarelli and F. Di Simone is gratefully acknowledged.

Author information

Authors and Affiliations

  1. Dipartimento Scienze degli Alimenti, Università degli Studi di Teramo, Via C.R. Lerici 1, Mosciano Sant’Angelo, 64023, (TE), Italy

    Rosanna Tofalo, Maria Schirone, Gianluca Ciro Telera, Anna Chiara Manetta, Aldo Corsetti & Giovanna Suzzi

Authors
  1. Rosanna Tofalo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Maria Schirone
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Gianluca Ciro Telera
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Anna Chiara Manetta
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Aldo Corsetti
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Giovanna Suzzi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Rosanna Tofalo.

Additional information

This paper is part of the special issue “Wine microbiology and safety: from the vineyard to the bottle (Microsafetywine)”, 19–20 November 2009, Martina Franca (Italy).

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tofalo, R., Schirone, M., Telera, G.C. et al. Influence of organic viticulture on non-Saccharomyces wine yeast populations. Ann Microbiol 61, 57–66 (2011). https://doi.org/10.1007/s13213-010-0102-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13213-010-0102-8

Keywords

Search

Navigation