European Food Research and Technology

, Volume 243, Issue 1, pp 41–48 | Cite as

Lactic acid bacteria communities in must, alcoholic and malolactic Tempranillo wine fermentations, by culture-dependent and culture-independent methods

  • Lucía González-Arenzana
  • Pilar Santamaría
  • Ana Rosa Gutiérrez
  • Rosa López
  • Isabel López-AlfaroEmail author
Original Paper


The lactic acid bacteria (LAB) communities from must and through alcoholic (AF) and malolactic fermentations (MLF) of Tempranillo red wines were studied in ten wineries from the Designation of Origin Rioja during three consecutive vintages. A statistical study with data from both methods, PCR-DGGE and plating, was performed. Results showed that the LAB community in the D.O. Rioja was highly determined by the type of fermentation and also by the different stages within the winemaking, while other factors such as year, winery, or sampling subzones had not significant effect on the LAB species distribution. Three microbial families, seven genera, and 25 species were described in this research, and Lactobacillus was the most commonly detected genus before MLF. Curiously, genera and species not frequently detected in wines as Weissella, Fructobacillus, and Oenococcus kitaharae were identified during AF, and no-Oenococcus oeni species were described in some MLF by both methods. For the first time, two new O. oeni allelic groups were determined by 16S rDNA/DGGE being randomly adapted to the wine environment. Further studies targeted to understand the implication of the novel species, and O. oeni allelic groups in Rioja wine fermentations could be really interesting.


Lactic acid bacteria DGGE O. oeni alleles Distribution Ecology 



This work was supported by funding and predoctoral grant (B.O.R. 6th March, 2009) of the Government of La Rioja, the I.N.I.A. Project RTA2007-00104-00-00 and it was possible thanks to the collaborating wineries.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Compliance with Ethics Requirements

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

Supplementary material

217_2016_2720_MOESM1_ESM.doc (86 kb)
Online resource 1. Consensus dendrogram obtained by composite data set combining the results of the LAB species detected with culture-dependent methods and independent methods (PCR-16S rDNA/DGGE and PCR-rpoB/DGGE) showing cophenetic correlation values. Samples were labelled with a number corresponding with the isolation stage (1: must, 2: middle AF, 3: final AF, 4: initial MLF, 5: middle MLF and 6: final MLF), with a capital letter indicative of the winery (from A to J) and Roman numbers placed in parenthesis representing the consecutive isolation years (I, II and III). The identified species were labelled with lower Latin letters placed at the top of each representative band (a: O. oeni, b: O. kitaharae, c: L. citreum, d: L. mesenteroides, e: L. pseudomesenteroides, f: L. fallax, g: F, ficulneus, h: F. tropaeoli, i: W. cibaria, j: W. paramesenteroides, k: Weisella sp., l: W. solis, m: L. brevis, n: L. buchneri, o: L. coryniformis, p: L. mali, q: L. hilgardii, r: L. plantarum, s: L. pentosus, t: L. rhamnosus, u: Lactobacillus sp., v: L. uvarum, w: P. pentosaceus, x: P. parvulus, y: L. lactis). Supplementary material 1 (DOC 85 kb)
217_2016_2720_MOESM2_ESM.doc (28 kb)
Online resource 2. Phylogram for the 110 sequences identified as species belonging to genus Oenococcus, obtained from 16S rDNA PCR/DGGE. Each sequence is referred with the most accurate identification and the identity percentage (%), with the given accession number and with a code that means the isolation stage (from 1 to 6), winery (from A to J) and year (I, II or III). The evolutionary distances are in the units of the number of base substitutions per site. Supplementary material 2 (DOC 28 kb)


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Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Lucía González-Arenzana
    • 1
  • Pilar Santamaría
    • 1
  • Ana Rosa Gutiérrez
    • 1
    • 2
  • Rosa López
    • 1
  • Isabel López-Alfaro
    • 1
    • 2
    Email author
  1. 1.CVV, Instituto de Ciencias de la Vid y del Vino(Gobierno de La Rioja, Universidad de La Rioja, CSIC)LogroñoSpain
  2. 2.Departamento de Agricultura y AlimentaciónUniversidad de La RiojaLogroñoSpain

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