Characterization of the “viable but nonculturable” (VBNC) state in the wine spoilage yeast Brettanomyces

Virginie Serpaggia, Fabienne Remizea, 1, Ghislaine Recorbetb, Eliane Gaudot-Dumasb, Anabelle Sequeira-Le Grandc, Hervé Alexandrea, , a Institut Universitaire de la Vigne et du Vin Jules Guyot, Université de Bourgogne, Rue Claude Ladrey, BP 27877, F-21078 Dijon Cedex, France

b UMR 1088 INRA/CNRS 5184/UB Plante-Microbe-Environnement, INRA-CMSE, Dijon, France, c Plate-forme de Cytométrie, IFR100 Santé STIC, Fac. Médécine, Université de Bourgogne, 7 Bd Jeanne D'Arc, 21000-Dijon, France

Received 4 October 2011. Revised 22 December 2011. Accepted 23 December 2011. Available online 8 January 2012.

http://dx.doi.org/10.1016/j.fm.2011.12.020, How to Cite or Link Using DOI

Abstract:

Although the viable but not culturable (VBNC) state has been studied in detail in bacteria, it has been suggested that maintenance of viability with loss of culturability also exists in eukaryotic cells, such as in the wine spoilage yeast Brettanomyces. To provide conclusive evidence for the existence of a VBNCstate in this yeast, we investigated its capacity to become viable and nonculturable after sulfite stress, and its ability to recover culturability after stressor removal. Sulfite addition induced loss of culturability but maintenance of viability. Increasing the medium pH to decrease the concentration of toxic SO2 allowed yeast cells to become culturable again, thus demonstrating the occurrence of a VBNC state in Brettanomyces upon SO2 exposure. Relative to culturable Brettanomyces, VBNC yeast cells were found to display a 22% decrease in size on the basis of laser granulometry. Assays for 4-ethylguaiacol and 4-ethylphenol, volatile phenols produced by Brettanomyces, indicated that spoilage compound production could persist in VBNC cells. These morphological and physiological changes in VBNC Brettanomyces were coupled to extensive protein pattern modifications, as inferred by comparative two-dimensional electrophoresis and mass spectrometric analyses. Upon identification of 53 proteins out of the 168 spots whose abundance was significantly modified in treated cells relative to control, we propose that the SO2-induced VBNC state in Brettanomyces is characterized by a reduced glycolytic flux coupled to changes in redox homeostatis/protein turnover-related processes. This study points out the existence of common mechanisms between yeast and bacteria upon entry to the VBNC state.

Voir le document, ci-joint: Characterization_viable_nonculturable_wine

Voir: http://www.sciencedirect.com/science/article/pii/S0740002011003030

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