Articles producció científica> Bioquímica i Biotecnologia

Viability-PCR allows monitoring yeast population dynamics in mixed fermentations including viable but non-culturable yeasts

  • Dades identificatives

    Identificador: imarina:8996754
    Autors:
    Navarro, YurenaTorija, Maria-JesusMas, AlbertBeltran, Gemma
    Resum:
    © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). The use of controlled mixed inocula of Saccharomyces cerevisiae and non-Saccharomyces yeasts is a common practice in winemaking, with Torulaspora delbrueckii, Lachancea thermotolerans and Metschnikowia pulcherrima being the most commonly used non-Saccharomyces species. Although S. cerevisiae is usually the dominant yeast at the end of mixed fermentations, some non-Saccharomyces species are also able to reach the late stages; such species may not grow in culture media, which is a status known as viable but non-culturable (VBNC). Thus, an accurate methodology to properly monitor viable yeast population dynamics during alcoholic fermentation is required to understand microbial interactions and the contribution of each species to the final product. Quantitative PCR (qPCR) has been found to be a good and sensitive method for determining the identity of the cell population, but it cannot distinguish the DNA from living and dead cells, which can overestimate the final population results. To address this shortcoming, viability dyes can be used to avoid the amplification and, therefore, the quantification of DNA from non-viable cells. In this study, we validated the use of PMAxx dye (an optimized version of propidium monoazide (PMA) dye) coupled with qPCR (PMAxx-qPCR), as a tool to monitor the viable population dynamics of the most common yeast species used in wine mixed fermentations (S. cerevisiae, T. delbrueckii, L. thermotolerans and M. pulcherrima), comparing the results with non-dyed qPCR and colony counting on differential medium. Our re
  • Altres:

    Autor segons l'article: Navarro, Yurena; Torija, Maria-Jesus; Mas, Albert; Beltran, Gemma
    Departament: Bioquímica i Biotecnologia
    Autor/s de la URV: Beltran Casellas, Gemma / Mas Baron, Alberto / Navarro García, Yurena de los Ángeles / Torija Martínez, María Jesús
    Paraules clau: Wine yeast Viable but non culturable Viability qpcr Saccharomyces cerevisiae Propidium monoazide Non-saccharomyces yeast Non-saccharomyces Listeria-monocytogenes Hanseniaspora-uvarum Ethidium monoazide Cerevisiae Cell-cell contact Alcoholic fermentation Acid bacteria
    Resum: © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). The use of controlled mixed inocula of Saccharomyces cerevisiae and non-Saccharomyces yeasts is a common practice in winemaking, with Torulaspora delbrueckii, Lachancea thermotolerans and Metschnikowia pulcherrima being the most commonly used non-Saccharomyces species. Although S. cerevisiae is usually the dominant yeast at the end of mixed fermentations, some non-Saccharomyces species are also able to reach the late stages; such species may not grow in culture media, which is a status known as viable but non-culturable (VBNC). Thus, an accurate methodology to properly monitor viable yeast population dynamics during alcoholic fermentation is required to understand microbial interactions and the contribution of each species to the final product. Quantitative PCR (qPCR) has been found to be a good and sensitive method for determining the identity of the cell population, but it cannot distinguish the DNA from living and dead cells, which can overestimate the final population results. To address this shortcoming, viability dyes can be used to avoid the amplification and, therefore, the quantification of DNA from non-viable cells. In this study, we validated the use of PMAxx dye (an optimized version of propidium monoazide (PMA) dye) coupled with qPCR (PMAxx-qPCR), as a tool to monitor the viable population dynamics of the most common yeast species used in wine mixed fermentations (S. cerevisiae, T. delbrueckii, L. thermotolerans and M. pulcherrima), comparing the results with non-dyed qPCR and colony counting on differential medium. Our results showed that the PMAxx-qPCR assay used in this study is a reliable, specific and fast method for quantifying these four yeast species during the alcoholic fermentation process, being able to distinguish between living and dead yeast populations. Moreover, the entry into VBNC status was observed for the first time in L. thermotolerans and S. cerevisiae during alcoholic fermentation. Further studies are needed to unravel which compounds trigger this VBNC state during alcoholic fermentation in these species, which would help to better understand yeast interactions.
    Àrees temàtiques: Plant science Microbiology Health professions (miscellaneous) Health (social science) Food science & technology Food science
    Accès a la llicència d'ús: https://creativecommons.org/licenses/by/3.0/es/
    Adreça de correu electrònic de l'autor: yurenadelosangeles.navarro@urv.cat gemma.beltran@urv.cat mjesus.torija@urv.cat albert.mas@urv.cat
    Identificador de l'autor: 0000-0002-7071-205X 0000-0001-6419-0745 0000-0002-0763-1679
    Data d'alta del registre: 2024-10-12
    Versió de l'article dipositat: info:eu-repo/semantics/publishedVersion
    URL Document de llicència: https://repositori.urv.cat/ca/proteccio-de-dades/
    Referència a l'article segons font original: Foods. 9 (10): 1373-
    Referència de l'ítem segons les normes APA: Navarro, Yurena; Torija, Maria-Jesus; Mas, Albert; Beltran, Gemma (2020). Viability-PCR allows monitoring yeast population dynamics in mixed fermentations including viable but non-culturable yeasts. Foods, 9(10), 1373-. DOI: 10.3390/foods9101373
    Entitat: Universitat Rovira i Virgili
    Any de publicació de la revista: 2020
    Tipus de publicació: Journal Publications
  • Paraules clau:

    Food Science,Food Science & Technology,Health (Social Science),Health Professions (Miscellaneous),Microbiology,Plant Science
    Wine yeast
    Viable but non culturable
    Viability qpcr
    Saccharomyces cerevisiae
    Propidium monoazide
    Non-saccharomyces yeast
    Non-saccharomyces
    Listeria-monocytogenes
    Hanseniaspora-uvarum
    Ethidium monoazide
    Cerevisiae
    Cell-cell contact
    Alcoholic fermentation
    Acid bacteria
    Plant science
    Microbiology
    Health professions (miscellaneous)
    Health (social science)
    Food science & technology
    Food science
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