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Comparison of DNA purification methods for high‐throughput sequencing of fungal communities from wine fermentation

High‐throughput sequencing approaches, which target a taxonomically discriminant locus, allow for in‐depth insight into microbial communities’ compositions. Although microorganisms are historically investigated by cultivation on artificial culture media, this method presents strong limitations, sinc...

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Autores principales: Gobert, Antoine, Evers, Marie Sarah, Morge, Christophe, Sparrow, Céline, Delafont, Vincent
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9593259/
https://www.ncbi.nlm.nih.gov/pubmed/36314746
http://dx.doi.org/10.1002/mbo3.1321
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author Gobert, Antoine
Evers, Marie Sarah
Morge, Christophe
Sparrow, Céline
Delafont, Vincent
author_facet Gobert, Antoine
Evers, Marie Sarah
Morge, Christophe
Sparrow, Céline
Delafont, Vincent
author_sort Gobert, Antoine
collection PubMed
description High‐throughput sequencing approaches, which target a taxonomically discriminant locus, allow for in‐depth insight into microbial communities’ compositions. Although microorganisms are historically investigated by cultivation on artificial culture media, this method presents strong limitations, since only a limited proportion of microorganisms can be grown in vitro. This pitfall appears even more limiting in enological and winemaking processes, during which a wide range of molds, yeasts, and bacteria are observed at the different stages of the fermentation course. Such an understanding of those dynamic communities and how they impact wine quality therefore stands as a major challenge for the future of enology. As of now, although high‐throughput sequencing has already allowed for the investigation of fungal communities, there is no available comparative study focusing on the performance of microbial deoxyribonucleic acid (DNA) extraction in enological matrixes. This study aims to provide a comparison of five selected extraction methods, assayed on both must and fermenting must, as well as on finished wine. These procedures were evaluated according to their extraction yields, the purity of their extracted DNA, and the robustness of downstream molecular analyses, including polymerase chain reaction and high‐throughput sequencing of fungal communities. Altogether, two out of the five assessed microbial DNA extraction methods (DNeasy PowerSoil Pro Kit and E.Z.N.A.® Food DNA Kit) appeared suitable for robust evaluations of the microbial communities in wine samples. Consequently, this study provides robust tools for facilitated upcoming studies to further investigate microbial communities during winemaking using high‐throughput sequencing.
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spelling pubmed-95932592022-10-26 Comparison of DNA purification methods for high‐throughput sequencing of fungal communities from wine fermentation Gobert, Antoine Evers, Marie Sarah Morge, Christophe Sparrow, Céline Delafont, Vincent Microbiologyopen Commentary High‐throughput sequencing approaches, which target a taxonomically discriminant locus, allow for in‐depth insight into microbial communities’ compositions. Although microorganisms are historically investigated by cultivation on artificial culture media, this method presents strong limitations, since only a limited proportion of microorganisms can be grown in vitro. This pitfall appears even more limiting in enological and winemaking processes, during which a wide range of molds, yeasts, and bacteria are observed at the different stages of the fermentation course. Such an understanding of those dynamic communities and how they impact wine quality therefore stands as a major challenge for the future of enology. As of now, although high‐throughput sequencing has already allowed for the investigation of fungal communities, there is no available comparative study focusing on the performance of microbial deoxyribonucleic acid (DNA) extraction in enological matrixes. This study aims to provide a comparison of five selected extraction methods, assayed on both must and fermenting must, as well as on finished wine. These procedures were evaluated according to their extraction yields, the purity of their extracted DNA, and the robustness of downstream molecular analyses, including polymerase chain reaction and high‐throughput sequencing of fungal communities. Altogether, two out of the five assessed microbial DNA extraction methods (DNeasy PowerSoil Pro Kit and E.Z.N.A.® Food DNA Kit) appeared suitable for robust evaluations of the microbial communities in wine samples. Consequently, this study provides robust tools for facilitated upcoming studies to further investigate microbial communities during winemaking using high‐throughput sequencing. John Wiley and Sons Inc. 2022-10-25 /pmc/articles/PMC9593259/ /pubmed/36314746 http://dx.doi.org/10.1002/mbo3.1321 Text en © 2022 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd. https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Commentary
Gobert, Antoine
Evers, Marie Sarah
Morge, Christophe
Sparrow, Céline
Delafont, Vincent
Comparison of DNA purification methods for high‐throughput sequencing of fungal communities from wine fermentation
title Comparison of DNA purification methods for high‐throughput sequencing of fungal communities from wine fermentation
title_full Comparison of DNA purification methods for high‐throughput sequencing of fungal communities from wine fermentation
title_fullStr Comparison of DNA purification methods for high‐throughput sequencing of fungal communities from wine fermentation
title_full_unstemmed Comparison of DNA purification methods for high‐throughput sequencing of fungal communities from wine fermentation
title_short Comparison of DNA purification methods for high‐throughput sequencing of fungal communities from wine fermentation
title_sort comparison of dna purification methods for high‐throughput sequencing of fungal communities from wine fermentation
topic Commentary
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9593259/
https://www.ncbi.nlm.nih.gov/pubmed/36314746
http://dx.doi.org/10.1002/mbo3.1321
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