High-throughput sequencing of amplicons for monitoring yeast biodiversity in must and during alcoholic fermentation

We compared pyrosequencing technology with the PCR-ITS-RFLP analysis of yeast isolates and denaturing gradient gel electrophoresis (DGGE). These methods gave divergent findings for the yeast population. DGGE was unsuitable for the quantification of biodiversity and its use for species detection was...

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Bibliographic Details
Published in:Journal of industrial microbiology & biotechnology 2014-05, Vol.41 (5), p.811-821
Main Authors: David, Vanessa, Terrat, Sébastien, Herzine, Khaled, Claisse, Olivier, Rousseaux, Sandrine, Tourdot-Maréchal, Raphaëlle, Masneuf-Pomarede, Isabelle, Ranjard, Lionel, Alexandre, Hervé
Format: Article
Language:English
Subjects:
alcoholic fermentation
Analysis
Biochemistry
Biodiversity
Bioinformatics
Biological and medical sciences
Biomedical and Life Sciences
Biotechnology
Chemical and Process Engineering
Denaturing Gradient Gel Electrophoresis
Engineering Sciences
Environmental Microbiology
Environmental Sciences
Ethanol - metabolism
Fermentation
Food engineering
Fundamental and applied biological sciences. Psychology
Genetic Engineering
grapes
High-Throughput Nucleotide Sequencing
Inorganic Chemistry
Life Sciences
Microbiology
monitoring
Polymerase Chain Reaction
Population
population dynamics
sequence analysis
Studies
Vegetal Biology
Vitaceae
Vitis - microbiology
Wineries & vineyards
Wines
Yeast
Yeasts
Yeasts - classification
Yeasts - genetics
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Description
Summary:We compared pyrosequencing technology with the PCR-ITS-RFLP analysis of yeast isolates and denaturing gradient gel electrophoresis (DGGE). These methods gave divergent findings for the yeast population. DGGE was unsuitable for the quantification of biodiversity and its use for species detection was limited by the initial abundance of each species. The isolates identified by PCR-ITS-RFLP were not fully representative of the true population. For population dynamics, high-throughput sequencing technology yielded results differing in some respects from those obtained with other approaches. This study demonstrates that 454 pyrosequencing of amplicons is more relevant than other methods for studying the yeast community on grapes and during alcoholic fermentation. Indeed, this high-throughput sequencing method detected larger numbers of species on grapes and identified species present during alcoholic fermentation that were undetectable with the other techniques.
ISSN:1367-5435
1476-5535
DOI:10.1007/s10295-014-1427-2