title: High‑throughput sequencing of amplicons for monitoring yeast biodiversity in must and during alcoholic fermentation.
abstract: 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-ITSRFLP 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.
authors.forename: Vanessa
authors.initials: V
authors.lastname: David
authors.forename: Sébastien
authors.initials: S
authors.lastname: Terrat
authors.forename: Khaled
authors.initials: K
authors.lastname: Herzine
authors.forename: Olivier
authors.initials: O
authors.lastname: Claisse
authors.forename: Sandrine
authors.initials: S
authors.lastname: Rousseaux
authors.forename: Raphaëlle
authors.initials: R
authors.lastname: Tourdot-Maréchal
authors.forename: Isabelle
authors.initials: I
authors.lastname: Masneuf-Pomarede
authors.forename: Lionel
authors.initials: L
authors.lastname: Ranjard
authors.forename: Hervé
authors.initials: H
authors.lastname: Alexandre
mesh_terms: D044822:Biodiversity; D058645:Denaturing Gradient Gel Electrophoresis; D000431:Ethanol / Q000378:metabolism; D005285:Fermentation; D059014:High-Throughput Nucleotide Sequencing; D016133:Polymerase Chain Reaction; D027843:Vitis / Q000382:microbiology; D015003:Yeasts / Q000145:classification* / Q000235:genetics
chemical_list: D000431:Ethanol