OENO IVAS 2019 banner
IVES 9 IVES Conference Series 9 OENO IVAS 9 OENO IVAS 2019 9 Grape and wine microorganisms: diversity and adaptation 9 Application of high-throughput sequencing tools for characterisation of microbial communities during alcoholic fermentation

Application of high-throughput sequencing tools for characterisation of microbial communities during alcoholic fermentation

Abstract

Developments in high-throughput sequencing (HTS) technologies allow us to obtain large amounts of microbial information from wine and must samples. Thus approaches, that are aimed at characterising the microbial diversity during fermentation, can be enhanced, or possibly even replaced, with HTS-based metabarcoding. To reduce experimental biases and increase data reproducibility, we compared 3 DNA extraction methods by evaluating differences in the fungal diversity with Riesling alcoholic fermentation samples at four different vineyards. The fungal diversity profiling was done using the genetic markers ITS2 and D2 using metabarcoding. The extraction methods compared consisted of a commercial kit, a recently published protocol that includes a DNA enhancer, and a protocol based on a buffer containing common inhibitor removal reagents. All methods were able to distinguish vineyard effects on the fungal diversity, but the results differed quantitatively. 

From the results of extraction methods, we applied the chosen methods and further combined the HTS tools of metabarcoding and metagenomics, to characterise how microbial communities of those samples, and their subsequent spontaneously fermented derivatives, vary. We specifically explored microbial community variation related to vineyard level, and during alcoholic fermentation. The vineyard was shown to be strongly influencing the microbial communities. Functional analyses were additionally included to investigate the microbial interactions. An increase in non-Saccharomycetaceae fungal functions and a decrease in bacterial functions were also observed during the early fermentation stage. Overall, our results highlight the importance of standardizing DNA extraction methods when characterising fungal diversity from wine and related samples, and showcase how metagenomic functional analysis offer possibilities to improve our insights into the wine alcoholic fermentation process, including highlighting microbe interactions.

DOI:

Publication date: June 23, 2020

Issue: OENO IVAS 2019

Type: Article

Authors

Sarah Siu Tze Mak, Kimmo Sirén, Christian Carøe, Ulrich Fischer, M. Thomas P. Gilbert 

Section for Evolutionary Genomics, Natural History Museum of Denmark, University of Copenhagen, Copenhagen, Denmark 
Institute for Viticulture and Oenology, Dienstleistungszentrum Ländlicher Raum Rheinpfalz, Neustadt an der Weinstraße, Germany 

Contact the author

Keywords

Riesling, Metabarcoding, Metagenomics, DNA extraction

Tags

IVES Conference Series | OENO IVAS 2019

Citation

Related articles…

Multivariate strategies for red wines classification using stilbenes and flavonols content

Bioactive polyphenols from grapes and wines, like stilbenes and flavonols (SaF), are often determined to nutritional evaluation, but also for many other purposes. The objective of this study was to quantify SaF in red wines from “Campanha Gaúcha”, a large and young viticultural region from South Brazil. Moreover, through statistical analysis, evaluate the influence of these compounds according to varieties, production process, harvest years and micro-regions of cultivation. A total of 58 samples of red wines were analyzed by high-performance liquid chromatography coupled to diode array detector (HPLC-DAD) for determination of trans-resveratrol (R), quercetin (Q), myricetin (M), kaempferol (K), trans-e-viniferin (V) and their precursor, cinnamic acid (C).

Validation of a method for the determination of volatile compounds in in spirituous beverages using contained ethanol as a reference substance

The results of experimental studies of the method based on the usage of ethyl alcohol as an internal standard for the direct determination of volatile compounds in wines and others alcohol contained products are presented. The method was validated in terms of precision, accuracy, limits of detection and quantification (lod and loq), linearity, and robustness.

Importanza del monitoraggio micro-meteorologico nella caratterizzazione del terroir

Le variabili meteorologiche e micro-meteorologiche ricoprono un importante ruolo sulla risposta vegeto-produttiva della vite e di conseguenza sulla qualità delle produzioni. Utilizzando una rete wireless di sensori sono stati monitorati i parametri meteorologici e micro-meteorologici di 4 vigneti del territorio toscano e in differenti condizioni di gestione agronomica.

A look back at 20 years of exploring the future of the vines and wines sector

What if, in 25 years, most wines were dealcoholized and flavored ? What if vines were only cultivated to combat erosion, store carbon, and provide anthocyanins…? What if climate change completely changed the list of vine varieties cultivable for wine production in France? What if food stores had completely disappeared in favor of virtual platforms? And if… because the long-term future is not predetermined and therefore not knowable, because the future is open to several possibilities, because the future does not emerge from nothing but from the present which conceals heavy trends and weak signals, prospective approaches make it possible to consider the room for maneuver that actors have to promote the advent of a future, which we can hope to be chosen, at least in part.

Release and perception of γ-nonalactone and massoia lactone in the red wine matrix: impact of ethanol and acidity

Climate change (CC) is altering grape/wine composition, challenging wine sensory quality. Rising temperatures increase grape sugar levels, with higher wine ethanol (EtOH) contents, reduce total acidity (TA) converging with increased pH and lead to the accumulation of CC odorous markers such as γ-nonalactone (γ-C9) and massoia lactone (ML).