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…

INFLUENCE OF GRAPE RIPENESS ON MACROMOLECULES EXTRACTABILITY FROM GRAPE SKIN TISSUES AND GRAPE SEEDS DURING WINEMAKING

A consequence of climate change is the modification of grape harvest quality and physico-chemical parameters of the obtained wine: increase in alcoholic degree, decrease in pH, and modification of the extractability of macromolecules, which leads to problems of microbiological, tartaric, colour and colloidal stability. In order to respond to these problems, the winemaking processes must be anticipated and adapted with a better knowledge of macromolecule extractability in grapes and their evolution, according to the grape variety, vintage and winemaking process. The purpose of this study was to understand 1) how the harvest date can influence the extractability of macromolecules, polysaccharides and phenolic compounds, which are responsible for wine stability 2) how to adapt the winemaking process to the harvest date in order to optimise wine quality.

Raman spectroscopy as a rapid method to assess grape polyphenolic maturation and wine malolactic fermentation on site

Wineries can increase their economic and environmental sustainability by optimizing the winemaking procedures, from harvest to wine maturation and conservation. Based on analytical data of the chemical composition and wine sensory evaluation, the enologist makes his own decision regarding the enological interventions at the harvest date selection, winemaking and post-winemaking.

Using atmospheric and statistical models to understand local climate and assess spatial temperature variability at a fine scale over the Stellenbosch wine district, South Africa

Atmospheric and statistical models were used to increase understanding of potential climatic impacts, resulting from mesoscale physical processes that cause significant temperature variability for viticulture within the Stellenbosch Wine of Origin district. Hourly temperature values from 16 automatic weather stations and 40 tinytag data loggers located in the vineyards were analysed.

BORDEAUX RED WINES WITHOUT ADDED SULFITES SPECIFICITIES: COMPOSITIONAL AND SENSORY APPROACHES TOWARDS HIGHLIGHTING AND EXPLAI-NING THEIR SPECIFIC FRUITINESS AND COOLNESS

With the development of naturality expectations, wines produced without any addition of sulfur dioxide (SO₂) become very popular for consumers and such wines are increasingly present on the market. Recent studies also showed that Bordeaux red wines without added SO₂ could be differentiated from a sensory point of view from similar wines produced with SO₂¹. Thus, the aim of the current study was to characterize from a sensory point of view, specific aromas of wines without added SO₂ and to identify compounds involved.

Sustaining wine identity through intra-varietal diversification

With contemporary climate change, cultivated Vitis vinifera L. is at risk as climate is a critical component in defining ecologically fitted plant materiel. While winegrowers can draw on the rich diversity among grapevine varieties to limit expected impacts (Morales-Castilla et al., 2020), replacing a signature variety that has created a sense of local distinctiveness may lead to several challenges. In order to sustain wine identity in uncertain climate outcomes, the study of intra-varietal diversity is important to reflect the adaptive and evolutionary potential of current cultivated varieties. The aim of this ongoing study is to understand to what extent can intra-varietal diversity be a climate change adaptation solution. With a focus on early (Sauvignon blanc, Riesling, Grolleau, Pinot noir) to moderate late (Chenin, Petit Verdot, Cabernet franc) ripening varieties, data was collected for flowering and veraison for the various studied accessions (from conservatory plots) and clones. For these phenological growing stages, heat requirements were established using nearby weather stations (adapted from the GFV model, Parker et al., 2013) and model performances were verified. Climate change projections were then integrated to predict the future behaviour of the intra-varietal diversity. Study findings highlight the strong phenotypic diversity of studied varieties and the importance of diversification to enhance climate change resilience. While model performances may require improvements, this study is the first step towards quantifying heat requirements of different clones and how they can provide adaptation solutions for winegrowers to sustain local wine identity in a global changing climate. As genetic diversity is an ongoing process through point mutations and epigenetic adaptations, perspective work is to explore clonal data from a wide variety of geographic locations.