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…

Spatial variability of temperature is linked to grape composition variability in the Saint-Emilion winegrowing area

Elevated temperature during the grape maturation period is a major threat for grape quality and thus wine quality. Therefore, characterizing the grape composition response to temperature at a larger scale would represent a crucial step towards adaptation to climate change. In response to changes in temperature, various physiological mechanisms regulate grape composition. Primary and secondary metabolisms are both involved in this response, with well-known effects, for example on anthocyanins, and lesser known effects, for example on aromas or aroma precursors. At the field scale or at the regional scale, however, numerous environmental or plant-specific factors intervene to make the effects of temperature difficult to distinguish from overall variability. In this study, it was attempted to overcome this difficulty by selecting well-characterized situations with differing temperatures. A long-term study of air temperature variability across several Merlot vineyards in the Saint-Emilion and Pomerol wine producing area found significant temperature differences and gradients at various time scales linked to environmental factors. From this study area, a few sites were selected with similar age, soil and training system conditions, and with repeated and contrasted temperature differences during the maturation period. The average temperature difference during the maturation period was about 2°C between cooler and warmer sites, a difference similar to that expected under future climate change scenarios. In close vicinity to the temperature sensors at each site, grape berries were sampled at different times until full maturity during 2019 and 2020. Also, berries from bunches on either side of the row were analyzed separately, allowing an investigation of bunch exposure effect associated with the coupling of berry temperature and solar radiation. Four replicates of pooled berries for each time – site – bunch exposure combination were obtained and analyzed for biochemical composition. Analyses of variance of the biochemical composition data collected at different sampling times reveal significant effects associated with temperature, site, and bunch azimuth. For instance, anthocyanins in grape skins are clearly influenced by temperature and solar radiation exposure, with up to 30% reduction in warmer conditions.

Biodiversidad de levaduras no-Saccharomyces aisladas de viñedos uruguayos: Lachancea thermotolerans y su potencial en la industria de bebidas fermentadas

Non-saccharomyces yeasts play a crucial role in fermentation, producing a variety of secondary metabolites and enzymes that contribute to aromatic and sensory complexity compared to saccharomyces yeasts. It is crucial to understand and control the dynamics of non-saccharomyces yeasts to produce distinctive and high-quality fermented beverages.

THE EFFECT OF PRE-FERMENTATIVE GLYPHOSATE ADDITION ON THE METABOLITE PROFILE OF WINE

The synthetic herbicide glyphosate has been used extensively in viticulture over many decades to combat weeds. Despite this, the possible influence of residual glyphosate on both the alcoholic fermentation of grape juice and the subsequent metabolite profile of wines has not been investigated. In this study, Pinot noir juice supplemented with different concentrations of glyphosate (0 µg L-1, 10 µg L-1 and 1000 µg L-1) was fermented with commercial Saccharomyces cerevisiae yeast strains. Using a combination of analytical methods, 80 metabolites were quantified in the resulting wines.

Berry shrivel causes – summarizing current hypotheses

Diverse ripening disorders affect grapevine resulting in high economic losses worldwide. The common obvious symptom is shriveling berries, however the shriveling pattern and the consequences for berry quality traits are distinct in each disorder. Among them, the disorder berry shrivel is characterized by a reduced sugar accumulation short after the onset of berry ripening leaving the clusters unsuitable for wine processing. Although our knowledge on BS increased recently, potential internal or external triggers contributing to the induction of BS are yet to be explored.

Terroir influence on growth, grapes and grenache wines in the AOC priorat, northeast Spain

The Mediterranean climate of The Priorat AOC, situated behind the coastal mountain range of Tarragona, tends towards continentality with very little precipitation during the vegetation cycle. The soil is poor, dry and rocky, largely composed of slate schist, known as “llicorella”. Vines primarily grow on steep slopes and terraces.