Macrowine 2021
IVES 9 IVES Conference Series 9 Assessment of wine non-Saccharomyces yeast strains as promising producers of glutathione

Assessment of wine non-Saccharomyces yeast strains as promising producers of glutathione

Abstract

AIM: Glutathione (GSH) is a non-protein thiol naturally present in grape berries and produced by yeasts during fermentation. It has a strong antioxidant activity, thus can be added during winemaking to limit the oxidative phenomena of wine, preserving sensory characteristics and stability, ultimately promoting a healthier product by reducing the need for SO2 addition. A promising alternative approach could be the use of yeast starter cultures high-producers of this compound in situ, during the fermentation process, in substitution of external GSH addition. Since this activity in non-Saccharomyces yeasts is currently poorly investigated, the aim of this research was to evaluate the ability of three strains of non-Saccharomyces (NS) yeasts to produce GSH, both in synthetic media and in fresh grape must.

METHODS: Lachancea thermotolerans SOL13, Metschnikowia sp. FIANO12, and Starmerella bacillaris MALV45 were tested in single or sequential inoculations,with Saccharomyces cerevisiae EC1118, in synthetic grape juice (SGJ) or in Pinot Grigio grape must, under static conditions, or in a medium optimized for GSH production (MGSH) in agitation (200 rpm). GSH concentration was determined using the Glutathione Assay Kit (Sigma-Aldrich). Population dynamics was evaluated by plate count and biomass dry weight, and fermentation kinetics through weight loss measurement.

RESULTS: A variability in GSH production was found among individual strains and growth conditions. Metschnikowia sp. FIANO12 showed the highest intracellular accumulation of GSH when cultivated alone in both synthetic media, and, as expected, higher levels in the optimized MGSH than in SGJ (4.59 vs. 0.19 nmol GSH/mg cells, respectively). In wine, fermentations with S. bacillaris MALV45 had the highest concentration of intracellular GSH, but the lowest content of wine-dissolved GSH. The maximum level of extracellular GSH (21 mg/L) was obtained in the sequential fermentation with L. thermotolerans SOL13, a significant increase compared to the control singly inoculated with S. cerevisiae.

CONCLUSIONS:

This study highlights a new potential interesting feature of NS yeasts to positively modify wine composition. The tested native strains, with previously demonstrated interesting oenological traits, showed a good capacity to accumulate GSH and to increase the concentration of this antioxidant compound in wine. Thus, the strategy of multi-starter fermentation can be a valuable tool to achieve a lower input winemaking. Future investigations are needed to assess the long-term stability of wine made from multi-starter fermentations with NS yeasts producers of GSH.ACKNOWLEDGEMENTS: We thank Nicolò Bersani for laboratory assistance.

DOI:

Publication date: September 3, 2021

Issue: Macrowine 2021

Type: Article

Authors

Renato L. Binati, Wilson J.F. LEMOS JUNIOR, Sandra TORRIANI

Department of Biotechnology, University of Verona, Italy, Department of Biotechnology, University of Verona, Italy, Department of Biotechnology, University of Verona, Italy

Contact the author

Keywords

Glutathione production, multi-starter fermentation, non-saccharomyces yeasts, saccharomycescerevisiae, winemaking, wine quality

Citation

Related articles…

Comparison of imputation methods in long and varied phenological series. Application to the Conegliano dataset, including observations from 1964 over 400 grape varieties

A large varietal collection including over 1700 varieties was maintained in Conegliano, ITA, since the 1950s. Phenological data on a subset of 400 grape varieties including wine grapes, table grapes, and raisins were acquired at bud break, flowering, veraison, and ripening since 1964. Despite the efforts in maintaining and acquiring data over such an extensive collection, the data set has varying degrees of missing cases depending on the variety and the year. This is ubiquitous in phenology datasets with significant size and length. In this work, we evaluated four state-of-the-art methods to estimate missing values in this phenological series: k-Nearest Neighbour (kNN), Multivariate Imputation by Chained Equations (mice), MissForest, and Bidirectional Recurrent Imputation for Time Series (BRITS). For each phenological stage, we evaluated the performance of the methods in two ways. 1) On the full dataset, we randomly hold-out 10% of the true values for use as a test set and repeated the process 1000 times (Monte Carlo cross-validation). 2) On a reduced and almost complete subset of varieties, we varied the percentage of missing values from 10% to 70% by random deletion. In all cases, we evaluated the performance on the original values using normalized root mean squared error. For the full dataset we also obtained performance statistics by variety and by year. MissForest provided average errors of 17% (3 days) at budbreak, 14% (4 days) at flowering, 14.5% (7 days) at veraison, and 17% (3 days) at maturity. We completed the imputations of the Conegliano dataset, one of the world’s most extensive and varied phenological time series and a steppingstone for future climate change studies in grapes. The dataset is now ready for further analysis, and a rigorous evaluation of imputation errors is included.

Modelling vine water stress during a critical period and potential yield reduction rate in European wine regions: a retrospective analysis

Most European vineyards are managed under rainfed conditions, where seasonal water deficit has become increasingly important. The flowering-veraison phenophase represents an important period for vine response to water stress, which is seldomly thoroughly evaluated. Therefore, we aim to quantify the flowering-veraison water stress levels using Crop Water Stress Indicator (CWSI) over 1986–2015 for important European wine regions, and to assess the respective potential Yield Lose Rate (YLR). Additionally, we also investigate whether an advanced flowering-veraison phase may help alleviating the water stress with improved yield. A process-based grapevine model STICS is employed, which has been extensively calibrated for flowering and veraison stages using observed data at 38 locations with 10 different grapevine varieties. Subsequently, the model is being implemented at the regional level, considering site-specific calibration results and gridded climate and soil datasets. The findings suggest wine regions with stronger flowering-veraison CWSI tend to have higher potential YLR. However, contrasting patterns are found between wine regions in France-Germany-Luxembourg and Italy-Portugal-Spain. The former tends to have slight-to-moderate drought conditions (CWSI<0.5) and a negligible-to-moderate YLR (<30%), whereas the latter possesses severe-to-extreme CWSI (>0.5) and substantial YLR (>40%). Wine regions prone to a high drought risk (CWSI>0.75) are also identified, which are concentrated in southern Mediterranean Europe. An advanced flowering-veraison phase may have benefited from cooler temperatures and a higher fraction of spring precipitation in wine regions of Italy-Portugal-Spain, resulting in alleviated CWSI and moderate reductions of YLR. For those of France-Germany-Luxembourg, this can have reduced flowering-veraison precipitation, but prevalent alleviations of YLR are also found, possibly because of shifted phase towards a cooler growing season with reduced evaporative demands. Overall, such a retrospective analysis might provide new insights towards better management of seasonal water deficit for conventionally vulnerable Mediterranean wine regions, but also for relatively cooler and wetter Central European regions.

δ13C : A still underused indicator in precision viticulture  

The first demonstration of the interest of carbon isotope composition of sugars in grapevine, as an integrated indicator of vineyard water status, dates back to 2000 (Gaudillère et al., 1999; Van Leeuwen et al., 2001). Thanks to the isotopic discrimination of Carbon that takes place during plant photosynthesis, under hydric stress conditions, it is possible to accurately estimate the photosynthetic activity. Ever since, δ13C has been widely applied with success to zonation, terroir studies and vine physiology research, but is still not widely used by viticulturists. This is quite astonishing by considering the impact of global warming on viticulture and the need to improve water management, that would justify a widespread use of δ13C.
The lack of private laboratories proposing the analysis, the cost of the technology, as well as the long analytical delays, have been detrimental to its development. Some laboratories tried to overcome the analytical difficulties of isotopic analysis by using fourier transformed infrared spectroscopy, as a fast and cheap alternative to the official OIV method (IRMS). These claimed FTIR models have never been published or peer reviewed and cannot be considered robust. In this work, thanks to the recent acquisition of IRMS technology, new modern and robust applications of δ13C for viticulture are proposed. This includes the use of the analysis to make parcel separations at harvesting, the possibility to increase the precision of hydric stress cartography and the potential cost reduction when compared with Scholander pressure bomb analysis.

Permanent cover cropping with reduced tillage increased resiliency of wine grape vineyards to climate change

Majority of California’s vineyards rely on supplemental irrigation to overcome abiotic stressors. In the context of climate change, increases in growing season temperatures and crop evapotranspiration pose a risk to adaptation of viticulture to climate change. Vineyard cover crops may mitigate soil erosion and preserve water resources; but there is a lack of information on how they contribute to vineyard resiliency under tillage systems. The aim of this study was to identify the optimum combination of cover crop sand tillage without adversely affecting productivity while preserving plant water status. Two experiments in two contrasting climatic regions were conducted with two cover crops, including a permanent short stature grass (P. bulbosa hybrid), barley (Hordeum spp), and resident vegetation under till vs. no-till systems in a Ruby Cabernet (V. vinifera spp.) (Fresno) and a Cabernet Sauvingon (Napa) vineyard. Results indicated that permanent grass under no-till preserved plant available water until E-L stage 17. Consequently, net carbon assimilation of the permanent grass under no-till system was enhanced compared to those with barley and resident vegetation. On the other hand, the barley under no-till system reduced grapevine net carbon assimilation during berry ripening that led to lower content of nonstructural carbohydrates in shoots at dormancy. Components of yield and berry composition including flavonoid profile at either site were not adversely affected by factors studied. Switching to a permanent cover crop under a no-till system also provided a 9% and 3% benefit in cultural practices costs in Fresno and Napa, respectively. The results of this work provides fundamental information to growers in preserving resiliency of vineyard systems in hot and warm climate regions under context of climate change.

Heatwaves and grapevine yield in the Douro region, crop model simulations

Heatwaves or extreme heat events can be particularly harmful to agriculture. Grapevines grown in the Douro winemaking region are particularly exposed to this threat, due to the specificities of the already warm and dry climatic conditions. Furthermore, climate change simulations point to an increase in the frequency of occurrence of these extreme heat events, therefore posing a major challenge to winegrowers in the Mediterranean type climates. The current study focuses on the application of the STICS crop model to assess the potential impacts of heatwaves in grapevine yields over the Douro valley winemaking region. For this purpose, STICS was applied to grapevines using high-resolution weather, soil and terrain datasets over the Douro. To assess the impact of heatwaves, the weather dataset (1989-2005) was artificially modified, generating periods with anomalously high temperatures (+5 ºC), at certain onset dates and with specific durations (from 5 to 9 days). The model was run with this modified weather dataset and results were compared to the original unmodified runs. The results show that heatwaves can have a very strong impact on grapevine yields, strongly depending on the onset dates and duration of the heatwaves. The highest negative impacts may result in a decrease in the yield by up to -35% in some regions. Despite some uncertainties inherent to the current modelling assessment, the present study highlights the negative impacts of heatwaves on viticultural yields in the Douro region, which is critical information for stakeholders within the winemaking sector for planning suitable adaptation measures.