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…

Making sense of available information for climate change adaptation and building resilience into wine production systems across the world

Effects of climate change on viticulture systems and winemaking processes are being felt across the world. The IPCC 6thAssessment Report concluded widespread and rapid changes have occurred, the scale of recent changes being unprecedented over many centuries to many thousands of years. These changes will continue under all emission scenarios considered, including increases in frequency and intensity of hot extremes, heatwaves, heavy precipitation and droughts. Wine companies need tools and models allowing to peer into the future and identify the moment for intervention and measures for mitigation and/or avoidance. Previously, we presented conceptual guidelines for a 5-stage framework for defining adaptation strategies for wine businesses. That framework allows for direct comparison of different solutions to mitigate perceived climate change risks. Recent global climatic evolution and multiple reports of severe events since then (smoke taint, heatwave and droughts, frost, hail and floods, rising sea levels) imply urgency in providing effective tools to tackle the multiple perceived risks. A coordinated drive towards a higher level of resilience is therefore required. Recent publications such as the Australian Wine Future Climate Atlas and results from projects such as H2020 MED-GOLD inform on expected climate change impacts to the wine sector, foreseeing the climate to expect at regional and vineyard scale in coming decades. We present examples of practical application of the Climate Change Adaptation Framework (CCAF) to impacts affecting wine production in two wine regions: Barossa (Australia) and Douro (Portugal). We demonstrate feasibility of the framework for climate adaptation from available data and tools to estimate historical climate-induced profitability loss, to project it in the future and to identify critical moments when disruptions may occur if timely measures are not implemented. Finally, we discuss adaptation measures and respective timeframes for successful mitigation of disruptive risk while enhancing resilience of wine systems.

Organic recycled mulches in sustainable viticulture: assessment of spontaneous plants communities and weed coverage

In recent years, developing more efficient and sustainable viticulture management has been essential due to the impact of climate change in semiarid regions. For this reason, the use of recycled organic mulching (ROM) in the vineyard has become an interesting strategy to cope with water stress, isolated soil from extreme temperatures and improving soil humidity, control the presence of weeds and therefore reduce the inputs of herbicides and improve soil fertility. This work aimed to analyse the effect of three different organic mulches [straw (S), grape pruning debris (GPD) and spent mushroom compost (SMC)] and two traditional soil management techniques [herbicide (H) and interrow (IN)] on weed coverage and the spontaneous plant communities’ presence. Data sampling was collected throughout the vine vegetative cycle of 2021 in La Rioja, Spain. The different soil management techniques had a clear effect on weed coverage and his development during the vine vegetative cycle. SMC and H were the treatments with the highest and the lowest coverage percentage, respectively. IN had a delayed weed emergence at the beginning of the vine vegetative cycle, but finally it reached maximum values nearby SMC. GPD and S had similar effects on weed emergence, reaching 25-30% of the maximum coverage values. A total of 29 herbaceous species were identified during the vegetative cycle, some of them very isolated and occasional. Principal component analysis (PCAs) showed a good association between spontaneous species and treatments, furthermore, specific species-treatment associations were found. Moreover, three clear groups of herbaceous communities were identified by cluster analysis. This study provides interesting information about the effect of different alternative soil management on herbaceous plant coverage and weed species communities which could contribute to making more sustainable viticulture.

Phenolic composition of Tempranillo Blanco grapes changes after foliar application of urea

Our research aimed to determine the effect and efficiency of foliar application of urea on the phenolic composition of Tempranillo Blanco grapes. The field experiment was carried out in 2019 and 2020 seasons and the plot was located in D.O.Ca Rioja (North of Spain). The vineyard was Vitis vinifera L. Tempranillo Blanco and grafted on Richter-110 rootstock. The treatments were control (C), whose plants were sprayed with water and three doses of urea: plants were sprayed with urea 3 kg N/ha (U3), 6 kg N/ha (U6) and 9 kg N/ha (U9). The applications were performed in two phenological stages, pre-veraison (Pre) and veraison (Ver). Also, each of the treatments was repeated one week later. Control and treatments were performed in triplicate and arranged in a randomised block design. Grapes were harvested at optimum ripening stage. High-performance liquid chromatography was used to analyse the phenolic composition of the grapes. Finally, the results obtained from the analytical determinations – flavonols, flavanols and non-flavonoid (hydroxybenzoic acids, hydroxycinnamic acids and stilbenes) – were studied statistically by analysis of variance. The results showed that, in 2019, U6-Pre and U9-Pre treatments increased the hydroxybenzoic acid content in grapes, and also all foliar treatments applied at Pre enhanced the stilbene concentration. Moreover, U3-Ver was the only treatment that rose flavonol and stilbene contents in the Tempranillo Blanco grapes. In 2020, all treatments applied at Pre enhanced the flavonol concentration in grapes. Furthermore, U3-Pre and U9-Pre treatments increased stilbene content in grapes. Nevertheless, the hydroxybenzoic acid content was improved by U6-Ver and U9-Ver and besides, hydroxycinnamic acid concentration in grapes was increased by all treatments applied at Ver. In conclusion, the lower and highest dose of urea (U3 and U9), applied at pre-veraison, were the best treatments to improve the Tempranillo Blanco grape phenolic composition.

Co-design and evaluation of spatially explicit strategies of adaptation to climate change in a Mediterranean watershed

Climate change challenges differently wine growing systems, depending on their biophysical, sociological and economic features. Therefore, there is a need to locally design and evaluate adaptation strategies combining several technical options, and considering the local opportunities and constraints (e.g. water access, wine typicity). The case study took place in a typical and heterogeneous Mediterranean vineyard of 1,500 ha in the South of France. We developed a participatory modeling approach to (1) conceptualize local climate change issues and design spatially explicit adaptation strategies with stakeholders, (2) numerically evaluate their effects on phenology, yield and irrigation needs under the high-emissions climate change scenario RCP 8.5, and (3) collectively discuss simulation results. We organized five sets of workshops, with in-between modeling phases. A process-based model was developed that allowed to evaluate the effects of six technical options (late varieties, irrigation, water saving by reducing canopy size, adjusting cover cropping, reducing density, and shading) with various distributions in the watershed, as well as vineyard relocation. Overall, we co-designed three adaptation strategies. Delay harvest strategy with late varieties showed little effects on decreasing air temperature during ripening. Water constraint limitation strategy would compensate for production losses if disruptive adaptations (e.g. reduced density) were adopted, and more land got access to irrigation. Relocation strategy would foster high premium wine production in the constrained mountainous areas where grapevine is less impacted by climate change. This research shows that a spatial distribution of technical changes gives room for adaptation to climate change, and that the collaboration with local stakeholders is a key to the identification of relevant adaptation. Further research should explore the potential of adaptation strategies based on soil quality improvement and on water stress tolerant varieties.

Combining effect of leaf removal and natural shading on grape ripening under two irrigation strategies in Manto negro (Vitis vinifera L.)

The increasingly frequent heat waves during grape ripening pose challenges for high quality wine grape production. Defoliation is a common practice that can improve the control of diseases in bunches, but also it increases the exposure to sunlight. Grapes exposed to solar radiation reach temperatures over the optimum for berry development and maturation. This makes the development of irrigation and canopy management techniques of great importance to maximize yield and grape quality. A field experiment was carried out during 2021 using Manto negro wine grapes to study the effect of applied irrigation and different light exposure levels on grape quality. Two irrigation treatments were imposed based on the frequency and amount of water doses in a four-block experimental vineyard at Bodega Ribas (Mallorca). Three light exposure treatments were randomly applied in each irrigation plot. The light treatments included exposed clusters from pea size, non-exposed clusters, and shaded clusters after softening. Leaf area index and canopy porosity was estimated every 2 weeks. Midday leaf water potential was measured weekly. Additionally, apparent electrical conductivity was measured between rows to estimate the soil water content variability. Light and temperature sensors were installed at the bunch level to quantify the differences in bunch temperature and light intensity among treatments. The effect of irrigation and cluster light exposure on berry weight, TSS, TA, malic acid, tartaric acid, K+, and pH were analysed at 5 moments along grape ripening. During different heat waves, the natural shading technique decreased the maximum bunch temperature around 10 °C respect to the exposed bunches in both irrigation strategies. The combination of defoliation and shading techniques after softening decreased TSS at harvest and affected most of the quality parameters during the last stages of ripening, showing an interesting technique to delay ripening in warm viticulture areas.