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…

Deconstructing the soil component of terroir: from controversy to consensus

Wine terroir describes the collectively recognized relation between a geographical area and the distinctive organoleptic characteristics of the wines produced in it. The overriding objective in terroir studies is therefore to provide scientific proof relating the properties of terroir components to wine quality and typicity. In scientific circles, the role of climate (macro-, meso- and micro-) on grape and wine characteristics is well documented and accepted as the most critical. Moreover, there has been increasing interest in recent years about new elements with possible importance in shaping wine terroir like berry/leaf/soil microbiology or even aromatic plants in proximity to the vineyard conferring flavors to the grapes. However, the actual effect of these factors is also dependent on complex interactions with plant material (variety/clone, rootstock, vine age) and with human factors.
The contribution of soil, although a fundamental component of terroir and extremely popular among wine enthusiasts, remains a much-debated issue among researchers. The role of geology is probably the one mostly associated by consumers with the notion of terroir with different parent rocks considered to give birth to different wine styles. However, the relationship between wine properties and the underlying parent material raises a lot of controversy especially regarding the actual existence of rock-derived flavors in the wine (e.g. minerality). As far as the actual soil properties are concerned, the effect of soil physical properties is generally regarded as the most significant (e.g sandy soils being associated with lighter wines while those on clay with colored and tannic ones) mostly through control of water availability which ultimately modifies berry ripening conditions either directly by triggering biosynthetic pathways, or indirectly by altering vigor and yield components. The role of soil chemistry seems to be weakly associated to wine sensory characteristic, although N, K, S and Ca, but also soil pH, are often considered important in the overall soil effect.
Recently, in the light of evidence provided by precision agriculture studies reporting a high variability of vineyard soils, the spatial scale should also be taken into consideration in the evaluation of the soil effects on wines. While it is accepted that soil effects become more significant than climate on a local level, it is not clear whether these micro-variations of vineyard soils are determining in the terroir effect. Moreover, as terroir is not a set of only natural factors, the magnitude of the contribution of human-related factors (irrigation, fertilization, soil management) to the soil effect still remains ambiguous. Lastly, a major shortcoming of the majority of works about soil effects on wine characteristics is the absence of connection with actual vine physiological processes since all soil effects on grape and wine chemistry and sensorial properties are ultimately mediated through vine responses.
This article attempts to breakdown the main soil attributes involved in the terroir effect to suggest an improved understanding about soil’s true contribution to wine sensory characteristics. It is proposed that soil parameters per se are not as significant determining factors in the terroir effect but rather their mutual interactions as well as with other natural and human factors included in the terroir concept. Consequently, similarly to bioclimatic indices, composite soil indices (i.e. soil depth, water holding capacity, fertility, temperature etc), incorporating multiple soil parameters, might provide a more accurate and quantifiable means to assess the relative weight of the soil component in the terroir effect.

Water deficit differentially impacts the performances and the accumulation of grape metabolites of new varieties tolerant to fungi

The use of resistant varieties is a long-term but promising solution to reduce chemical input in viticulture. Several important breeding programs in Europe and abroad are now releasing a range of new hybrids performing well regarding fungi susceptibility and producing good quality wines. Unfortunately, insufficient attention is paid by the breeders to the adaptation of these varieties to climatic changes, notably to the increased climatic demand and water deficit (WD). Thus, prior to the adoption of such varieties by the wine industry in Mediterranean regions, there is a need to consider their suitability to WD. This study aimed to characterize the different drought-strategies adopted by 6 new resistant varieties selected by INRAE in comparison to Syrah. To allow the assessment of long-term impacts of WD, field-grown vines were exposed to contrasted WD from 2018 to 2021 under a semi-arid Mediterranean climate. A gradient of WD was applied in the field and controlled through plant measurements at the single plant level. Grape development was non-destructively monitored to determine the arrest of berry phloem unloading. The impacts of WD on berry composition, including water, primary metabolites (sugars, organic acids), secondary metabolites (anthocyanins, thiols precursors) and main cations contents, were assessed at this specific stage. Results showed different varietal responses during the year and inter-annual acclimation in terms of plant water use efficiency, biomass accumulation, as well as yield components and berry composition. WD differentially reduced the accumulation of primary metabolites at plant and berry levels, but it little changed their concentrations in the fruits at the ripe stage. Moreover, WD differentially impacted the accumulation of secondary metabolites and major cations between the varieties. In the talk, we’ll present the main results regarding the WD impacts on fruit metabolites and enlarge the reflection about the practical assessment of the grapevine acclimation to WD.

Exploring resilience and competitiveness of wine estates in Languedoc-Roussillon in the recent past: a multi-level perspective

The Languedoc-Roussillon wineries are facing a decline in wine yields particularly PGI yields due to many factors. Climate change is just ones, but is expected to increase in the future. There is also structurally a large heterogeneity of yield profiles among terroirs, varieties and strategies. This work investigates the link between yield, competitiveness and resilience to explore how resilient winegrowers have been in the recent past. To this end two approaches have been combined; (i) an accountancy database analysis at estate scale and (ii) municipality level competitiveness analysis. A new resilience indicator that characterizes the capacity of an estate to absorb yield variation is also defined. The FADN database between 2000 and 2018 of ex-Languedoc-Roussillon (France) and other data are used to analyse the current situation and the past evolution of competitiveness and resilience by type of estate (type of farm: PGI and/or PDO & type of commercialization: bulk and/or bottles). The net margin, which defines competitiveness, is not correlated to yield for all types but depends on the type of commercialization and the level of specialisation. The resilience indicator shows that the net margin of estates specialized in PGI is particularly sensitive to yield declines. We also show that price evolutions seem to compensate the effect of yield losses for the majority of types. Municipality scale analysis shows the links between local pedoclimate, yield, commercialization strategies and price. Overlapping a PDO with a PGI does not always increase a municipality’s PGI competitiveness. It is difficult to make links between causes and effects due to the complexity of the wine production system. Production diversification may be a solution. Resorting to the two level of analysis helps resolving the data gap that is necessary to explore the links between yield and economic performance of the wine estates in the long term.

Grapevine yield estimation in a context of climate change: the GraY model

Grapevine yield is a key indicator to assess the impacts of climate change and the relevance of adaptation strategies in a vineyard landscape. At this scale, a yield model should use a number of parameters and input data in relation to the information available and be able to reproduce vineyard management decisions (e.g. soil and canopy management, irrigation). In this study, we used data from six experimental sites in Southern France (cv. Syrah) to calibrate a model of grapevine yield limited by water constraint (GraY). Each yield component (bud fertility, number of berries per bunch, berry weight) was calculated as a function of the soil water availability simulated by the WaLIS water balance model at critical phenological phases. The model was then evaluated in 10 grapegrowers’ plots, covering a diversity of biophysical and technical contexts (soil type, canopy size, irrigation, cover crop). We identified three critical periods for yield formation: after flowering on the previous year for the number of bunches and berries, around pre-veraison and post-veraison of the same year for mean berry weight. Yields were simulated with a model efficiency (EF) of 0.62 (NRMSE = 0.28). Bud fertility and number of berries per bunch were more accurately simulated (EF = 0.90 and 0.77, NRMSE = 0.06 and 0.10, respectively) than berry weight (EF = -0.31, NRMSE = 0.17). Model efficiency on the on-farm plots reached 0.71 (NRMSE = 0.37) simulating yields from 1 to 8 kg/plant. The GraY model is an original model estimating grapevine yield evolution on the basis of water availability under future climatic conditions.  It allows to evaluate the effects of various adaptation levers such as planting density, cover crop management, fruit/leaf ratio, shading and irrigation, in various production contexts.

Underpinning terroir with data: rethinking the zoning paradigm

Agriculture, natural resource management and the production and sale of products such as wine are increasingly data-driven activities. Thus, the use of remote and proximal crop and soil sensors to aid management decisions is becoming commonplace and ‘Agtech’ is proliferating commercially; mapping, underpinned by geographical information systems and complex methods of spatial analysis, is widely used. Likewise, the chemical and sensory analysis of wines draws on multivariate statistics; the efficient winery intake of grapes, subsequent production of wines and their delivery to markets relies on logistics; whilst the sales and marketing of wines is increasingly driven by artificial intelligence linked to the recorded purchasing behaviour of consumers. In brief, there is data everywhere!

Opinions will vary on whether these developments are a good thing. Those concerned with the ‘mystique’ of wine, or the historical aspects of terroir and its preservation, may find them confronting. In contrast, they offer an opportunity to those interested in the biophysical elements of terroir, and efforts aimed at better understanding how these impact on vineyard performance and the sensory attributes of resultant wines. At the previous Terroir Congress, we demonstrated the potential of analytical methods used at the within-vineyard scale in the development of Precision Viticulture, in contributing to a quantitative understanding of regional terroir. For this conference, we take this approach forward with examples from contrasting locations in both the northern and southern hemispheres. We show how, by focussing on the vineyards within winegrowing regions, as opposed to all of the land within those regions, we might move towards a more robust terroir zoning than one derived from a mixture of history, thematic mapping, heuristics and the whims of marketers. Aside from providing improved understanding by underpinning terroir with data, such methods should also promote improved management of the entire wine value chain.