IVAS 2022 banner
IVES 9 IVES Conference Series 9 IVAS 9 IVAS 2022 9 Applications of FTIR microspectroscopy in oenology: shedding light on Saccharomyces cerevisiae cell wall composition and autolytic capacity

Applications of FTIR microspectroscopy in oenology: shedding light on Saccharomyces cerevisiae cell wall composition and autolytic capacity

Abstract

Many microbial starters for the alcoholic and malolactic fermentation processes are commercially available, indicated for diverse wine styles and quality goals. The screening protocols cover a wide range of oenologically relevant features, although some characteristics could also be studied using underexplored powerful techniques. In this study, we applied Fourier Transform Infrared (FTIR) microspectroscopy [1,2] to compare the cell wall biochemical composition and monitor the autolytic process in several wine strains of Saccharomyces cerevisiae. After cell death, autolysis trigger the release of mannoproteins and β-glucans, major components of yeast cell walls, influencing color, aroma, body, foaming properties, and stability of wine. Moreover, they can stimulate the metabolism of malolactic bacteria, while some fatty acids, also released during autolysis, act as inhibitors. Analysis of the cell wall structure was carried out both on cells grown in liquid medium and on cell walls previously separated from the other components. The autolytic capacity was assayed by sampling the cells at different times during induced autolysis in clarified and pasteurized must. From five to ten infrared spectra were acquired on each sample in the 4,000-700 cm-1 range in Attenuated Total Reflection on a 50×50 μm2 area. Spectra were analyzed after pretreatment through Hierarchical Cluster Analysis and Principal Component Analysis. Preliminary results were evaluated in relation to conventional spectrophotometric methods to quantify mannoproteins and β-glucans. The thickness of the cell walls was determined by means of scanning (SEM) and transmission electron microscopy (TEM). From the point of view of cell wall composition two groups of yeasts were distinguished by multivariate statistical analysis on the FTIR spectra since the strains EC1118, MY11 and PEDRO2000E showed higher absorption bands of mannoproteins and β-glucans. With conventional methods, the cell walls of the first two strains, alongside K1 and MY8, displayed a higher content of parietal polysaccharides, while the latter had the thickest wall among all the tested yeasts. The strains BM45 and D47 have a thinner surface structure. Regarding the autolytic process, again two different clusters were found distinguishing the behavior of the strains EC1118 and FRB with a similar timing of autolysis on one side from CH and Q20 on the other side. Furthermore, the latter strain presented a higher absorption in the spectral zone related to lipids, which can be correlated with a greater release of fatty acids in the medium. In conclusion, FTIR microspectroscopy proved to be an accurate and informative technique, suitable to highlight profound differences among S. cerevisiae strains as concerns both the content of parietal polysaccharides and the evolution of autolysis. Thus, this technique may become an option for the selection of starter cultures with properties fo great interest for the wine sector.

References

[1] Burattini, E., Cavagna, M., Dell’Anna, R., Malvezzi Campeggi, F., Monti, F., Rossi, F., & Torriani, S. (2008). A FTIR microspectroscopy study of autolysis in cells of the wine yeast Saccharomyces cerevisiae. Vibrational Spectroscopy, 47(2), 139-147. https://doi.org/10.1016/j.vibspec.2008.04.007.
[2] Cavagna, M., Dell’Anna, R., Monti, F., Rossi, F., & Torriani, S. (2010). Use of ATR-FTIR microspectroscopy to monitor autolysis of Saccharomyces cerevisiae cells in a base wine. Journal of Agricultural and Food Chemistry, 58(1), 39–45. https://doi.org/10.1021/jf902369s.

DOI:

Publication date: June 27, 2022

Issue: IVAS 2022

Type: Poster

Authors

Martelli Francesco1, Binati Renato Leal1, Monti Francesca2, Felis Giovanna1 and Torriani Sandra1

1Department of Biotechnology, University of Verona 
2Department of Computer Science, University of Verona

Contact the author

Keywords

FTIR microspectroscopy; starter cultures; Saccharomyces cerevisiae; autolysis; wine quality

Tags

IVAS 2022 | IVES Conference Series

Citation

Related articles…

Estimating bulk stomatal conductance of grapevine canopies

In response to changes in their environment, grapevines regulate transpiration using various physiological mechanisms that alter conductance of water through the soil-plant-atmosphere continuum. Expressed as bulk stomatal conductance at the canopy scale, it varies diurnally in response to changes in vapor pressure deficit and net radiation, and over the season to changes in soil water deficits and hydraulic conductivity of both soil and plant. It is necessary to characterize the response of conductance to these variables to better model how vine transpiration also responds to these variables. Furthermore, to be relevant for vineyard-scale modeling, conductance is best characterized using data collected in a vineyard setting. Applying a crop canopy energy flux model developed by Shuttleworth and Wallace, bulk stomatal conductance was estimated using measurements of individual vine sap flow, temperature and humidity within the vine canopy, and estimates of net radiation absorbed by the vine canopy. These measurements were taken on several vines in a non-irrigated vineyard in Bordeaux France, using equipment that did not interfere with ongoing vineyard operations. An inverted Penman-Monteith equation was then used to calculate bulk stomatal conductance on 15-minute intervals from July to mid-September 2020. Time-series plots show significant diurnal variation and seasonal decreases in conductance, with overall values similar to those in the literature. Global sensitivity analysis using non-parametric regression found transpiration flux and vapor pressure deficit to be the most important input variables to the calculation of bulk stomatal conductance, with absorbed net radiation and bulk boundary layer conductance being much less important. Conversely, bulk stomatal conductance was one of the most important inputs when calculating vine transpiration, further emphasizing the need for characterizing its response to environmental changes for use in vineyard water use modeling.

Genotypic variability in root architectural traits and putative implications for water uptake in grafted grapevine

Root system architecture (RSA) is important for soil exploration and edaphic resources acquisition by the plant, and thus contributes largely to its productivity and adaptation to environmental stresses, particularly soil water deficit. In grafted grapevine, while the degree of drought tolerance induced by the rootstock has been well documented in the vineyard, information about the underlying physiological processes, particularly at the root level, is scarce, due to the inherent difficulties in observing large root systems in situ. The objectives of this study were to determine genetic differences in the root architectural traits and their relationships to water uptake in two Vitis rootstocks genotypes (RGM, 140Ru) differing in their adaptation to drought. Young rootstocks grafted upon the Riesling variety were transplanted into cylindrical tubes and in 2D rhizotrons under two conditions, well watered and moderate water stress. Root traits were analyzed by digital imaging and the amount of transpired water was measured gravimetrically twice a week. Root phenotyping after 30 days reveal substantial variation in RSA traits between genotypes despite similar total root mass; the drought-tolerant 140Ru showed higher root length density in the deep layer, while the drought-sensitive RGM was characterised by shallow-angled root system development with more basal roots and a larger proportion of fine roots in the upper half of the tube. Water deficit affected canopy size and shoot mass to a greater extent than root development and architectural-related traits for both 140Ru and RGM, suggesting vertical distribution of roots was controlled by genotype rather than plasticity to soil water regime. The deeper root system of 140Ru as compared to RGM correlated with greater daily water uptake and sustained stomata opening under water-limited conditions but had little effect on above-ground growth. Our results highlight that grapevine rootstocks have constitutively distinct RSA phenotypes and that, in the context of climate change, those that develop an extensive root network at depth may provide a desirable advantage to the plant in coping with reduced water resources.

Delaying irrigation initiation linearly reduces yield with little impact on maturity in Pinot noir

When to initiate irrigation is a critical annual management decision that has cascading effects on grapevine productivity and wine quality in the context of climate change. A multi-site trial was begun in 2021 to optimize irrigation initiation timing using midday stem water potential (ψstem) thresholds characterized as departures from non-stressed baseline ψstemvalues (Δψstem). Plant material, vine and row spacing, and trellising systems were concomitant among sites, while vine age, soil type, and pruning systems varied. Five target Δψstem thresholds were arranged in an RCBD and replicated eight times at each site: 0.2, 0.4, 0.6, 0.8, and 1.0 MPa (T1, T2, T3, T4, and T5, respectively). When thresholds were reached, plots were irrigated weekly at 70% ETc. Yield components and berry composition were quantified at harvest. To better generalize inferences across sites, data were analyzed by ANOVA using a mixed model including site as a random factor. Across sites, irrigation was initiated at Δψstem = 0.24, 0.50, 0.65, 0.93, and 0.98 MPa for T1, T2, T3, T4, and T5, respectively. Consistent significant negative linear trends were found for several key yield and berry composition variables. Yield decreased by 12.9, 15.9, 19.5, and 27.4% for T2, T3, T4, and T5, respectively, compared to T1 (p < 0.0001) across sites that were driven by similarly linear reductions in berry weight (p < 0.0001). Comparatively, berry composition varied little among treatments. Juice total soluble solids decreased linearly from T1 to T5 – though only ranged 0.9 Brix (p = 0.012). Because producers are paid by the ton, and contracts simply stipulate a target maturity level, first-year results suggest that there is no economic incentive to induce moderate water deficits before irrigation initiation, regardless of vineyard site. Subsequent years will further elucidate the carryover effects of delaying irrigation initiation on productivity over the long term.

Using δ13C and hydroscapes as a tool for discriminating cultivar specific drought response

Measurement of carbon isotope discrimination in berry juice sugars at maturity (δ13C) provides an integrated assessment of water use efficiency (WUE) during the period of berry ripening, and when collected over multiple seasons can be used as an indication of drought stress response. Berry juice δ13C measurements were carried out on 48 different varieties planted in a common garden experiment in Bordeaux, France from 2014 through 2021 and were paired with midday and predawn leaf water potential measurements on the same vines in a subset of six varieties. The aim was to discriminate a large panel of varieties based on their stomatal behaviour and potentially identify hydraulic traits characterizing drought tolerance by comparing δ13C and hydroscapes (the visualisation of plant stomatal behaviour as a response to predawn water potential). Cluster analysis found that δ13C values are likely affected by the differing phenology of each variety, resulting in berry ripening of different varieties taking place under different stress conditions within the same year. We accounted for these phenological differences and found that cluster analysis based on specific δ13C metrics created a classification of varieties that corresponds well to our current empirical understanding of their relative drought tolerances. In addition, we analysed the water potential regulation of the subset of six varieties (using the hydroscape approach) and found that it was well correlated with some δ13C metrics. Surprisingly, a variety’s water potential regulation (specifically its minimum critical leaf water potential under water deficit) was strongly correlated to δ13C values under well-watered conditions, suggesting that base WUE may have a stronger impact on drought tolerance than WUE under water deficit. These results give strong insights on the innate WUE of a very large panel of varieties and suggest that studies of drought tolerance should include traits expressed under non-limiting conditions.

Different soil types and relief influence the quality of Merlot grapes in a relatively small area in the Vipava Valley (Slovenia) in relation to the vine water status

Besides location and microclimatic conditions, soil plays an important role in the quality of grapes and wine. Soil properties influence…