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…

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.

VineyardFACE: Investigation of a moderate (+20%) increase of ambient CO2 level on berry ripening dynamics and fruit composition

Climate change and rising atmospheric carbon dioxide concentration is a concern for agriculture, including viticulture. Studies on elevated carbon dioxide have already been on grapevines, mainly taking place in greenhouses using potted plants or using field grown vines under higher CO2 enrichment, i.e. >650 ppm. The VineyardFACE, located at Hochschule Geisenheim University, is an open field Free Air CO2 Enrichment (FACE) experimental set-up designed to study the effects of elevated carbon dioxide using field grown vines (Vitis vinifera L. cvs. Riesling and Cabernet Sauvignon). As the carbon dioxide fumigation started in 2014, the long term effects of elevated carbon dioxide treatment can be investigated on berry ripening parameters and fruit metabolic composition.
The present study aims to investigate the effect on fruit composition under a moderate increase (+20%; eCO2) of carbon dioxide concentration, as predicted for 2050 on both Riesling and Cabernet Sauvignon. Berry composition was determined for primary (sugars, organic acids, amino acids) and secondary metabolites (anthocyanins). Special focus was given on monitoring of berry diameter and ripening rates throughout three growing seasons. Compared to previous results of the early adaptative phase of the vines [1], our results show little effect of eCO2 treatment on primary metabolites composition in berries. However, total anthocyanins concentration in berry skin was lower for eCO2 treatment in 2020, although the ratio between anthocyanins derivatives did not differ.
[1] Wohlfahrt Y., Tittmann S., Schmidt D., Rauhut D., Honermeier B., Stoll M. (2020) The effect of elevated CO2 on berry development and bunch structure of Vitis vinifera L. cvs. Riesling and Cabernet Sauvignon. Applied Science Basel 10: 2486

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.

Modeling the suitability of Pinot Noir in Oregon’s Willamette Valley in a changing climate

Air temperature is the key driver of grapevine phenology and a significant environmental factor impacting yield and quality for a winegrape growing region. In this study the optimal downscaled CMIP5 ensemble for computing thegrowing season average temperature (GST) viticulture climate classification index was determined to spatially compute on a decadal basis predictions of the GST climate index and the grapevine sugar ripeness (GSR) model for Pinot Noir throughout the Willamette Valley (WV) American Viticultural Area (AVA). Forecasts for average temperature and a 220 g/L target sugar concentration level were computed using daily Localized Constructed Analogs (LOCA) downscaled CMIP5 historic and Representative Concentration Pathways (RCP) future climate projections of minimum and maximum daily temperature. We explore spatiotemporal trends of the GST climate classification index and Pinot Noir specific applications of the GSR phenology model for the WV AVA. Spatiotemporal computations of the GST climate index and Pinot Noir specific applications of the GSR model enable the opportunity to explore relationships between their computed values with one intent being to provide updated GST ranges that better align with current temperature-based modeling understanding of Pinot Noir grapevine phenology and the viticultural application of LOCA CMIP5 climate projections for the WV AVA. The Pinot Noir specific applications of the GSR model or the GST index with updated bounds indicate that the percent of the WV AVA area suitable for Pinot Noir production is currently at or near its peak value in the upper 80s to lower 90s of this century.

Ecophysiological performance of Vitis rootstocks under water stress

The use of rootstocks tolerant to soil water deficit is an interesting strategy to cope with limited water availability. Currently, several nurseries are breeding new genotypes, but the physiological basis of its responses under water stress are largely unknown. To this end, an ecophysiological assessment of the conventional 110-Richter (110R) and SO4, and the new M1 and M4 rootstocks was carried out in potted ungrafted plants. During one season, these Vitis genotypes were grown under greenhouse conditions and subjected to two water regimes, well-watered and water deficit. Water potentials of plants under water deficit down to < -1.4 MPa, and net photosynthesis (AN) <5 μmol m-2 s-1 did not cause leaf oxidative stress damage compared to well-watered conditions in any of the genotypes. The antioxidant capacity was sufficient to neutralize the mild oxidative stress suffered. Under both treatments, gravimetric differences in daily water use were observed among genotypes, leading to differences in the biomass of root, shoot and leaf. Under well-watered conditions, SO4 and 110R were the most vigorous and M1 and M4 the least. However, under water stress, SO4 exhibited the greatest reduction in biomass while M4 showed the lowest. Remarkably, under these conditions, SO4 reached the least negative stem water potential (Ψstem), while M1 reduced stomatal conductance (gs) and AN the most. In addition, SO4 and M1 genotypes also showed the highest and lowest hydraulic conductance values, respectively. Our results suggest that there are differences in water use regulation among genotypes, not only attributed to differences in stomatal regulation or intrinsic water use efficiency at the leaf level. Therefore, because no differences in canopy-to-root ratio were achieved, it is hypothesized that xylem vessel anatomical differences may be driving the reported differences among rootstocks performance. Results demonstrate that each Vitis rootstock differs in its ecophysiological responses under water stress.