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…

Long-term drought resilience of traditional red grapevine varieties from a semi-arid region

In recent decades, the scarcity of water resources in agriculture in certain areas has been aggravated by climate change, which has caused an increase in temperatures, changes in rainfall patterns, as well as an increase in the frequency of extreme phenomena such as droughts and heat waves. Although the vine is considered a drought-tolerant specie, it has to satisfy important water requirements to complete its cycle, which coincides with the hottest and driest months. Achieving sustainable viticulture in this scenario requires high levels of efficiency in the use of water, a scarce resource whose use is expected to be severely restricted in the near future. In this regard, the use of drought-tolerant varieties that are able to maintain grape yield and quality could be an effective strategy to face this change. During three consecutive seasons (2018-2020) the behavior in rainfed regime of 13 traditional red grapevine varieties of the Spain central region was studied. These varieties were cultivated in a collection at Centro de Investigación de la Vid y el Vino de Castilla-La Mancha (IVICAM-IRIAF) located in Tomelloso (Castilla-La Mancha, Spain). Yield components (yield, mean bunch and berry weight, pruning weight), physicochemical parameters of the musts (brix degree, total acidity, pH) and some physiological parameters related with water stress during ripening period (δ13C, δ18O) were analysed. The application of different statistical techniques to the results showed the existence of significant differences between varieties in their response to stressful conditions. A few varieties highlighted for their high ability to adapt to drought, being able to maintain high yields due to their efficiency in the use of water. In addition, it was possible quantify to what extent climate can be a determinant in the δ18O of musts under severe water stress conditions.

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.

Effect of multi-level and multi-scale spectral data source on vineyard state assessment

Currently, the main goal of agriculture is to promote the resilience of agricultural systems in a sustainable way through the improvement of use efficiency of farm resources, increasing crop yield and quality under climate change conditions. This last is expected to drastically modify plant growth, with possible negative effects, especially in arid and semi-arid regions of Europe on the viticultural sector. In this context, the monitoring of spatial behavior of grapevine during the growing season represents an opportunity to improve the plant management, winegrowers’ incomes, and to preserve the environmental health, but it has additional costs for the farmer. Nowadays, UAS equipped with a VIS-NIR multispectral camera (blue, green, red, red-edge, and NIR) represents a good and relatively cheap solution to assess plant status spatial information (by means of a limited set of spectral vegetation indices), representing important support in precision agriculture management during the growing season. While differences between UAS-based multispectral imagery and point-based spectroscopy are well discussed in the literature, their impact on plant status estimation by vegetation indices is not completely investigated in depth. The aim of this study was to assess the performance level of UAS-based multispectral (5 bands across 450-800nm spectral region with a spatial resolution of 5cm) imagery, reconstructed high-resolution satellite (Sentinel-2A) multispectral imagery (13 bands across 400-2500 nm with spatial resolution of <2 m) through Convolutional Neural Network (CNN) approach, and point-based field spectroscopy (collecting 600 wavelengths across 400-1000 nm spectral region with a surface footprint of 1-2 cm) in a plant status estimation application, and then, using Bayesian regularization artificial neural network for leaf chlorophyll content (LCC) and plant water status (LWP) prediction. The test site is a Greco vineyard of southern Italy, where detailed and precise records on soil and atmosphere systems, in-vivo plant monitoring of eco-physiological parameters have been conducted.

Measurement of redox potential as a new analytical winegrowing tool

Excell laboratory has initiated the development of an analytical method based on electrochemistry to evaluate the ability of wines to undergo or resist to oxidative phenomena. Electrochemistry is a powerful tool to probe reactions involving electron transfers and offers possibility of real-time measurements. In that context, the laboratory has implemented electrochemical analysis to assess oxidation state of different wine matrices but also in order to evaluate oxidative or reduced character of leaf and soil. Initially, our laboratory focused on dosage of compounds involved in responses of plant stresses and we were also interested in microbiological activity of soils. These analyses were compared with the measurement of redox potential (Eh) and pH which are two fundamental variables involved in the modulation of plant metabolism. Indeed, the variation of redox states of the plant reflects its biological activity but also its capacity to absorb nutriments. The Eh-pH conditions mainly determine metabolic processes involved in soil and leaf and our goal is to determine if this combined analytical approach will be sufficiently precise to detect biological evolutions (plant health, parasitic attack…).

Effect of partial net shading on the temperature and radiation in the grapevine canopy, consequences on the grape quality of cv. Gros Manseng in PDO Pacherenc-du-vic-Bilh

As elsewhere, southwestern France vineyards face more recurrent summer heat waves these last years. Among the possibilities of adaptation to this climate changing parameter, the use of net shading is a technique that allow for limiting canopy exposure to radiations. In this trial, we tested net shading installed on one face of the canopy, on a north-south row-oriented plot of cv. Gros Manseng trained on VSP system in the PDO Pacherenc-du-Vic-Bilh. The purpose was to characterize the effects on the ambient canopy temperatures and radiations during the season and to observe the consequences on the composition of grapes and wines. Two sorts of net were used with two levels of obstruction (50% and 75%) of the photosynthesis active radiation (PAR). They have been installed on the west side of the canopy and compared to a netless control. Temperature and PAR sensors registered hourly data during the season. On specific summer day (hot and sunny) manual measurements took also place on bunches (temperature) and in different spots of the canopy (PAR). The results showed that, on clear days, the radiation is lowered by the shade nets respecting the supplier criteria. The effects on the ambient canopy temperature were inconstant on this plot when we observed the data from the global period of shading between fruit set and harvest. However, during hot days (>30°C), the temperature in the canopy was reduced during afternoon and the temperature of the bunch surface was reduced as well comparing to the control. A decrease of the maturity parameters of the berries, sugar and acidity, was also observed. Concerning the wine aromatic potential, no differences clearly appeared.