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…

Vineyards and clay minerals: multi-technique analytical approach and correlations with soil properties

Purpose of this research is to quantitatively assess the mineral component of vineyard soils, with particular attention to the mineralogical analysis of clays, which represent an element of high importance in the vineyard culture as well as in general agriculture. An X-ray diffraction (XRD) / thermogravimetric (TG) multi-technique analytical approach was developed, tested on soil samples taken from vineyards around the world. This codified analytical procedure was necessary to obtain precise qualitative and quantitative mineralogical data, globally comparable to distinguish the geopedological identity of the vineyards. Soil samples from vineyards of various locations were analysed, in very different geological conditions. The bulk-rock quantitative phase analysis (QPA) was obtained by the Rietveld method while the detailed composition of the clay-sized fraction was determined by modelling of the oriented X-ray diffraction patterns. The research provided a precise classification of the mineral component of soils, distinguishing the mineral phases of the clays and the so-called mixed-layer clay minerals. We found that the content in mixed layers can be directly correlated with the water retention and the cation exchange capacity ​​of the soil, while the presence of other clayey minerals and phyllosilicates in this research did not affect this CEC parameter, which codes the fertility level of the soils. The study demonstrates that terroir, in particular soils formed in complex or very different geological conditions, can only be effectively interpreted by properly analysing its mineral phases, in particular the mixed-layer clay component. These are characteristic abiotic ecological indicators, which may have specific eco-physiological influences on the plant.

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.

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…

Effect of regulated deficit irrigation regime on amino acids content of Monastrell (Vitis vinifera L.) grapes

Irrigation is an important practice to influence vine quality, especially in Mediterranean regions, characterized by hot summers and severe droughts during the growing season. This study focused on deficit irrigation regime influence on amino acids composition of Monastrell grapevines under semiarid conditions (Albacete, Southeastern of Spain). In 2019, two treatments were applied: non-irrigation (NI) and regulated deficit irrigation (RDI), watered at 30% of the estimated crop evapotranspiration from fruit set to onset of veraison. Grape amino acids content was analyzed by HPLC. Berries from non-irrigated vines showed higher concentration of several amino acids, such as tryptophan (73%), arginine (70%), lysine (36%), isoleucine (27%), and leucine (21%), compared to RDI grapes. Arginine is, together with ammonium ion, the principal nitrogen source for yeasts during the alcoholic fermentation; while isoleucine, tryptophan, and leucine are precursors of fermentative volatile compounds, key compounds for wine quality. Moreover, NI treatment increased in a 14% the total amino acids content in grapes compared to RDI treatment. The reported effects might be because yield was 70% higher in RDI vines than in the NI ones and, therefore, the sink demand was increased in the irrigated vines. In addition, NI vines suffered more severe water stress and it is known that the amino acids synthesis and accumulation can be influenced by the plant response to stress. According to the results, the irrigation regime showed effect on amino acids concentration in Monastrell grapes under semiarid conditions. Grapes from non-irrigated vines showed a higher content of several amino acids relevant to the fermentative process and to the wine aroma compounds formation. It is demonstrated that the final content of nitrogen-related components in grapes is influenced by the irrigation regime. The convenience of the irrigation strategy to suggest will depend on the desired wine style and the target yield levels.

Diagnosis of soil quality and evaluation of the impact of viticultural practices on soil biodiversity in a vineyard in southwestern France

Viticulture is facing two major changes – climate change and agroecological transition. In both cases, soil quality is seen as a lever to move towards a more sustainable viticulture. However, soil biological quality is little considered in the implementation of viticultural practices. Gascogn’Innov (2017-2022) is an Operational Group funded by the European Innovation Partnership for Agriculture. As such, it brings together winegrowers from the south-west of France, scientists, advisors and technicians, around a project focused on viticultural soil biological functioning and the design of technical routes more respectful toward soil heritage. To achieve this, the project aims to acquire references on the impact of viticultural practices on soil biology from a dynamic way, and to test a methodology to integrate information provided by the soil bioindicators to manage farming systems. A set of indicators of soil biological quality are evaluated in the project: microorganisms (bacteria and fungi abundance and diversity), fauna (abundance and diversity of nematodes and earthworms), physico-chemical characteristics, soil structure assessment and degradation rate of organic matter. Based on a network of 13 plots that have been subject to an initial diagnosis in 2017, several agronomical practices to restore soil fertility are experimented to redesign the cropping system (for instance plant cover, organic matter inputs, reduction of herbicides, mineral fertilizers). System redesign was made in collaboration by winegrowers and an interdisciplinary group of experts (agronomists, biologists). Several indicators are measured on vine and soil at each vintage to assess vine health and productivity. At the end of the project (2021), a final diagnosis was carried out. Gascogn’Innov allowed to create a regional database on the quality of wine-growing soils, which permitted to evaluate the effect of practices according to soil types. Especially, decreasing the intensity of tillage and increasing the duration and diversity of grass coverage tends to increase the abundance of all the organisms studied. This project confirmed the value of soil biological quality indicators to drive the sustainability of practices, but also highlighted the key-role of expertise, in both agronomy and soil biology, to help winegrowers understand and appropriate their soil quality diagnoses.