OENO IVAS 2019 banner
IVES 9 IVES Conference Series 9 OENO IVAS 9 OENO IVAS 2019 9 Grape and wine microorganisms: diversity and adaptation 9 Influence of cell-cell contact on yeast interactions and exo-metabolome

Influence of cell-cell contact on yeast interactions and exo-metabolome

Abstract

Alcoholic fermentation is the main step for winemaking, mainly performed by the yeast Saccharomyces cerevisiae. But other wine yeasts called non-Saccharomyces may contribute to alcoholic fermentation and modulate the wine aroma complexity. The recurrent problem with the use of these non-Saccharomyces yeasts is their trend to die off prematurely during alcoholic fermentation, leading to a lack of their interesting aromatic properties searched in the desired wine. This phenomenon appears to be mainly due to interactions with S. cerevisiae. These interactions are often negatives but remain unclear because of the species and strain specific response. Among the non-Saccharomyces yeasts, Lachancea thermotolerans is a wine yeast naturally found in grape must and well known as a great L-lactic acid producer and an aromatic molecules enhancer, but its behavior during alcoholic fermentation can be completely different in co-fermentation with S. cerevisiae in function of strain used. Thus, S. cerevisiae/L. thermotolerans couple was used to unravel interactions between these two species during alcoholic fermentation. Thanks to a modified S. cerevisiae strain expressing a GFP allowing discrimination between yeast populations, both yeast viability was monitored by flow cytometry in pure and sequential fermentations of grape must with or without cell-cell contact. This reveal a decrease in cell viability for S. cerevisiae in both conditions with a greater decrease in case of cell-cell contact. Investigating the nature of the interactions, our results demonstrate a competition between species for nitrogen compounds, oxygen and for the first time a competition for must sterols. Volatile compounds analysis revealed changes in sequential fermentations compared to pure fermentations and showed also that cell-cell contact modify yeast metabolism since the volatile compound profile was significantly different from sequential fermentation without cell-cell contact. Yeast metabolism modifications associated with cell-cell contact were confirmed further by analyzing the exo-metabolome of all fermentations by FT-ICR-MS analysis. These analyses show for the first time a specific metabolite production and quantitative metabolite changes linked to each fermentation condition. This study shows that cell-cell contact not only impact cell viability as already reported but deeply changes the yeast metabolism.

DOI:

Publication date: June 23, 2020

Issue: OENO IVAS 2019

Type: Article

Authors

Clément Petitgonnet, Géraldine L.Klein, Chloé Roullier-Gall, Philippe Schmitt-Kopplin, Beatriz Quintanilla-Casas, Stefania Vichi, Diane Julien-David, Hervé Alexandre

Helmholtz Zentrum Muenchen, Research unit Analytical BioGeoChemistry, Ingolstaedter Landstrasse 1, 85764 Neuherberg, Germany 
Technische Universität Muenchen, Analytical Food Chemistry, Alte Akademie 10, 85354 Freising, Germany 
Nutrition, Food Science and Gastronomy Department, INSA – XaRTA (Catalonian Reference Network on Food Technology), University of Barcelona, Santa Coloma de Gramenet, Spain 
Université de Strasbourg, CNRS, IPHC UMR 7178, F-67000 Strasbourg, France

Contact the author

Keywords

Interactions, Yeast, Flow cytometry, Exo-metabolome 

Tags

IVES Conference Series | OENO IVAS 2019

Citation

Related articles…

Local ancient grapevine cultivars to face future viticulture

Among the different strategies to cope with the negative impacts of climate change on viticulture, the exploitation of genetic diversity is one of the most promising to adapt to new conditions and maintain wine production and quality. One of the biggest concerns in the context of climate change is to improve water use efficiency (WUE). In this way, the use of genotypes that present a better response to drought and high WUE is a key issue. In this work, physiological performance analysis was conducted to compare the water deficit stress (WDS) responses of local and widespread grapevines cultivars. Leaf gas exchange, water use efficiency (WUE) at different levels (leaf and long-term WUE (∆13C)), leaf osmotic adjustment and other water relations parameters were determined in plants under well-watered and WDS conditions alongside assessment of the levels of foliar hormones concentrations. Results denote that local cultivars displayed better physiological performance under WDS as compared to the widely-distributed ones. he results corroborate the hypothesis that better stomatal control allows increasing leaf WUE under drought as occurred in the local Callet cv.; but the minority local cultivar Escursac cv. showed high WUE under both treatments. In this case, high WUE can be related to maintaining higher photosynthetic activity under drought. The different mechanisms underlying the better performance under WDS and high WUE of minority local cultivars are discussed.

How distinctive are single vineyard Gewürztraminer musts and wines from Alto Adige (Italy) based on untargeted analysis, sensory profiling, and chemometric elaboration?

Vitis vinifera L. ‘Gewürztraminer’ is a historical grape variety of Alto Adige (Südtirol), Italy, which is widely grown in the area of Tramin an der Weinstraße, but is also grown globally. It produces highly aromatic wines that are strongly influenced by the terroir of the vineyard sites where they are grown. This study looked at musts and young wines from ‘Gewürztraminer’ grapes harvested in seven distinct vineyards near Tramin and then processed at Cantina di Termeno, minimizing winemaking protocol variability. Samples were profiled using bidimensional gas chromatography–time-of-flight mass spectrometry, liquid chromatography coupled to electrochemical detection, and near-IR spectrometry. The data were subjected to Principle Component Analysis and Hierarchical Clustering Analysis. Sensory discriminant testing was undertaken using the sorting method with a semi-trained panel, and the data were processed using Multidimensional Scaling. Seven must/wine pairs could be distinguished based on their untargeted volatilome profiles and on sensory evaluation. As expected, there were greater differences in the volatile compounds between the wines than between the musts. The wines from vineyards 4 and 5 were nonetheless quite homogenous in terms of chemical and sensory analyses, as were the wines from vineyards 1 and 3. For the phenolic profile, differences were noted between the musts and wines of vineyards 2, 3, and 4, but the musts from vineyards 5 and 7 were similar. Sensory analysis showed the wines from vineyards 6 and 7 to be distinct from the rest. These results reinforce that the composition of ‘Gewürztraminer’ musts and wines is strongly determined by vineyard site, even in a small geographic area with high variability of the terroir (soil and microclimate), and that these differences are apparent in the flavours and aromas of the finished wines. Further confirmation would require a larger sample of wines, preferably from several vintages.

The use of rootstock as a lever in the face of climate change and dieback of vineyard

As viticulture faces challenges such as climate change or vineyard dieback, the choice of the variety and rootstock becomes more and more crucial. To study rootstock levers in the Bordeaux region, a parcel of Cabernet Sauvignon (CS) was planted with four rootstocks in 2014. Twenty repetitions of each of the following four rootstocks were set up: 101-14 MGt, Nemadex AB, 420A MGt and Gravesac. The number of bunches, yields and pruning weights of the vine shoots were measured individually on 240 vines from 2017 to 2021. Since 2020, nitrogen status assessed by assimilable nitrogen level, hydric status assessed by δ13C and berry maturity were measured on 80 samples taken from 20 repetitions of the four rootstocks. A lower yield was measured for CS grafted onto Nemadex AB due to the lower number of bunches and the lower weight of berries. The differences between the other three rootstocks are small, but CS grafted onto 420A MGt was the most productive. The CS grafted onto Nemadex AB had the lowest pruning weight while 101-14 MGt had the highest. In 2020, δ13C showed a more moderate water stress with 101-14 MGt and 420A MGt than with Nemadex AB. Surprisingly, the Gravesac was under more stress than the 101-14 MGt. The nitrogen status in the berries was better for Nemadex AB but this was perhaps due to the significantly lower weight of the berries.Rootstock 101-14 MGt attained the highest accumulation of sugars in the berries while 420A MGt allows to preserve higher acidity. The parcel is still young which may explain some of the results. These measures must therefore be continued over the next several years to fully assess the effects of these rootstocks on the development of the vines and the quality of the production under new climatic conditions.

Downscaling of remote sensing time series: thermal zone classification approach in Gironde region

In viticulture, the challenges of local climate modelling are multiple: taking into account the local environment, fine temporal and spatial scales, reliable time series of climate data, ease of implementation and reproducibility of the method. At the local scale, recent studies have demonstrated the contribution of spatialization methods for ground-based climate observation data considering topographic factors such as altitude, slope, aspect, and geographic coordinates (Le Roux et al, 2017; De Rességuier et al, 2020). However, these studies have shown questions in terms of the reproducibility and sustainability of this type of climate study. In this context, we evaluated the potential of MODIS thermal satellite images validated with ground-based climate data (Morin et al, 2020). Previous studies have been encouraging, but questions remain to be explored at the regional scale, particularly in the dynamics of the massive use of bioclimatic indices to classify the climate of wine regions. The results at the local scale were encouraging, but this approach was tested in the current study at the regional scale. Several objectives were set: 1) to evaluate the downscaling method for land surface temperature time series, 2) to identify regional thermal structure variations. We used weekly minimum and maximum surface temperature time series acquired by MODIS satellites at a spatial resolution of 1000 m and downscaled at 500 m using topographical variables. Two types of analyses were performed:

Metabolomic discrimination of grapevine water status for Chardonnay and Pinot noir

Water status impact in viticulture has been widely explored, as it strongly affects grapevine physiology and grape chemical composition. It is considered as a key component of vitivinicultural terroir. Most of the studies concerning grapevine water status have focused on either physiological traits, or berry compounds, or traits involved in wine quality. Here, the response of grapevine to water availability during the ripening period is assessed through non-targeted metabolomics analysis of grape berries by ultra-high resolution mass spectrometry. The grapevine water status has been assessed during 2 consecutive years (2019 & 2020), through carbon isotope discrimination on juices from berries collected at maturity (21.5 brix approx.) for 2 Vitis vinifera cv. Pinot noir (PN) and Chardonnay (CH). A total of 220 grape juices were collected from 5 countries worldwide (Italy; Argentina; France; Germany; Portugal). Measured δ13C (‰) varied from -28.73 to -22.6 for PN, and from -28.79 to -21.67 for CH. These results also clearly revealed higher water stress for the 2020 vintage. The same grape juices have been analysed by Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (FT-ICR-MS) and Liquid Chromatography coupled to Mass Spectrometry (LC-qTOF-MS), leading to the detection of up to 4500 CHONS containing elemental compositions, and thus likely tens of thousands of individual compounds, which include fatty acids, organic acids, peptides, phenolics, also with high levels of glycosylation. Multivariate statistical analysis revealed that up to 160 elemental compositions, covering the whole range of detected masses (100 –1000 m/z), were significantly correlated to the observed gradients of water status. Examples of chemical markers, which are representative of these complex fingerprints, include various derivatives of the known abscisic acid (ABA), such as phaesic acid or abscisic acid glucose ester, which are significantly correlated with higher water stress, regardless of the variety. Cultivar-specific behaviours could also be identified from these fingerprints. Our results provide an unprecedented representation of the metabolic diversity, which is involved in the water status regulation at the grape level, and which could contribute to a better knowledge of the grapevine mitigation strategy in a climate change context.