Macrowine 2021
IVES 9 IVES Conference Series 9 Response of different nitrogen supplementation on Saccharomyces cerevisiae metabolic response and wine aromatic profile

Response of different nitrogen supplementation on Saccharomyces cerevisiae metabolic response and wine aromatic profile

Abstract

The wine yeast Saccharomyces cerevisiae can highly affect wine aromatic profile by producing and/or mediating the release of a whole range of metabolites (such as thiols, esters, and terpenes), which in turn contribute to enhanced aroma and flavor. These metabolites depend on yeast metabolism activated during fermentation which can constitute the ‘’metabolic footprint’’ of the yeast strain that carried out the process. The aim of the present research was to study the metabolism of S. cerevisiae under two different nitrogen supplementation status and to investigate the relative expression of specific genes, that are directly related to the biosynthesis of specific potent odornats such as, terpenes and esters. 

The commercial yeast strains 1X (S.cerevisiae) and 2X (S.cerevisiae x S.bayanus) were inoculated in Moschofilero (Vitis Vinifera L. cv) grape must under two different concentrations of yeast assimilable nitrogen (YAN), a  low at 150 mg/L and a high at 300 mg/L. The produced wines were analyzed for their standard enological parameters, their volatile composition by SPE/GC–MS analysis as well as for their sensory profile. Totally 8 fermentations trials, were realised in triplicate. The fermentation was monitored by measuring the optical density and sugar consumption. Metabolic response was tested through real-time RT-PCR of genes implicated in aroma production of esters and terpenes such as ATF1, ATF2, EEB1, EHT1, IAH1, BGL2, EXG1. Sampling for metabolites and gene expression analysis were taken at the time of inoculation, after 48 hours, when two thirds of the sugars were depleted and at the end of the alcoholic fermentation (< 2g/L rs).

In terms of the volatile characterization of the wines, esters, linalool and nerol appeared to be clearly distinct between the different levels of YAN, which confirms the specialization in volatile compounds production among different nitrogen concentration levels. For instance, linalool was found to be at 0.05 mg/L for low nitrogen concentration, while high nitrogen levels resulted to a concentration of 0.12 mg/L. Real-time-PCR results revealed that, in both cases of nitrogen implementation, the analyzed genes were found to be expressed mainly before the fermentation of the 70% of the sugars. In addition, an overexpression of the BGL2 gene, corresponded well to the linalool concentration found, was observed in case of high nitrogen condition. Also, the EHT1 was expressed five times higher in case of high nitrogen concentration. Finally, correlations between ethyl esters and EEB1, acetate esters and ATF2 (p<0.05) were also found in both cases. 

Our study revealed the impact of different nitrogen implementations on the volatile compounds and the relative expression of specific genes. Metabolic analysis of selected volatile components of the wine aroma in conjunction with transcriptional analyses provide a great approach to orient the fermentation process towards a desirable wine aromatic profile.

DOI:

Publication date: September 3, 2021

Issue: Macrowine 2021

Type: Article

Authors

Despina Lola, Chrysanthi KALLONIATI, Maria DIMOPOULOU, Maria Ioanna XENIA, Emmanouil FLEMETAKIS, Yorgos KOTSERIDIS

Laboratory of Oenology and Alcoholic Drinks (LEAD), Agricultural University of Athens, 75 Iera Odos, 11855 Athens, Greece, Laboratory of Molecular Biology, Agricultural University of Athens, 75 Iera Odos, 11855 Athens, Greece, Department of Wine, Vine and Beverage Sciences, School of Food Science, University of West Attica, Greece, Laboratory of Oenology and Alcoholic Drinks  (LEAD), Agricultural University of Athens, 75 Iera Odos, 11855 Athens, Greece Laboratory of Molecular Biology, Agricultural University of Athens, 75 Iera Odos, 11855 Athens, Greece, Laboratory of Enology and Alcoholic Drinks (LEAD), Agricultural University of Athens, 75 Iera Odos, 11855 Athens, Greece

Contact the author

Keywords

yeast metabolism, yan, nitrogen supplementation, volatile profile, gene expression

Citation

Related articles…

Optimizing stomatal traits for future climates

Stomatal traits determine grapevine water use, carbon supply, and water stress, which directly impact yield and berry chemistry. Breeding for stomatal traits has the strong potential to improve grapevine performance under future, drier conditions, but the trait values that breeders should target are unknown. We used a functional-structural plant model developed for grapevine (HydroShoot) to determine how stomatal traits impact canopy gas exchange, water potential, and temperature under historical and future conditions in high-quality and hot-climate California wine regions (Napa and the Central Valley). Historical climate (1990-2010) was collected from weather stations and future climate (2079-99) was projected from 4 representative climate models for California, assuming medium- and high-emissions (RCP 4.5 and 8.5). Five trait parameterizations, representing mean and extreme values for the maximum stomatal conductance (gmax) and leaf water potential threshold for stomatal closure (Ψsc), were defined from meta-analyses. Compared to mean trait values, the water-spending extremes (highest gmax or most negative Ysc) had negligible benefits for carbon gain and canopy cooling, but exacerbated vine water use and stress, for both sites and climate scenarios. These traits increased cumulative transpiration by 8 – 17%, changed cumulative carbon gain by -4 – 3%, and reduced minimum water potentials by 10 – 18%. Conversely, the water-saving extremes (lowest gmax or least negative Ψsc) strongly reduced water use and stress, but potentially compromised the carbon supply for ripening. Under RCP 8.5 conditions, these traits reduced transpiration by 22 – 35% and carbon gain by 9 – 16% and increased minimum water potentials by 20 – 28%, compared to mean values. Overall, selecting for more water-saving stomatal traits could improve water-use efficiency and avoid the detrimental effects of highly negative canopy water potentials on yield and quality, but more work is needed to evaluate whether these benefits outweigh the consequences of minor declines in carbon gain for fruit production.

Sustaining wine identity through intra-varietal diversification

With contemporary climate change, cultivated Vitis vinifera L. is at risk as climate is a critical component in defining ecologically fitted plant materiel. While winegrowers can draw on the rich diversity among grapevine varieties to limit expected impacts (Morales-Castilla et al., 2020), replacing a signature variety that has created a sense of local distinctiveness may lead to several challenges. In order to sustain wine identity in uncertain climate outcomes, the study of intra-varietal diversity is important to reflect the adaptive and evolutionary potential of current cultivated varieties. The aim of this ongoing study is to understand to what extent can intra-varietal diversity be a climate change adaptation solution. With a focus on early (Sauvignon blanc, Riesling, Grolleau, Pinot noir) to moderate late (Chenin, Petit Verdot, Cabernet franc) ripening varieties, data was collected for flowering and veraison for the various studied accessions (from conservatory plots) and clones. For these phenological growing stages, heat requirements were established using nearby weather stations (adapted from the GFV model, Parker et al., 2013) and model performances were verified. Climate change projections were then integrated to predict the future behaviour of the intra-varietal diversity. Study findings highlight the strong phenotypic diversity of studied varieties and the importance of diversification to enhance climate change resilience. While model performances may require improvements, this study is the first step towards quantifying heat requirements of different clones and how they can provide adaptation solutions for winegrowers to sustain local wine identity in a global changing climate. As genetic diversity is an ongoing process through point mutations and epigenetic adaptations, perspective work is to explore clonal data from a wide variety of geographic locations.

VINIoT – Precision viticulture service

The project VINIoT pursues the creation of a new technological vineyard monitoring service, which will allow companies in the wine sector in the SUDOE space to monitor plantations in real time and remotely at various levels of precision. The system is based on spectral images and an IoT architecture that allows assessing parameters of interest viticulture and the collection of data at a precise scale (level of grape, plant, plot or vineyard) will be designed. In France, three subjects were specifically developed: evaluation of maturity, of water stress, and detection of flavescence dorée. For the evaluation of maturity, it has been decided first to work at the berry scale in the laboratory, then at the bunch scale and finally in the vineyard. The acquisition of the spectral hyperstal image as well as the reference analyzes to measure the maturity, were carried out in the laboratory after harvesting the berries in a maturity monitoring context. This work focuses on a case study to predict sugar content of three different grape varieties: Syrah, Fer Servadou and Mauzac. A robust method called Roboost-PLSR, developed in the framework of this work (Courand et al., 2022), to improve prediction model performance was applied on spectra after the acquirement of hyperspectral images. Regarding the evaluation of water stress, to work with a significant variability in terms of water status, it has been worked first with potted plants under 2 different water regimes. The facilities have allowed the supervision of irrigation and micro-climatic conditions. The regression models on agronomic variables (stomatal conductance, water potential, …) are studied. To detect flavescence dorée, the experimental plan has consisted of work at leaf scale in the laboratory first, and then in the field. To detect the disease from hyper-spectral imaging, a combination of multivariate curve resolution-alternating least squares (MCR-ALS) and factorial discriminant analysis (FDA) was proposed. This strategy proved the potential towards the discrimination of healthy and infected leaves by flavescence dorée based on the use of hyperspectral images (Mas Garcia et al., 2021).

Impact of changes in pruning practices on vine growth and yield

A gradual decline in vineyards has been observed over the past twenty years worldwide. This might be explained by the climate change, practices change or the increase of dieback diseases. To increase the longevity of vines, we studied the impact of different pruning strategies in four adult and four young vineyards located in France and Spain. In France, vineyards were planted with Cabernet franc on 3309C while Spanish trials were planted with Tempranillo grafted on 110R. Vegetative expression, yield, quality of berries and wood vessels conductivity were measured. The distribution of vegetative expression, yield and berry composition between primary and secondary vegetation were quantified. Finally, tomography was used to evaluate the implication of the treatments on sap flows.
First results show that i) the respectful pruning leads to an increase of 30 to 50% more secondary shoots than the aggressive pruning in France and between 15 and 20% in Spain, ii) there is no major effect on the yield over the first two years following the implementation of the new pruning practices, although the proportion of clusters from suckers is higher on the respectful pruning method. On young vines, the development of the trunk according to a respectful pruning leads to a loss of harvest 2 years after planting. This is due to the removal, on the future trunk, of the green suckers which carrying bunches. This operation carried out in spring rather than during winter pruning, would promote a better leaf / fruit balance when the plant comes into production, and could lead to better hydraulic conduction in the vessels of the trunk. Maintaining these trials for several years will provide more robust data to assess the impact of these practices on the vines over the long term.

Climate and the evolving mix of grape varieties in Australia’s wine regions

The purpose of this study is to examine the changing mix of winegrape varieties in Australia so as to address the question: In the light of key climate indicators and predictions of further climate change, how appropriate are the grape varieties currently planted in Australia’s wine regions? To achieve this, regions are classified into zones according to each region’s climate variables, particularly average growing season temperature (GST), leaving aside within-region variations in climates. Five different climatic classifications are reported. Using projections of GSTs for the mid- and late 21st century, the extent to which each region is projected to move from its current zone classification to a warmer one is reported. Also shown is the changing proportion of each of 21 key varieties grown in a GST zone considered to be optimal for premium winegrape production. Together these indicators strengthen earlier suggestions that the mix of varieties may be currently less than ideal in many Australian wine regions, and would become even less so in coming decades if that mix was not altered in the anticipation of climate change. That is, grape varieties in many (especially the warmest) regions will have to keep changing, or wineries will have to seek fruit from higher latitudes or elevations if they wish to retain their current mix of varieties and wine styles.