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…

Combining effect of leaf removal and natural shading on grape ripening under two irrigation strategies in Manto negro (Vitis vinifera L.)

The increasingly frequent heat waves during grape ripening pose challenges for high quality wine grape production. Defoliation is a common practice that can improve the control of diseases in bunches, but also it increases the exposure to sunlight. Grapes exposed to solar radiation reach temperatures over the optimum for berry development and maturation. This makes the development of irrigation and canopy management techniques of great importance to maximize yield and grape quality. A field experiment was carried out during 2021 using Manto negro wine grapes to study the effect of applied irrigation and different light exposure levels on grape quality. Two irrigation treatments were imposed based on the frequency and amount of water doses in a four-block experimental vineyard at Bodega Ribas (Mallorca). Three light exposure treatments were randomly applied in each irrigation plot. The light treatments included exposed clusters from pea size, non-exposed clusters, and shaded clusters after softening. Leaf area index and canopy porosity was estimated every 2 weeks. Midday leaf water potential was measured weekly. Additionally, apparent electrical conductivity was measured between rows to estimate the soil water content variability. Light and temperature sensors were installed at the bunch level to quantify the differences in bunch temperature and light intensity among treatments. The effect of irrigation and cluster light exposure on berry weight, TSS, TA, malic acid, tartaric acid, K+, and pH were analysed at 5 moments along grape ripening. During different heat waves, the natural shading technique decreased the maximum bunch temperature around 10 °C respect to the exposed bunches in both irrigation strategies. The combination of defoliation and shading techniques after softening decreased TSS at harvest and affected most of the quality parameters during the last stages of ripening, showing an interesting technique to delay ripening in warm viticulture areas.

Influence of climatic conditions on grape composition of Tempranillo in La Mancha DO (Spain)

The aim of this work was to analyze the variability in grape composition of the Tempranillo cultivar related to climatic conditions, in La Mancha Designation of Origin. Grape composition (sugar content, total acidity, pH, malic acid, and total and extractable anthocyanins) recorded during ripening, were analysed for the period 2000-2019. The weather conditions at daily time scale, recorded during the same period, were also evaluated. The relationships between grape parameters with climatic variables related to temperature and to water deficits, referring different periods between phenological events along the growing cycle, were evaluated using regression analysis. High variability in grape composition was observed in the period analysed. Total acidity varied between 3.7 and 7.3 gL-1 while malic acid varied between 1.2 and 4 gL-1. The extractable anthocyanins ranged between 526 and 972 mgL-1, and total anthocyanins ranged between 922 and 1388 mgL-1, being the lowest values recorded in the hottest year (2017). Total acidity decreased 0.77 gL-1 for an increase of 100 GDD, while malic acid decrease in 0.42 gL-1 for the same GDD increase, being the period between veraison and harvest the one that seemed to have higher influence on acidity. In addition, it was confirmed that increasing water deficits decreased acidity. Total and extractable anthocyanins increased in about 210 and 105 mgL-1, respectively, with an increase of 100 GDD from veraison to harvest, and the increase in water deficits favour the increase of anthocyanins, both total and extractable anthocyanins. Total and extractable anthocyanins concentration increased in 35 and 22 mgL-1 per an increase of 10 mm in the water deficit. These results can be of interest to understand the potential changes that grapes composition may suffer under future warmer climates.

Use of a new, miniaturized, low-cost spectral sensor to estimate and map the vineyard water status from a mobile 

Optimizing the use of water and improving irrigation strategies has become increasingly important in most winegrowing countries due to the consequences of climate change, which are leading to more frequent droughts, heat waves, or alteration of precipitation patterns. Optimized irrigation scheduling can only be based on a reliable knowledge of the vineyard water status.

In this context, this work aims at the development of a novel methodology, using a contactless, miniaturized, low-cost NIR spectral tool to monitor (on-the-go) the vineyard water status variability. On-the-go spectral measurements were acquired in the vineyard using a NIR micro spectrometer, operating in the 900–1900 nm spectral range, from a ground vehicle moving at 3 km/h. Spectral measurements were collected on the northeast side of the canopy across four different dates (July 8th, 14th, 21st and August 12th) during 2021 season in a commercial vineyard (3 ha). Grapevines of Vitis vinifera L. Graciano planted on a VSP trellis were monitored at solar noon using stem water potential (Ψs) as reference indicators of plant water status. In total, 108 measurements of Ψs were taken (27 vines per date).

Calibration and prediction models were performed using Partial Least Squares (PLS) regression. The best prediction models for grapevine water status yielded a determination coefficient of cross-validation (r2cv) of 0.67 and a root mean square error of cross-validation (RMSEcv) of 0.131 MPa. This predictive model was employed to map the spatial variability of the vineyard water status and provided useful, practical information towards the implementation of appropriate irrigation strategies. The outcomes presented in this work show the great potential of this low-cost methodology to assess the vineyard stem water potential and its spatial variability in a commercial vineyard.

Using δ13C and hydroscapes as a tool for discriminating cultivar specific drought response

Measurement of carbon isotope discrimination in berry juice sugars at maturity (δ13C) provides an integrated assessment of water use efficiency (WUE) during the period of berry ripening, and when collected over multiple seasons can be used as an indication of drought stress response. Berry juice δ13C measurements were carried out on 48 different varieties planted in a common garden experiment in Bordeaux, France from 2014 through 2021 and were paired with midday and predawn leaf water potential measurements on the same vines in a subset of six varieties. The aim was to discriminate a large panel of varieties based on their stomatal behaviour and potentially identify hydraulic traits characterizing drought tolerance by comparing δ13C and hydroscapes (the visualisation of plant stomatal behaviour as a response to predawn water potential). Cluster analysis found that δ13C values are likely affected by the differing phenology of each variety, resulting in berry ripening of different varieties taking place under different stress conditions within the same year. We accounted for these phenological differences and found that cluster analysis based on specific δ13C metrics created a classification of varieties that corresponds well to our current empirical understanding of their relative drought tolerances. In addition, we analysed the water potential regulation of the subset of six varieties (using the hydroscape approach) and found that it was well correlated with some δ13C metrics. Surprisingly, a variety’s water potential regulation (specifically its minimum critical leaf water potential under water deficit) was strongly correlated to δ13C values under well-watered conditions, suggesting that base WUE may have a stronger impact on drought tolerance than WUE under water deficit. These results give strong insights on the innate WUE of a very large panel of varieties and suggest that studies of drought tolerance should include traits expressed under non-limiting conditions.

Geospatial trends of bioclimatic indexes in the topographically complex region of Barolo DOCG

Barolo DOCG is an economically important wine producing region in Northwest Italy. It is a small region of approximately 70 km2 gross area. The topography is very complex with steep sloped hills ranging in elevation from below 200 m to 550 m. Barolo DOCG wine is made exclusively from the Nebbiolo grape. Bioclimatic indexes are often used in viticulture to gain a better understanding of broader climate trends which can be compared temporally and geographically. These indexes are also used for identifying potential phenological timing, growing region suitability, and potential risks associated with expected climatic changes. Understanding how topography influences bioclimatic indexes can help with understanding of mesoscale climate behaviour leading to improved decision making and risk management strategies. The average monthly maximum and minimum temperatures, the Cool Night Index, the Huglin Index, and the monthly diurnal range (from July to October) were calculated using data from 45 weather stations within a 40 km radius of the Barolo DOCG growing area between the years 1996 and 2019. Linear and multiple regression models were developed using independent variables (elevation, aspect, slope) extracted from a digital elevation model to identify significant relationships. Bioclimatic indexes were then kriged with external drift using independent variables that showed significant relationships with the bioclimatic index using a 100 m resolution grid. The maximum monthly temperatures and the Huglin Index showed consistent significant negative relationships with elevation in all years. The minimum monthly temperatures showed no relationship with elevation but in some months a small but significant relationship was observed with aspect. Due to the lack of a relationship between minimum monthly temperatures and elevation compared to the significant relationship between maximum monthly temperatures and elevation, monthly diurnal range had a negative relationship with elevation.