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…

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.

Sustainable fertilisation of the vineyard in Galicia (Spain)

Excessive fertilization of the vineyard leads to low quality grapes, increased costs and a negative impact on the environment. In order to establish an integrated management system aimed at a sustainable fertilization of the vineyards, nutritional reference levels were established. For this purpose, 30 representative vineyards of the Albariño variety were studied, in which soil and petiole analyses were carried out for two years and grape yield and quality at harvest were measured. In both years of study, soil pH, calcium, sodium and cation exchange capacity were positively correlated with calcium content and negatively correlated with manganese in grapes. Irrigated vineyards had higher levels of aluminium in soil and lower levels of calcium in petiole. Climatic conditions were very different in the years of the study. The year 2019 was colder than usual, in 2020 there was a marked water stress with high summer temperatures. This resulted in medium-high acidity in grapes in 2019 and low acidity in 2020, with sugar levels being similar both years. A very marked decrease in must amino nitrogen was observed in 2020, with ammonia nitrogen remaining stable. The correlation of acidity and sugar values in grapes with soil and petiole analysis data made it possible to establish reference levels for the nutritional diagnosis of the Albariño variety in this region. Based on these results, an easy-to-use TIC application is currently being created for grapegrowers, aimed at improving the sustainability of the vineyard through reasoned fertilization. This study has now been extended to other Galician vine varieties.

Anthocyanin profile is differentially affected by high temperature, elevated CO2 and water deficit in Tempranillo (Vitis vinifera L.) clones

Anthocyanin potential of grape berries is an important quality factor in wine production. Anthocyanin concentration and profile differ among varieties but it also depends on the environmental conditions, which are expected to be greatly modified by climate change in the future. These modifications may significantly modify the biochemical composition of berries at harvest, and thus wine typicity. Among the diverse approaches proposed to reduce the potential negative effects that climate change may have on grape quality, genetic diversity among clones can represent a source of potential candidates to select better adapted plant material for future climatic conditions. The effects of individual and combined factors associated to climate change (increase of temperature, rise of air CO2 concentration and water deficit) on the anthocyanin profile of different clones of Tempranillo that differ in the length of their reproductive cycle were studied. The aim was to highlight those clones more adapted to maintain specific Tempranillo typicity in the future. Fruit-bearing cuttings were grown in controlled conditions under two temperatures (ambient temperature versus ambient temperature + 4ºC), two CO2 levels (400 ppm versus 700 ppm) and two water regimes (well-watered versus water deficit), both in combination or independently, in order to simulate future climate change scenarios. Elevated temperature increased anthocyanin acylation, whereas elevated CO2 and water deficit favoured the accumulation of malvidin derivatives, as well as the acylation and tri-hydroxylation level of anthocyanins. Although the changes in anthocyanin profile observed followed a common pattern among clones, such impact of environmental conditions was especially noticeable in one of the most widely distributed Tempranillo clones, the accession RJ43.

The impact of leaf canopy management on eco-physiology, wood chemical properties and microbial communities in root, trunk and cordon of Riesling grapevines (Vitis vinifera L.)

In the last decades, climate change required already adaptation of vineyard management. Increase in temperature and unexpected weather events cause changes in all phenological stages requiring new management tools. For example, defoliation can be a useful tool to reduce the sugar content in the berries creating differences in the wine profiles. In a ten-year field experiment using Riesling (Vitis vinifera L, planted 1986, Geisenheim, Germany), various mechanical defoliation strategies and different intensities were trialed until 2016 before the vineyard was uprooted. Wood was sampled from the plant compartments root, trunk, cordon and shoot for analyses of physicochemical properties (e.g. lignin and element content, pH, diameter), nonstructural carbohydrates and the microbial communities. The aim of the study was to investigate the influence of reduced canopy leaf area on the sink-source allocation into different compartments and potential changes of the fungal and prokaryotic wood-inhabiting community using a metabarcoding approach. Severe summer pruning (SSP) of the canopy and mechanical defoliation (MDC) above the bunch zone decreased the leaf area by 50% compared to control (C). SSP reduced the photosynthetic capacity, which resulted in an altered source-sink allocation and carbohydrate storage. With lower leaf area, less carbohydrates are allocated. This for example resulted in a decreased trunk diameter. Further, it affected the composition of the grapevine wood microbiota. SSP and MDC management changed significantly the prokaryotic community composition in wood of the root samples, but had no effect in other compartments. In general, this study found strong compartment and less management effects of the microbial community composition and associated physicochemical properties. The highest microbial diversities were identified in the wood of the trunk, and several species were recorded the first time in grapevine.

Pruned vine biomass exclusion from a clay loam vineyard soil – examining the impact on physical/chemical properties

The wine industry worldwide faces increasing challenges to achieve sustainable levels of carbon emission mitigation. This project seeks to establish the feasibility of harvesting winter pruned vineyard biomass (PVB) for potential use in carbon footprint reduction, through its use as a renewable biofuel for energy production. In order to make this recommendation, technical issues such as the potential environmental impact, chemical composition and fuel suitability, and logistical challenges of harvesting biomass needs to be understood to compare with the results from similar studies. Of particular interest is the role PVB plays as a carbon source in vineyard soils and what effect annual removal might have on soil carbon sequestration. A preliminary trial was established in the Waite Campus vineyard (University of Adelaide) to test current management strategies. Vines are grown in a Eutrophic, Red Dermosol clay loam soil with well managed midrow swards. A comparison was undertaken of mid-row treatments in two 0.25 Ha blocks (Shiraz and Semillon), including annual cultivation for seed bed preparation, the deliberate exclusion of PVB (25 years) and incorporation of PVB (13 years) at an average of 3.4 and 5.5 Mg/Ha-1 for Shiraz and Semillon respectively. In both 0-10cm and 10-30cm soil core sample depths, combined soil carbon % measures in the desired range of 1.80 to 3.50, were not significantly different between treatments or cultivars and yielded an estimated 42 Mg/ha-1 of sequestered soil carbon. Other key physical and chemical measures were likewise not significantly different between treatments. Preliminary results suggest that in a temperate zone vineyard, managed such as the one used in this study, there is no long term negative impact on soil carbon sequestration through removing PVB. This implies that growers could confidently harvest PVB for use in several end fates including as a bio fuel.