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…

δ13C : A still underused indicator in precision viticulture  

The first demonstration of the interest of carbon isotope composition of sugars in grapevine, as an integrated indicator of vineyard water status, dates back to 2000 (Gaudillère et al., 1999; Van Leeuwen et al., 2001). Thanks to the isotopic discrimination of Carbon that takes place during plant photosynthesis, under hydric stress conditions, it is possible to accurately estimate the photosynthetic activity. Ever since, δ13C has been widely applied with success to zonation, terroir studies and vine physiology research, but is still not widely used by viticulturists. This is quite astonishing by considering the impact of global warming on viticulture and the need to improve water management, that would justify a widespread use of δ13C.
The lack of private laboratories proposing the analysis, the cost of the technology, as well as the long analytical delays, have been detrimental to its development. Some laboratories tried to overcome the analytical difficulties of isotopic analysis by using fourier transformed infrared spectroscopy, as a fast and cheap alternative to the official OIV method (IRMS). These claimed FTIR models have never been published or peer reviewed and cannot be considered robust. In this work, thanks to the recent acquisition of IRMS technology, new modern and robust applications of δ13C for viticulture are proposed. This includes the use of the analysis to make parcel separations at harvesting, the possibility to increase the precision of hydric stress cartography and the potential cost reduction when compared with Scholander pressure bomb analysis.

Grapevine xylem embolism resistance spectrum reveals which varieties have a lower mortality risk in a future dry climate

Wine growing regions have recently faced intense and frequent droughts that have led to substantial economical losses, and the maintenance of grapevine productivity under warmer and drier climate will rely notably on planting drought-resistant cultivars. Given that plant growth and yield depend on water transport efficiency and maintenance of photosynthesis, thus on the preservation of the vascular system integrity during drought, a better understanding of drought-related hydraulic traits that have a significant impact on physiological processes is urgently needed. We have worked towards this end by assessing vulnerability to xylem embolism in 30 grapevine commercial varieties encompassing red and white Vitis vinifera varieties, hybrid varieties characterized by a polygenic resistance for powdery and downy mildew, and commonly used rootstocks. These analyses further allowed a global assessment of wine regions with respect to their varietal diversity and resulting vulnerability to stem embolism. Hybrid cultivars displayed the highest vulnerability to embolism, while rootstocks showed the greatest resistance. Significant variability also arose among Vitis vinifera varieties, with Ψ12 and Ψ50 values ranging from -0.4 to -2.7 MPa and from -1.8 to -3.4 MPa, respectively. Cabernet franc, Chardonnay and Ugni blanc featured among the most vulnerable varieties while Pinot noir, Merlot and Cabernet Sauvignon ranked among the most resistant. In consequence, wine regions bearing a significant proportion of vulnerable varieties, such as Poitou-Charentes, France and Marlborough, New Zealand, turned out to be at greater risk under drought. These results highlight that grapevine varieties may not respond equally to warmer and drier conditions, outlining the importance to consider hydraulic traits associated with plant drought tolerance into breeding programmes and modeling simulations of grapevine yield maintenance under severe drought. They finally represent a step forward to advise the wine industry about which varieties and regions would have the lowest risk of drought-induced mortality under climate change.

Teasing apart terroir: the influence of management style on native yeast communities within Oregon wineries and vineyards

Newer sequencing technologies have allowed for the addition of microbes to the story of terroir. The same environmental factors that influence the phenotypic expression of a crop also shape the composition of the microbial communities found on that crop. For fermented goods, such as wine, that microbial community ultimately influences the organoleptic properties of the final product that is delivered to customers. Recent studies have begun to study the biogeography of wine-associated microbes within different growing regions, finding that communities are distinct across landscapes. Despite this new knowledge, there are still many questions about what factors drive these differences. Our goal was to quantify differences in yeast communities due to management style between seven pairs of conventional and biodynamic vineyards (14 in total) throughout Oregon, USA. We wanted to answer the following questions: 1) are yeast communities distinct between biodynamic vineyards and conventional vineyards? 2) are these differences consistent across a large geographic region? 3) can differences in yeast communities be tied to differences in metabolite profiles of the bottled wine? To collect our data we took soil, bark, leaf, and grape samples from within each vineyard from five different vines of pinot noir. We also collected must and a 10º brix sample from each winery. Using these samples, we performed 18S amplicon sequencing to identify the yeast present. We then used metabolomics to characterize the organoleptic compounds present in the bottled wine from the blocks the year that we sampled. We are actively in the process of analysing our data from this study.

The impact of sustainable management regimes on amino acid profiles in grape juice, grape skin flavonoids, and hydroxycinnamic acids

One of the biggest challenges of agriculture today is maintaining food safety and food quality while providing ecosystem services such as biodiversity conservation, pest and disease control, ensuring water quality and supply, and climate regulation. Organic farming was shown to promote biodiversity and carbon sequestration, and is therefore seen as one possibility of environmentally friendly production. Consumers expect organically grown crops to be free from chemical pesticides and mineral fertilizers and often presume that the quality of organically grown crops is different or higher compared to conventionally grown crops. Integrated, organic, and biodynamic viticulture were compared in a replicated field trial in Geisenheim, Germany (Vitis vinifera L. cv. Riesling). Amino acid profiles in juice, grape skin flavonoids, and hydroxycinnamic acids were monitored over three consecutive seasons beginning 7 years after conversion to organic and biodynamic viticulture, respectively. In addition, parameters such as soil nutrient status, yield, vigor, canopy temperature, and water stress were monitored to draw conclusions on reasons for the observed changes. Results revealed that the different sustainable management regimes highly differed in their amino acid profiles in juice and also in their skin flavonol content, whereas differences in the flavanol and hydroxycinnamic acid content were less pronounced. It is very likely that differences in nutrient status and yield determined amino acid profiles in juice, although all three systems showed similar amounts of mineralized nitrogen in the soil. Canopy structure and temperature in the bunch zone did not differ among treatments and therefore cannot account for the observed differences in favonols. A different light exposure of the bunches in the respective systems due to differences in vigor together with differences in berry size and a different water status of the vines might rather be responsible for the increase in flavonol content under organic and biodynamic viticulture.

Extreme canopy management for vineyard adaptation to climate change: is it a good idea?

Climate change constitutes an enormous challenge for humankind and for all human activities, viticulture not being an exception. Long-term strategic changes are probably needed the most, but growers also need to deal with short-term changes: summers that are getting progressively warmer, earlier harvest dates and higher pH in musts and wines. In the last 10-15 years, a relevant corpus of research is being developed worldwide in order to evaluate to which extent extreme canopy management operations, aimed at reducing leaf area and, thus, limiting the source to sink ratio, could be useful to delay ripening. Although extreme canopy management can result in relevant delays in harvest dates, longer term studies, as well as detailed analysis of their implications on carbohydrate reserves, bud fertility and future yield are desirable before these practices can be recommended.