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 Nitrogen metabolism in Kluyveromyces marxianus and Saccharomyces cerevisiae: towards a better understanding of fermentation aroma production

Nitrogen metabolism in Kluyveromyces marxianus and Saccharomyces cerevisiae: towards a better understanding of fermentation aroma production

Abstract

During wine alcoholic fermentation, yeasts produce volatile aroma compounds from sugar and nitrogen metabolism. Some of the metabolic pathways leading to these compounds have been known for more than a century. Yet, the differences in compound yield and nature between species remain poorly understood. Using a two-pronged approach of isotopic filiation and transcriptome analysis, this study endeavoured to shed new light on the utilisation of nitrogen sources by two wine-related yeast species, Saccharomyces cerevisiae Lalvin EC1118® (Lallemand) and Kluyveromyces marxianus IWBT Y885. 

The data showed that, although the order and intensity of uptake of nitrogen sources was broadly similar, those of ammonium and arginine differed. Furthermore, the utilisation of assimilated amino acids also differed significantly. While S. cerevisiae redistributed the nitrogen in these amino acids evenly for the production of other amino acids, K. marxianus clearly favoured specific amino acids. As for amino acids used as substrates for the production of aroma compounds, the fate of leucine and valine did not differ significantly between the two species. However, phenylalanine metabolism differed, and a larger proportion of phenylalanine was channelled through the Ehrlich pathway in K. marxianus, resulting in increased production of phenylethanol. Transcriptome data suggest that this shift can be explained by the higher expression of aromatic amino transferases in K. marxianus. Taken together, the data show that metabolic pathways are broadly conserved, but that individual nitrogen sources are not always assimilated and metabolised in identical ways. The study also provides new insights on the modulation of fermentative aroma profiles by yeast species of commercial interest.

DOI:

Publication date: June 10, 2020

Issue: OENO IVAS 2019

Type: Article

Authors

Benoit Divol, Stephanie Rollero, Audrey Bloem, Anne Ortiz-Julien, Florian Bauer, Carole Camarasa

Institute for Wine Biotechnology, Department of Viticulture and Oenology, Stellenbosch University, Private Bag X1, Matieland 7602, South Africa

Contact the author

Keywords

Nitrogen metabolism, Kluyveromyces marxianus, Saccharomyces cerevisiae, Fermentative aroma compounds 

Tags

IVES Conference Series | OENO IVAS 2019

Citation

Related articles…

What are the optimal ranges and thresholds for berry solar radiation for flavonoid biosynthesis?

In wine grape production, canopy management practices are applied to control the source-sink balance and improve the cluster microclimate to enhance berry composition. The aim of this study was to identify the optimal ranges of berry solar radiation exposure (exposure) for upregulation of flavonoid biosynthesis and thresholds for their degradation, to evaluate how canopy management practices such as leaf removal, shoot thinning, and a combination of both affect the grapevine (Vitis vinifera L. cv. Cabernet Sauvignon) yield components, berry composition, and flavonoid profile under context of climate change. First experiment assessed changes in the grape flavonoid content driven by four degrees of exposure. In the second experiment, individual grape berries subjected to different exposures were collected from two cultivars (Cabernet Sauvignon and Petit Verdot). The third experiment consisted of an experiment with three canopy management treatments (i) LR (removal of 5 to 6 basal leaves), (ii) ST (thinned to 24 shoots per vine), and (iii) LRST (a combination of LR and ST) and an untreated control (UNT). Berry composition, flavonoid content and profiles, and 3-isobutyl 2-methoxypyrazine were monitored during berry ripening. Although increasing canopy porosity through canopy management practices can be helpful for other purposes, this may not be the case of flavonoid compounds when a certain proportion of kaempferol was achieved. Our results revealed different sensitivities to degradation within the flavonoid groups, flavonols being the only monitored group that was upregulated by solar radiation. Within different canopy management practices, the main effects were due to the ST. Under environmental conditions given in this trial, ST and LRST hastened fruit maturity; however, a clear improvement of the flavonoid compounds (i.e., greater anthocyanin) was not observed at harvest. Methoxypyrazine berry content decreased with canopy management practices studied. Although some berry traits were improved (i.e. 2.5° Brix increase in berry total soluble solids) due to canopy management practices (ST), this resulted in a four-fold increase in labor operations cost, two-fold decrease in yield with a 10-fold increase in anthocyanin production cost per hectare that should be assessed together as the climate continues to get hot.

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.

austrianvineyards.com: online viewer of all designations of Austrian wine

To digitally record and present all the origins of Austrian wines in the same perfect and clear way was the motivation for the Austrian Wine Marketing Board (Austrian Wine) to start with the project in 2018. In June 2021 the results were presented to the public in an online viewer showing all the designations of Austrian wine, available at https://austrianvineyards.com in a largely barrier-free manner. The online viewer provides tailored individual maps fitted to the respective zoom level. The smallest unit of wine-origins in Austria is called Ried and is displayed in a plot-specific manner highlighting areas under vine. Information on the Ried include administrative district, winegrowing municipality, cadastral municipality, large collective vineyard site, specific winegrowing region, generic winegrowing region, winegrowing area and, in many cases, an illustrative picture. Complementary data on the size, elevation (minimum-maximum), orientation (in 8 sectors plus flat) and gradient (minimum, maximum, average) are based on the area under vine according to the EU’s Integrated Administration and Control System. Additional information covers climate data. The diagrams are taken from the monthly breakdown of data in the annals of the Central Institute for Meteorology and Geodynamics, Austria provide a display of values for air temperature, precipitation, and sunshine hours for the reference year and the long-term average. Seasonal aggregated data on temperature, precipitation, and sunshine hours complete the display. Short descriptions with emphasis on geology and soil, field name in historical maps, etymology of the denomination, and main planted variety complements the available information for the main designations in the online viewer. These descriptions are compiled by winegrowers, geologists, historians, and journalists. All the information and data can be extracted to a pdf-file. Printed vineyard maps are also available. Missing content regarding wine origins in Styria will be completed in winter 2021/22.

The rootstock, the neglected player in the scion transpiration even during the night

Water is the main limiting factor for yield in viticulture. Improving drought adaptation in viticulture will be an increasingly important issue under climate change. Genetic variability of water deficit responses in grapevine partly results from the rootstocks, making them an attractive and relevant mean to achieve adaptation without changing the scion genotype. The objective of this work was to characterize the rootstock effect on the diurnal regulation of scion transpiration. A large panel of 55 commercial genotypes were grafted onto Cabernet Sauvignon. Three biological repetitions per genotype were analyzed. Potted plants were phenotyped on a greenhouse balance platform capable of assessing real-time water use and maintaining a targeted water deficit intensity. After a 10 days well-watered baseline period, an increasing water deficit was applied for 10 days, followed by a stable water deficit stress for 7 days. Pruning weight, root and aerial dry weight and transpiration were recorded and the experiment was repeated during two years. Transpiration efficiency (ratio between aerial biomass and transpiration) was calculated and δ13C was measured in leaves for the baseline and stable water deficit periods. A large genetic variability was observed within the panel. The rootstock had a significant impact on nocturnal transpiration which was also strongly and positively correlated with maximum daytime transpiration. The correlations with growth and water use efficiency related traits will be discussed. Transpiration data were also related with VPD and soil water content demonstrating the influence of environmental conditions on transpiration. These results highlighted the role of the rootstock in modulating water deficit responses and give insights for rootstock breeding programs aimed at identifying drought tolerant rootstocks. It was also helpful to better define the mechanisms on which the drought tolerance in grapevine rootstocks is based on.

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.