IVAS 2022 banner
IVES 9 IVES Conference Series 9 IVAS 9 IVAS 2022 9 Unravelling Saccharomyces cerevisiae biosynthethic pathways of melatonin, serotonin and hydroxytyrosol  by UPLC-HRMS Isotopic labelling analysis

Unravelling Saccharomyces cerevisiae biosynthethic pathways of melatonin, serotonin and hydroxytyrosol  by UPLC-HRMS Isotopic labelling analysis

Abstract

The main objective is to unravel the yeast biosynthetic pathways for MEL, SER and HT by using the respective labelled amino acids precursors: 15N2-L tryptophan and 13C-tyrosine.
The alcoholic fermentation experiments are performed with two different commercial S cereviseae yeasts using synthetic must with the addition of the labelled compounds and the bioactive compounds were followed during the fermentation process. Six biological replicates of the fermentations were considered. MEL, SER and HT were analysed by UHPLC coupled to High Resolution Mass Spectrometry (HRMS). Accurate mass determination allowed to unequivocally distinguishing labelled and unlabelled compounds. The analytical determination was performed using external calibration curves with 10 points, which were freshly prepared at every analytical session. Potential intermediates in the synthesis of MEL are detected as labelled metabolites following a time sequence that fits with the pathway described for the synthesis of MEL in mammals. At day 3 all the initial 15N2 L-TRP has been consumed. However, L-TRP as such was detected despite it was not added to the fermentation medium, thus demonstrating de novo tryptophan formation which can be used to synthetize MEL thereafter. Similarly, the results obtained in the fermentation carried out with 13-C TYR verify that HT is formed both from labelled tyrosine and from intermediates of the Erlich pathway.

DOI:

Publication date: June 24, 2022

Issue: IVAS 2022

Type: Poster

Authors

Troncoso Ana M.1, Gallardo-Fernández Marta1, Valls-Fonayet Josep2, Valero Eva3, Hornedo-Ortega Ruth1, Richard Tristan2 and García-Parrilla M Carmen1 

1Universidad de Sevilla
2Université de Bordeaux
3Universidad Pablo de Olavide

Contact the author

Keywords

yeast, alcoholic fermentation, bioactive compounds, isotopically labelled compounds

Tags

IVAS 2022 | IVES Conference Series

Citation

Related articles…

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).

Late frost protection in Champagne

Probably one of the most counterintuitive impacts of climate change on vine is the increased frequency of late frost. Champagne, due to its septentrional position is historically and regularly affected by this meteorological hazard. Champagne has therefore developed a strong experience in frost protection with first experiments dating from the end of 19th century. Frost protection can be divided in two parts: passive and active. Passive protection includes all the methods that do not seek to modify the vine’s environment or resistance at the time of frost. The most iconic passive protection in Champagne is the establishment of the individual reserve. This reserve allows to stock a certain quantity of clear wine during a surplus year to compensate a meteorological hazard like frost during the following years. Other common passive methods are the control of planting area (walls, bushes, topography), the choice of grape variety, late pruning, or the impact of grass cover and tillage. Active frost protection is also divided in two parts. Most of the existing techniques tend to modify vine’s environment. Most of the time they provide warmth (candles, heaters, windmills, heating cables…), or stabilise bud’s temperature above a lethal threshold (water sprinkling). The other way to actively fight is to enhance the resistance of buds to frost (elicitors). The Comité Champagne evaluates frost protection methods following three main axes: the efficiency, the profitability, and the environmental impact through a lifecycle assessment. This study will present the results on both passive and active protection following these three axes.

How can historical cultivars mitigate the effects of climate change?

IFV, INRAe and the national network “Partenaires de la Sélection Vigne” representing 37 organizations from the different wine regions, have been working increasingly closely over the last 2 decades towards the preservation of the French varietal patrimony. There are approximately 600 patrimonial varieties according to INRAe and SupAgro Montpellier experts, including ancient cultivars (400) and intravarietal crossbreeds obtained since the 19th century. In the context of a drastic reduction in such varieties from the mid 1980’s in favor of mainstream varieties, it was essential to carry out an inventory of old vines and vineyards. INRAe Vassal collection plays a key role here as it holds the largest diversity available, along with a rich bibliography and herbariums, offering us the opportunity to document and double check the identity of a cultivar, consolidating the expertise of ampelographers. The work is carried out in several stages, from verifying the existence of a variety in a small region, through to rehabilitation. During this session, the authors present the process that leads to the official registration of a variety. After this, IFV selection center takes over to initiate the process of selection and propagation. A specific focus within regions such as the Alps, Champagne and the South-West will provide details of the full procedure. Bia, Bouysselet, Chardonnay rose, Mecle and the aptly named Tardif, are some of the cultivars that have followed this procedure. Furthermore, a recent regulation established by INAO on “varieties of interest for adaptation purposes” might boost uptake by growers. Since 2006, 36 historical cultivars have been registered. Most of these have been neglected in the past due to late maturity, lack of sugar and high titratable acidity at harvest time. Such characteristics are today considered as positive qualities, not only in mitigation of the effects of climate change, but also as an opportunity for restoring diversity…

Grapevine yield estimation in a context of climate change: the GraY model

Grapevine yield is a key indicator to assess the impacts of climate change and the relevance of adaptation strategies in a vineyard landscape. At this scale, a yield model should use a number of parameters and input data in relation to the information available and be able to reproduce vineyard management decisions (e.g. soil and canopy management, irrigation). In this study, we used data from six experimental sites in Southern France (cv. Syrah) to calibrate a model of grapevine yield limited by water constraint (GraY). Each yield component (bud fertility, number of berries per bunch, berry weight) was calculated as a function of the soil water availability simulated by the WaLIS water balance model at critical phenological phases. The model was then evaluated in 10 grapegrowers’ plots, covering a diversity of biophysical and technical contexts (soil type, canopy size, irrigation, cover crop). We identified three critical periods for yield formation: after flowering on the previous year for the number of bunches and berries, around pre-veraison and post-veraison of the same year for mean berry weight. Yields were simulated with a model efficiency (EF) of 0.62 (NRMSE = 0.28). Bud fertility and number of berries per bunch were more accurately simulated (EF = 0.90 and 0.77, NRMSE = 0.06 and 0.10, respectively) than berry weight (EF = -0.31, NRMSE = 0.17). Model efficiency on the on-farm plots reached 0.71 (NRMSE = 0.37) simulating yields from 1 to 8 kg/plant. The GraY model is an original model estimating grapevine yield evolution on the basis of water availability under future climatic conditions.  It allows to evaluate the effects of various adaptation levers such as planting density, cover crop management, fruit/leaf ratio, shading and irrigation, in various production contexts.

Frost risk projections in a changing climate are highly sensitive in time and space to frost modelling approaches

Late spring frost is a major challenge for various winegrowing regions across the world, its occurrence often leading to important yield losses and/or plant failure. Despite a significant increase in minimum temperatures worldwide, the spatial and temporal evolution of spring frost risk under a warmer climate remains largely uncertain. Recent projections of spring frost risk for viticulture in Europe throughout the 21st century show that its evolution strongly depends on the model approach used to simulate budburst. Furthermore, the frost damage modelling methods used in these projections are usually not assessed through comparison to field observations and/or frost damage reports.
The present study aims at comparing frost risk projections simulated using six spring frost models based on two approaches: a) models considering a fixed damage threshold after the predicted budburst date (e.g BRIN, Smoothed-Utah, Growing Degree Days, Fenovitis) and b) models considering a dynamic frost sensitivity threshold based on the predicted grapevine winter/spring dehardening process (e.g. Ferguson model). The capability of each model to simulate an actual frost event for the Vitis vinifera cv. Chadonnay B was previously assessed by comparing simulated cold thermal stress to reports of events with frost damage in Chablis, the northernmost winegrowing region of Burgundy. Models exhibited scores of κ > 0.65 when reproducing the frost/non-frost damage years and an accuracy ranging from 0.82 to 0.90.
Spring frost risk projections throughout the 21st century were performed for all winegrowing subregions of Bourgogne-Franche-Comté under two CMIP5 concentration pathways (4.5 and 8.5) using statistically downscaled 8×8 km daily air temperature and humidity of 13 climate models. Contrasting results with region-specific spring frost risk trends were observed. Three out of five models show a decrease in the frequency of frost years across the whole study area while the other two show an increase that is more or less pronounced depending on winegrowing subregion. Our findings indicate that the lack of accuracy in grapevine budburst and dehardening models makes climate projections of spring frost risk highly uncertain for grapevine cultivation regions.