qNMR metabolomics a tool for wine authenticity and winemaking processes discrimination

In Champagne, the vine adaptation to different climatic and technical changes during these last 20 years can be seen through physiological balance disruptions. These disruptions emphasize the general grapevine decline. Since the 2000s, among other nitrogen stress indicators, the must nitrogen has been decreasing. The combination of restricted mineral fertilizers and herbicide use, the growing variability of spring rainfall, the increasing thermal stress as well as the soil type heterogeneity are only a few underlying factors that trigger loss of physiological balance in the vineyards. It is important to weigh and quantify the impact of these factors on the vine. In order to do so, the Comité Champagne uses two key-tools: networking and modelization. The use of quantitative and harmonized ecophysiological indicators is necessary, especially in large spatial scales such as the Champagne appellation. A working group with different professional structures of Champagne has been launched by the Comité Champagne in order to create a common ecophysiology protocol and thus monitor the vine physiology, yearly, around 100 plots, with various cultural practices and types of soil. The use of crop modelling to follow the vine physiological balance within different pedoclimatic conditions enables to understand the present balance but also predict the possible disruptions to come in future climatic scenarios. The physiological references created each year through the working group, benefit the calibration of the STICS model used in Champagne. In return, the model delivers ecophysiology indicators, on a daily scale and can be used on very different types of soils. This study will present the bottom-up method used to give accurate information on the impacts of soil, climate and cultural practices on vine physiology.

The aim of this study was to evaluate the effects of different soil types on the chemical composition of Mediterranean red wines, through untargeted and targeted 1H-NMR metabolomics. One milliliter of raw wine was analyzed by means of a Bruker Avance II 400 spectrometer operating at 400.15 MHz. The spectra were recorded by applying the NOESYGPPS1D pulse sequency, to achieve water and ethanol signals suppression. No modification of the pH was performed to avoid any chemical alteration of the matrix. The generation of input variables for untargeted analysis was done via bucketing the spectra. The resulting dataset was preprocessed prior to perform unsupervised PCA, by means of MetaboAnalyst web-based tool suite. The identification of compounds for the targeted analysis was performed by comparison to pure compounds spectra by means of SMA plug-in of MNova 14.2.3 software. The dataset containing the concentrations (%) of identified compounds was subjected to one-way analysis of variance (ANOVA) to highlight significant differences among the wines. The untargeted analysis, carried out through the PCA, revealed a clear differentiation among the wines. The fragments of the spectra contributing mostly to the separation were attributed to flavonoids, aroma compounds and amino acids. The targeted analysis leaded to the identification of 68 compounds, whose concentrations were significant different among the wines. The results were related to soils physical-chemical analysis and showed that: 1) high concentrations of flavan-3-ols and flavonols are correlated with high clay content in soils; 2) high concentrations of anthocyanins, amino acids, and aroma compounds are correlated with neutral and moderately alkaline soil pH; 3) low concentrations of flavonoids and aroma compounds are correlated with high soil organic matter content and acidic pH. The 1H-NMR metabolomic analysis proved to be an excellent tool to discriminate between wines originating from grapes grown on different soil types and revealed that soils in the Mediterranean area exert a strong impact on the chemical composition of the wines.

The main innovation in the VINIoT service is the joint use of two technologies that are currently used separately: vineyard monitoring using multispectral imaging and deployed terrain sensors. One part of the system is based on the development of artificial intelligence algorithms that are feed on the images of the multispectral camera and IoT sensors, high-level information on water stress, grape ripening status and the presence of diseases. In order to obtain algorithms to determine the state of ripening of the grapes and avoid losing information due to the diversity of the grape berries, it was decided to work along the first year 2020 at berry scale in the laboratory, during the second year at the cluster scale and on the last year at plot scale. Different varieties of white and red grapes were used; in the case of Galicia we worked with the white grape variety Treixadura and the red variety Mencía. During the 2020 and 2021 campaigns, multispectral images were taken in the visible and infrared range of: 1) sets of 100 grapes classifying them by means of densimetric baths, 2) individual bunches. The images taken with the laboratory analysis of the ripening stage were correlated. Technological maturity, pH, probable degree, malic acid content, tartaric acid content and parameters for assessing phenolic maturity, IPT, anthocyanin content were determined. It has been calculated for each single image the mean value of each spectral band (only taking into account the pixels of interest) and a correlation study of these values with laboratory data has been carried out. These studies are still provisional and it will be necessary to continue with them, jointly with the training of the machine learning algorithms. Processed data will allow to determine the sensitivity of the multispectral images and select bands of interest in maturation.

Climate is the major factor influencing the dynamics of the vegetative cycle and can determine the timing of phenological periods. Knowledge of the phenology of varieties, their chronological duration, and thermal requirements, allows not only for the better management of interventions in the vineyard, but also to predict the varieties’ behaviour in a scenario of climate change, giving the wine producer the possibility of selecting the grape varieties that are best adapted to the climatic conditions of a certain terroir. In 2014, Symington Family Estates, Vinhos, established two grape variety libraries in two different places with distinctive climate conditions (Douro Superior, and Cima Corgo), with the commitment of contributing to a deeper agronomic and oenological understanding of some grape varieties, in hot climate conditions. In these research vineyards are represented local varieties that are important in the regional and national viticulture, but also others that have over time been forgotten — as well as five international reference cultivars. From 2017 to 2021, phenological observations have been made three times a week, following a defined protocol, to determine the average dates of budbreak, flowering and veraison. With the climate data of each location, the thermal requirements of each variety and the chronological duration of each phase have been calculated. During maturation, berry samples have been gathered weekly to study the dynamics of sugar accumulation, between other parameters. The data was analysed applying phenological and sugar accumulation models available in literature. The results obtained show significant differences between the varieties over several parameters, from the chronological duration and thermal requirements to complete the various stages of development, to the differences between the two locations, confirming the influence of the climate on phenology and the stages of maturation, in these specific conditions.

The effects of climate change are seriously affecting the quality of wine grapes. High temperatures and drought cause imbalances in the chemical composition of grapes. The result is overripe grapes with low acidity and high sugar content, which produce wines with excessive alcohol content, lacking in freshness and not very aromatic. As a consequence, the search of varieties with capacity of produce quality grapes in adverse climate conditions is a good alternative to preserve the sustainability of vineyards. In this work, quality parameters of seven Vitis vinifera L. cultivars (five whites and two reds) recently recovered from extinction and grown under two different hydric regimes (rainfed and irrigated) were analyzed during the 2020 vintage. At harvest time, weight of 100 berries, must physicochemical parameters (brix degree, total acidity, malic acid, pH), and carbon and oxygen isotope ratios (δ13C, δ18O) were determined. Subsequently, varietal aroma potential index (IPAv) and total polyphenol index (TPI) were analyzed. Quality parameters, IPAv and TPI, showed significant differences between varieties and water regimes. Both red varieties, Moribel and Tinto Fragoso, stood out for their high aromatic and phenolic potential, which was higher under rainfed regime. Regarding to white varieties, Montonera del Casar and Jarrosuelto stood out in terms of varietal aroma potential. Montonera del Casar high acidity in its musts and Jarrosuelto showed the highest berry weights.