Targeted UHPLC-QqQ-MS/MS metabolomics for phenol identification in grapevine and wine: study of a Tempranillo clone with a dark-blue berry colour

Our research aimed to determine the effect and efficiency of foliar application of urea on the phenolic composition of Tempranillo Blanco grapes. The field experiment was carried out in 2019 and 2020 seasons and the plot was located in D.O.Ca Rioja (North of Spain). The vineyard was Vitis vinifera L. Tempranillo Blanco and grafted on Richter-110 rootstock. The treatments were control (C), whose plants were sprayed with water and three doses of urea: plants were sprayed with urea 3 kg N/ha (U3), 6 kg N/ha (U6) and 9 kg N/ha (U9). The applications were performed in two phenological stages, pre-veraison (Pre) and veraison (Ver). Also, each of the treatments was repeated one week later. Control and treatments were performed in triplicate and arranged in a randomised block design. Grapes were harvested at optimum ripening stage. High-performance liquid chromatography was used to analyse the phenolic composition of the grapes. Finally, the results obtained from the analytical determinations – flavonols, flavanols and non-flavonoid (hydroxybenzoic acids, hydroxycinnamic acids and stilbenes) – were studied statistically by analysis of variance. The results showed that, in 2019, U6-Pre and U9-Pre treatments increased the hydroxybenzoic acid content in grapes, and also all foliar treatments applied at Pre enhanced the stilbene concentration. Moreover, U3-Ver was the only treatment that rose flavonol and stilbene contents in the Tempranillo Blanco grapes. In 2020, all treatments applied at Pre enhanced the flavonol concentration in grapes. Furthermore, U3-Pre and U9-Pre treatments increased stilbene content in grapes. Nevertheless, the hydroxybenzoic acid content was improved by U6-Ver and U9-Ver and besides, hydroxycinnamic acid concentration in grapes was increased by all treatments applied at Ver. In conclusion, the lower and highest dose of urea (U3 and U9), applied at pre-veraison, were the best treatments to improve the Tempranillo Blanco grape phenolic composition.

The wine sector is of great relevance and tradition in Mediterranean countries, however, it may be most susceptible to climate change. In recent years, wine production is facing changes worldwide, both at environmental as well as commercial levels, due to global warming and the shift in consumers’ preferences. Wine growers and wine makers are in search of solutions that allow to face these new challenges. One of the most promising initiatives in the long term is the introduction of new plant materials, specifically intraspecific hybridizations between premium varieties that may improve traditional germplasm in its adaptation to climate change. These inter-varietal crosses have the potential to generate quality wines, whilst maintaining the regional typicity, and constitute an attractive alternative for the consumer due to their sensory attributes. In this study, we have evaluated wines from 29 intraspecific Garnacha x Tempranillo hybrids in two different locations, with the aim to assess their oenological potential and sensory attributes. Thirteen of the selections were white and 16 were red. Microvinifications were conducted with two or three replications depending on grape availability. Conventional oenological parameters were determined for all wines. The sensory evaluation and hedonic scores were given by five experts. Red selections obtained higher quality scores than white ones. Among the white selections with higher quality scores, GT-41 Varea and GT-159 Varea outstand, due to their high total acidity and high malic acid content. Regarding red selections, GT-57 Varea and GT-57 UR were perceived as higher in quality, highlighted for their moderate alcoholic and high anthocyanin content. Our results indicate that intraspecific hybridization may be a powerful tool for adapting traditional cultivars to climate change in Rioja.

Water status impact in viticulture has been widely explored, as it strongly affects grapevine physiology and grape chemical composition. It is considered as a key component of vitivinicultural terroir. Most of the studies concerning grapevine water status have focused on either physiological traits, or berry compounds, or traits involved in wine quality. Here, the response of grapevine to water availability during the ripening period is assessed through non-targeted metabolomics analysis of grape berries by ultra-high resolution mass spectrometry. The grapevine water status has been assessed during 2 consecutive years (2019 & 2020), through carbon isotope discrimination on juices from berries collected at maturity (21.5 brix approx.) for 2 Vitis vinifera cv. Pinot noir (PN) and Chardonnay (CH). A total of 220 grape juices were collected from 5 countries worldwide (Italy; Argentina; France; Germany; Portugal). Measured δ13C (‰) varied from -28.73 to -22.6 for PN, and from -28.79 to -21.67 for CH. These results also clearly revealed higher water stress for the 2020 vintage. The same grape juices have been analysed by Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (FT-ICR-MS) and Liquid Chromatography coupled to Mass Spectrometry (LC-qTOF-MS), leading to the detection of up to 4500 CHONS containing elemental compositions, and thus likely tens of thousands of individual compounds, which include fatty acids, organic acids, peptides, phenolics, also with high levels of glycosylation. Multivariate statistical analysis revealed that up to 160 elemental compositions, covering the whole range of detected masses (100 –1000 m/z), were significantly correlated to the observed gradients of water status. Examples of chemical markers, which are representative of these complex fingerprints, include various derivatives of the known abscisic acid (ABA), such as phaesic acid or abscisic acid glucose ester, which are significantly correlated with higher water stress, regardless of the variety. Cultivar-specific behaviours could also be identified from these fingerprints. Our results provide an unprecedented representation of the metabolic diversity, which is involved in the water status regulation at the grape level, and which could contribute to a better knowledge of the grapevine mitigation strategy in a climate change context.

The OIV[i] defines terroir as a concept referring to an area in which collective knowledge of the interactions between the physical and biological environment (soil, topography, climate, landscape characteristics and biodiversity features) and vitivinicultural practices develops, providing distinctive wine characteristics. Those are perceptible in the taste of wine, which drives consumer preference and, therefore, wine’s value in the marketplace. Geographical indications (GI) are recognized regulatory constructs formalizing and protecting the nexus between wine taste and the terroir generating it. Despite considering updates, GIs do not consider the nexus as a dynamic one and do not anticipate change, namely of climate. Being climate a fundamental feature of terroir, it strongly impacts wine characteristics, such as taste. According to IPCC[ii], many widespread, rapid and unprecedented changes of climate occurred, some being irreversible over hundreds to thousands of years. Climatic shifts and atmospheric-driven extreme events have been widely reported worldwide. Recent climatic trends are projected to strengthen in upcoming decades, whereas extremes are expected to increase in frequency and intensity, forcing wines away from GI definitions. Geographical shifts of viticultural suitability are projected, often moving into regions and countries different from current ones. Some authors propose adaptation in viticulture, winemaking and product innovation. We show evidence of climate changing wine characteristics in the Douro valley, home of 270-year-old Port GI. We discuss herein resist or adapt stances for when climate changes the nexus between terroir and wine characteristics. Using the MED-GOLD[iii] dashboard, a tool allowing for easy visual navigation of past and future climates, we demonstrate how policymakers can identify future moments, throughout the 21st century under different emission scenarios, when GI specifications will likely need updates (e.g., boundaries, varieties) to reduce climate-change impacts.

Purpose of this research is to quantitatively assess the mineral component of vineyard soils, with particular attention to the mineralogical analysis of clays, which represent an element of high importance in the vineyard culture as well as in general agriculture. An X-ray diffraction (XRD) / thermogravimetric (TG) multi-technique analytical approach was developed, tested on soil samples taken from vineyards around the world. This codified analytical procedure was necessary to obtain precise qualitative and quantitative mineralogical data, globally comparable to distinguish the geopedological identity of the vineyards. Soil samples from vineyards of various locations were analysed, in very different geological conditions. The bulk-rock quantitative phase analysis (QPA) was obtained by the Rietveld method while the detailed composition of the clay-sized fraction was determined by modelling of the oriented X-ray diffraction patterns. The research provided a precise classification of the mineral component of soils, distinguishing the mineral phases of the clays and the so-called mixed-layer clay minerals. We found that the content in mixed layers can be directly correlated with the water retention and the cation exchange capacity ​​of the soil, while the presence of other clayey minerals and phyllosilicates in this research did not affect this CEC parameter, which codes the fertility level of the soils. The study demonstrates that terroir, in particular soils formed in complex or very different geological conditions, can only be effectively interpreted by properly analysing its mineral phases, in particular the mixed-layer clay component. These are characteristic abiotic ecological indicators, which may have specific eco-physiological influences on the plant.