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

The objective of the work described here is the elaboration of a map of the different types of vineyard soils that to guide the famers in the choice of the most productive vine rootstocks and varieties. 90 vineyard soils profiles were analysed in the entire territory of the Origen Denominations of Valdepeñas. The sampling was carried out in 2018 (June to October) by making a sampling grid, followed by photointerpretation and control in the field. The studied soils can be grouped into 9 different soil types (according to FAO 2006 classification): Leptosols, Regosols, Fluvisols, Gleysols, Cambisols, Calcisols, Luvisols and Anthrosols. A map showing the soil distribution with different type of soils has been made with the ArcGIS program. Regarding to the choice of rootstock, Calcisoles are soils with a high active limestone content, so the rootstocks used in these soils must be resistant to this parameter; Luvisols are deep soils with high clay content, so they will support vigorous rootstocks. Because the cartographic units are composed of two or more subgroups, with are associated in variable proportions, 9 different soil associations have been established; Unit 1: Leptosols, Cambisols and Luvisols (80%, 15% and 5% respectively); Unit 2: Cambisols with Regosols and Luvisols (40%, 30% and 30% respectively); Unit 3: Cambisols and Gleysols with Regosols (40%, 40% and 20% respectively); Unit 4: Regosols with Cambisols, Leptosols and Calcisols (40%, 30%, 15% and 15% respectively); Unit 5: Cambisols, Leptosols, Calcisols and Regosols (25% each of them); Unit 6: Luvisols with Cambisol and Calcisols (80%, 10% and 10% respectively); Unit 7: Luvisols and Calcisols with Cambisols (40%, 40% and 20% respectively); Unit 8: Calcisols with, Cambisols and Luvisols (80%, 10% and 10% respectively); Unit 9: Anthrosols. These study allow to elaborate the first map of vineyard soils of this Protected Designation of Origin in Castilla-La Mancha.

In the last decades, climate change required already adaptation of vineyard management. Increase in temperature and unexpected weather events cause changes in all phenological stages requiring new management tools. For example, defoliation can be a useful tool to reduce the sugar content in the berries creating differences in the wine profiles. In a ten-year field experiment using Riesling (Vitis vinifera L, planted 1986, Geisenheim, Germany), various mechanical defoliation strategies and different intensities were trialed until 2016 before the vineyard was uprooted. Wood was sampled from the plant compartments root, trunk, cordon and shoot for analyses of physicochemical properties (e.g. lignin and element content, pH, diameter), nonstructural carbohydrates and the microbial communities. The aim of the study was to investigate the influence of reduced canopy leaf area on the sink-source allocation into different compartments and potential changes of the fungal and prokaryotic wood-inhabiting community using a metabarcoding approach. Severe summer pruning (SSP) of the canopy and mechanical defoliation (MDC) above the bunch zone decreased the leaf area by 50% compared to control (C). SSP reduced the photosynthetic capacity, which resulted in an altered source-sink allocation and carbohydrate storage. With lower leaf area, less carbohydrates are allocated. This for example resulted in a decreased trunk diameter. Further, it affected the composition of the grapevine wood microbiota. SSP and MDC management changed significantly the prokaryotic community composition in wood of the root samples, but had no effect in other compartments. In general, this study found strong compartment and less management effects of the microbial community composition and associated physicochemical properties. The highest microbial diversities were identified in the wood of the trunk, and several species were recorded the first time in grapevine.

The current climate changes are directly threatening the balance of the vineyard at harvest time. The maturation period of the grapes is shifted to the middle of the summer, at a time when radiation and air temperature are at their maximum. In this context, the implementation of corrective practices becomes problematic. Unfortunately, our knowledge of the climate effect on the quality of different grape varieties remains very incomplete to guide these choices. During the Innovine project, original experiments were carried out on Syrah to study the combined effects of normal or high air temperature and varying degrees of exposure of the berries to the sun. Berries subjected to these different conditions were sampled and analyzed throughout the maturation period. Several quality characteristics were determined, including anthocyanin content. The objective of the experiments was to investigate which climatic determinants were most important for anthocyanin accumulation in the berries. Temperature and irradiance data, observed over time with a very thin discretization step, are called functional data in statistics. We developed the procedure SpiceFP (Sparse and Structured Procedure to Identify Combined Effects of Functional Predictors) to explain the variations of a scalar response variable (a grape berry quality variable for example) by two or three functional predictors (as temperature and irradiance) in a context of joint influence of these predictors. Particular attention was paid to the interpretability of the results. Analysis of the data using SpiceFP identified a negative impact of morning combinations of low irradiance (lower than about 100 μmol m−2 s−1 or 45 μmol m−2 s−1 depending on the advanced-delayed state of the berries) and high temperature (higher than 25oC). A slight difference associated with overnight temperature occurred between these effects identified in the morning.

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.

Irrigation is an important practice to influence vine quality, especially in Mediterranean regions, characterized by hot summers and severe droughts during the growing season. This study focused on deficit irrigation regime influence on amino acids composition of Monastrell grapevines under semiarid conditions (Albacete, Southeastern of Spain). In 2019, two treatments were applied: non-irrigation (NI) and regulated deficit irrigation (RDI), watered at 30% of the estimated crop evapotranspiration from fruit set to onset of veraison. Grape amino acids content was analyzed by HPLC. Berries from non-irrigated vines showed higher concentration of several amino acids, such as tryptophan (73%), arginine (70%), lysine (36%), isoleucine (27%), and leucine (21%), compared to RDI grapes. Arginine is, together with ammonium ion, the principal nitrogen source for yeasts during the alcoholic fermentation; while isoleucine, tryptophan, and leucine are precursors of fermentative volatile compounds, key compounds for wine quality. Moreover, NI treatment increased in a 14% the total amino acids content in grapes compared to RDI treatment. The reported effects might be because yield was 70% higher in RDI vines than in the NI ones and, therefore, the sink demand was increased in the irrigated vines. In addition, NI vines suffered more severe water stress and it is known that the amino acids synthesis and accumulation can be influenced by the plant response to stress. According to the results, the irrigation regime showed effect on amino acids concentration in Monastrell grapes under semiarid conditions. Grapes from non-irrigated vines showed a higher content of several amino acids relevant to the fermentative process and to the wine aroma compounds formation. It is demonstrated that the final content of nitrogen-related components in grapes is influenced by the irrigation regime. The convenience of the irrigation strategy to suggest will depend on the desired wine style and the target yield levels.