OENO IVAS 2019 banner
IVES 9 IVES Conference Series 9 OENO IVAS 9 OENO IVAS 2019 9 Analytical developments from grape to wine, spirits : omics, chemometrics approaches… 9 D-wines: use of LC-MS metabolomic space to discriminate italian mono-varietal red wines

D-wines: use of LC-MS metabolomic space to discriminate italian mono-varietal red wines

Abstract

Studying wine metabolome through multiple targeted methods is complicated and limitative; since grapes, yeasts, bacteria, oxygen, enological techniques and wine aging collaborate to deliver one of the richest metabolomic fingerprint. Therefore, untargeted metabolomics, that developed and evolved as a consequence of the need to obtain a comprehensive characterization of the organic molecules in any biological sample, is the current methodology offering the best coverage of wine metabolome. Taking into account the large genetic diversity, the diversity of the climate and of the agronomical practices, and the wide winemaking culture characterizing the Italian wines, the metabolomic untargeted approach appears as an appropriate analytical tool to study such metabolic space. 

According to the national project D-Wines, 110 single-cultivar red wines from the 2016 vintage were collected directly from wineries across different regions of Italy: Sangiovese from Tuscany and Romagna, Nebbiolo from Piemont, Aglianico from Campania, Nerello Mascalese from Sicily, Primitivo from Apulia, Raboso and Corvina from Veneto, Cannonau from Sardinia, Teroldego from Trentino, Sagrantino from Umbria, and Montepulciano from Abruzzo. The wines were analyzed according to a well-defined RP-UPLC-HRMS-QTOF-MS protocol. 

The results of the data analysis, after their validation: a) confirmed untargeted LC-MS-based metabolomics as a powerful authenticity tool; b) provided indications about the similarity between the cultivars, clustering the wines in three major groups (Primitivo – Nebbiolo, Corvina, Raboso, Sangiovese – Teroldego, Sagrantino, Cannonau, Nerello, Aglianico, Montepulciano); c) furnished a rich list of putative markers characterizing each cultivar, where Primitivo, Teroldego and Nebbiolo had the maximum number of unique putative markers; d) revealed that the putative markers were not only phenolic metabolites; and e) pointed out rt/mz chromatographic sections helpful to distinguish each cultivar from the others. 

This study, together with other D-Wines analytical results, is directed to understand the diversity of Italian red wines and to characterize them in term of metabolic space coverage/variability and taste and in consequence comprehend better their quality. 

Acknowledgements

MIUR project N. 20157RN44Y. A. Curioni, A. Gambuti, V. Gerbi, S. Giacosa, G.P. Parpinello, D. Perenzoni, P. Piombino, A. Rinaldi, S. Río Segade, B. Simonato, G. Tornielli, S. Vincenzi

DOI:

Publication date: June 19, 2020

Issue: OENO IVAS 2019

Type: Article

Authors

Panagiotis Arapitsas, Maurizio Ugliano, Matteo Marangon, Luigi Moio, Luca Rolle, Andrea Versari, Fulvio Mattivi

Department of Food Quality and Nutrition, Research and Innovation Centre, Fondazione Edmund Mach, San Michele all’Adige (Italy)
Department of Biotechnology, University of Verona (Italy)
Department of Agronomy, Food, Natural Resources, Animals and Environment, University of Padova (Italy)
Department of Agricultural Sciences, University of Naples Federico II, Avellino (Italy); Dipartimento di Scienze Agrarie, Forestali e Alimentari, Universitàdi Torino (Italy)
Department of Agricultural and Food Sciences, University of Bologna (Italy); Centre Agriculture Food Environment, University of Trento (Italy)

Contact the author

Keywords

mass spectrometry, wine authenticity, bioinformatics, metabolomics 

Tags

IVES Conference Series | OENO IVAS 2019

Citation

Related articles…

Grapevine xylem embolism resistance spectrum reveals which varieties have a lower mortality risk in a future dry climate

Wine growing regions have recently faced intense and frequent droughts that have led to substantial economical losses, and the maintenance of grapevine productivity under warmer and drier climate will rely notably on planting drought-resistant cultivars. Given that plant growth and yield depend on water transport efficiency and maintenance of photosynthesis, thus on the preservation of the vascular system integrity during drought, a better understanding of drought-related hydraulic traits that have a significant impact on physiological processes is urgently needed. We have worked towards this end by assessing vulnerability to xylem embolism in 30 grapevine commercial varieties encompassing red and white Vitis vinifera varieties, hybrid varieties characterized by a polygenic resistance for powdery and downy mildew, and commonly used rootstocks. These analyses further allowed a global assessment of wine regions with respect to their varietal diversity and resulting vulnerability to stem embolism. Hybrid cultivars displayed the highest vulnerability to embolism, while rootstocks showed the greatest resistance. Significant variability also arose among Vitis vinifera varieties, with Ψ12 and Ψ50 values ranging from -0.4 to -2.7 MPa and from -1.8 to -3.4 MPa, respectively. Cabernet franc, Chardonnay and Ugni blanc featured among the most vulnerable varieties while Pinot noir, Merlot and Cabernet Sauvignon ranked among the most resistant. In consequence, wine regions bearing a significant proportion of vulnerable varieties, such as Poitou-Charentes, France and Marlborough, New Zealand, turned out to be at greater risk under drought. These results highlight that grapevine varieties may not respond equally to warmer and drier conditions, outlining the importance to consider hydraulic traits associated with plant drought tolerance into breeding programmes and modeling simulations of grapevine yield maintenance under severe drought. They finally represent a step forward to advise the wine industry about which varieties and regions would have the lowest risk of drought-induced mortality under climate change.

Elucidating vineyard site contributions to key sensory molecules: Identification of correlations between elemental composition and volatile aroma profile of site-specific Pinot noir wines

The reproducibility of elemental profile in wines produced across multiple vintages has been previously reported using grapes from a single scion clone of Vitis vinifera L. cv. Pinot noir. The grapevines were grown on fourteen different vineyard sites, from Oregon to southern California in the U.S.A., which span distances from approximately hundreds of meters to 1450 km, while elevations range from near sea level to nearly 500 m. In addition, sensorial (i.e. aroma, taste, and mouthfeel) and chemical (i.e. polyphenolic and volatile) differences across the different vineyard sites have also been observed among these wines at two aging time points. While strong evidence exists to support that grapes grown in different regions can produce wines with unique chemical and sensorial profiles, even when a single clone is used, the understanding of growing site characteristics that result in this reproducible differentiation continues to emerge. One hypothesis is that the elemental profile that a vineyard site imparts to the grape berries and the resulting wine is an important contributor to this differentiation in chemistry and sensory of wines. For example, various classes of enzymes that catalyze the formation of key aroma compounds or their precursors require specific metals. In this work, we begin to report correlations between elemental and volatile aroma profiles of site-specific Pinot noir wines, made under standardized winemaking conditions, that have been previously shown to be distinguished separately by these chemical analyses.

Mobile device to induce heat-stress on grapevine berries

Studying heat stress response of grapevine berries in the field often relies on weather conditions during the growing season. We constructed a mobile heating device, able to induce controlled heat stress on grapes in vineyards. The heater consisted of six 150 W infrared lamps mounted in a profile frame. Heating power of the lamps could be controlled individually by a control unit consisting of a single board computer and six temperature sensors to reach a pre-set temperature. The heat energy applied to individual berries within a cluster decreases by the squared distance to the heat source, enabling the establishment of temperature profiles within individual clusters. These profiles can be measured by infrared thermography once a steady state has been reached. Radiant flux density received by a berry depending on the distance was calculated based on a view factor and measured lamp surface temperature and resulted to 665 Wm-2 at 7cm. Infrared thermography of the fruit surface was in good agreement with measurements conducted with a thermocouple inserted at epidermis level. In combination with infrared thermography, the presented device offers possibilities for a wide range of applications like phenotyping for heat tolerance in the field to proceed in the understanding of the complex response of plants to heat stress. Sunburn necrosis symptoms were artificially induced with the aid of the device for cv. Bacchus and cv. Sylvaner in the 2020 and 2021 growing season. Threshold temperatures for sunburn induction (LT5030min) were derived from temperature data of single berries and visual sunburn assessment, applying logistic regression. A comparison of threshold temperatures for the occurrence of sunburn necrosis confirmed the higher susceptibility of cv. Bacchus. The lower susceptibility of cv. Sylvaner did not seem to be related to its phenolic composition, rendering a thermoprotective role of berry phenolic compounds unlikely.

Investigating the impact of grape exposure and UV radiations on rotundone in Vitis vinifera L. Tardif grapes under field trial conditions

Rotundone is the main aroma compound responsible for peppery notes in wines whose biosynthesis is negatively affected by heat and drought. Through the alteration of precipitation regime and the increase in temperature during maturation, climate change is expected to affect wine peppery typicality. In this context there is a demand for developing sustainable viticultural strategies to enhance rotundone accumulation or limit its degradation. It was recently proposed that ultraviolet (UV) radiations could stimulate rotundone production. The aim of this study was to investigate under field trial conditions the impact of grape exposure and UV treatments on rotundone in Vitis vinifera L. Tardif, an almost extinct grape variety from south-west France that can express particularly high rotundone levels. Four different treatments were compared in 2021 to a control treatment using a randomised complete block design with three replications per treatment. Grape exposure was manipulated through early or late defoliation. Leaf and laterals shoots were removed at Eichorn Lorenz growth stages 32 or 34 on the morning-sun side of the canopy. During grape maturation, UV radiations were either reduced by 99% by installing UV radiation-shielding sheets, or applied four times using the Boxilumix™ non thermal device (Asclepios Tech, Tournefeuille) with the aim of activating plant signalling pathway. Loggers displayed in solar radiation shields were used to assess the effect of such shielding sheets on air temperature within the bunch zone. The composition of grapes subjected to these treatments will be soon analysed for their rotundone content and basic classical laboratory analyses. Grapes will be harvested to elaborate wines under standardized small-scale vinification conditions (60kg) that will be assessed by a trained sensory panel.

Metabolomic discrimination of grapevine water status for Chardonnay and Pinot noir

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.