Macrowine 2021
IVES 9 IVES Conference Series 9 Macrowine 9 Macrowine 2021 9 Grapevine diversity and viticultural practices for sustainable grape growing 9 Targeted UHPLC-QqQ-MS/MS metabolomics for phenol identification in grapevine and wine: study of a Tempranillo clone with a dark-blue berry colour

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

Abstract

Grapevine vegetative multiplication allows the accumulation of spontaneous mutations and increase intra-cultivar genetic diversity that can be exploited to maintain grape wine quality, tipicity and adaptation to different climate conditions. Non-volatile phenolic compounds are intrinsic components of grape fruits and derived products, particularly wine. They constitute a heterogeneous family of compounds and play an important role on the sensorial attributes of wine because they are responsible for some of important organoleptic properties as colour, flavour, bitterness and astringency. In the present study, we used a targeted metabolomics approach based on ultra-high performance liquid chromatography with tandem mass spectrometry (UHPLC-QqQ-MS/MS) to study the anthocyanin and non-coloured phenol profiles of a singular Tempranillo clone (Tempranillo negro or VN21), characterized by a dark-blue color in grape berry skin, as compared to RJ43, one of the most cultivated clones in D.O.Ca. Rioja (Spain). In addition, we investigated differences between VN21 and RJ43 clones, in the phenolic transference from grape to wine at different phases of the winemaking process. The results showed that anthocyanin and non-colored phenol content was higher in VN21 grape skin and seeds than in RJ43. With respect to anthocyanins, the singular color of grape skin in VN21 could be explained by higher concentrations of peonidin and cyanidin derivatives. Regarding non-colored phenols, the main differences were observed for proanthocyanidins and stilbenes concentration in grape skin and more importantly in seeds. Those content differences observed in berries were enhanced in the VN21 wines, displaying significantly higher concentrations of anthocyanins, as well as significantly increased contents of mainly proanthocyanidins and stilbenes. The results manifest the importance of intra-cultivar genetic diversity to obtain red wines with a high phenolic content, responsible of key quality aspects of the wine such as organoleptic properties, stability, complexity and health benefits. Moreover, this study exemplifies how spontaneous somatic variation can be used through grapevine clonal selection combining metabolomic analyses.

FUNDING SOURCES

This work was partially supported by project BIO2017-86375-R from the Spanish Ministry of Economy and Competitiveness (co-funded by the European Social Fund, European Union); YF was supported by a grant from Government of La Rioja; M.J. Motilva thanks to CSIC for partial funding through the “Ayudas incorporación a escalas científicas CSIC, 2018” (Reference 201870I129).

DOI:

Publication date: September 2, 2021

Issue: Macrowine 2021

Type: Article

Authors

Yolanda Ferradás 

Instituto de Ciencias de la Vid y del Vino (Consejo Superior de Investigaciones Científicas, Universidad de La Rioja, Gobierno de La Rioja), Finca La Grajera, Ctra. de Burgos Km. 6 (LO-20 – salida 13). 26007 Logroño (La Rioja), Spain,Carolina ROYO, José Miguel MARTÍNEZ-ZAPATER and María José MOTILVA  Instituto de Ciencias de la Vid y del Vino (Consejo Superior de Investigaciones Científicas, Universidad de La Rioja, Gobierno de La Rioja), Finca La Grajera, Ctra. de Burgos Km. 6 (LO-20 – salida 13). 26007 Logroño (La Rioja), Spain

Contact the author

Keywords

anthocyanins, berry phenolic composition, wine phenolic composition, somatic variation, grapevine, phenolic compounds, stilbenes, tempranillo

Citation

Related articles…

The use of rootstock as a lever in the face of climate change and dieback of vineyard

As viticulture faces challenges such as climate change or vineyard dieback, the choice of the variety and rootstock becomes more and more crucial. To study rootstock levers in the Bordeaux region, a parcel of Cabernet Sauvignon (CS) was planted with four rootstocks in 2014. Twenty repetitions of each of the following four rootstocks were set up: 101-14 MGt, Nemadex AB, 420A MGt and Gravesac. The number of bunches, yields and pruning weights of the vine shoots were measured individually on 240 vines from 2017 to 2021. Since 2020, nitrogen status assessed by assimilable nitrogen level, hydric status assessed by δ13C and berry maturity were measured on 80 samples taken from 20 repetitions of the four rootstocks. A lower yield was measured for CS grafted onto Nemadex AB due to the lower number of bunches and the lower weight of berries. The differences between the other three rootstocks are small, but CS grafted onto 420A MGt was the most productive. The CS grafted onto Nemadex AB had the lowest pruning weight while 101-14 MGt had the highest. In 2020, δ13C showed a more moderate water stress with 101-14 MGt and 420A MGt than with Nemadex AB. Surprisingly, the Gravesac was under more stress than the 101-14 MGt. The nitrogen status in the berries was better for Nemadex AB but this was perhaps due to the significantly lower weight of the berries.Rootstock 101-14 MGt attained the highest accumulation of sugars in the berries while 420A MGt allows to preserve higher acidity. The parcel is still young which may explain some of the results. These measures must therefore be continued over the next several years to fully assess the effects of these rootstocks on the development of the vines and the quality of the production under new climatic conditions.

A better understanding of the climate effect on anthocyanin accumulation in grapes using a machine learning approach

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.

VineyardFACE: Investigation of a moderate (+20%) increase of ambient CO2 level on berry ripening dynamics and fruit composition

Climate change and rising atmospheric carbon dioxide concentration is a concern for agriculture, including viticulture. Studies on elevated carbon dioxide have already been on grapevines, mainly taking place in greenhouses using potted plants or using field grown vines under higher CO2 enrichment, i.e. >650 ppm. The VineyardFACE, located at Hochschule Geisenheim University, is an open field Free Air CO2 Enrichment (FACE) experimental set-up designed to study the effects of elevated carbon dioxide using field grown vines (Vitis vinifera L. cvs. Riesling and Cabernet Sauvignon). As the carbon dioxide fumigation started in 2014, the long term effects of elevated carbon dioxide treatment can be investigated on berry ripening parameters and fruit metabolic composition.
The present study aims to investigate the effect on fruit composition under a moderate increase (+20%; eCO2) of carbon dioxide concentration, as predicted for 2050 on both Riesling and Cabernet Sauvignon. Berry composition was determined for primary (sugars, organic acids, amino acids) and secondary metabolites (anthocyanins). Special focus was given on monitoring of berry diameter and ripening rates throughout three growing seasons. Compared to previous results of the early adaptative phase of the vines [1], our results show little effect of eCO2 treatment on primary metabolites composition in berries. However, total anthocyanins concentration in berry skin was lower for eCO2 treatment in 2020, although the ratio between anthocyanins derivatives did not differ.
[1] Wohlfahrt Y., Tittmann S., Schmidt D., Rauhut D., Honermeier B., Stoll M. (2020) The effect of elevated CO2 on berry development and bunch structure of Vitis vinifera L. cvs. Riesling and Cabernet Sauvignon. Applied Science Basel 10: 2486

Heatwaves and grapevine yield in the Douro region, crop model simulations

Heatwaves or extreme heat events can be particularly harmful to agriculture. Grapevines grown in the Douro winemaking region are particularly exposed to this threat, due to the specificities of the already warm and dry climatic conditions. Furthermore, climate change simulations point to an increase in the frequency of occurrence of these extreme heat events, therefore posing a major challenge to winegrowers in the Mediterranean type climates. The current study focuses on the application of the STICS crop model to assess the potential impacts of heatwaves in grapevine yields over the Douro valley winemaking region. For this purpose, STICS was applied to grapevines using high-resolution weather, soil and terrain datasets over the Douro. To assess the impact of heatwaves, the weather dataset (1989-2005) was artificially modified, generating periods with anomalously high temperatures (+5 ºC), at certain onset dates and with specific durations (from 5 to 9 days). The model was run with this modified weather dataset and results were compared to the original unmodified runs. The results show that heatwaves can have a very strong impact on grapevine yields, strongly depending on the onset dates and duration of the heatwaves. The highest negative impacts may result in a decrease in the yield by up to -35% in some regions. Despite some uncertainties inherent to the current modelling assessment, the present study highlights the negative impacts of heatwaves on viticultural yields in the Douro region, which is critical information for stakeholders within the winemaking sector for planning suitable adaptation measures.

Variations of soil attributes in vineyards influence their reflectance spectra

Knowledge on the reflectance spectrum of soil is potentially useful since it carries information on soil chemical composition that can be used to the planning of agricultural practices. If compared with analytical methods such as conventional chemical analysis, reflectance measurement provides non-destructive, economic, near real-time data. This paper reports results from reflectance measurements performed by spectroradiometry on soils from two vineyards in south Brazil. The vineyards are close to each other, are on different geological formations, but were subjected to the same management. The objective was to detect spectral differences between the two areas, correlating these differences to variations in their chemical composition, to assess the technique’s potential to predict soil attributes from reflectance data.To that end, soil samples were collected from ten selected vine parcels. Chemical analysis yield data on concentration of twenty-one soil attributes, and spectroradiometry was performed on samples. Chemical differences significant to a 95% confidence level between the two studied areas were found for six soil attributes, and the average reflectance spectra were separated by this same level along most of the observed spectral domain. Correlations between soil reflectance and concentrations of soil attributes were looked for, and for ten soil traits it was possible to define wavelength domains were reflectance and concentrations are correlated to confidence levels from 95% to 99%. Partial Least Squares Regression (PLSR) analyses were performed comparing measured and predicted concentrations, and for fifteen out of 21 soil traits we found Pearson correlation coefficients r > 0.8. These preliminary results, which have to be validated, suggest that variations of concentration in the investigated soil attributes induce differences in reflectance that can be detected by spectroradiometry. Applications of these observations include the assessment of the chemical content of soils by spectroradiometry as a fast, low-cost alternative to chemical analytical methods.