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…

What are the optimal ranges and thresholds for berry solar radiation for flavonoid biosynthesis?

In wine grape production, canopy management practices are applied to control the source-sink balance and improve the cluster microclimate to enhance berry composition. The aim of this study was to identify the optimal ranges of berry solar radiation exposure (exposure) for upregulation of flavonoid biosynthesis and thresholds for their degradation, to evaluate how canopy management practices such as leaf removal, shoot thinning, and a combination of both affect the grapevine (Vitis vinifera L. cv. Cabernet Sauvignon) yield components, berry composition, and flavonoid profile under context of climate change. First experiment assessed changes in the grape flavonoid content driven by four degrees of exposure. In the second experiment, individual grape berries subjected to different exposures were collected from two cultivars (Cabernet Sauvignon and Petit Verdot). The third experiment consisted of an experiment with three canopy management treatments (i) LR (removal of 5 to 6 basal leaves), (ii) ST (thinned to 24 shoots per vine), and (iii) LRST (a combination of LR and ST) and an untreated control (UNT). Berry composition, flavonoid content and profiles, and 3-isobutyl 2-methoxypyrazine were monitored during berry ripening. Although increasing canopy porosity through canopy management practices can be helpful for other purposes, this may not be the case of flavonoid compounds when a certain proportion of kaempferol was achieved. Our results revealed different sensitivities to degradation within the flavonoid groups, flavonols being the only monitored group that was upregulated by solar radiation. Within different canopy management practices, the main effects were due to the ST. Under environmental conditions given in this trial, ST and LRST hastened fruit maturity; however, a clear improvement of the flavonoid compounds (i.e., greater anthocyanin) was not observed at harvest. Methoxypyrazine berry content decreased with canopy management practices studied. Although some berry traits were improved (i.e. 2.5° Brix increase in berry total soluble solids) due to canopy management practices (ST), this resulted in a four-fold increase in labor operations cost, two-fold decrease in yield with a 10-fold increase in anthocyanin production cost per hectare that should be assessed together as the climate continues to get hot.

Differential responses of red and white grape cultivars trained to a single trellis system – the VSP

Commercial grape production relies on training grapevine cultivars onto a variety of trellis systems. Training allows for well-lit leaves and clusters, maximizing fruit quality in addition to facilitating cultivation, harvesting, and diseases control. Although grapevines can be trained onto an infinite variety of trellis systems, most red and white cultivars are trained to the standard VSP (Vertical Shoot Positioning) system. However, red and white cultivars respond differently to VSP in fruit composition and growth characteristics, which are yet to be fully understood. Therefore, the objective of this study was to examine the influence of the VSP trellis system on fruit composition of three red, Cabernet Sauvignon, Merlot and Syrah, and three white, Chardonnay, Riesling, and Gewurztraminer cultivars grown under uniform growing conditions in the same vineyard. All cultivars were monitored for maturity and harvested at their physiologically maximum possible sugar concentration to compare various fruit quality attributes such as Brix, pH, TA, malic and tartaric acids, glucose and fructose, potassium, YAN, and phenolic compounds including total anthocyanins, anthocyanin profile, and tannins. A distinct pattern in fruit composition was observed in each cultivar. In regards to growth characteristics, Syrah grew vigorously with the highest cluster weight. Although all cultivars developed pyriform seeds, the seed size and weight varied among all cultivars. Also varied were mesocarp cell viability, brush morphology, and cane structure. This knowledge of the canopy architectural characteristics assessed by the widely employed fruit compositional attributes and growth characteristics will aid the growers in better management of the vines in varied situations.

Local ancient grapevine cultivars to face future viticulture

Among the different strategies to cope with the negative impacts of climate change on viticulture, the exploitation of genetic diversity is one of the most promising to adapt to new conditions and maintain wine production and quality. One of the biggest concerns in the context of climate change is to improve water use efficiency (WUE). In this way, the use of genotypes that present a better response to drought and high WUE is a key issue. In this work, physiological performance analysis was conducted to compare the water deficit stress (WDS) responses of local and widespread grapevines cultivars. Leaf gas exchange, water use efficiency (WUE) at different levels (leaf and long-term WUE (∆13C)), leaf osmotic adjustment and other water relations parameters were determined in plants under well-watered and WDS conditions alongside assessment of the levels of foliar hormones concentrations. Results denote that local cultivars displayed better physiological performance under WDS as compared to the widely-distributed ones. he results corroborate the hypothesis that better stomatal control allows increasing leaf WUE under drought as occurred in the local Callet cv.; but the minority local cultivar Escursac cv. showed high WUE under both treatments. In this case, high WUE can be related to maintaining higher photosynthetic activity under drought. The different mechanisms underlying the better performance under WDS and high WUE of minority local cultivars are discussed.

Impact on leaf morphology of Vitis vinifera L. cvs Riesling and Cabernet Sauvignon under Free Air Carbon dioxide Enrichment (FACE)

Atmospheric carbon dioxide (CO2) concentration has continuously increased since pre-industrial times from 280 ppm in 1750, and is predicted to exceed 700 ppm by the end of 21st century. For most of C3 plant species elevated CO2 (eCO2) improve photosynthetic apparatus results in an increased plant biomass production. To investigate the effects of eCO2 on morphological leaf characteristics the two Vitis vinifera L. cultivars, Riesling and Cabernet Sauvignon, grown in the Geisenheim VineyardFACE (Free Air Carbon dioxide Enrichment) system were used. The FACE site is located at Geisenheim University (49° 59′ N, 7° 57′ E, 94 m above sea level), Germany and was implemented in 2014 comparing future atmospheric CO2-concentrations (eCO2, predicted for the mid-21st century) with current ambient CO2-conditions (aCO2). Experiments were conducted under rain-fed conditions for two consecutive years (2015 and 2016). Six leaves per repetition of the CO2 treatment were sampled in the field and immediately fixed in a FAA solution (ethanol, H2O, formaldehyde and glacial acetic acid). After 24 h leaf samples were transferred and stored in an ethanol solution. Subsequently, leaf tissue was dehydrated using ethanol series and embedded in paraffin. By using a rotary microtomesections of 5 µm were prepared and fixed on microscopic slides. Subsequent the samples were stained using consecutive staining and washing solutions. Afterwards pictures of the leaf cross-sections were taken using a light microscope and consecutive measurements were conducted with an open source image software. Differences found in leaf cross-sections of the two CO2 treatments were detected for the palisade parenchyma. Leaf thickness, upper and lower epidermis and spongy parenchyma remained less affected under eCO2 conditions. The observed results within grapevine leaf tissues can provide first insights to seasonal adaptation strategies of grapevines under future elevated CO2 concentrations.

Adapting the vineyard to climate change in warm climate regions with cultural practices

Since the 1980s global regime shift, grape growers have been steadily adapting to a changing climate. These adaptations have preserved the region-climate-cultivar rapports that have established the global trade of wine with lucrative economic benefits since the middle of 17th century. The advent of using fractions of crop and actual evapotranspiration replacement in vineyards with the use of supplemental irrigation has furthered the adaptation of wine grape cultivation. The shift in trellis systems, as well as pruning methods from positioned shoot systems to sprawling canopies, as well as adapting the bearing surface from head-trained, cane-pruned to cordon-trained, spur-pruned systems have also aided in the adaptation of grapevine to warmer temperatures. In warm climates, the use of shade cloth or over-head shade films not only have aided in arresting the damage of heat waves, but also identified opportunities to reduce the evapotranspiration from vineyards, reducing environmental footprint of vineyard. Our increase in knowledge on how best to understand the response of grapevine to climate change was aided with the identification of solar radiation exposure biomarker that is now used for phenotyping cultivars in their adaptability to harsh environments. Using fruit-based metrics such as sugar-flavonoid relationships were shown to be better indicators of losses in berry integrity associated with a warming climate, rather than solely focusing on region-climate-cultivar rapports. The resilience of wine grape was further enhanced by exploitation of rootstock × scion combinations that can resist untoward droughts and warm temperatures by making more resilient grapevine combinations. Our understanding of soil-plant-atmosphere continuum in the vineyard has increased within the last 50 years in such a manner that growers are able to use no-till systems with the aid of arbuscular mycorrhiza fungi inoculation with permanent cover cropping making the vineyard more resilient to droughts and heat waves. In premium wine grape regions viticulture has successfully adapted to a rapidly changing climate thus far, but berry based metrics are raising a concern that we may be approaching a tipping point.