Stilbene Composition Research Articles

Effects of Growing Areas, Pruning Wound Protection Products, and Phenological Stage on the Stilbene Composition of Grapevine (Vitis vinifera L.) Canes.

Applying plant protection products (PPP) on grapevine pruning wounds is a viticultural practice used to mitigate the spread of grapevine tuck disease, which is posing serious economic losses in the vine-wine industry. However, the impact of PPP on woody tissues remains unclear. Our study, conducted in two European vineyards, investigated the effects of Cuprocol, Tessior, Esquive, and Bentogran on stilbenes, in canes of Cabernet sauvignon and Syrah, at three phenological stages. Main stilbenes, quantified by HPLC-UV-DAD (1260 Agilent Infinity System) and identified by HPLC-ESI/MS (Thermo Scientific LCQ FLEET system), included E-resveratrol, E-ε-viniferin, E-piceatannol, and E-polydatin. Canes exhibited varying proportions of individual stilbenes, reflecting differences based on climatic conditions and phenological phases, rather than on the application of specific PPP. Vines grown in cool-climate conditions exhibited higher levels of E-resveratrol, whereas vines from the Mediterranean climate area exhibited higher levels of E-ε-viniferin. We also observed divergences in the accumulation trend of wood stilbenes throughout the season in canes collected in the two different growing areas.

Characterization of Stilbene Composition in Grape Berries from Wild Vitis Species in Year-To-Year Harvest.

Asian and American Vitis species possess a strong potential for crossbreeding programs, owing to their several resistance properties. Stilbenes are phenolic compounds present in grape berries and are well-known for their main role as phytoalexins and resistance to biotic stresses in plants. However, their identification and quantification in grape berries from wild Vitis remains unexplored. A mass spectrometry multiple reaction monitoring method combined with the analysis of pure standards allowed for the unambiguous characterization of 20 stilbenes in the grape berry skin extracts of nine native Vitis species and one cultivated Vitis vinifera species (cv. Cabernet Sauvignon). A main occurrence of monomeric (Z-piceid, E-piceid, E-isorhapontin, and E-astringin), dimeric (E-ε-viniferin, Z-ε-viniferin, and pallidol), and oligomeric (isohopeaphenol and r-viniferin) stilbenes was highlighted. Some stilbenes were clearly characterized for the first time in grape berries, such as the dimers ampelopsin A, E-vitisinol C, and parthenocissin A as well as the tetramers r2-viniferin and r-viniferin. Stilbene composition and content varied widely among several Vitis species and vintage years.

Grapevine Cane Extracts: Raw Plant Material, Extraction Methods, Quantification, and Applications.

Grapevine canes are viticulture waste that is usually discarded without any further use. However, recent studies have shown that they contain significant concentrations of health-promoting compounds, such as stilbenes, secondary metabolites of plants produced as a response to biotic and abiotic stress from fungal disease or dryness. Stilbenes have been associated with antioxidant, anti-inflammatory, and anti-microbial properties and they have been tested as potential treatments of cardiovascular and neurological diseases, and even cancer, with promising results. Stilbenes have been described in the different genus of the Vitaceae family, the Vitis genera being one of the most widely studied due to its important applications and economic impact around the world. This review presents an in-depth study of the composition and concentration of stilbenes in grapevine canes. The results show that the concentration of stilbenes in grapevine canes is highly influenced by the Vitis genus and cultivar aspects (growing conditions, ultraviolet radiation, fungal attack, etc.). Different methods for extracting stilbenes from grapevine canes have been reviewed, and the extraction conditions have also been studied, underlining the advantages and disadvantages of each technique. After the stilbenes were extracted, they were analyzed to determine the stilbene composition and concentration. Analytical techniques have been employed with this aim, in most cases using liquid chromatography, coupled with others such as mass spectrometry and/or nuclear magnetic resonance to achieve the individual quantification. Finally, stilbene extracts may be applied in multiple fields based on their properties. The five most relevant are preservative, antifungal, insecticide, and biostimulant applications. The current state-of-the-art of the above applications and their prospects are discussed.

Stilbenes in grapes and wines of Tannat, Marselan and Syrah from Uruguay

Aim: The aim of the study was to investigate the stilbene composition of grapes and wines of the Vitis vinifera cultivars Tannat, Marselan and Syrah cultivated in Uruguay. The effects of delaying the harvest on stilbene concentrations were determined, and the stability of stilbenes during wine storage was assessed.Methods and results: Stilbene concentrations were determined in the grapes and wines of two vintages (2015 and 2016) and two harvest dates for each cultivar. Vinification was carried out by traditional maceration, and samples of the wines of each vintage were analysed in the period from 3 months after devatting to up to 24 months later. After solid-phase extraction, stilbenes were identified and quantified by HPLC-ESI-MS/MS using a multiple reaction-monitoring approach. In the grape berries, stilbene concentrations were between 1.6 and 7.7 mg/kg, depending on grape cultivar, growing season, and in Syrah, harvest date. In the wines, stilbene concentrations were initially between 0.9 and 5.0 mg/L, being highest in Syrah, lowest in Marselan, and intermediate in Tannat. Stilbene concentrations in the Marselan wines were lower than expected based on stilbene concentrations in the grapes from which they were produced, suggesting poor extraction during winemaking. Total stilbene concentrations remained very stable during the analytical period.Conclusions: Delaying the harvest does not necessarily increase the stilbene content of grapes, but it can do so significantly, as shown for Syrah. For some grape cultivars, such as Marselan, poor extraction of stilbenes during winemaking can limit their concentrations in the resulting wines.Significance and impact of the study: The results of this study show the relevance of grape cultivar, degree of maturity and storage time may have into stilbenes. They provide reference data on the stilbene composition of grapes and wines produced under Uruguayan winegrowing conditions. The high stability of stilbenes during wine storage is relevant for consumers interested in red wine as a source of bioactive compounds.
 In memory of Prof. Isidro Hermosín-Gutiérrez.

Changes in stilbene composition during postharvest ozone treatment of 'Moscato bianco' winegrapes.

Stilbenes, including trans-resveratrol and its derivatives, are compounds naturally present in grapes and have gained a growing interest due to reported health-promoting properties. The production of resveratrol-enriched table grapes has promoted recent research on stress-induced synthesis of stilbenes. The oxidizing properties of ozone have been successfully exploited to its use as sanitizing agent and stilbene elicitor during table grapes storage. In winegrapes, this study represents the first research focused on the effect of postharvest ozone treatments on the accumulation of stilbene compounds. The study was carried out on Moscato bianco winegrapes (Vitis vinifera L.) and several gaseous ozone treatments were investigated differing in ozone dose (30 and 60 μL/L), exposure time (24 h, 48 h, and several days until 30% of weight loss), and delay time until processing (just after and several days after treatment). The stilbene production induced by ozone exposure was assessed in fresh and partially dehydrated winegrapes up to 5, 10, 15, 20, and 30% of weight loss aiming to evaluate the single and combined effect of oxidative and osmotic stresses. The results obtained showed that short-term exposure of fresh winegrapes at 60 μL/L of ozone for 48 h was not effective in inducing resveratrol accumulation just after treatment, but it had an elicitor effect on total stilbenes (+36%) in grapes subsequently dehydrated up to 20% of weight loss with a significant overproduction of trans-resveratrol and trans-piaceatannol. In addition, long-term and continuous treatments under ozone-enriched atmosphere can be also used during dehydration to sanitize winegrapes without affecting negatively the concentration of stilbenes. Therefore, the use of gaseous ozone during storage and dehydration could be indicated to reduce the use of sulfur dioxide and, depending on ozone dose and exposure time, the synthesis of stilbene compounds could increase.

Effect of the Sequential Inoculation of Non-Saccharomyces/Saccharomyces on the Anthocyans and Stilbenes Composition of Tempranillo Wines.

The phenolic compounds of red wines are responsible for their color, astringency, and antioxidant properties. The fermentative yeasts might be used to modulate wines in terms of their color, aroma and probably healthy properties. In this study, six non-Saccharomyces species were tested because they might enhance the properties of red Tempranillo wines from Rioja. The results confirmed that the anthocyanins and stilbenes composition of wine can be modulated with the use of a specific fermentation starter. Metschnikowia pulcherrima, Zygosaccharomyces bailii, Candida zeylanoides, and Torulaspora delbrueckii achieved the greatest improvements of the monomeric anthocyanin composition, and the latter three yeast species achieved the best results of stilbene composition when compared to S. cerevisiae and the other non-Saccharomyces yeasts. Overall, results suggested that the use of M. pulcherrima, Z. bailii, C. zeylanoides and T. delbrueckii as fermentation starters could be of great interest to achieve wines with better color and likely healthy properties.

Polyphenolic profile and stilbene content of Albanian “Kallmet” monovarietal wine

Aim: The aim of this work was to characterize the phenolic content of wines from the most promising autochthonous red grape variety in Albania, the Kallmet.Methods and results: To make this first survey, 16 commercial monovarietal Kallmet wines were analyzed with special attention to proanthocyanidin (condensed tannins), anthocyanin and stilbene composition. The average content of total proanthocyanidins in these wines was 3877 mgL-1 and the anthocyanin content averaged 242 mgL-1. The mean degree of polymerization (mDP) varied from 8 subunits in young wine to 31 subunits in 6-year-old wine, with a mean value of 14.5 subunits. The total free anthocyanin content ranged from 85 mgL-1 in old wine to 609 mgL-1 in young wine, with a mean value of 242.5 mgL-1. Trans-piceid was the most abundant stilbene (11 mgL-1), followed by trans-resveratrol (1.8 mgL-1). It was evidenced that cis-forms of resveratrol and piceid presented a lower content than their trans-forms. The ratio between piceid (glycoside form) and resveratrol (aglycone form) was 8.2, showing a similar stilbene pattern as in Mediterranean wines.Conclusions: Kallmet wines show proanthocyanidin polymerization characteristics and stilbene profiles very similar to red wines deriving from other varieties grown under warm climates.Significance and impact of the study: In spite of the strong interest in Kallmet grape, there is still a need for detailed studies to know the range of the different components of interest found in the produced wine.

Resveratrol and Related Stilbenoids, Nutraceutical/Dietary Complements with Health-Promoting Actions: Industrial Production, Safety, and the Search for Mode of Action.

This paper reviews the potential of stilbenoids as nutraceuticals. Stilbenoid compounds in wine are considered key factors in health-promoting benefits. Resveratrol and resveratrol-related compounds are found in a large diversity of vegetal products. The stilbene composition varies from wine to wine and from one season to another. Therefore, the article also reviews how food science and technology and wine industry may help in providing wines and/or food supplements with efficacious concentrations of stilbenes. The review also presents results from clinical trials and those derived from genomic/transcriptomic studies. The most studied stilbenoid, resveratrol, is a very safe compound. On the other hand, the potential benefits of stilbene intake are multiple and are apparently due to downregulation more than upregulation of gene expression. The field may take advantage from identifying the mechanism of action(s) and from providing useful data to show evidence for specific health benefits in a given tissue or for combating a given disease.

Insights on the stilbenes in Raboso Piave grape (Vitis vinifera L.) as a consequence of postharvest vs on-vine dehydration.

Grape withering is a process used to produce reinforced wines and raisins. Dehydration is usually carried out postharvest by keeping ripe grapes in special warehouses in controlled conditions of temperature, relative humidity (RH) and air flow. Alternatively, grape clusters can be left on the vines after the canes have been pruned. In general, dehydration increases stilbenes in grape, but there are few studies on the effects of on-vine withering. The stilbene profiles of Raboso Piave grape during postharvest and on-vine dehydration were studied here. High-resolution mass spectrometry (MS) was used to identify 19 stilbenes, including resveratrol monomers, dimers (viniferins), oligomers and glucoside derivatives. The two dehydration methods generally had different effects on the above nutraceuticals in grape. The samples kept in warehouses revealed significant increases in Z-ω-viniferin, E-ϵ-viniferin, δ-viniferin and another resveratrol dimer which were not observed in the plants. Trans-Resveratrol increased significantly only in samples dehydrated in the warehouse at 21 °C and 60-70% RH. The findings increase knowledge of stilbene composition in grapes subjected to withering on-vine. The choice of dehydration method affects the contents of these nutraceuticals in the grape and consequently in wines. Reasonably, it could also affect other secondary metabolites important for wine quality. © 2017 Society of Chemical Industry.

Regulation of stilbene biosynthesis in plants.

This review analyzes the advances in understanding the natural signaling pathways and environmental factors regulating stilbene biosynthesis. We also discuss the studies reporting on stilbene content and repertoire in plants. Stilbenes, including the most-studied stilbene resveratrol, are a family of phenolic plant secondary metabolites that have been the subject of intensive research due to their valuable pharmaceutical effects and contribution to plant disease resistance. Understanding the natural mechanisms regulating stilbene biosynthesis in plants could be useful for both the development of new plant protection strategies and for commercial stilbene production. In this review, we focus on the environmental factors and cell signaling pathways regulating stilbene biosynthesis in plants and make a comparison with the regulation of flavonoid biosynthesis. This review also analyzes the recent data on stilbene biosynthetic genes and summarizes the available studies reporting on both stilbene content and stilbene composition in different plant families.

Elicitors used as a tool to increase stilbenes in grapes and wines

Two preharvest treatments (methyl jasmonate or cell wall yeast) of grapevines (Monastrell and Tempranillo) were applied during two vintages (2014 and 2015) to check whether these elicitors enhanced stilbene accumulation in berries at the moment of harvest and in the corresponding wines elaborated with them. The main objective was checking the effect of treatment, variety and year on stilbene composition due to the interesting health-related properties of these compounds in both grapes and wines. The results pointed to inter-varietal and inter-annual differences, and that the treatments generally enhanced the stilbene composition of grapes and, particularly, of wines. The increase was more evident in Monastrell variety than in Tempranillo variety and in their wines more than their grapes during 2014 vintage but not during 2015 vintage.

Grapevine cane’s waste is a source of bioactive stilbenes

Grapevine canes represent a large source of waste derived from the wine industry with low added-value. In the current work, the cane stilbene composition of twenty two grape cultivars (including Vitis vinifera sativa, Vitis vinifera sylvestris and hybrid direct producers) was investigated. Ten stilbenes were identified and quantified: hopeaphenol, ampelopsin A, isohopeaphenol, piceatannol, trans-piceid, trans-resveratrol, miyabenol C, ε-viniferin, r-viniferin and ω-viniferin. Total stilbene concentration ranged from 2400 to 5800mg/kg dry weight. Principal component analysis applied to stilbene concentration showed a separation of cultivars in five groups. Antioxidant capacity of the stilbene extracts varied from 1700 to 5300μmol Trolox equivalent/g dry weight. No relationship was found between stilbene content and ORAC values. Finally, the effect of year on cane stilbene content was also studied in V. vinifera sylvestris grapevine. The effect of the year depended on the cultivar. Data reported sustain the use of grapevine cane byproducts as a low cost source of bioactive stilbenes for the development of natural fungicides as well as nutraceutical, as strategy for sustainable viticulture.

In Planta Localization of Stilbenes within Picea abies Phloem.

Phenolic stilbene glucosides (astringin, isorhapontin, and piceid) and their aglycons commonly accumulate in the phloem of Norway spruce (Picea abies). However, current knowledge about the localization and accumulation of stilbenes within plant tissues and cells remains limited. Here, we used an innovative combination of novel microanalytical techniques to evaluate stilbenes in a frozen-hydrated condition (i.e. in planta) and a freeze-dried condition across phloem tissues. Semiquantitative time-of-flight secondary ion-mass spectrometry imaging in planta revealed that stilbenes were localized in axial parenchyma cells. Quantitative gas chromatography analysis showed the highest stilbene content in the middle of collapsed phloem with decreases toward the outer phloem. The same trend was detected for soluble sugar and water contents. The specimen water content may affect stilbene composition; the glucoside-to-aglycon ratio decreased slightly with decreases in water content. Phloem chemistry was correlated with three-dimensional structures of phloem as analyzed by microtomography. The outer phloem was characterized by a high volume of empty parenchyma, reduced ray volume, and a large number of axial parenchyma with porous vacuolar contents. Increasing porosity from the inner to the outer phloem was related to decreasing compactness of stilbenes and possible secondary oxidation or polymerization. Our results indicate that aging-dependent changes in phloem may reduce cell functioning, which affects the capacity of the phloem to store water and sugar, and may reduce the defense potential of stilbenes in the axial parenchyma. Our results highlight the power of using a combination of techniques to evaluate tissue- and cell-level mechanisms involved in plant secondary metabolite formation and metabolism.

Terroir and variety: Two key factors for obtaining stilbene-enriched grapes

Grapes and wine are among the main sources of bioactive stilbenes in the diet. The stilbene composition of grapes depends on both variety and terroir. The effect of terroir on grape stilbenes has not been widely studied. In this research the study of the stilbene content (resveratrol, piceatannol and viniferins) in four different red grape varieties (Syrah, Merlot, Cabernet sauvignon and Pinot noir) and from four different Andalusian terroirs (Jerez, Cabra, Cadiar and Ronda) was carried out during the 2009 vintage. In addition, stilbene induction capacity was studied in each terroir and variety after ultraviolet-C (UVC) postharvest treatment. Data were analysed by multivariate statistical methods. Principal component analysis applied to the oenological parameters and stilbene data classified varieties into two terroir groups satisfactorily: Jerez and Cabra on the one hand, and Ronda and Cadiar on the other hand. Terroir effect is stronger than variety on stilbene induction capacity after UVC treatment. Syrah from Cabra stood out due to both the high basal (13.07mg/kg fw) and induced (33.14mg/kg fw) stilbene concentration attained. Therefore, it can be affirmed that Cabra and Syrah are the most suitable terroir and variety, respectively, for obtaining a stilbene-enriched wine.

Analysis of some stilbenes in Italian wines by liquid chromatography/tandem mass spectrometry

Stilbenes from grapes and wines play a central role in the human diet because of their antioxidant, antimutagenic and anticarcinogenic properties. We describe a method for the direct determination of some stilbenes (cis- and trans-resveratrol, cis- and trans-resveratrol glucoside, cis- and trans-piceatannol, and cis- and trans-piceatannol glucoside) in wine by high-performance liquid chromatography/mass spectrometry using a triple quadrupole (QqQ) mass spectrometer, in multiple reaction monitoring (MRM) mode, acquiring two diagnostic product ions from the chosen precursor. All the target analytes were separated on a C-18 column using gradient elution, in a single run. Electrospray ionization (ESI) in negative ion mode gives higher sensitivity for all the target compounds than atmospheric pressure chemical ionization (APCI). For the identification of piceatannol glucoside (astringin), because of the lack of a suitable standard, an HPLC/TOFMS method was used. The method permits direct injection of samples and it is time-saving, removing the need for sample pre-treatment. The detection limits were 48.0 ng mL(-1) for cis- and trans-resveratrol and for cis- and trans-resveratrol glucoside, and 50.0 ng mL(-1) for cis- and trans-piceatannol. The procedure proved to be simple and suitable for routine and confirmatory purposes. A total of 19 red and 3 white Italian wines were analyzed and differences in the stilbene composition were found among these samples.

Stilbenes: Quantitative extraction from grape skins, contribution of grape solids to wine and variation during wine maturation

With the objective of studying the relationship between stilbenic composition in grape skins and that in corresponding red wine, we have firstly optimized a method for quantitative extraction and purification of stilbenes from grape skins, used for posterior HPLC analysis. Stilbene levels in grape skins of three Vitis vinifera varieties (Castelão, Syrah and Tinta Roriz) and in their corresponding wines made by traditional winemaking technologies were analyzed. Stilbene composition in grape skin varied considerably, depending on the grapevine variety. The contents of trans-resveratrol in grape skins of the three varieties were not significantly different. However, their contents of trans-piceid on dry skin were very significantly different (67.24, 10.43 and 11.57 mg kg −1, respectively). Moreover, cis-piceid, which was absent in Syrah and Tinta Roriz grape skins, was detected in Castelão grape skin with an important amount (58.9 mg kg −1 on dry skin). The high concentrations of stilbenes were found in stem tissue (cv. Castelão): 145.52 mg kg −1 of trans-resveratrol, 61.43 mg kg −1 of trans-piceid and 143.85 mg kg −1 of cis-piceid (on dry stem), while only trans-resveratrol was quantified in the grape seeds with a low concentration (6.8 mg kg −1 on dry seed). Simulated maceration of each solid part of grapes using model wine solution indicated that both stem and seed have little contribution of stilbenes to wines while skins are the major source of these compounds in wine. On the other hand, cis-piceid was not detected in the corresponding wines made by these grapevine varieties, but its isomer trans-piceid was detected in an important concentration, suggesting that cis-piceid, once extracted, isomerized to its trans-form. The evolution of these compounds during the elaboration of wines (from the end of alcoholic fermentation until second racking, i.e., just before bottling) showed that in general, trans-resveratrol concentration decreases gradually, trans-piceid kept constant, while cis-resveratrol increases slightly. The important finding of this work is that there was a significant correlation between the concentration of total piceid in grape skins and that in the respective wine. On the contrary, no such relationship was found for resveratrol.