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Article

Rootstock Effect Influences the Phenolic and Sensory Characteristics of Syrah Grapes and Wines in a Mediterranean Climate

by
Florencia Heller-Fuenzalida
1,
Italo F. Cuneo
1,
Nathalie Kuhn
1,
Álvaro Peña-Neira
2 and
Alejandro Cáceres-Mella
1,*
1
School of Agronomy, Faculty of Food and Agronomical Sciences, Pontificia Universidad Católica de Valparaíso, San Francisco s/n, Quillota 2260000, Chile
2
Faculty of Agronomical Sciences, University of Chile, Santa Rosa 11315, La Pintana, Santiago 8820808, Chile
*
Author to whom correspondence should be addressed.
Agronomy 2023, 13(10), 2530; https://doi.org/10.3390/agronomy13102530
Submission received: 5 September 2023 / Revised: 22 September 2023 / Accepted: 29 September 2023 / Published: 30 September 2023
(This article belongs to the Section Horticultural and Floricultural Crops)
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Abstract

:
Wine quality depends on grape quality, which is affected by many factors such as edaphic, climatic and genetic, i.e., cultivar and rootstock. Rootstocks have been selected in worldwide viticulture to confer to vines some adaptation to several types of soil conditions in vineyards, but this adaptation may affect vine physiology and consequently may affect the chemical compounds of grapes, especially their phenolic compounds. Thus, this study compares the chemical composition of grapes and wines, and the sensory profile of wines from vines of the cv. Syrah grafted on two rootstocks, 5C and Gravesac, grown under a biodynamical management system. The results showed higher skin total phenols and skin total tannins in grapes from 5C rootstock. In the resulting wines, the same results were observed. The multivariate analysis demonstrated that the 5C wines presented a higher relationship with all the families of the low-molecular-weight phenolic compounds, while the Gravesac wines showed a strong relationship with acetylated and p-coumaroylated anthocyanins. The sensory analysis showed that the 5C wines presented more color intensity, more astringency and more meaty aromas compared with the Gravesac wines. The results proved that it was possible to obtain grapes and wines of different qualities depending on the rootstocks used under the same climatic and agronomical condition.

1. Introduction

Wine quality depends on grape quality, and the latter is determined by at least three major factors, including soil, climate, agricultural management and genetics [1]. Wine quality is closely related to various compounds, such as organic acids, aromatic compounds and especially phenolic compounds. Phenolic compounds are organic molecules, belonging to the secondary metabolism of the vine, and are classified as non-flavonoids or flavonoids, which differ in their structure. Among the non-flavonoid compounds, phenolic acids such as hydroxycinnamic and hydroxybenzoic acids are important in the copigmentation process in red wines and stilbenes such as the most recognized trans-resveratrol have interesting antioxidant properties for human health. Regarding flavonoids, there are flavonols, flavononols, flavones, flavanols and anthocyanins [2]. Flavanols, or commonly called condensed tannins, are the ones that produce taste sensations like body, astringency and bitterness in the wine [3]. Anthocyanins, located in grape skins, are responsible for delivering color to red wines [4]. The quality of a wine is based on its chemical composition, thus the accumulation of phenolic compounds in the grape is a turning point.
The use of rootstocks today depends on different needs and is relevant for the productive success of a vineyard. Rootstocks are used to generate resistance to nematodes, adapt to soil conditions such as problems of carbonates and saline soils and because they resist conditions of water deficit at the root level [5,6,7]. Rootstocks might also alter the vegetative and reproductive growth of the vines, either accelerating or postponing their stages of ripening [8,9,10]. By influencing the microclimate of the canopy and the technological ripening of the berries, rootstocks may modulate the production of phenolic compounds in grapes [7,11].
Furthermore, Zombardo et al. [12] show that, even without severe stress that can intensify differences, rootstocks can exert significant influence on the characteristics of the grape, affecting the overall quality of the berry. As has been raised before, phenolic compounds depend on different factors, and they are fundamental in the determination of wine’s organoleptic and sensory characteristics. Therefore, any modification in the accumulation of these compounds in the grape can impact on wine quality [13]. Most of the existing research related to rootstocks focuses on roots, photosynthesis and productivity, and studies on the chemical characteristics of the grape and later of the wine are scarce [14]. The foregoing highlights the importance of conducting more research on how rootstocks can influence the composition and concentration of various chemical compounds in grapes, such as phenolic compounds, and how this can also interfere with wine quality. Therefore, the objective of this study is to compare the influence of two rootstocks, 5C and Gravesac, on the chemical characteristics of grapes and wines and sensory parameters in cv. Syrah.

2. Materials and Methods

2.1. Site of Study, Experimental Design and Treatments

The assay was carried out on a 2021 vintage from Hacienda Valdeperillo (33°03′49″ S, 71°13′16″ W), a biodynamic winery located in Limache, Central Chile. Limache has a warm, temperate Mediterranean climate with a mean summer temperature of 18.4 °C, a mean winter temperature of 9.9 °C and a mean annual rainfall of 408 mm [15]. The plant material had cv. Syrah, clone 470, grafted in two rootstocks that were planted in 2013. The treatments consisted of two rootstocks, 5C and Gravesac, in two vineyard plots on which the plants were grafted. The grapevines were trained using a vertical trellis system and pruned used the double Guyot method with vines spaced 1.8 m between rows and 1 m over the rows in both treatments. The average yield was approximately 7.7 ton ha−1. Both rootstocks received the same vineyard management practices, following biodynamic applications, and a drip irrigation system was used for watering the plants. The two vineyard plots were separated by a 2 m wide path, so the soil material was similar in both vineyard plots with a clay loam texture in the first 60 cm and a high presence of porosity. The soil in the deeper (>60 cm) section showed a presence of granitic and quartz. The soil presents a neutral pH (6.9–7.0).
The design of the experiment followed a completely randomized design comprising five replicates, with each replicate consisting of seven consecutive vines, excluding those located at the borders of the vineyard or showing obvious symptoms of diseases. The berry sampling was realized at the commercial harvest period (23–24 °Brix, March 31st) and consisted of 100 berries per replicate, where 50 berries were randomly selected from each side of the canopy. The samples that were collected were weighed, frozen and stored at −40 °C until they were processed. The following physical and chemical parameters were assessed in accordance with the OIV protocol: the weight of 50 berries, the skin weight of 50 berries, the seed weight of 50 berries, titratable acidity (measured in grams of tartaric acid per liter), pH and total soluble solids (measured in °Brix) [16].

2.2. Extraction of Phenolic Compounds from Grape Berries

The skins and seeds of 50 berries were manually separated. Two consecutive extractions under mechanical agitation were performed, one using a methanol–water solution (80:20 v/v) and the other using an acetone–water solution (80:20 v/v). In each case, we agitated 100 mL of solution on an orbital shaker at 20 °C for 60 min. We combined both solutions and, to separate the solid from the liquid portion, we subjected the samples to centrifugation at 4000 rpm for 10 min at 4 °C using a K2015R refrigerated centrifuge (Centurion Scientific, Stoughton, UK). After that, the organic fraction was evaporated at 30°C using a rotary evaporator. The resulting solution was adjusted to a final volume of 100 mL with distilled water, filtered through a 0.45 µm PVDF membrane and then stored in amber-colored bottles [17].

2.3. Winemaking Procedure

Due to limitations in grape volume, four replicates were created by combining adjacent vineyard replicates for the winemaking process (n = 4). The grapes were harvested by hand and fermented following the protocol of the winery. In brief, the clusters were destemmed and crushed with the use of a semiautomatic crusher machine and placed in 100 L stainless steel vats for each replicate. Pre-fermentative maceration for three days at 10 °C was performed. Then, the fermentation was carried out using native yeast at 24 °C for two weeks with two punch downs per day. When the fermentation was completed, we pressed the replicates, and the free-run fractions from each deposit were racked, cold-stabilized and adjusted to a level of 30 mg L−1 of free SO2. Afterward, the wine was immediately bottled in 750 mL dark-green glass bottles (CristalChile, Chile) and stored at 15 °C for subsequent analyses. We measured the following chemical parameters in accordance with the OIV protocol: alcohol content (% v/v), titratable acidity (measured in grams of tartaric acid per liter), pH, free and total SO2 as well as reducing sugars [16].

2.4. Spectrophotometric Characterization of Grape Berries and Wine

Grape berries and wine phenolic compounds were analyzed via spectrophotometry. We determined the total phenol concentration by measuring UV absorption at 280 nm with gallic acid serving as the standard and expressed the results as mg GAE (gallic acid equivalent) per liter [18]. The concentration of total tannins was measured using methylcellulose as a precipitating agent [19]. The total anthocyanins were characterized using sodium bisulfite, following the Ribereau-Gayon and Stonestreet methods [20]. Color intensity (CI) was assessed following the Gloried method by measuring UV absorption at 420 nm (yellow), 520 nm (red) and 620 nm (blue) [18]. The determination of the color coordinates lightness (L*), chroma (C*) and hue (h*) was performed using the methodology described by Ayala et al. [21]. The spectrophotometry characterization was conducted using a UV−visible spectrophotometer, specifically the UV/Vis 1700 Pharmaspec model (Shimadzu, Kyoto, Japan).

2.5. High-Performance Liquid Chromatography (HPLC-DAD) Analyses in Grapes and Wines

2.5.1. HPLC-DAD Analysis of Organic Acids in Grape Berries

Five grams of berries without the seeds were weighed, crushed and homogenized. Subsequently, the resulting must was centrifuged for 5 min at 1800 rpm using a refrigerated centrifuge (Centurion Scientific, Stoughton, UK). We obtained one milliliter of supernatant and diluted it in nine milliliters of Milli-Q water. Afterwards, we stirred it on an orbital shaker for 30 seconds to achieve homogenization and, finally, filtered through a 0.22 µm pore membrane. Organic acids were analyzed via HPLC-DAD, following Gil I Cortiella et al.’s [22] methodology. An Agilent 1260 Infinity Series chromatograph (Agilent Technologies, Santa Clara, CA, USA) equipped with a quaternary pump, an autosampler, a column oven and a diode array detector (DAD) was utilized for the chromatographic separation of the sample. A Supelcogel H column (250 mm × 4.6 mm, Sigma-Aldrich, Santiago, Chile) was used under isocratic conditions. Samples were diluted 1:5 with a 0.1% (v/v) phosphoric acid aqueous solution (mobile phase). For analysis, we injected twenty microliters of each sample into the column, and the elution of organic acids was monitored at 210 nm. Tartaric, malic and citric acids (Sigma-Aldrich, Chile) were both identified and quantified using external standards.

2.5.2. HPLC-DAD Analysis of Low-Molecular-Weight Phenolic Compounds of Grape Berries and Wine

A 50 mL portion of the skins, seeds extracts and wine was further subjected to additional extraction with diethyl ether (3 × 20 mL) and ethyl acetate (3 × 20 mL) to concentrate the phenolic compounds. The organic fractions were then combined, subjected to dehydration with 2.5 g of anhydrous sodium sulfate and evaporated to dryness under vacuum conditions at 35 °C. The resulting solid residue was dissolved in 2 mL of a methanol/water solution (1:1, v/v) and filtered through a 0.22 µm pore size membrane. Individual phenolic compounds were identified using a chromatographic system comprising an Agilent Technologies 1100 Series instrument (Agilent Technologies, Santa Clara, CA, USA), which included a diode-array detector (DAD, Model G1315B), a quaternary pump (Model QuatPump G1311A), a degasser (Model G1379A) and an autosampler (Model G1329A). We introduced 25 µL aliquots of the final solution for reverse-phase chromatographic separation at 20 °C utilizing a reverse-phase Nova Pack C18 column (4 µm, 3.9 mm inner diameter × 300 mm; Waters Corp., Milford, MA, USA). The photodiode array detector (DAD) was configured with a range from 210 to 360 nm. Two mobile phases were employed as follows: Phase A consisted of water/acetic acid (98:2 v/v) and Phase B consisted of water/acetonitrile/acetic acid (78:20:2 v/v/v). A gradient elution was applied at a flow rate of 1.0 mL/min from 0 to 55 min and 1.2 mL/min from 55 to 90 min with the following proportions: 100% to 20% A, followed by a transition from 20% to 10% A between 55 and 57 min, and finally from 10% to 0% A from 57 to 90 min. Each major peak observed in the HPLC chromatograms of the extracts was based on both its retention time and absorption spectrum (ranging from 210 to 360 nm), following the methodology of Peña-Neira et al. [23]. Quantitative assessments were carried out using the external standard method employing commercial standards. Calibration curves were generated by injecting standard solutions into the column before the extraction process, using the same conditions employed for sample analysis, spanning the observed concentrations range (r2 ≥ 0.93). The quantification of flavonol glycosides was performed using the quercetin curve, while flavan-3-ols were quantified using the (+)-catechin curve.

2.5.3. HPLC-DAD Analysis of Anthocyanins from Grape Skins and Wine

Two milliliters of the aqueous solution from grape skins or wine underwent filtration through a 0.22 μm PVDF pore size membrane. Then, a 150 μL aliquot was subjected to reversed-phase chromatographic separation at 20 °C using a LiChro Cart 100 RP-18 column (5 μm, 4.0 mm × 250 mm, Agilent Technologies, USA) within an Agilent 1200 Series system (Agilent Technologies, USA). A gradient consisting of solvent A (water/formic acid, 90:10, v/v) and solvent B (acetonitrile) was applied at a flow rate of 1.1 mL/min from 0 to 22 min, followed by 1.5 mL/min from 22 to 35 min. The gradient was as follows: 96% to 85% A and 4% to 15% B from 0 to 12 min, maintained at 85% A and 15% B from 12 to 22 min and 85% to 70% A and 15% to 30% B from 22 to 35 min. This was followed by a final wash with 100% methanol and re-equilibration of the column. Diode array detection covered the range from 210 to 600 nm, and quantification was based on peak area measurements at 520 nm. Anthocyanin content was expressed using malvidin-3-glucoside chloride as a standard for the calibration curve (r2 = 0.99). Calibration curves at 520 nm were established by injecting different volumes of standard solutions under the same conditions as the analyzed samples [24].

2.6. Wine Sensory Evaluation

The descriptive analysis [25], conducted one month after bottling, involved a sensory panel comprising 10 judges who had received prior training in the sensory evaluation of wines. The evaluation session took place in individual tasting booths at a room temperature of 20 °C. Over a period of 3 h, the judges assessed four wines, corresponding to each treatment (5C and Gravesac), with each wine being evaluated in duplicate. Thirteen attributes were considered, including color intensity, vegetable aroma, red fruits aroma, spice aroma, tobacco aroma and meaty aroma, as well as acidity, astringency, bitterness, red fruits, body, persistence and overall quality. The assessments were made on a 15 cm unstructured scale anchored between “low” and “high” intensity. Prior to evaluation, 20 mL of wine was served at a temperature of 18–19 °C in technical glasses (Viticole, Arcoroc, France) with the wines labeled with a three-digit code in a completely randomized order. Between samples, the judges used water and unsalted crackers to cleanse their palates. Data were collected in a paper ballot.

2.7. Statistics

For the chemical analyses, we employed analysis of variance (ANOVA) to separate means, utilizing a significance level of 95% (p < 0.05). In the case of the descriptive analysis, ANOVA and the LSD test were used with a significance level of 90% (p < 0.1). To visualize the relationship between rootstocks and phenolic composition in wines, we conducted a principal component analysis (PCA). All the chemical and sensory analyses were carried out using R statistical software version 4.1.1 (R Foundation for Statistical Computing, Vienna, Austria). The PCA analysis was performed using Statgraphic Centurion XV (Statpoint Technologies, Warrenton, VA, USA).

3. Results

3.1. General Physical and Chemical Composition in Syrah Grapes from Different Rootstocks

Table 1 shows the chemical and physical analyses of Syrah grapes from two rootstocks, 5C and Gravesac. The weight of 50 berries varied from 56.10 to 62.08 g with clear differences between rootstocks. The weight of the skins and seeds only showed differences in the skins weight. The Gravesac rootstock showed a higher skins weight and berry weight. The soluble solids ranged from 23.01 to 23.40 °Brix and the titratable acidity varied from 5.55 to 6.15 g tartaric acid L−1, showing that 5C had the higher titratable acidity. The pH of the grapes varied from 2.80 to 2.85 with no differences between rootstocks. In terms of the organic acids measured via liquid chromatography, there were only differences in the concentration of malic acid, showing a higher concentration in the grapes from the 5C rootstock.
Regarding the global phenolic composition in grape skins, skins total phenols ranged from 5.67 to 7.25 mg GAE g−1 and skins tannins ranged from 8.33 to 10.26 mg catechin g−1; in both analyses, 5C showed the higher concentration. With respect to the global phenolic composition in grape seeds, the seeds’ total phenols varied from 15.47 to 17.96 mg GAE g−1 and the seeds’ tannins ranged from 65.41 to 77.70 mg catechin g−1. In this case, there were no differences in both grape parameters between rootstocks. The analyses of total anthocyanins varied from 11.30 to 11.47 mg malvidin g−1 with no differences between rootstocks.
Figure 1 shows the profile of the low-molecular-weight phenolic compounds in grape skins and seeds from two rootstocks in cv. Syrah. In grape skins, the total phenolic acids varied from 18.99 to 19.48 mg kg−1 with no differences between rootstocks. For the total flavanols, the concentration ranged from 88.65 to 106.12 mg kg−1 with no significant differences between rootstocks. In the case of the total flavonols, the values ranged from 205.45 to 392.18 mg kg−1, showing that the Gravesac rootstock had a higher concentration. The same behavior was observed for stilbenes, where Gravesac reached a higher concentration value compared with the 5C rootstock. For the grape skins, the profile of the phenolic compounds can be observed in Table S1.
The phenolic composition in grape seeds shows that the phenolic acids varied from 30.48 to 30.89 mg kg−1 and the flavanols varied from 1428.19 to 1577.43 mg kg−1. In this case, there were no differences in the phenolic composition of grape seeds between the rootstocks (Table S2).
The anthocyanin profile of grape skins shows thirteen anthocyanins; five glucosylated anthocyanins such as delphinidin-3-glucoside, cyanidin-3-glucoside, petunidin-3-glucoside, peonidin-3-glucoside and malvidin-3-glucoside; four acetylated anthocyanins such as delphinidin-3-acetylglucoside, peonidin-3-acetylglucoside, peonidin-3-acetylglucoside and malvidin-3-acetylglucoside; and four coumaroylated anthocyanins, cyanidin-3-p-coumaroylglucoside, petunidin-3-p-coumaroylglucoside, peonidin-3-p-coumaroylglucoside and malvidin-3-p-coumaroylglucoside. In this analysis, there were only differences in malvidin-3-p-coumaroylglucoside with a higher concentration in grape skins from the Gravesac rootstock (Figure 2).

3.2. Chemical Composition and Color Parameters in Syrah Wines

In Table 2, we present the chemical analyses and color parameters in Syrah wine from grapes obtained from rootstocks 5C and Gravesac. The global analyses in the wines only showed differences in terms of the titratable acidity, alcohol content, fermentable sugars and reducing sugars. The total phenols ranged from 715.47 to 1268.74 mg GAE L−1 and showed that the content of total phenols in wines from the 5C rootstock was approximately 43% higher than the concentration in Gravesac wines. The same behavior was observed for total tannins, showing a variation from 1834.62 to 2587.21 mg (-)-epicatechin L−1 with 5C presenting the greatest concentration, about 29% higher than Gravesac wines. The total anthocyanins varied from 764.01 to 786.72 mg malvidin L−1 with no differences between rootstocks. The color parameters only showed differences in h*, with a higher value in 5C wines.
Figure 3 shows the principal component analysis illustrating the relationship between the wines from both rootstocks and the phenolic compounds measured via spectrophotometry and liquid chromatography. PC1 and PC2 accounted for 82.7% of the total variation (PC1, 56.3% and PC2, 26.4%). PC1 was characterized by coumaroylated anthocyanins, flavanols, flavonols, phenolic acids, total anthocyanins, total phenols and total tannins. PC2 was characterized by acetylated anthocyanins, glucosylated anthocyanins and stilbenes. The overall results indicate that the wines from grapes of the rootstock 5C are related with glucosylated anthocyanins, total phenols, total anthocyanins, total tannins, flavanols, flavonols, phenolic acids and stilbenes, whereas the wines from the Gravesac rootstock correlate with acetylated anthocyanins and coumaroylated anthocyanins. The profiles of the low-molecular-weight phenolic compounds and the profile of anthocyanins are shown in Tables S3 and S4.

3.3. Sensory Analysis of Wines

Figure 4 shows the terms evaluated by the judges, with their scores used to determine if the rootstocks produced differences in the sensory profiles of the wines. The results obtained showed that there were differences in three parameters, color, meaty aroma and astringency, which were found in a greater proportion in the wines of the 5C rootstock.

3.4. Relationships between Chemical and Sensory Parameters in Syrah Wines

Correlations were determined for some analytical parameters and some sensory attributes, especially the color and mouthfeel sensation of the wines to understand the relationships of the wines’ components (Table 3). Total phenols were positively correlated with total tannins, flavanols, glucosilated anthocyanins and astringency. Total tannins were positively correlated with flavanols, flavonols, color intensity and astringency. There was a good positive correlation between flavanols and astringency and a negative correlation between flavanols and bitterness. Flavanols and flavonols showed a positive relationship. Moreover, flavonols were negatively correlated with p-coumaroylated anthocyanins and bitterness and positively correlated with color intensity and astringency. Finally, acetylated anthocyanins showed a negative relation with body and p-coumaroylated anthocyanins showed a negative relationship with color, astringency and body.

4. Discussion

The phenolic content of grapes and wines depends on several factors, such as parameters specific to a particular location, for example, humidity, temperature, row orientation and soil type, as well as genetic factors, such as variety, clone and cultural labor, that is, the work of people in the field to achieve quality grapes. Rootstocks have been used for quite some time, first to combat phylloxera and the problems it caused in Europe and then to adapt plants to different soil realities, i.e., hydric stress, salinity and diseases, among other aspects. In this investigation, the effect of two rootstocks on the chemical composition and some organoleptic properties of Syrah grapes and wines were analyzed under the same agricultural climate and growing conditions. The results showed that the Gravesac rootstock had higher berry and skin weights compared with 5C, which could be explained by genetic differences. One explanation could be that although the same controlled water stress was applied to both Syrah quarters (i.e., plants on rootstock) 15 days prior to harvest, abiotic stresses such as extreme temperature and water deficit do not only influence plant development, and it is know that Gravesac has a higher drought tolerance than 5C [26], for which reason it could have had a greater amount of water in its tissues, producing berries with a higher weight. Another explanation could be that perhaps the grapes from the vines grafted on Gravesac rootstock had skins with more cells. A future observation through microscopy could be very interesting to corroborate this hypothesis, which would mean obtaining findings on how the rootstocks can morphologically modulate grape cells.
The same behavior was observed in terms of the concentration of organic acids, especially malic acid, which had a greater concentration in 5C grapes, and in terms of phenolic compounds, such as skins total phenols and skins tannins, with a greater concentration in 5C (Table 1). Similar results were found by Li et al. [27], where 5C rootstock significantly raised the grape skin phenols content of cv. Cabernet Sauvignon, and other authors [7]. These findings are interesting since small berries could be convenient for wine production, producing a concentration effect. Similar findings were reported by Gutierrez-Gamboa et al. [28], who investigated the impact of eight different rootstocks on grape anthocyanins, skin and seed proanthocyanidins, wine color and phenolic compounds using the same clone of the Merlot grape variety. The results showed that the rootstocks utilized modify the proanthocyanidins. In general, grapes from vines grafted on SO4 had a higher concentration of total proanthocyanidins in skins and seeds compared with the other rootstocks, while those on Gravesac had the lowest concentration. In the study carried out by Riaz et al. [29] on the genetic diversity and parentage analysis of grape rootstocks, results indicated that SO4 and 5C share the same parents (V. berlandieri × V. riparia) and possess the same behavior and performance. Therefore, in the study mentioned above, 5C could be represented by SO4. It should be noted that Gravesac’s parents are V. berlandieri × V. riparia × V. Rupestris.
The resulting wines produced from the grapes of the different rootstocks corroborated the results obtained for the grapes, especially the global phenolic characterization. Regarding the titratable acidity, Gravesac presented the higher concentration, but in the grapes, it presented the lowest concentration. This discrepancy could be due to the presence of mineral compounds such as potassium and calcium, which can form salts with the organic acids in the wine, reducing its acidity [30,31]. Furthermore, studies have shown that potassium can be regulated in grapevines through rootstocks due to their different capacities to take potassium from the soil and transport it between roots and shoots [32]. In this context, it was also reported that the SO4 rootstock can accumulate a higher potassium content in the vine [33], with SO4 again representing 5C.
In terms of the alcoholic strength, the 5C wine presented a higher concentration of ethanol compared with the Gravesac wine. The same results were observed in global phenolic analyses, for example the 5C wines presented 43% more total phenols than the Gravesac wines and, in the same way, the 5C had 29% more tannins than the Gravesac wines. The changes in tannin concentrations during fermentation depend on the percentage of alcohol and the duration of contact between grape solids (seeds, pulp and skins) and the wine. As the alcohol increases, grape pomace will be more depectinated and release larger amounts of phenolic compounds [34]. Since the contact time was the same in both vinification processes, the higher alcohol content in the 5C wines resulted in more tannin extraction during vinification, especially from the grape skins.
The profile of low-molecular-weight phenolic compounds in seeds and skins showed that there were no differences in the seeds between grapes from the different rootstocks, and there were some differences in the skins, especially in the concentrations of flavonols and stilbenes, with a greater concentration in the Gravesac rootstock (Figure 1 and Figure 2). The same results were observed in the profile of anthocyanins with only differences in malvidin-3-p-coumaroylglucoside (Figure 3). These results disagreed somewhat with the phenolic profile of the resulting wines, where 5C showed a higher concentration of some compounds compared with Gravesac (Tables S1 and S3). Winemaking is a complex process that encompassed numerous procedures that can significantly alter the chemical composition of the grape berries. This complexity often poses challenges when attempting to extrapolate information from grape berries to the chemical composition of the resulting wine [35]. Previous studies have shown that the extraction of anthocyanins from grape skins can be constrained by various factors, such as cell wall components, including polysaccharide compounds [36,37]. Likewise, it could be possible that the cell skins in Gravesac are more fragile, delivering compounds much more easily, especially when using strong solvents such as methanol and acetone, which were used for the extraction of compounds from the skins and seeds. It would be highly intriguing to validate this hypothesis in a future study.
In the same way, the multivariate analysis allowed us to observe the relationship between the resulting wines and the phenolic composition, and it was noted that the 5C wines are related with most of the phenolic compounds analyzed in this study, but in contrast, Gravesac wines are only related with a minority of anthocyanins, such as acetylated and coumaroylated fractions. These results corroborated the profiles of the low-molecular-weight phenolic compounds listed in Table S3 and the anthocyanin profiles in Table S4. The above-mentioned results indicate that under the same agroclimatic conditions and with the same vinification method, the rootstocks analyzed can modulate the chemical composition of the wines produced, so they can not only serve to adapt to environmental conditions but also allow wines with different phenolic compositions to be obtained.
In terms of the descriptive analysis, the 5C wines presented more color intensity, more astringency and more meaty aromas (leather, ham ad bacon). The color intensity can be explained by the greater presence of malvidin-3-glucoside in the 5C wines. The Gravesac wines presented acetylated and coumaroylated anthocyanins. Most of those compounds were petunidin and peonidin, which are characterized by being blue-purple and purple, respectively [38], while 5C was dominated by malvidin, which has a reddish-purple color, explaining the higher color intensity in the 5C wines. Moreover, the negative correlation of p-coumaroylated anthocyanins with color supports the above results. Sherman et al. [39] discovered that wine color was positively influenced by higher ethanol concentrations, which promoted the formation of polymeric pigments, resulting in darker wines. The results showed that the 5C wines presented more tannins from the grape skins. Proanthocyanidins in the skins were present in a greater amount, which produces a greater sensation of astringency in the mouth [40,41]; therefore, the higher concentration of alcohol, flavanols and skins tannins in the 5C wines could explain their color intensity and astringency. These results are corroborated by observing that the total tannins and flavanols in wines presented a positive correlation with the astringency measure perceived by a sensory panel. Regarding the difference in the meaty aroma, it is known that the temperature can influence the concentration of aromatic compounds in wine [42,43]; here, the geographical area where the vines were grown presents warm conditions, leading to the 5C wine having this type of aroma. Therefore, future studies should analyze these aromatic compounds and how they can be modulated according to the rootstocks used.

5. Conclusions

The results showed differences in the chemical and sensory characteristics of the grapes, especially in grape skins, and the resulting wines between rootstocks under the same edaphoclimatic and cultural conditions. The application of multivariate analysis supported the findings of the phenolic analyses, ultimately leading us to the conclusion that the 5C rootstock is distinguished by a higher concentration of phenolic compounds. In the same way, the sensory analysis showed that the 5C wines presented a higher color intensity, meaty aromas and astringency. In this case, we demonstrated that the rootstocks not only serve to adapt the plant to several conditions in the field but can modulate the chemical composition of grapes and wines and also the sensory characteristics of the resulting wines.
Further investigations are needed to examine other essential compounds, including aromatic compounds. Additionally, repeating the study over two or more years is essential to account for seasonal variations and to evaluate the influence of the rootstocks on both the chemical and sensory attributes of the resulting wines.

Supplementary Materials

The following supporting information can be downloaded at https://www.mdpi.com/article/10.3390/agronomy13102530/s1, Table S1: Low-molecular-weight phenolic compounds in Syrah grape skins from rootstocks 5C and Gravesac; Table S2: Low-molecular-weight phenolic compounds in Syrah grape seeds from rootstocks 5C and Gravesac; Table S3: Low-molecular-weight phenolic compounds in Syrah wines from rootstocks 5C and Gravesac; Table S4: Anthocyanins in Syrah wines from rootstocks 5C and Gravesac.

Author Contributions

F.H.-F.: Data Curation; Formal Analysis; Investigation; Writing—original draft. I.F.C.: Investigation; Formal Analysis; Writing—Review and Editing; Visualization. N.K.: Investigation; Visualization. Á.P.-N.: Resources; Visualization. A.C.-M.: Conceptualization; Funding Acquisition; Project Administration; Resources; Supervision; Writing—Original Draft; Writing—Review and Editing. All authors have read and agreed to the published version of the manuscript.

Funding

This research received funding from the Agencia Nacional de Investigación y Desarrollo (ANID-Chile), through the Fondecyt Iniciación Fund, under grant number 11180265.

Data Availability Statement

The data presented in this study can be obtained upon request from the corresponding author.

Acknowledgments

The authors would like to express their gratitude to Hacienda Valdeperillo (Limache, Chile) for their collaboration in providing the plant material and field support.

Conflicts of Interest

The authors declare that this research was conducted in the absence of any commercial or financial relationships that could be construed as potential conflicts of interest.

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Agronomy 13 02530 g001
Figure 1. Low-molecular-weight phenolic compounds in Syrah grape skins (A) and seeds (B) from rootstocks 5C and Gravesac. * denotes significant differences between rootstocks according to ANOVA (p < 0.05).
Figure 1. Low-molecular-weight phenolic compounds in Syrah grape skins (A) and seeds (B) from rootstocks 5C and Gravesac. * denotes significant differences between rootstocks according to ANOVA (p < 0.05).
Agronomy 13 02530 g001
Agronomy 13 02530 g002
Figure 2. Anthocyanin profile in Syrah grape skins. * denotes significant differences between rootstocks according to ANOVA (p < 0.05). D: delphinidin, C: cyanidin, P: peonidin, Pe: petunidin, M: malvidin, G: glucoside, AcG: acetylglucoside, CouG: p-coumaroylglucoside.
Figure 2. Anthocyanin profile in Syrah grape skins. * denotes significant differences between rootstocks according to ANOVA (p < 0.05). D: delphinidin, C: cyanidin, P: peonidin, Pe: petunidin, M: malvidin, G: glucoside, AcG: acetylglucoside, CouG: p-coumaroylglucoside.
Agronomy 13 02530 g002
Agronomy 13 02530 g003
Figure 3. Principal component analysis was used to analyze phenolic composition of Syrah wines from rootstocks 5C and Gravesac.
Figure 3. Principal component analysis was used to analyze phenolic composition of Syrah wines from rootstocks 5C and Gravesac.
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Agronomy 13 02530 g004
Figure 4. Descriptive sensory analysis of Syrah wine from rootstocks 5C and Gravesac. * denotes significant differences between rootstocks according to ANOVA (p < 0.05).
Figure 4. Descriptive sensory analysis of Syrah wine from rootstocks 5C and Gravesac. * denotes significant differences between rootstocks according to ANOVA (p < 0.05).
Agronomy 13 02530 g004
Table 1. Chemical and physical analyses of grapes of cv. Syrah from rootstocks 5C and Gravesac.
Table 1. Chemical and physical analyses of grapes of cv. Syrah from rootstocks 5C and Gravesac.
Parameters5CGRAVESAC
Weight of 50 berries (g)56.10 ± 1.57 b62.08 ± 1.13 a
Skins weight (g)6.30 ± 0.17 b7.04 ± 0.09 a
Seeds weight (g)2.08 ± 0.071.98 ± 0.14
Soluble solids (°Brix)23.40 ± 0.4023.01 ± 0.22
Titratable acidity (g tartaric acid L−1)6.15 ± 0.16 a5.55 ± 0.20 b
pH
Tartaric acid (mg kg−1)		2.80 ± 0.03
2608.30 ± 43.50		2.85 ± 0.02
2425.38 ± 217.21
Malic acid (mg kg−1)1154.34 ± 92.96 a723.88 ± 72.43 b
Citric acid (mg kg−1)51.84 ± 2.5846.00 ± 4.54
Skins total phenols (mg GAE g−1)7.25 ± 0.49 a5.67 ± 0.20 b
Seeds total phenols (mg GAE g−1)15.47 ± 1.1617.96 ± 2.21
Skins tannins mg (-)-epicatechin g−1)10.26 ± 0.53 a8.33 ± 0.61 b
Seeds tannins mg (-)-epicatechin g−1)65.41 ± 5.0777.70 ± 9.28
Total anthocyanins (mg malvidin g−1)11.47 ± 0.6111.30 ± 0.47
The values are presented as mean ± standard error (n = 5). Distinct lowercase letters are used to denote significant differences between rootstocks according to ANOVA (p < 0.05). GAE: gallic acid equivalent.
Table 2. Chemical analyses and color parameters of Syrah wines from rootstocks 5C and Gravesac.
Table 2. Chemical analyses and color parameters of Syrah wines from rootstocks 5C and Gravesac.
Parameters5CGRAVESAC
pH3.16 ± 0.043.10 ± 0.02
Titratable acidity (g tartaric acid L−1)1.50 ± 0.19 b2.47 ± 0.20 a
Alcohol content (% v/v)14.23 ± 0.10 a13.55 ± 0.12 b
Volatile acidity (g acetic acid L−1)0.46 ± 0.030.39 ± 0.04
Free SO2 (mg L−1)27.25 ± 2.6630.75 ± 1.89
Total SO2 (mg L−1)136.50 ± 8.01125.75 ± 16.98
Fermentable sugars (g L−1)0.23 ± 0.03 a0.10 ± 0.03 b
Reducing sugars (g L−1)4.01 ± 0.43 a2.52 ± 0.18 b
Total phenols (mg GAE L−1)1268.74 ± 19.63 a715.47 ± 12.81 b
Total tannins (mg (-)-epicatechin L−1)2587.21 ± 64.16 a1834.62 ± 243.12 b
Total anthocyanins (mg malvidin L−1)786.72 ± 8.36764.01 ± 9.49
Color intensity14.48 ± 0.5112.86 ± 0.80
Tone0.54 ± 0.000.51 ± 0.01
L*61.28 ± 1.0263.98 ± 1.83
C*44.99 ± 1.0145.41 ± 2.63
h*357.95 ± 0.18 a355.33 ± 0.23 b
The values are presented as mean ± standard error (n = 4). Distinct lowercase letters denote significant differences between rootstocks according to ANOVA (p < 0.05). GAE: gallic acid equivalent.
Table 3. Correlation matrix of chemical analyses with descriptive analysis of Syrah wines from rootstocks 5C and Gravesac.
Table 3. Correlation matrix of chemical analyses with descriptive analysis of Syrah wines from rootstocks 5C and Gravesac.
TPTTTAFlavanolsFlavonolsGlu-AntAce-AntCou-AntCIColorAstrinBitterBody
TP1.00------------
TT0.74 *1-----------
TA0.650.071----------
Flavanols0.95 *0.78 *0.641---------
Flavonols0.70.72 *0.290.84 *1--------
Glu-ant0.71 *0.630.550.680.241-------
Ace-ant−0.020.18−0.16−0.12−0.460.581------
Cou-ant−0.58−0.42−0.14−0.56−0.78 *0.120.551-----
CI0.510.76 *−0.050.630.79 *0.28−0.05−0.621----
Color0.70.250.610.680.680.1−0.55−0.81 *0.31---
Astrin0.78 *0.83 *0.170.79 *0.86 *0.31−0.28−0.76 *0.70.551--
Bitter−0.53−0.37−0.55−0.71 *−0.72 *−0.310.350.38−0.3−0.63−0.451-
Body0.410.310.060.420.67−0.28−0.72 *−0.83 *0.30.770.65−0.391
* Significance at p < 0.05. TP, total phenols; TT, total tannins; TA, total anthocyans; Ant, anthocyanins; Glu, glucosilated; Ace, acetylated; Cou, p-coumaroylated; CI, color intensity; Astrin, astringency; Bitter, bitterness.
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Heller-Fuenzalida, F.; Cuneo, I.F.; Kuhn, N.; Peña-Neira, Á.; Cáceres-Mella, A. Rootstock Effect Influences the Phenolic and Sensory Characteristics of Syrah Grapes and Wines in a Mediterranean Climate. Agronomy 2023, 13, 2530. https://doi.org/10.3390/agronomy13102530

AMA Style

Heller-Fuenzalida F, Cuneo IF, Kuhn N, Peña-Neira Á, Cáceres-Mella A. Rootstock Effect Influences the Phenolic and Sensory Characteristics of Syrah Grapes and Wines in a Mediterranean Climate. Agronomy. 2023; 13(10):2530. https://doi.org/10.3390/agronomy13102530

Chicago/Turabian Style

Heller-Fuenzalida, Florencia, Italo F. Cuneo, Nathalie Kuhn, Álvaro Peña-Neira, and Alejandro Cáceres-Mella. 2023. "Rootstock Effect Influences the Phenolic and Sensory Characteristics of Syrah Grapes and Wines in a Mediterranean Climate" Agronomy 13, no. 10: 2530. https://doi.org/10.3390/agronomy13102530

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