Assessment of the volatile and non-volatile profile of Savatiano PGI wines as affected by various terroirs in Attica, Greece.

Establishing the concept of terroir in wines, combined with the exploitation of native grape varieties, is considered a viable alternative to produce quality wines for increasing interest in the wine market. The aim of this study is the characterization of Protected Geographical Indication (PGI) Savatiano white wines from different regions of Attica (Greece), through the chemical and sensory description of the aroma of wines and the determination of their phenolic profiles. The wines produced with the same vinification protocol were evaluated using the descriptive sensory analysis method while they underwent profiling of volatile and phenolic compounds by gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) respectively. The presence of regional differences within the wines was also explored, elaborated with variables that contribute to their differentiation, such as soil type, monthly average temperature, and rainfall. Volatile and sensory profiles of the wines separated the regions and confirmed that sub-regional differences attributed mainly to soil characteristics affect wine aroma. The wines from the east part of Attica were higher in esters, terpenes and higher alcohols with high scores in fruity and blossoms attributes while wines from the north part of Attica presented higher intensity of mineral, nutty and herbaceous attributes. The separations based solely on the phenolic compounds concentration were less clear but a relationship was found between the content of phenolic acids and flavonoids and the studied regions. To our knowledge this is the first characterization of Savatiano PGI wines of Attica generating a fingerprint including chemical composition and sensory aroma characteristics to differentiate wines, combining this pattern with particular sub-regions.

Chemical and Sensory Characterization of Xinomavro Red Wine Using Grapes from Protected Designations of Northern Greece.

Despite Xinomavro (Vitis vinifera L.) being a well-known noble red grape variety of northern Greece, little is known about its ''bouquet'' typicity. Volatile compounds of Xinomavro wines produced using a common vinification protocol were analyzed by gas chromatography-mass spectrometry and sensory descriptive analysis was carried out with a trained panel. Wines were characterized by the presence of fatty acids, ethyl and acetate esters, and alcohols, with contributions from terpenes and a volatile phenol. The most active aroma compounds were determined to be 3-methylbutyl acetate, ß-damascenone, ethyl esters of octanoic and hexanoic acids, and eugenol. Those compounds positively correlated with fruity and spicy odor descriptors, with the wines being mostly characterized by five typical aroma terms: strawberry, berry fruit, spices, tomato, and green bell pepper. Partial least squares regression (PLSR) analysis was used to visualize relationship between the orthonasal sensory attributes and the volatile aroma compounds with calculated OAVs > 1. Key aroma-active volatiles in the wines were identified using GC-MS/olfactometry, providing a list of 40 compounds, among which 13 presented a modified detection frequency > 70%. This study is the first of its kind and provided strong indications regarding the aroma compounds defining the sensory characteristics of Xinomavro wines.

Benzothiadiazole Affects Grape Polyphenol Metabolism and Wine Quality in Two Greek Cultivars: Effects during Ripening Period over Two Years.

Grape berries are one of the most important sources of phenolic compounds, either consumed fresh or as wine. A pioneer practice aiming to enrich grape phenolic content has been developed based on the application of biostimulants such as agrochemicals initially designed to induce resistance against plant pathogens. A field experiment was conducted in two growing seasons (2019-2020) to investigate the effect of benzothiadiazole on polyphenol biosynthesis during grape ripening in Mouhtaro (red-colored) and Savvatiano (white-colored) varieties. Grapevines were treated at the stage of veraison with 0.3 mM and 0.6 mM benzothiadiazole. The phenolic content of grapes, as well as the expression level of genes involved in the phenylpropanoid pathway were evaluated and showed an induction of genes specifically engaged in anthocyanins and stilbenoids biosynthesis. Experimental wines deriving from benzothiadiazole-treated grapes exhibited increased amounts of phenolic compounds in both varietal wines, as well as an enhancement in anthocyanin content of Mouhtaro wines. Taken together, benzothiadiazole can be utilized to induce the biosynthesis of secondary metabolites with oenological interest and to improve the quality characteristics of grapes produced under organic conditions.

Impact of Assimilable Nitrogen Supplementation on Saccharomyces cerevisiae Metabolic Response and Aromatic Profile of Moschofilero Wine.

The concentration of nitrogen in must is critical to yeast fermentation efficiency and wine aroma profile. The present work determined the effect of the amount of yeast assimilable nitrogen (YAN) on fermentation kinetics, aroma production, and gene expression patterns of the wine yeast Saccharomyces cerevisiae. Fermentations were performed under two different YAN concentrations of must. Acetate esters, linalool, and nerol appeared to be clearly affected by the different YAN levels. Real-time-PCR results revealed that the genes involved in ethyl and acetate esters production recorded, in general, higher transcript levels under high nitrogen supplementation. In addition, an up-regulation of the BGL2 and EXG1 genes, which are related to terpenes production, was observed in the case of high nitrogen content and it is well corresponded to the terpenol concentration found. Our study revealed the impact of nitrogen supplementation on yeast metabolism and its importance to adjust wine's aromatic composition and sensory profile.

Abscisic Acid and Chitosan Modulate Polyphenol Metabolism and Berry Qualities in the Domestic White-Colored Cultivar Savvatiano.

During the last decade, several studies demonstrated the effect of biostimulants on the transcriptional and metabolic profile of grape berries, suggesting their application as a useful viticultural practice to improve grape and wine quality. Herein, we investigated the impact of two biostimulants-abscisic acid (0.04% w/v and 0.08% w/v) and chitosan (0.3% w/v and 0.6% w/v)-on the polyphenol metabolism of the Greek grapevine cultivar, Savvatiano, in order to determine the impact of biostimulants' application in the concentration of phenolic compounds. The applications were performed at the veraison stage and the impact on yield, berry quality traits, metabolome and gene expression was examined at three phenological stages (veraison, middle veraison and harvest) during the 2019 and 2020 vintages. Results showed that anthocyanins increased during veraison after treatment with chitosan and abscisic acid. Additionally, stilbenoids were recorded in higher amount following the chitosan and abscisic acid treatments at harvest. Both of the abscisic acid and chitosan applications induced the expression of genes involved in stilbenoids and anthocyanin biosynthesis and resulted in increased accumulation, regardless of the vintage. Alterations in other phenylpropanoid gene expression profiles and phenolic compound concentrations were observed as well. Nevertheless, they were mostly restricted to the first vintage. Therefore, the application of abscisic acid and chitosan on the Greek cultivar Savvatiano showed promising results to induce stilbenoid metabolism and potentially increase grape defense and quality traits.

The Effect of Water Deficit on Two Greek Vitis vinifera L. Cultivars: Physiology, Grape Composition and Gene Expression during Berry Development.

Plants are exposed to numerous abiotic stresses. Drought is probably the most important of them and determines crop distribution around the world. Grapevine is considered to be a drought-resilient species, traditionally covering semiarid areas. Moreover, in the case of grapevine, moderate water deficit is known to improve the quality traits of grape berries and subsequently wine composition. However, against the backdrop of climate change, vines are expected to experience sustained water deficits which could be detrimental to both grape quality and yield. The influence of water deficit on two Greek Vitis vinifera L. cultivars, 'Agiorgitiko' and 'Assyrtiko', was investigated during the 2019 and 2020 vintages. Vine physiology measurements in irrigated and non-irrigated plants were performed at three time-points throughout berry development (green berry, veraison and harvest). Berry growth and composition were examined during ripening. According to the results, water deficit resulted in reduced berry size and increased levels of soluble sugars, total phenols and anthocyanins. The expression profile of specific genes, known to control grape color, aroma and flavor was altered by water availability during maturation in a cultivar-specific manner. In agreement with the increased concentration of phenolic compounds due to water deficit, genes of the phenylpropanoid pathway in the red-skinned Agiorgitiko exhibited higher expression levels and earlier up-regulation than in the white Assyrtiko. The expression profile of the other genes during maturation or in response to water deficit was depended on the vintage.

Explore the Rare-Molecular Identification and Wine Evaluation of Two Autochthonous Greek Varieties: "Karnachalades" and "Bogialamades".

Wines produced from autochthonous Vitis vinifera varieties have an essential financial impact on the national economy of Greece. However, scientific data regarding characteristics and quality aspects of these wines is extremely limited. The aim of the current study is to define the molecular profile and to describe chemical and sensory characteristics of the wines produced by two autochthonous red grapevine varieties-"Karnachalades" and "Bogialamades"-grown in the wider area of Soufli (Thrace, Greece). We used seven microsatellites to define the molecular profile of the two varieties, and then we compared their profile to similar molecular data from other autochthonous as well as international varieties. Grape berries were harvested at optimum technological maturity from a commercial vineyard for two consecutive vintages (2017-2018) and vilification was performed using a common vinification protocol: the 2017 vintage provided wines, from both varieties, with greater rates of phenolics and anthocyanins than 2018, whereas regarding the sensory analysis, "Bogialamades" wine provided a richer profile than "Karnachalades". To our knowledge, this is the first study that couples both molecular profiling and exploration of the enological potential of the rare Greek varieties "Karnachalades" and "Bogialamades"; they represent two promising varieties for the production of red wines in the historic region of Thrace.

Assessment of Antioxidant and Antimutagenic Properties of Red and White Wine Extracts In Vitro.

Wine is an alcoholic beverage of complex composition obtained through the fermentation of grape must. The consumption of wine has already been associated with a multitude of beneficial effects due to its high polyphenolic content. In this study, four Greek emblematic wines from two red (i.e., Xinomavro and Agiorgitiko) and two white (i.e., Assyrtiko and Malagouzia) varieties were analyzed for the estimation of their antioxidant profiles. To address this question, we assessed their ability to scavenge both synthetic and endogenous free radicals, such as DPPH•, ABTS+•, OH•, O2-, their potential reducing power, and their antimutagenic and antigenotoxic properties. All varieties exhibited potent antioxidant activity, as indicated by the results of methods above, with the red wines appearing more effective than the white ones regarding antioxidant capacity. Our small-scale study is the first to reveal that these wine varieties may have the ability to scavenge the most reactive endogenous radicals. In the future, this finding must be accompanied by larger studies to fill a knowledge gap in the scientific literature concerning a holistic approach of the in vitro antioxidant action of plant polyphenolic compounds. Conclusively, we believe that wines possess high bioactivity that allow them to settle in the industry of food additives and medicinal products.

Odor Characterization of White Wines Produced from Indigenous Greek Grape Varieties Using the Frequency of Attribute Citation Method with Trained Assessors.

The aim of this study was to investigate the sensory aroma profiles of white wines of the indigenous Greek grape varieties Assyrtiko, Malagousia, Moschofilero, and Roditis. Twenty-three panelists evaluated 17 wines of the aforementioned varieties using the frequency of attribute citation method. Three indices were calculated to assess panel performance in terms of reproducibility. Correspondence analysis and cluster analysis were employed to investigate the sensory space of the wines. Samples of the Roditis variety were characterized mainly by Banana and Vanilla odors; Assyrtiko samples had Earthy, Mushroom, and Nutty odors, as well as Lemon and Honey for some of the samples. Malagousia wines were described as having Lemon, Grapefruit, and Citrus blossom character, and they shared some descriptors with Assyrtiko wines, such as Mushroom and Earthy, and some with Moschofilero samples, i.e., floral and citrus notes. All Moschofilero wines exhibited a floral odor profile: specifically, Rose, Jasmine, or more Citrus blossom-like. Moreover, some Moschofilero samples also revealed a Grapefruit, Lemon, and/or Earthy character, while others expressed Honey notes. In conclusion, despite common characteristics found within varieties, some samples of different varieties exhibited overlapping profiles, and in some cases, samples of the same variety were quite different from each other.

Proanthocyanidin content as an astringency estimation tool and maturation index in red and white winemaking technology.

Selecting the appropriate type of barrel for wine maturation but also deciding on the optimum maturation length, is a challenge for winemakers. As different types of barrel woods emerge, it is of great importance for a guideline to be established, which could facilitate winemaking decisions. Since the sensory perception of the finished wine, and particularly the intensity of astringency, is a decisive factor for the quality of a barrel-aged wine, in this experiment, the structural characteristics of wine proanthocyanidins were determined and their correlation with astringency was established. According to the results obtained, the proanthocyanidin content and the type of subunit that is dominant in tannin chains could be used to construct an astringency estimation model. The findings could provide winemakers with a useful tool when deciding how long to mature a specific type of wine in a specific wood container without making it appear coarse and astringent.

Changes in Tannin Composition of Syrah Grape Skins and Seeds during Fruit Ripening under Contrasting Water Conditions.

Tannin accumulation and composition were determined in skins and seeds isolated from Vitis vinifera cv. Syrah grapes submitted to contrasting water regimes under semiarid climatic conditions. Three irrigation treatments were conducted, starting at berry set through harvest of two growing seasons, 2011 and 2012: irrigation at 100% of crop evapotranspiration ETc (FI), irrigation at 50% of ETc (DI) and non-irrigated (NI). Seed total tannins did not vary with maturity but those of skins underwent a progressive decline (especially in 2011), expressed both on a fresh weight and on a per berry basis. Skin total tannin concentration and content per berry were increased under NI and DI conditions, mainly in 2012. In contrast, seed total tannins (in 2012) and flavan-3-ol monomers and tannin oligomers (both years) were higher in the fully irrigated vines (FI). Skin polymer size increased during ripening, NI and DI skins showing higher mean degree of polymerization (mDP) compared to FI at harvest. NI was also associated with a lower percentage of galloylation (%G) in skin oligomeric fraction (in 2012) and a lower percentage of prodelphinidins in the skin polymeric fraction (both years) at harvest. The mDP and %G of seed extracts did not vary during ripening and were higher in NI but only in 2012. According to the results, management of vine water status was shown to influence tannin amount and composition of Syrah grapes grown under semiarid conditions.

Effect of irrigation regime on perceived astringency and proanthocyanidin composition of skins and seeds of Vitis vinifera L. cv. Syrah grapes under semiarid conditions.

In this work, the effect of water availability on astringency of seed and skin extracts of Vitis vinifera cv. Syrah berries under the typical semiarid conditions of Greece was investigated. Moreover, astringency was assessed in relation to proanthocyanidin composition. For this purpose, three irrigation treatments were applied starting at berry set through harvest of 2011 and 2012: full irrigation (FI) at 100% of crop evapotranspiration, deficit irrigation (DI) at 50% and non-irrigated (NI). FI skin and seed extracts were perceived significantly more astringent than NI. Total phenol, total tannin, (+)-catechin, (-)-epicatechin and procyanidin C1 concentrations were positively correlated with astringency. Positive correlations were also obtained among astringency and average degree of polymerization and proportion of the extension units of shorter tannins while astringency of larger tannins was correlated with the proportion of terminal units. On the contrary, total anthocyanin and epigallocatechin contents were negatively correlated with astringency.

Effect of irrigation regime on anthocyanin content and antioxidant activity of Vitis vinifera L. cv. Syrah grapes under semiarid conditions.

BACKGROUND: Irrigation management is a powerful technique to control grape and wine colour. However, water deficit effects on anthocyanins are often contrasting, depending on the severity of water restriction. In addition, the effect of irrigation on the antiradical capacity of grapes has not been extensively studied. In this work the effect of water availability on anthocyanin profile and content as well as the antioxidant activity of Syrah grapes was investigated in an irrigation trial under semiarid climate conditions. RESULTS: Three irrigation treatments were applied in a 15-year-old Vitis vinifera cv. Syrah vineyard, starting at berry set through harvest of 2011 and 2012: full irrigation (FI) at 100% of crop evapotranspiration (ETc), deficit irrigation (DI) at 50% of ETc and non-irrigated (NI). NI grapes were characterized by increased individual anthocyanin content. However, differences among irrigation treatments were significant only around 18-24 days after veraison, when anthocyanin accumulation was maximum. The antioxidant activity of the skin extracts was not affected by irrigation. CONCLUSION: Irrigation proved to be an effective technique to control anthocyanin content in Syrah grapes under semiarid conditions. However, anthocyanin accumulation pattern should be considered by winemakers to appropriately select harvest time for improved wine colour.

Differentiation of Wines Treated with Wood Chips Based on Their Phenolic Content, Volatile Composition, and Sensory Parameters.

The effects of both wood chips addition and contact time on phenolic content, volatile composition, color parameters, and organoleptic character of red wine made by a native Greek variety (Agiorgitiko) were evaluated. For this purpose, chips from American, French, Slavonia oak, and Acacia were added in the wine after fermentation. A mixture consisting of 50% French and 50% Americal oak chips was also evaluated. In an attempt to categorize wine samples, various chemical parameters of wines and sensory parameters were studied after 1, 2, and 3 mo of contact time with chips. The results showed that regardless of the type of wood chips added in the wines, it was possible to differentiate the samples according to the contact time based on their phenolic composition and color parameters. In addition, wood-extracted volatile compounds seem to be the critical parameter that could separate the samples according to the wood type. The wines that were in contact with Acacia and Slavonia chips could be separated from the rest mainly due to their distinct sensory characters.

Direct and simultaneous quantification of tannin mean degree of polymerization and percentage of galloylation in grape seeds using diffuse reflectance fourier transform-infrared spectroscopy.

The direct and simultaneous quantitative determination of the mean degree of polymerization (mDP) and the degree of galloylation (%G) in grape seeds were quantified using diffuse reflectance infrared Fourier transform spectroscopy and partial least squares (PLS). The results were compared with those obtained using the conventional analysis employing phloroglucinolysis as pretreatment followed by high performance liquid chromatography-UV and mass spectrometry detection. Infrared spectra were recorded in solid state samples after freeze drying. The 2nd derivative of the 1832 to 1416 and 918 to 739 cm(-1) spectral regions for the quantification of mDP, the 2nd derivative of the 1813 to 607 cm(-1) spectral region for the degree of %G determination and PLS regression were used. The determination coefficients (R(2) ) of mDP and %G were 0.99 and 0.98, respectively. The corresponding values of the root-mean-square error of calibration were found 0.506 and 0.692, the root-mean-square error of cross validation 0.811 and 0.921, and the root-mean-square error of prediction 0.612 and 0.801. The proposed method in comparison with the conventional method is simpler, less time consuming, more economical, and requires reduced quantities of chemical reagents and fewer sample pretreatment steps. It could be a starting point for the design of more specific models according to the requirements of the wineries.

Methoxypyrazine analysis and influence of viticultural and enological procedures on their levels in grapes, musts, and wines.

This review discusses the factors that affect the concentrations of methoxypyrazines (MPs) and the techniques used to analyze MPs in grapes, musts, and wines. MPs are commonly studied pyrazines in food science due to their contribution of aroma and flavor to numerous vegetables such as peas and asparagus. They are described as highly odorous compounds with a very low olfactory threshold. The grape varietals that exhibit green or herbaceous aromas that are characteristic of MPs are predominantly Vitis vinifera cv. Cabernet Sauvignon and Sauvignon Blanc, but include others. The most extensively studied MPs include 3-isobutyl-2-methoxypyrazine, 3-isopropyl-2-methoxypyrazine, and 3-sec-butyl-2-methoxypyrazine. It outlines the significance of methoxypyrazines in grapes, musts, and wines in terms of the concentrations that are capable of contributing their sensory characteristics to wines. This review discusses methods for analyzing MPs including gas chromatography-mass spectroscopy (one or two dimension) and high-performance liquid chromatography, the appropriate extraction techniques, and the efficacy of these methods. Additionally, this review explores factors that affect pyrazine content of grapes, must, and wines, such as the effects of different viticultural practices, effects of light exposure and grape maturation, climate, soil, the multi-colored Asian lady beetle and the effects of different vinification processes.

Development of a rapid method for the quantitative analysis of four methoxypyrazines in white and red wine using multi-dimensional Gas Chromatography-Mass Spectrometry.

Alkyl-methoxypyrazines (MPs) are important odour-active constituents of many grape cultivars and their wines. Recently, a new MP - 2,5-dimethyl-3-methoxypyrazine (DMMP) - has been reported as a possible constituent of wine. This study sought to develop a rapid and reliable method for quantifying DMMP, isopropyl methoxypyrazine (IPMP), secbutyl methoxypyrazine (SBMP) and isobutyl methoxypyrazine (IBMP) in wine. The proposed method is able to rapidly and accurately resolve all 4 MPs in a range of wine styles, with limits of detection between 1 and 2 ng L(-1) for IPMP, SBMP and IBMP and 5 ng L(-1) for DMMP. Analysis of a set of 11 commercial wines agrees with previously published values for IPMP, SBMP and IBMP, and shows for the first time that DMMP may be an important and somewhat common odorant in red wines. To our knowledge, this is the first analytical method developed for the quantification of DMMP in wine.

Occurrence and contribution of alkyl methoxypyrazines in wine tainted by Harmonia axyridis and Coccinella septempunctata.

BACKGROUND: Ladybug taint (LBT) is a wine fault caused by the inadvertent incorporation of ladybeetles to the wine during the winemaking process. Harmonia axyridis, also known as the multicolored Asian lady-beetle (MALB), was the only species considered responsible for causing the taint. A second species, Coccinella septempunctata (7 Spot), has recently also been implicated. The main objectives of this study were to identify and quantify the compounds associated with ladybug taint of white and red wine from these two Coccinellidae species (Harmonia axyridis and Coccinella septempunctata), and determine the most odor-active compounds in LBT-affected wines. RESULTS: Results show that Coccinellidae can contribute dimethyl, isopropyl, secbutyl and isobutyl methoxypyrazine to wine when they are processed with grapes, and that the methoxypyrazine composition varies between MALB and 7 Spot. Isopropyl methoxypyrazine is the major contributor to LBT in wines produced with these ladybeetle species, although secbutyl and isobutyl methoxypyrazine may contribute in MALB- and 7 Spot-affected wines, respectively. Finally, dimethyl methoxypyrazine is reported for the first time in wines not affected by Coccinellidae; the global significance and prevalence of this compound in wine remains to be determined. CONCLUSION: Both Harmonia axyridis and Coccinella septempunctata can contribute alkyl methoxypyrazines, and particularly isopropyl methoxypyrazine, to wine at concentrations that can have a negative impact on wine quality. These findings will allow grape growers and winemakers to be more aware of the risks posed by 7 Spot beetles, and to take the appropriate preventative and remedial actions. Furthermore, this is the first time dimethyl methoxypyrazine is reported in wine, either as an endogenous or Coccinellidae-derived odorant.

Effects of severity of post-flowering leaf removal on berry growth and composition of three red Vitis vinifera L. cultivars grown under semiarid conditions.

The effects of the severity of post-flowering leaf removal on the growth and phenolic composition of berry skin and seeds were studied in three Vitis vinifera L. genotypes over two consecutive seasons, 2007 and 2008. The study was conducted in a commercial vertical shoot positioned (VSP)-trained nonirrigated vineyard of northern Greece, planted with cultivars Merlot, Cabernet Sauvignon, and Sangiovese. Three different severities of leaf removal in the fruit zone were applied manually at berry set: nondefoliated (ND), removal of the lateral shoots of the first six basal nodes (LR), and full removal of the total leaf area (main leaves and lateral shoots) of the first six basal nodes (FR). Grape samples were obtained at commercial harvest. Leaf removal decreased yield per vine and cluster weight in Merlot and Sangiovese. Cluster compactness was reduced with the severity of defoliation only in Merlot, due to a decrease in berry number per cluster; berry fresh weight was unaffected in both cultivars. On the contrary, in Cabernet Sauvignon, yield was unaffected but berry size was restrained by leaf removal. Skin and seed mass followed variations in berry mass (except for seed mass in Sangiovese). Fruit zone leaf removal did not affect must soluble solids and increased titratable acidity only in Merlot. Defoliation increased skin anthocyanins in Merlot and Cabernet Sauvignon in the order FR > LR > ND but significantly reduced seed flavan-3-ols mainly as a result of the reduction in catechin and epicatechin amount. For these varieties, FR had lower seed flavan-3-ols than ND in both varieties, whereas LR had intermediate values. However, in Sangiovese, the highest seed phenolic content was recorded in LR. The results showed that post-flowering leaf removal improved the overall berry composition in Merlot and Cabernet Sauvignon but had limited effect in Sangiovese.