The Role of Terroir on the Ripening Traits of V. vinifera cv 'Glera' in the Prosecco Area.

The grapevine (Vitis vinifera L.) is widely cultivated worldwide owing to the substantial commercial value of the grapes and other products derived from their processing, wines in particular. The grapevine is characterized by a remarkable phenotypic plasticity within the same variety, which shapes the final berry quality attributes hence reflecting the complex interactions between the plant and the environment leading to the expression of wine typicity. In this study, we explored the metabolomic and transcriptomic basis of the plasticity of Glera, a white berry grapevine variety particularly renowned for the production of wine Prosecco. The two selected vineyards varied for site altitude and pedoclimatic conditions. We highlighted that these environments determined different berry ripening dynamics at the level of both technological parameters and the total abundance and intrafamily distribution of phenolic compounds. Moreover, a clear impact on the grape aroma profile was observed. The genome-wide gene expression analysis of the berries revealed remarkable differences in the ripening transcriptomic program, reflecting the differences in water status, light exposure, and temperature experienced by the plants while growing at the two sites. Overall, this survey portrayed how the quality attributes of the cv 'Glera' grape berries may be affected by different environmental conditions within the typical area of Prosecco wine production.

Metabolomics Insight into the Variety-Mediated Responses to Aspergillus carbonarius Infection in Grapevine Berries.

Limited knowledge regarding the susceptibility of grape varieties to ochratoxin A (OTA)-producing fungi is available to date. This study aimed to investigate the susceptibility of different grape varieties to Aspergillus carbonarius concerning OTA contamination and modulation at the metabolome level. Six grape varieties were selected, sampled at early veraison and ripening, artificially inoculated with A. carbonarius, and incubated at two temperature regimes. Significant differences were observed across cultivars, with Barbera showing the highest incidence of moldy berries (around 30%), while Malvasia and Ortrugo showed the lowest incidence (about 2%). OTA contamination was the lowest in Ortrugo and Malvasia, and the highest in Croatina, although it was not significantly different from Barbera, Merlot, and Sauvignon Blanc. Fungal development and mycotoxin production changed with grape variety; the sugar content in berries could also have played a role. Unsupervised multivariate statistical analysis from metabolomic fingerprints highlighted cultivar-specific responses, although a more generalized response was observed by supervised OPLS-DA modeling. An accumulation of nitrogen-containing compounds (alkaloids and glucosinolates), phenylpropanoids, and terpenoids, in addition to phytoalexins, was observed in all samples. A broader modulation of the metabolome was observed in white grapes, which were less contaminated by OTA. Jasmonates and oxylipins were identified as critical upstream modulators in metabolomic profiles. A direct correlation between the plant defense machinery and OTA was not observed, but the information was acquired and can contribute to optimizing preventive actions.

Moderate Wine Consumption and Health: A Narrative Review.

Although it is clearly established that the abuse of alcohol is seriously harmful to health, much epidemiological and clinical evidence seem to underline the protective role of moderate quantities of alcohol and in particular of wine on health. This narrative review aims to re-evaluate the relationship between the type and dose of alcoholic drink and reduced or increased risk of various diseases, in the light of the most current scientific evidence. In particular, in vitro studies on the modulation of biochemical pathways and gene expression of wine bioactive components were evaluated. Twenty-four studies were selected after PubMed, Scopus and Google Scholar searches for the evaluation of moderate alcohol/wine consumption and health effects: eight studies concerned cardiovascular diseases, three concerned type 2 diabetes, four concerned neurodegenerative diseases, five concerned cancer and four were related to longevity. A brief discussion on viticultural and enological practices potentially affecting the content of bioactive components in wine is included. The analysis clearly indicates that wine differs from other alcoholic beverages and its moderate consumption not only does not increase the risk of chronic degenerative diseases but is also associated with health benefits particularly when included in a Mediterranean diet model. Obviously, every effort must be made to promote behavioral education to prevent abuse, especially among young people.

Impact of Climatic Conditions on the Resveratrol Concentration in Blend of Vitis vinifera L. cvs. Barbera and Croatina Grape Wines.

The aim of this work was to investigate the effect of meteorological conditions on resveratrol concentration of red wines produced in Piacenza viticultural region (Italy). In this regard, six representative estates producing Colli Piacentini Gutturnio DOC (a blend of V. vinifera L. cvs. Barbera and Croatina) vintage wines were analysed for trans- and cis-resveratrol over an 8-year period (1998-2005). Grapes were taken from the same vineyard in each estate by using the same enological practices over the entire investigated period. The meteorological conditions corresponding to the production areas were recorded, and bioclimatic indices were calculated as well. Overall, cis-resveratrol concentration was negatively correlated to Huglin index and August mean temperature, whilst positive correlation coefficients were found when considering the Selianinov index and the rainfall of September.

Assessment of Resistance Components for Improved Phenotyping of Grapevine Varieties Resistant to Downy Mildew.

Grapevine varieties showing partial resistance to downy mildew, caused by Plasmopara viticola, are a promising alternative to fungicides for disease control. Resistant varieties are obtained through breeding programs aimed at incorporating Rpv loci controlling the quantitative resistance into genotypes characterized by valuable agronomic and wine quality traits by mean of crossing. Traditional phenotyping methods used in these breeding programs are mostly based on the assessment of the resistance level after artificial inoculation of leaf discs in bioassays, by using the visual score proposed in the 2nd Edition of the International Organization of Vine and Wine (OIV) Descriptor List for Grape Varieties and Vitis species (2009). In this work, the OIV score was compared with an alternative approach, not used for the grapevine-downy mildew pathosystem so far, based on the measurement of components of resistance (RCs); 15 grapevine resistant varieties were used in comparison with the susceptible variety 'Merlot'. OIV scores were significantly correlated with P. viticola infection frequency (IFR), the latent period for the downy mildew (DM) lesions to appear (LP50), and the number of sporangia produced per lesion (SPOR), so that when the OIV score increased (i.e., the resistance level increases), IFR and SPOR decreased, while LP50 increased. The relationship was linear for LP50, monomolecular for IFR and hyperbolic for SPOR. No significant correlation was found between OIV score and DM lesion size, sporangia produced per unit area of lesion, length of infectious period, and infection efficiency of the sporangia produced on DM lesions. The correlation between OIV score and area under the disease progress curve (AUDPC) calculated by using the RCs and a simulation model was significant and fit an inverse exponential function. Based on the results of this study, the measurement of the RCs to P. viticola in grapevine varieties by means of monocyclic, leaf disc bioassays, as well as their incorporation into a model able to simulate their effect on the polycyclic development of DM epidemics in vineyards, represents an improved method for phenotyping resistance level.

Phenolic profiles and anti-inflammatory activities of sixteen table grape (Vitis vinifera L.) varieties.

Several studies have shown that fresh grape and its derivatives contain phenolic compounds exhibiting antioxidant and health promoting effects, particularly in relation to the cardiovascular system. In this study, two methods were developed to characterize sixteen varieties of table and wine grapes: (1) a LC-MS method to identify major and minor phenolic compounds; and (2) a HPLC-DAD method to quantify the most representative compounds. Sixty-seven molecules belonging to different classes of phenolic compounds were identified: anthocyanins, flavan-3-ols, flavonols, stilbenes and organic acids. In parallel, the free radical scavenging activity and anti-inflammatory activities of the 16 grape varieties were evaluated. The results showed a good correlation between the total phenolic content and the biological activity. Extracts from Exalta and Albarossa grape varieties were the most active in reducing IL-8 release by gastric epithelial cells (IC50 = 8.48 µg mL-1 and 6.68 µg mL-1, respectively), a biomarker of inflammatory processes. The observed biological activities were mainly associated with skin and seed extracts/portions. The interest in studying table grapes and their non-fermented derivatives as sources of healthy compounds has increased in the last few years and our findings suggest that table grapes and their fresh derivatives, in addition to wine, could be involved in the health promoting effects of the Mediterranean diet.

Chitosan treatment elicited defence mechanisms, pentacyclic triterpenoids and stilbene accumulation in grape (Vitis vinifera L.) bunches.

The stimulation of the plant response to pathogen attack by the application of resistance inducers, called elicitors, could represent an environmentally and commercially viable alternative or complement to existing pathogen control methods. In this work, the elicitor chitosan was sprayed on grape (Vitus vinifera L.) berries growing on the vine to shed light into the elicitation mechanisms underlying its application, with untreated bunches as controls. To gain a more comprehensive picture of the complex molecular processes elicited by chitosan, a proteomic approach was complemented by target and untargeted mass spectrometric analyses. The treatment altered the regulation of reactive oxygen species, with Cu/Zn superoxide dismutase and glyoxal oxidase showing up-accumulation. This might lead to an increased lignification via hypersensitive response mechanisms. Furthermore, enzymes involved in anthocyanin rather than stilbene phytoalexins accumulated in treated bunches. Stilbenes increased from 1.6 times (resveratrol) up to 3.8 times (piceid) over untreated bunches. The up accumulation of hydroperoxide lyase might lead to accumulation of oxylipins. Furthermore, the pentacyclic triterpenoids ursolate, oleanoate and betulinate increased by 1.25, 1.47 and 3.68 times in treated grape bunches (p < 0.01). Hence, the main processes underlying the response of grape fruits to chitosan treatment involved the accumulation of phenylpropanoid and triterpenoids phytoalexins, as well as the modulation of oxidative stress-related enzymes.

Association between taste receptor (TAS) genes and the perception of wine characteristics.

Several studies have suggested a possible relationship between polymorphic variants of the taste receptors genes and the acceptance, liking and intake of food and beverages. In the last decade investigators have attempted to link the individual ability to taste 6-n-propylthiouracil (PROP) and the sensations, such as astringency and bitterness, elicited by wine or its components, but with contradictory results. We have used the genotype instead of the phenotype (responsiveness to PROP or other tastants), to test the possible relation between genetic variability and the perception of wine characteristic in 528 subjects from Italy and the Czech Republic. We observed several interesting associations, among which the association between several TAS2R38 gene single nucleotide polymorphisms (P = 0.002) and the TAS2R16-rs6466849 polymorphism with wine sourness P = 0.0003). These associations were consistent in both populations, even though the country of origin was an important factor in the two models, thus indicating therefore that genetics alongside cultural factors also play a significant role in the individual liking of wine.

Wine Resveratrol: From the Ground Up.

The ability of the grapevine to activate defense mechanisms against some pathogens has been shown to be linked to the synthesis of resveratrol and other stilbenes by the plant (inducible viniferins). Metabolized viniferins may also be produced or modified by extracellular enzymes released by the pathogen in an attempt to eliminate undesirable toxic compounds. Because of the important properties of resveratrol, there is increasing interest in producing wines with higher contents of this compound and a higher nutritional value. Many biotic and abiotic elicitors can trigger the resveratrol synthesis in the berries, and some examples are reported. Under the same elicitation pressure, viticultural and enological factors can substantially affect the resveratrol concentration in the wine. The production of high resveratrol-containing grapes and wines relies on quality-oriented viticulture (suitable terroirs and sustainable cultural practices) and winemaking technologies that avoid degradation of the compound. In general, the oenological practices commonly used to stabilize wine after fermentation do not affect resveratrol concentration, which shows considerable stability. Finally the paper reports on two sirtuin genes (SIRT) expressed in grapevine leaves and berries and the role of resveratrol on the deacetylation activity of the encoded enzymes.

Mediterranean Way of Drinking and Longevity.

The relation between alcohol consumption and mortality is a J-shaped curve in most of the many studies published on this topic. The Copenhagen Prospective Population Studies demonstrated in the year 2000 that wine intake may have a beneficial effect on all cause mortality that is additive to that of alcohol. Wine contains various poliphenolic substances which may be beneficial for health and in particular flavonols (such as myricetin and quercetin), catechin and epicatechin, proanthocyanidins, anthocyanins, various phenolic acids and the stilbene resveratrol. In particular, resveratrol seems to play a positive effect on longevity because it increases the expression level of Sirt1, besides its antioxidant, anti-inflammatory and anticarcinogenic properties. Moderate wine drinking is part of the Mediterranean diet, together with abundant and variable plant foods, high consumption of cereals, olive oil as the main (added) fat and a low intake of (red) meat. This healthy diet pattern involves a "Mediterranean way of drinking," that is a regular, moderate wine consumption mainly with food (up to two glasses a day for men and one glass for women). Moderate wine drinking increases longevity, reduces the risk of cardiovascular diseases and does not appreciably influence the overall risk of cancer.

Study of Grape Polyphenols by Liquid Chromatography-High-Resolution Mass Spectrometry (UHPLC/QTOF) and Suspect Screening Analysis.

Suspect screening analysis is a targeted metabolomics method in which the identification of compounds relies on specific available information, such as their molecular formula and isotopic pattern. This method, coupled to liquid chromatography-high-resolution mass spectrometry, is effective in the study of grape metabolomics, in particular for characterization of flavonols, stilbene derivatives, and anthocyanins. For identification of compounds expected in the samples, a new database of putative compounds was expressly constructed by using the molecular information on potential metabolites of grape and wine from the literature and other electronic databases. Currently, this database contains around 1,100 compounds. The method allows identification of several hundred grape metabolites with two analyses (positive and negative ionization modes), and performing of data reprocessing using "untargeted" algorithms also provided the identification of some flavonols and resveratrol trimers and tetramers in grape for the first time. This approach can be potentially used in the study of metabolomics of varieties of other plant species.

Profiling of grape monoterpene glycosides (aroma precursors) by ultra-high performance-liquid chromatography-high resolution mass spectrometry (UHPLC/QTOF).

A 'suspect screening analysis' method for grape metabolomics by ultra-high performance-liquid chromatography (UHPLC) and high-resolution quadrupole-time of flight (QTOF) mass spectrometry was recently developed. This method was applied to study grape monoterpene glycosides, the main grape aroma precursors. Since standard compounds were not available, they were tentatively identified by overlapping various analytical approaches, in agreement with the indications recommended in mass spectrometry (MS)-based metabolomics. Accurate mass and isotopic pattern, MS/MS fragmentation, correlation between fragments observed and putative structures and between liquid chromatography coupled with mass spectrometry (LC/MS) and gas chromatography/mass spectrometry signals were studied. Seventeen monoterpene glycosides were identified without performing the hydrolytic artifacts commonly used to study these compounds which may affect sample profile. This is the first time that a detailed study of these aroma precursors has been carried out by direct LC/MS analysis.

Genetic variability and geographic typicality of Italian former Prosecco grape variety using PCR-derived molecular markers.

This study uses PCR-derived marker systems to investigate the extent and distribution of genetic variability of 80 Italian Prosecco accessions coming from Prosecco DOC area (north-east area of Italy). The studied samples include genotypes from Veneto and Friuli Venezia Giulia region. In order to verify the varietal identity of the samples, analyses based on 22 SSR loci were performed, and two grape varieties were found: Prosecco tondo and Prosecco lungo. In addition to microsatellite analysis, intra-varietal variability study was performed using AFLP, SAMPL, ISSR, and M-AFLP molecular markers. This molecular approach could discriminate different Prosecco tondo accessions coming from Treviso hills, from Veneto plain, from Friuli Venezia Giulia region, and from Padua hills (Serprina samples). As concerning Prosecco lungo variety, it was possible to discriminate molecularly the accessions from Veneto region and those from Friuli Venezia Giulia region. The molecular analysis allowed a distinction of the Prosecco genotypes on the basis of their geographic origins with plant-specific markers able to differentiate all Prosecco accessions. In this paper, the studied grape variety is termed Prosecco and not Glera (which is the present name) because the sampled vineyards were established many years ago when the name of the variety was Prosecco.

Identification of saffron aroma compound ß-isophorone (3,5,5-trimethyl-3-cyclohexen-1-one) in some V. vinifera grape varieties.

Carotenoid-derived aroma compounds play an important role in the composition of aroma and grapes, and consequently of wine. The volatile composition of forty-five grape varieties harvested in 2011 and 2012 was investigated by solid phase extraction of samples and gas chromatography-mass spectrometry (GC-MS) analysis. In grape extracts of cultivars Barbera, Refosco dal Peducolo Rosso, Ribolla gialla and Rossese, the C9-norisoprenoid compound ß-isophorone (3,5,5-trimethyl-3-cyclohexen-1-one) was found and quantified. This carotenoid-derived compound is characterised by a saffron aroma and is here reported in grape for the first time.

Advanced knowledge of three important classes of grape phenolics: anthocyanins, stilbenes and flavonols.

Grape is qualitatively and quantitatively very rich in polyphenols. In particular, anthocyanins, flavonols and stilbene derivatives play very important roles in plant metabolism, thanks to their peculiar characteristics. Anthocyanins are responsible for the color of red grapes and wines and confer organoleptic characteristics on the wine. They are used for chemotaxonomic studies and to evaluate the polyphenolic ripening stage of grape. They are natural colorants, have antioxidant, antimicrobial and anticarcinogenic activity, exert protective effects on the human cardiovascular system, and are used in the food and pharmaceutical industries. Stilbenes are vine phytoalexins present in grape berries and associated with the beneficial effects of drinking wine. The principal stilbene, resveratrol, is characterized by anticancer, antioxidant, anti-inflammatory and cardioprotective activity. Resveratrol dimers and oligomers also occur in grape, and are synthetized by the vine as active defenses against exogenous attack, or produced by extracellular enzymes released from pathogens in an attempt to eliminate undesirable toxic compounds. Flavonols are a ubiquitous class of flavonoids with photo-protection and copigmentation (together with anthocyanins) functions. The lack of expression of the enzyme flavonoid 3',5'-hydroxylase in white grapes restricts the presence of these compounds to quercetin, kaempferol and isorhamnetin derivatives, whereas red grapes usually also contain myricetin, laricitrin and syringetin derivatives. In the last ten years, the technological development of analytical instrumentation, particularly mass spectrometry, has led to great improvements and further knowledge of the chemistry of these compounds. In this review, the biosynthesis and biological role of these grape polyphenols are briefly introduced, together with the latest knowledge of their chemistry.

Cancer prevention in Europe: the Mediterranean diet as a protective choice.

In the coming years, European death rates because of cancer will further decline, but the overall number of cases will increase, mostly as a consequence of the ageing of the population. The target for cancer prevention in Europe will remain a healthy diet and control of obesity in addition to a decrease in smoking. A healthy diet model in European countries is the traditional Mediterranean diet, which is based on abundant and variable plant foods, high consumption of cereals, olive oil as the main (added) fat, low intake of (red) meat and moderate consumption of wine. The Mediterranean diet is associated with a reduced risk of cardiovascular disease and cancer. The biological mechanisms for cancer prevention associated with the Mediterranean diet have been related to the favourable effect of a balanced ratio of omega 6 and omega 3 essential fatty acids and high amounts of fibre, antioxidants and polyphenols found in fruit, vegetables, olive oil and wine. The Mediterranean diet also involves a 'Mediterranean way of drinking', that is, regular, moderate consumption of wine mainly with food. This pattern of drinking increases longevity, reduces the risk of cardiovascular disease and does not appreciably influence the overall risk of cancer. However, heavy alcohol drinking is associated with digestive, upper respiratory tract, liver and breast cancers; therefore, avoidance or restriction of alcohol consumption to two drinks/day in men and one drink/day in women is a global public health priority.

Effects of elicitors, viticultural factors, and enological practices on resveratrol and stilbenes in grapevine and wine.

The ability of grapevine to activate defense mechanisms against some pathogens has been shown to be linked to the synthesis of stilbenes by the plant (inducible viniferins). Metabolized viniferins may also be produced or modified by extracellular enzymes released by the pathogen in an attempt to eliminate undesirable toxic compounds. Due to the important properties of resveratrol, there is increasing interest in producing foods with higher contents of this compound and higher nutritional value. The production of high resveratrol-containing grapes and wines relies on quality-oriented viticulture (suitable terroirs, sustainable cultural practices) and wine-making technologies which avoid degradation of the compound. The technique of skin extraction and enzymatic hydrolysis of glucoside forms in wine-making plays an important role in resveratrol wine concentration. Other factors affecting its final concentration and balance in wine are conditions for promoting trans-cis isomerization of aglycones, the type of yeast used, and the presence of lactic bacteria with ß-glucosidase activity. In general, the enological practices commonly used to stabilize wine after fermentation do not significantly affect resveratrol concentrations, which show considerable stability.

Alcohol and wine in relation to cancer and other diseases.

Heavy alcohol consumption is associated with increased overall mortality, cancer, liver, and cardiovascular diseases; but low doses of alcohol (up to one drink per day) are not associated with the risk of any cancer site with the exception of breast cancer and possibly of oral and pharyngeal cancers. Moreover, recent evidence indicates that moderate alcohol and specifically wine intake provides cardioprotection and neuroprotection and may increase longevity. Various experimental data hypothesize a potential cancer chemopreventive role of some grape extracts, and complete sequencing of the grapevine genome has revealed genes responsible for the synthesis of health-promoting compounds (resveratrol and other polyphenols), thus advocating the development of future potential nutraceutical strategies. This focuses on the pros and cons of moderate alcohol and wine consumption and opens a debate on this topic.

A strategy to investigate the intravarietal genetic variability in Vitis vinifera L. for clones and biotypes identification and to correlate molecular profiles with morphological traits or geographic origins.

Grapevine is the most economically important and widely cultivated fruit crop in the world. Molecular markers have been used on Vitis vinifera to distinguish among both varieties and clones. Microsatellites are used to fingerprint varieties and several other techniques, reported in many papers, are used to analyze the differences among clones, but it is not available in the literature as a well defined strategy to screen a large number of Vitis cultivars. In fact, it is often necessary to use different techniques to investigate the genetic variability in different grapevine varieties and a proposed technique is used to study a cultivar, which is often not suitable for either the study of another cultivar or compare the genetic relationship among various cultivars. We describe here a strategy used for the analysis of several grapevine cultivars to describe a universal method to obtain DNA polymorphisms of Vitis vinifera genotypes from the same cultivar by using amplified fragment length polymorphism (AFLP), selective amplification of microsatellite polymorphic loci (SAMPL), microsatellites AFLP (M-AFLP), and ISSR molecular markers. The strategy here adopted permitted both to identify different biotypes (i.e., Primitivo), accessions (i.e., Garnacha tinta), and clones (i.e., Callet, Manto Negro, Moll) among the variability of same variety and to correlate the genetic differences to their geographical origins (i.e., Garnacha tinta; Malvasia nera di Brindisi/Lecce) or morphological traits (i.e., Malvasia of Candia). Here is also described the application of the protocol that allows to highlight the genetic variability accumulated during centuries of cultivations and selections of the same variety in different environments by vine growers.