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Enchantment is a Vitis hybrid released from the University of Arkansas System Division of Agriculture (UA System) wine-grape breeding program in 2016. This new teinturier cultivar has potential for producing high-quality wines. The effects of oak addition (no oak, American oak, and French oak) and storage on Enchantment wine attributes were evaluated in 2017 and 2018. Enchantment grapes were harvested in August of both years for wine production. The 2017 and 2018 wines were analyzed initially (0-months) for basic chemistry, anthocyanin, color, and aroma attributes, and 2017 wines were analyzed during storage (0, 6, and 12 months at 15°C) for basic chemistry, anthocyanin, and color attributes. Regardless of oak additions, initial chemistries of wines in both years were typical for dry wines and remained stable during storage. In both years, malvidin-3-glucoside was the predominant anthocyanin in Enchantment wine, and malvidin-3-glucoside, petunidin-3-glucoside, and delphinidin-3-glucoside composed over 80% of total anthocyanin content. Although anthocyanins decreased during storage, the deep, red color of the wine remained stable. In 2018, wines had a deeper red, darker color than 2017 wines, and this corresponded with higher anthocyanin levels in 2018. There were about 50 volatile aroma compounds identified in Enchantment wines. There was minimal impact of oak treatment on basic chemistry and anthocyanin, but some impact on color attributes. However, oak addition greatly impacted aroma attributes, resulting in wines with oaky, roasted, and caramelized aroma compounds in both years. These results demonstrated the potential of Enchantment wine grapes for producing deeply red-colored single varietal wines and blends with oaking and storage potential.

Anthocyanins composed the major pigments which conferred sensory and chemical values to red grapes and wines. Multiple factors involved in grape anthocyanin biosynthesis have been greatly covered to date. However, the endophytes which closely associate with plants, their possible roles in plant coloration are still under clear. Our present research firstly investigated the diversity distributions of culturable fungal endophytes (CFE) in different colored grapevine leaves which harvested from same vineyard and cultivar, and then proposed method to analyze the possible effects of these fungal endophytes on anthocyanin concentrations of grape cells. In totally 532 endophytic fungal isolates, 19 OTUs belong to 13 genera were isolated from five color different grapevine leaves. Obvious leaf color specificity of CFE communities were observed distribution in different colored grapevine leaves in this experiment. Assessing the influences of those isolated CFEs on anthocyanin concentrations of a teinturier grape pulp cells via dual culture system revealed that anthocyanin promotion fungal strains were more possibly isolated from red and purple colored grape leaves. And elite CFE strains such as DQ53 and DQ55, both belong to the fungal genus Epicoccum, which conferred significant promotion effects on grape cellular anthocyanin contents were selected out. And the transcriptions of anthocyanin biosynthesis association genes MYBA1, UFGT and F3′5′H in grape cells, were significantly influenced by these endophytic fungi, furtherly implicated the involvements of these genes in fungal endophytes-mediated grape cell anthocyanin promotion process. Additionally, endophytic fungi triggered anthocyanin promotion in grape cells seem independently to that of the light mediated anthocyanin synthesis pathway. This work demonstrated that beside other environmental factors, endophytes may also involve in grape anthocyanin metabolisms. And the study provided methods and clues to explore fungal endophytes in colored grape leaves which possibly applied in grape pigmentation processing.

Downy mildew is a fungal disease that seriously threatens the normal growth and fruit quality of grapes; therefore, it is urgent to identify resistant germplasm resources. To identify the differences in the resistance rates of various Chinese wild grapevine plants to downy mildew, 120 grapevine germplasm resources were selected as experimental materials. Leaf disk in vitro testing was applied to evaluate the downy mildew resistance level of plants, and quantitative reverse transcription-polymerase chain reaction (RT-qPCR) was used to test the expression patterns of some key genes at different time courses after inoculation with Plasmopara viticola. Additionally, simple sequence repeat (SSR) markers were selected to identify resistance in different grapevine cultivars and to provide a basis for marker-assisted selection. The results showed that the disease index of 120 grapevine germplasm resources ranged from 0 to 78.70. Among the 120 grapevine varieties, 5 Vitis rotundifolia presented immunity to downy mildew; 3 wild grapevine strains, including Yunnan-Yuanmou 2, Yunnan-2, and Muzhaling-3, were highly resistant strains; 38 grapevine strains were resistant varieties, 71 grapevine strains were susceptible varieties, and 3 V. vinifera strains, namely Italian Riesling, Medoc Noir, and Gamay Teinturier were highly susceptible to downy mildew. After infection with P. viticola, the expression patterns of key genes in resistant and susceptible plants were different. Except for WRKY33, which showed a low expression level in all plants, all the genes presented an initial increasing trend followed by a decreasing trend over the inoculation period. Among the 30 pairs of SSR markers, 22 SSR markers, including CHr14V015, UDV-370 and GF09-46, could explain only 25.83–48.33% of the phenotypic variation, which means that they cannot be applied for downy mildew resistance identification. The RGA9 marker explained the highest phenotypic variation, at 80.83%, and was expected to be a standard marker that can be applied in disease-resistance production, thus providing the basis for both the rapid and mass identification of downy mildew resistance in grapevine germplasm resources and marker-assisted selection.

Teinturier cultivars of grape (Vitis vinifera) accumulate anthocyanins in both the skin and pulp of grape berries. RNA-seq and HPLC/MS were used to elucidate the mechanism of anthocyanin biosynthesis in whole grape berries of a new source of teinturier germplasm, ZhongShan-HongYu (ZS-HY). Berries were sampled at 3 days after anthesis (3 DAA, green-pulp), 6 days after anthesis (6 DAA, pulp-veraison), 22 days after anthesis (22 DAA, pulp-half-colored), and at maturity (77 DAA). Results indicated that total anthocyanin levels increased with berry development. The relative content of anthocyanins was different in the pulp vs. skin. Besides, Pelargonidin-3-O-monoglucoside, which unable to be produced in most varieties of Vitis vinifera, was detected in ZS-HY. RNA-seq analysis result that a total of 56 candidate genes, including 30 structural and 26 regulatory genes, were identified that putatively participate in biosynthesis pathway of anthocyanins in ZS-HY. A KEGG analysis indicated that photosynthesis – antenna proteins and photosynthesis, which is significantly enriched in response to light, is proposed to be responsible for the regulation of anthocyanin production in teinturier grapes by light. The present study provides the first transcriptomic analysis of teinturier grapes and provides a foundation for further studies that will assist in the breeding of high-quality teinturier grapes.

Anthocyanins are vital components of plant secondary metabolites, and are also the most important coloring substances in wine. Teinturier cultivars are rich in anthocyanins. However, the differences in anthocyanin accumulation and profiles between teinturier and non-teinturier cultivars have not been reported. In this study, Yan 73 and Dunkelfelder were selected as the experimental materials, and three non-teinturier cultivars were used for comparison. LC-MS and qRT-PCR were used to determine the individual anthocyanin contents and the relative gene expression. The results show that the total anthocyanin content of the teinturier cultivars was considerably higher than that in non-teinturier cultivars, and the levels of individual anthocyanins increased gradually during ripening. Lower ratios of modified anthocyanins were found in the teinturier cultivars, which was not only due to the high expression level of VvUFGT and VvGST4, but also due to the relatively low expression of VvOMT in these cultivars. Cluster analysis of gene expression and anthocyanin accumulation showed that VvUFGT is related to anthocyanin accumulation, and that AM1 is related to the synthesis and transport of methylated anthocyanins. Our results will be useful for further clarifying the pathways of anthocyanin synthesis, modification, and transport in teinturier cultivars.

In most red grape cultivars, anthocyanins accumulate exclusively in the berry skin, while ‘teinturier’ cultivars also accumulate anthocyanins in the pulp. Here, we investigated the teinturier cvs. ‘Gamay de Bouze’ and ‘Gamay Fréaux’ (two somatic variants of the white-fleshed cv. ‘Gamay’) through metabolic and transcript analysis to clarify whether these two somatic variants have the same anthocyanin accumulation pattern in the skin and pulp, and whether primary metabolites are also affected. The skin of the three cultivars and the pulp of ‘Gamay de Bouze’ begun to accumulate anthocyanins at the onset of berry ripening. However, the pulp of ‘Gamay Fréaux’ exhibited a distinct anthocyanin accumulation pattern, starting as early as fruit set with very low level of sugars. The highest level of anthocyanins was found in ‘Gamay Fréaux’ skin, followed by ‘Gamay de Bouze’ and ‘Gamay’. Consistently, the transcript abundance of genes involved in anthocyanin biosynthesis were in line with the anthocyanin levels in the three cultivars. Despite no evident differences in pulp sugar content, the concentration of glucose and fructose in the skin of ‘Gamay Fréaux’ was only half of those in the skin of ‘Gamay’ and ‘Gamay de Bouze’ throughout all berry ripening, suggesting an uncoupled accumulation of sugars and anthocyanins in ‘Gamay Fréaux’. The study provides a comprehensive view of metabolic consequences in grape somatic variants and the three almost isogenic genotypes can serve as ideal reagents to further uncover the mechanisms underlying the linkage between sugar and anthocyanin accumulation., This research was supported partly by the National Key R&D Program of China (2018YFD1000200) and was conducted as part of the LIA INNOGRAPE International Associated Laboratory.

Anthocyanins with important physiological functions mainly accumulate in grape berry, but teinturier grape cultivars can accumulate anthocyanins in both reproductive and vegetative tissues. The molecular regulatory mechanisms of anthocyanin biosynthesis in grapevine reproductive and vegetative tissues are different. Therefore, teinturier grapevine cultivar provides opportunities to investigate transcriptional regulation of vegetative anthocyanins, and to compare with mechanisms that regulate grape berry anthocyanins. Yan73 is a teinturier Vitis vinifera variety with vegetative tissues able to accumulate anthocyanins, but the anthocyanin pattern and the molecular mechanism regulating anthocyanin biosynthesis in these tissues remain uncharacterized. We analyzed the anthocyanin metabolic and transcriptome profiles of the vegetative tissues of Yan73 and its male parent with HPLC-ESI-MS/MS and RNA-sequencing technologies. Yan73 vegetative tissues had relatively high 3'-OH, acylated, and methoxylated anthocyanins. Furthermore, peonidin-3-O-(trans-6-coumaryl)-glucoside is the most abundant anthocyanin in Yan73 grapevine vegetative tissues. A total of 30,17 and 10 anthocyanin biosynthesis genes showed up-regulated expression in Yan73 leaf, stem and tendril, respectively, indicating anthocyanin biosynthesis in Yan73 vegetative tissues is regulated by transcription factors. The up-regulated expression of VvMYBA1 on chromosome 2 and VvMYBA5, VvMYBA6, and VvMYBA7 on chromosome 14 are responsible for the anthocyanin patterns of Yan73 vegetative tissues. The expression of a set of R2R3-MYB C2 repressor genes is activated and may negatively regulate anthocyanin biosynthesis in Yan73 vegetative tissues. These findings enhance our understanding of anthocyanin biosynthesis in grapevine.

In this work, matrix solid-phase dispersion (MSPD) is proposed for the first time as an efficient and rapid alternative to extract fatty acids (FAs) from grape seeds. In addition, the use of green solvents as limonene allowed obtaining similar yields to other longer extraction procedures using more unhealthy solvents. The optimization of the MSPD procedure was performed using experimental design strategies and considering the use of solvents such as ethyl lactate and limonene with interesting possibilities for the extraction of bioactive compounds from natural sources. Gas chromatography-mass spectrometry (GC-MS) was used to analyze the seed extracts. The MSPD-GC-MS method was applied to characterize the FA profile of 27 seed samples from grapes belonging to Vitis vinifera white cultivars (Albarino, Caino, Godello, Loureiro, Torrontes, Treixadura, Gewurtztraminer, Pinot blanc, and Pinot gris); a red cultivar (Mencia); a teinturier cultivar (Alicante Bouschet); and two hybrids producing white and rose grapes, respectively. Results showed that unsaturated FAs represented 82% of the total, with linoleic being the main acid (73.7%). The content of saturated FAs was 12.5%. Despite slight differences between cultivars, quite similar profiles were obtained independently of the cultivar, the type of sample (fresh grapes or distillation wastes), and the collection year, which is a very interesting conclusion for a potential exploitation of seeds as a source of good-quality oil. A simple technique such as MSPD using limonene as extraction solvent constituted an efficient and greener alternative for the extraction of grape seed FAs.

Background and Aims Yan73 and Kolor are Vitis vinifera grape cultivars with both red skin and red pulp. The aim of the present work was to determine the distribution of flavonoid compounds in the skin, pulp and seed, as well as the aromatic composition of the whole berry. Methods and Results Anthocyanin and flavonol were extracted from grape skin and pulp, while proanthocyanidin was isolated from skin, pulp and seeds; individual anthocyanins, flavonols and proanthocyanidins were then separated and determined by LC/MS. Anthocyanin was dominated by malvidin derivatives in the skin and by peonidin derivatives in the pulp. The distribution of flavonols in the skin and pulp of the two cultivars showed no consistent trend. Proanthocyanidins were unequally distributed in the skin, pulp and seeds. Proanthocyanidin content, composition and mean degree of polymerisation varied distinctly in the three tissues, especially the profile of the proanthocyanidin terminal and extension subunits. Yan73 and Kolor contained similar aroma compounds, both in free form and as glycosidically bound precursors, as determined by GC/MS, but their content varied significantly. Conclusions Flavonoid compounds were asymmetrically distributed in the skin, pulp and seeds. Epicatechin was the common predominant extension subunit in the three tissues, while epigallocatechin, catechin and epicatechin‐3‐O‐gallate were the characteristic extension subunits of the skin, pulp and seeds, respectively. Kolor accumulated more C6 aldehydes in the free fraction and more alcohols in bound fraction, while Yan73 was characterised by a higher content of terpenes. Significance of the Study The results define the distribution of flavonoid compounds in two teinturier cultivars.

Yan73 is a teinturier Vitis vinifera variety with red berry flesh, but the molecular mechanisms underlying its flesh coloration remain unclear. We analyzed the flavonoid metabolic and transcriptome profiles of Yan73 berry red and white flesh using HPLC-ESI-MS/MS and RNA-sequencing technologies. Anthocyanins are the main flavonoids responsible for Yan73 berry flesh color, and the coloration is coordinately regulated by the VvMYBA1 transcriptional activator and VvMYBC2-L1 transcriptional repressor. Furthermore, yeast one- and two-hybrid, dual luciferase, and bimolecular fluorescence complementation assays suggested that VvMYBA1 positively regulates Yan73 berry flesh color via interactions with VvWDR1 and the activation of the VvCHI3, VvOMT, and VvGST4 promoters, whereas VvMYBC2-L1 negatively regulates Yan73 berry flesh color, possibly by competing with the R2R3-MYB transcriptional activators for bHLH partners or by repressing VvOMT and VvGST4 expression. Our findings provide new insights into the molecular mechanisms regulating grape flesh color.

Successful cross-breeding requires the investigation into the inheritance pattern of desirable traits. The present study recorded the segregation in coloration and analyzed anthocyanin concentration and composition in berry skin and flesh of the F1 interspecific populations (‘Beifeng’ × ‘Yan-73’) during the three successive growing seasons. All the progeny presented color in berry skin, whereas segregation was observed in berry flesh coloration with the ratio of 1:1. Meanwhile, attentions were paid to anthocyanin mono-and di-glucosides for the first time. Di-glucosides were only detected in the maternal parent ‘Beifeng’ and around a half number of the progeny in berry skin and colored flesh. On the other hand, mono-glucosides were detected in both parents and all the progeny in berry skin and with colored flesh. Concentrations of total and major anthocyanins in the progeny were continuously distributed and skewed towards the low concentration area. Transgressive inheritance was also noted, indicated by the exceptionally high anthocyanin concentrations observed amongst some of the progeny. Positive correlations were observed between traits except the slightly negative correlations between mono-and di-glucosides in both skin and flesh. These results are of both practical and scientific interests for further breeding efforts and researches on different coloration mechanisms in berry skin and flesh.

Proanthocyanidins (PAs) are one of the most important polyphenolic compounds in wine. Among PAs, prodelphinidin (PD) dimers and trimers have not been widely detected in wines due to the lack of available commercial standards and the difficulty to detect and isolate them from natural sources. LC-ESI-MS (liquid chromatography-electrospray ionization-mass spectrometry) with the right chromatographic conditions has proven to be a powerful tool for PAs detection and identification in complex samples. This technique has been applied to an exhaustive study of PA composition of two Portuguese red wines made with teinturier grapes, especially for the identification of PD dimers and trimers. Tandem mass spectrometry (MS/MS) with ion trap provided additional information about the structures of these compounds through the fragmentation patterns of the pseudomolecular ions. A LC-ESI-MS method was optimized and 41 different compounds were found. Among them are included 8 PD dimers and 13 PD trimers.

Yan73, a teinturier (dyer) grape variety in China, is one of the few Vitis vinifera cultivars with red-coloured berry flesh. To examine the tissue-specific expression of genes associated with berry colour in Yan73, we analysed the differential accumulation of anthocyanins in the skin and flesh tissues of two red-skinned grape varieties with either red (Yan73) or white flesh (Muscat Hamburg) based on HPLC-MS analysis, as well as the differential expression of 18 anthocyanin biosynthesis genes in both varieties by quantitative RT-PCR. The results revealed that the transcripts of GST, OMT, AM3, CHS3, UFGT, MYBA1, F3′5′H, F3H1 and LDOX were barely detectable in the white flesh of Muscat Hamburg. In particular, GST, OMT, AM3, CHS3 and F3H1 showed approximately 50-fold downregulation in the white flesh of Muscat Hamburg compared to the red flesh of Yan73. A correlation analysis between the accumulation of different types of anthocyanins and gene expression indicated that the cumulative expression of GST, F3′5′H, LDOX and MYBA1 was more closely associated with the acylated anthocyanins and the 3′5′-OH anthocyanins, while OMT and AM3 were more closely associated with the total anthocyanins and methoxylated anthocyanins. Therefore, the transcripts of OMT, AM3, GST, F3′5′H, LDOX and MYBA1 explained most of the variation in the amount and composition of anthocyanins in skin and flesh of Yan73. The data suggest that the specific localization of anthocyanins in the flesh tissue of Yan73 is most likely due to the tissue-specific expression of OMT, AM3, GST, F3′5′H, LDOX and MYBA1 in the flesh.

Grapevine (Vitis vinifera) teinturier cultivars are characterized by their typical reddish leaves and red-fleshed berries due to ectopic anthocyanin formation. Wines of these varieties have economic importance as they can be used for blending to enhance the color of red wines. The unique and heritable mutation has been known for a long time but the underlying genetic mechanism still is not yet understood. Here we describe the association of the red-fleshed berry phenotype with a 408 bp repetitive DNA element in the promoter of the VvmybA1 gene (grapevine color enhancer, GCE). Three different clones of &lsquo, Teinturier&rsquo, were discovered with two, three and five allelic GCE repeats (MybA1t2, MybA1t3 and MybA1t5). All three clones are periclinal chimeras, these clones share the same L1 layer, but have distinct L2 layers with different quantities of GCE repeats. Quantitative real time PCR and HPLC analysis of leaf and berry samples showed that the GCE repeat number strongly correlates with an increase of the expression of VvmybA1 itself and the VvUFGT gene regulated by it and the anthocyanin content. A model is proposed based on autoregulation of VvmybA1t to explain the red phenotype which is similar to that of red-fleshed apples. This study presents results about the generation and modes of action of three MybA1t alleles responsible for the red-fleshed berry phenotype of teinturier grapevines.

The study investigated the profile of fatty acids (FA) in grape seed (GS) and grape seed oil (GSO) of a newly developed teinturier grape variety, named Manjari Medika (MM). FAs were identified and quantified by gas chromatography with flame ionisation detection. The identification of FAs was further confirmed using a GC with mass selective detector (GC-MS). The profiles of MM-GS and MM-GSO were compared with the corresponding products of two established varieties, viz. Cabernet Sauvignon (CS) and Sauvignon Blanc (SB). The unique FA profile of MM was differentiated from CS and SB through multivariate statistical analysis and pattern recognition of selective FAs. MM-GSO was found to be superior to SB-GSO and equivalent to CS-GSO in terms of FA profile. The presence of unsaturated FAs at a higher level and a favourable ratio of essential FAs (omega-6: omega-3) indicated the potential health benefits of MM- GS and GSO. The FA levels in MM-GSO were within or above the Codex standards for edible oil. The typical FA profile uniquely differentiated these three varieties of seeds and corresponding oils. The results formed the basis of a future endeavor in establishing their varietal authenticity.

International audience; Anthocyanin biosynthesis is regulated by environmental factors (such as light, temperature, and water availability) and nutrient status (such as carbon, nitrogen, and phosphate nutrition). Previous reports show that low nitrogen availability strongly enhances anthocyanin accumulation in non carbon-limited plant organs or cell suspensions. It has been hypothesized that high carbon-to-nitrogen ratio would lead to an energy excess in plant cells, and that an increase in flavonoid pathway metabolic fluxes would act as an "energy escape valve," helping plant cells to cope with energy and carbon excess. However, this hypothesis has never been tested directly. To this end, we used the grapevine Vitis vinifera L. cultivar Gamay Teinturier (syn. Gamay Freaux or Freaux Tintorier, VIVC #4382) cell suspension line as a model system to study the regulation of anthocyanin accumulation in response to nitrogen supply. The cells were sub-cultured in the presence of either control (25 mM) or low (5 mM) nitrate concentration. Targeted metabolomics and enzyme activity determinations were used to parametrize a constraint-based model describing both the central carbon and nitrogen metabolisms and the flavonoid (phenylpropanoid) pathway connected by the energy (ATP) and reducing power equivalents (NADPH and NADH) cofactors. The flux analysis (2 flux maps generated, for control and low nitrogen in culture medium) clearly showed that in low nitrogen-fed cells all the metabolic fluxes of central metabolism were decreased, whereas fluxes that consume energy and reducing power, were either increased (upper part of glycolysis, shikimate, and flavonoid pathway) or maintained (pentose phosphate pathway). Also, fluxes of flavanone 3 beta-hydroxylase, flavonol synthase, and anthocyanidin synthase were strongly increased, advocating for a regulation of the flavonoid pathway by alpha-ketoglutarate levels. These results strongly support the hypothesis of anthocyanin biosynthesis acting as an energy escape valve in plant cells, and they open new possibilities to manipulate flavonoid production in plant cells. They do not, however, support a role of anthocyanins as an effective mechanism for coping with carbon excess in high carbon to nitrogen ratio situations in grape cells. Instead, constraint-based modeling output and biomass analysis indicate that carbon excess is dealt with by vacuolar storage of soluble sugars.

In red wines, important sensory attributes such as color and astringency are the result of the accumulation of anthocyanins, which are located in the berry skin (and in the pulp of teinturier cultivars). Each cultivar has a unique set of anthocyanins and both qualitative and qualitative anthocyanin composition at harvest are crucial factors for winemaking. Anthocyanins are synthesized through the so-called phenylpropanoid pathway. Vitis vinifera L. ?Malbec? and ?Cabernet Sauvignon? are red cultivars adapted to similar environmental conditions. Both cultivars are usually cropped to have from medium to low yields to ensure the production of premium and ultra-premium wines. Although ?Malbec? has significant differences in the dihydroflavonol concentration, being 10 times higher than in other red cultivars such as ?Cabernet Sauvignon?, this increment could be partially explained by differences in the gene sequences coding for enzymes working between the naringenin chalcone and leucoanthocyanins steps. In order to investigate the dynamics of anthocyanin accumulation in grapevines, we sequenced cDNAs coding for enzymes of the anthocyanin biosynthetic pathway isolated from grape berry skin, sampled throughout ripening from two selected clones of ?Malbec? and ?Cabernet Sauvignon?. Specific primers were synthesized using EST sequences (NCBI) for flavonoid 3?5?-hydroxylase, flavanone 3-hydroxylase, flavonol synthase and dihydroflavonol 4-reductase. Total RNA was isolated from berry skins and cDNA was used to study these gene sequences. Finally, fragments were cloned and sequenced, and compared between cultivars. The comparison of the deduced aminoacidic sequences against the reference data base show identities higher than 99% with the cultivar ?Pinot Noir? for all the enzymes, and the identified mutations do not seem to have any effect on the protein structure. When a comparison between ?Malbec? and ?Cabernet Sauvignon? sequences was performed, the identity was also higher th

Amino acid composition of the grape berry at harvest is important for wine making. The present study investigates the complex interplay between tissue, cultivar and light conditions that determine berry amino acid content. Twenty amino acids were assessed in the berry skin and pulp of two grape cultivars (Gamay Noir and Gamay Fréaux), grown under either light exposure or cluster shading conditions. In all samples, cluster shading significantly reduced most amino acids, except gamma-aminobutyric acid (GABA) and phenylalanine. However, the magnitude of the decrease was stronger in the skin (67.0% decrease) than in the pulp (30.4%) and stronger in cv. Gamay Noir (69.7%) than in Gamay Fréaux (30.7%). Cluster shading also significantly modified amino acid composition by decreasing the proline content while increasing the GABA content. These results are of oenological interest for shaping the amino acid composition of the must and improving wine quality.

The grape is considered a major source of phenolic compounds when compared to other fruits and vegetables, however, there are many cultivars with distinct characteristics directly linked to phenolic profile. Thus, the present study aimed to identify and quantify, for the first time and in detail, the phenolic compounds present in the skin, flesh and seeds of BRS Violeta grape berry using combination of SPE methodologies and analytical HPLC–DAD–ESI-MS/MS. The study was extended to the different berry parts and the most important grape and wine phenolic families, and has revealed interesting features. Violeta grape has a very thick skin (46% of grape weight) that accumulated the most of grape phenolic compounds: great amount of anthocyanins (3930 mg/kg, as malvidin 3,5-diglucoside), together with also important amounts of flavonols (150 mg/kg, as quercetin 3-glucoside), hydroxycinnamic acid derivatives (HCAD; 120 mg/kg, as caftaric acid), and proanthocyanidins (670 mg/kg, as (+)-catechin); in contrast, it seems to be a low resveratrol producer. Violeta grape seeds accounted for similar proportions of low molecular weight flavan-3-ols (mainly monomers; 345 mg/kg, as (+)-catechin) and proanthocyanidins (480 mg/kg, as (+)-catechin). Violeta grape is a teinturier cultivar, but it only contained traces of anthocyanins and low amounts of all the other phenolic types in its red-colored flesh. The anthocyanin composition of Violeta grape was dominated by anthocyanidin 3,5-diglucosides (90%). Within flavonols, myricetin-type predominated and kaempferol-type was missing. In addition to expected hydroxycinnamoyl-tartaric acids, several isomeric esters of caffeic and p-coumaric acids with hexoses were tentatively identified, accounting for relevant proportions within the pool of HCAD. Although pending of further confirmation over successive vintages, the aforementioned results suggest that BRS Violeta grape cultivar could be considered an interesting candidate for the elaboration of highly colored and antioxidant-rich grape juices and wines.

The products and the related gene expression of flavonoid metabolism were examined in the skin and pulp of Yan73 (Vitis vinifera L. cv.) grapes, a teinturier variety cultivated in China, and the data were compared with those in the skin and pulp of Cabernet Sauvignon (Vitis vinifera L. cv.) grapes, a well-known red variety. The results showed that, in comparison with the skins of Cabernet Sauvignon, the skins of Yan73 berries were characterised by fewer types of flavonols and anthocyanins, a higher level of flavonoids, and a much lower percentage of 3′-hydroxylated flavonols. Flavonoid metabolism was also present in the pulp of Yan73, and the percentages of 3′-hydroxylated flavonols and 3′-hydroxylated flavan-3-ols were significantly higher than the percentages of their corresponding 3′,5′-hydroxylated forms. Only flavan-3- ols were found in the pulp of Cabernet Sauvignon. The expression of VvmybA1 exhibited a co-ordination with the accumulation of anthocyanins in the pulp of the teinturier cultivar, which showed great similarity to that in the skin. A good correlation was found between the expression of VvF3′H and 3′-hydroxylated flavonoids, as well as between the expression of VvF3′5′H and 3′,5′-hydroxylated flavonoids in the skin and pulp of the grape berries. It is suggested that tissue-specific accumulation of flavonoids in grape berries is related to the transcriptional expression of VvF3′H and VvF3′5′H.

The anthocyanin composition and concentration of various organs of teinturier grape Yan-73 were studied throughout the growing season. Nineteen anthocyanins were identified by HPLC-MS as monoglucosides and their derivatives. Anthocyanin composition and concentration varied among grape organs and by developmental stage. Skin anthocyanins were mainly composed of malvidin derivatives, while peonidin derivatives were the most dominant anthocyanins in the pulp. Both malvidin and peonidin derivatives were predominant components in carpopodia (swollen pedicel at point of berry attachment), berry pedicels, leaf lamina, vein and petioles, and living bark at the base of the shoot. Anthocyanins were very low before veraison, and then increased sharply at veraison in berry skin and pulp. Anthocyanins in carpopodia and berry pedicels also increased sharply, although occurring later than in berry skin and pulp. Anthocyanins were high in young and senescing leaf lamina and low in expanding and mature lamina. Anthocyanins did not vary much in leaf vein and petiole tissue, or in bark, throughout the growing season.

The detailed phenolic composition (anthocyanins, flavonols, hydroxycinnamic acid derivatives, stilbenes, and flavan-3-ols) in the skin and flesh of the new BRS Clara and BRS Morena seedless table grapes has been studied using HPLC-DAD-ESI-MS/MS. The two grapes, especially BRS Morena, contained high amounts of phenolic compounds, mainly located in their skins and qualitatively not different from those found in Vitis vinifera grapes. In addition, BRS Morena (a teinturier variety) showed qualitatively different phenolic compositions in its skin and flesh, mainly affecting the anthocyanin and flavonol profiles. Consistent with high phenolic contents, high antioxidant capacity values were registered for both grape varieties, especially for BRS Morena. Proanthocyanidins and hydroxycinnamoyl-tartaric acids were the major phenolic compounds found in BRS Clara and were also important in BRS Morena, although anthocyanins were the main phenolic compounds in the latter case. These results suggest that the entire grapes, including the skin, may potentially possess properties that are beneficial to human health. In this context, the BRS Morena grape can be considered as a high resveratrol producer.

Red wines made from Garnacha Tintorera grapes (also known as Alicante Bouschet and one of the few so-called teinturier Vitis vinifera grape cultivars, because its berry flesh is also red-colored) are usually blended with other red wines as a natural way for enhancing the color intensity of red wines. The phenolic composition of Garnacha Tintorera grapes showed already described interesting features such as the occurrence of unusual anthocyanins (pelargonidin-based anthocyanins and a peonidin dihexoside) that had not been previously described in V. vinifera cultivars. The isolation and further structure elucidation (GC-MS analysis of sugar moieties, UV-vis, LC-MS(n), and (1)H and (13)C NMR spectroscopic data) have allowed the identification of the aforementioned peonidin dihexoside as peonidin 3,7-β-O-diglucoside. This peonidin derivative, in combination with the previously reported pelargonidin-type anthocyanins, can be suggested as a suitable chemical marker for this singular V. vinifera grape cultivar and the wines made totally or partially from this grape cultivar.

Yan73 is a “teinturier” red wine variety cultivated in China and used in winemaking to strengthen red wine color. Here, the anthocyanin profile in both the skin and pulp of this grape variety was analyzed by HPLC-MS. The results showed that 18 anthocyanins were detected in both the skin and the pulp, and pelargonidin-3-O-glucoside, an anthocyanin compound hardly detected in most other Vitis viniferaberries, was found. However, the contents of individual anthocyanins in the skin and the pulp were significantly different. Compared with the skin, the pulp exhibited much lower ratio of 3’,5’-substituted to 3’-substituted anthocyanins and much higher ratio of methoxylation of anthocyanin B-ring to non methoxylation, and with regard to the aromatic acylated and aliphatic acylated anthocyanins, both their contents in the skin are higher than in the pulp. The findings will provide some new insight for the tissue-specific expression and regulation of the genes involving in anthocyanin biosynthesis in grape berries.

Garnacha Tintorera (also known as Alicante Bouschet) is one of the few V. vinifera grape cultivars with red-colored berry flesh. The study of the phenolic composition of both berry flesh and skin of Garnacha Tintorera grapes shows interesting findings. Anthocyanins were asymmetrically distributed within grape flesh and skins. Malvidin derivatives dominated in skin, followed by peonidin-type anthocyanins; in contrast, the flesh almost exclusively contained peonidin 3-glucoside. In addition, LC-UV-vis and LC-MS evidence suggest the presence of small amounts of peonidin 3,5-diglucoside and a second peonidin dihexoside derivative, and, very likely, the first report of the occurrence of pelargonidin 3-glucoside and its acetyl and p-coumaroyl derivatives in V. vinifera grapes. Flavonols also occurred in the flesh of Garnacha Tintorera grapes, but its flavonol profile showed lower contribution of trisubstituted flavonoid structures (myricetin, laricitrin, and syringetin) when compared to that of skin. The skin of Garnacha Tintorera grapes contained hydroxycinnamic acids in higher amounts than in flesh, caftaric acid being the main derivative found, and coutaric acid accounting for its highest proportion in the skin. The phenolic composition of the whole grape berries reflected the average of the differences described for the two aforementioned berry parts, and subsequently, the red wines made from these grapes had a phenolic composition closer to that shown by the whole berries. The formation of anthocyanin-derived pigments in Garnacha Tintorera red wines makes necessary the use of LC-MS for detecting the minor pelargonidin-based anthocyanins and peonidin dihexoside, which could be suggested as chemical markers for cultivar authentication of this grape cultivar and its wines.

Light exclusion reduces the concentration and modifies the composition of grape anthocyanins, by altering the expression of genes involved in anthocyanin biosynthesis and transport, in a cultivar- and tissue-specific manner. Unlike most grapes, teinturier grapes accumulate anthocyanins both in skin and flesh. However, the concentration and composition of anthocyanins in both tissues differ, providing a valuable system to study tissue-specific regulation of anthocyanin synthesis. Furthermore, little is known about the mechanisms controlling the sensitivity of anthocyanin accumulation to light. Here, light was excluded from Gamay (white-fleshed) and Gamay Freaux (teinturier mutant) berries throughout berry development. Under light-exposed conditions, the skin of Gamay Freaux accumulated the highest level of anthocyanins, followed by the skin of Gamay, while the pulp of Gamay Freaux had much lower anthocyanins than the skins. Network analysis revealed the same order on the number of significant correlations among metabolites and transcripts in the three colored tissues, indicating a higher connectivity that reflects a higher efficiency of the anthocyanin pathway. Compared to light conditions, light exclusion reduced the total amount of anthocyanins, most severely in the skin of Gamay and to a lesser extent in the flesh and skin of Gamay Freaux. Coordinated decrease in the transcript abundance of structural, regulatory and transporter genes by light exclusion correlated with the reduced anthocyanin concentration in a cultivar- and tissue-specific manner. Moreover, light exclusion increased the ratio of dihydroxylated to trihydroxylated anthocyanins, in parallel with F3'H and F3'5'H transcript amounts. Sugars and ABA only play a limited role in the control of anthocyanin synthesis in the berries, in contrast with what has been described in cell suspensions. This study provides novel insights into the regulation of anthocyanin in wild type and teinturier cultivars.

This paper deals with the results of studies on changes in the content of total anthocyanins depending on their fermentative maceration from grapes of traditional Moravian cultivars (Blauer Portugieser, Saint Laurent and Blaufrankisch). Colouring matters were macerated in the course of alcoholic fermentation either in a closed rotary tank or in an open tank with a periodically plunging pulp cap. The losses resulting from adsorption of anthocyanins on yeast biomass were quantified. Concentration losses of anthocyanins were also observed in the course of fermentation of musts made of teinturier grape cultivars Alibernet and Neronet, when the skins of berries were separated. The process of fermentative maceration of anthocyanins from grape skins consisted of three dynamically different stages. In the stage with the exponential increase in the content of anthocyanins no significant differences were observed between the closed rotary tank technique and the open tank technique with periodic plunging of pulp cap. It was demonstrated that the losses of anthocyanins took place during the whole process of fermentation. The correlation between the an - thocyanin losses and the production of yeast biomass was statistically highly significant.

International audience; Yan-73 (Vitis vinifera) is a teinturier grape cultivar, which accumulates anthocyanins in skin, pulp, pedicels, and rachis. The effects of sunlight on anthocyanin biosynthesis and regulation in various tissues of this teinturier cultivar were investigated. Light transmission measurements showed that covering with opaque boxes substantially reduced light intensity around clusters; ≤0.25% of incident light reached the berry skin and

Teinturier variety grape cell cultures produce anthocyanins and exhibit a red appearance, although the major cell population is composed of nonpigmented cells. A novel cell suspension culture was initiated from selected nonpigmented cells that were shown, to synthesize catechins. Anthocyanin synthesis has been triggered when cell division was reduced by Pi deficiency. Furthermore, the addition of dihydroquercetin resulted in a 10-fold increase in dihydroflavonol 4-reductase (DFR) activity. In nonpigmented cells a higher level of precursors or an elevated level of substrate availability would induce anthocyanin synthesis.

Yan73 is a 'teinturier' red wine variety cultivated in China and widely used in winemaking to strengthen red wine colour. The objective of this study was to evaluate the effect of exogenous abscisic acid (ABA) applied to the grapevine cluster on the antioxidant capacity and phenolic content of the wine made from Yan73. Two hundred mg/l ABA was applied on Yan73 grapevine cluster during veraison. As they mature, these ABA-treated and untreated grape berries were transformed into wines, respectively, and the phenolic content and antioxidant capacity of these wines were compared. The results showed that phenolic content (total phenolics, tannins, flavonoids and anthocyanins) and antioxidant capacity were higher in the wine produced with ABA-treated Yan73 grapes than those in the wine from untreated grapes. Compared to Cabernet Sauvignon wine, Yan73 wine had higher phenolic content and stronger antioxidant capacity. These strongly suggest that exogenously applied ABA to Yan73 grapes can enhance phenolic content and antioxidant capacity of its wine, and Yan73 wine has the higher utilization value and potential for development.

Sweet wines are traditionally elaborated in Galicia (the N.W. corner of Spain). The denomination of origin (DO) Valdeorras, one of the five DOs in Galicia, wants to promote the production and marketing of new sweets wines. The first one is made with dried red grapes Vitis vinifera L. Garnacha Tintorera (GNSW); this cultivar is a teinturier cultivar which has excellent potential to produce wines from raisined grapes. The second one, a fortified sweet wine aged in oak barrels (GFSW). Additionally a dry young wine (GBW) was produced from the same variety. Their aroma profiles and chromatic characteristics (determined by simple spectrophotometric methods) have been previously established. Now, proanthocyanidins, flava-3-ol monomers, anthocyanins, phenolic acids, flavonols and resveratrol were determined by HPLC, for the same three wines. The highest concentration of total proanthocyanidins (PAs) was evaluated in the GBW (525mgL(-1)), which was about 2-fold the concentration in the GNSW (236mgL(-1)) and about more 10-fold the concentration in the GFSW (44mgL(-1)). No apparent difference in the aDP (mean degree of polymerisation) was observed for the GBW (1.9) and the GNSW (2.1), whereas a slightly lower value was obtained for the GFSW (1.5). Total anthocyanin concentration was described as follow as GBW: 390mgL(-1)≫GNSW: 57mgL(-1)>GFSW: 25mgL(-1), which indicates that sweet wines were polymerised in great extent. Only vitisin A and B were found the main concentration in GFSW when compared to GBW by the ageing process. In sweet wines, phenolic acids (hydroxybenzoic and hydroxycinnamic acids) and flavonols were lowest when compared to GBW and resveratrol not was found in sweet wines.

Valdeorras (the N.W. corner of Spain) wants to promote the production and marketing of new sweet wines. The present work represents the first study on sweet wines manufactured with red grapes Vitis vinifera L. Garnacha Tintorera, a teinturier cultivar. Two different red sweet wines were elaborated: the first one was made with dried grapes; Vitis vinifera L. Garnacha Tintorera has excellent potential to produce wines from raisined grapes; the second one, a fortified sweet wine aged in oak barrels. Different red Garnacha Tintorera-based wines (a dry base wine, GBW; a naturally sweet wine, GNSW; and a fortified sweet wine, GFSW) were characterized. Chromatic characteristics and phenolic compounds were established by spectrophotometric methods in order to assess the technology of Garnacha Tintorera-based sweet wines. High molecular weight brown polymers, produced during the grape drying process and isolated from sweet wines by the dialysis process, were responsible for the brown colour of sweet wines. As a consequence, yellowness of sweet wines was also higher which was confirmed by colorimetric indexes. With respect to phenolic content, GFSW presented the lowest content because the maceration-alcoholic fermentation was stopped through the addition of alcohol before the diffusion of red pigments from skins to must was complete. GNSW presented the highest phenolic content due to the concentration effect resulting from evaporation of water from the grapes. Anthocyanins of sweet wines were polymerised in great extent. The percentage of polymerised tannins was sufficient to guarantee the aging process of sweet wines.

A cell suspension of Vitis vinifera L. cv. Gamay Frédaux var. teinturier composed of 50% pigmented cells was grown in Gamborg B5 medium supplemented with (per litre) 250 mg casein hydrolysate, 0.1 mg α-naphthalenacetic acid, 0.2 mg kinetin, and either 20, 30, 50, or 60 g sucrose. In the presence of 20 and 30 g sucrose/L, growth of cells was characterized by a typical sigmoid pattern and maximum cell density was obtained in 30 g sucrose/L. In both media, the anthocyanin content of pigmented cells did not change significantly throughout the growth cycle. In the presence of 50 g sucrose/L, cell growth was characterized by an extended lag phase. During this period, there was a significant (P