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A study on Sauvignon blanc (SB) cultivar in France showed that curettage had an effect on the resilience of GTD grapevines. No experiments, however, have been conducted on its effects on wine quality, particularly on white Sauvignon blanc cultivar wines. Methods and results: Grapevines from Sauvignon blanc cultivar that had expressed Esca-foliar symptoms were used for the study, with some of them having been curetted in 2014. Subsequently, bunches from Control (asymptomatic), Curetted and Esca-symptomatic vines were harvested in 2017 and 2018 in order to produce white wine. Technical and chemical results on both must and wine showed that wines from curetted plants were similar to those from asymptomatic vines. There were differences, however, for Esca-diseased vines, where the alcoholic fermentation of musts was faster than for the other modalities. Olfactometry results showed that, for the one-year-old 2017 vintage wines, no differences were detected, although they were for the 2018 vintage. Conclusion: The results of the chemical analyses and wine tasting showed that the wines from curetted and asymptomatic grapevines were similar, and that their quality was the same. Significance of the study: The quality of wines from curetted vines compared to asymptomatic ones was confirmed and validated through chemical and sensory analyses of the must and the one-year-old wines. k e y w o r d s curettage, Esca, Sauvignon blanc, musts, chemical analyses, wine quality

International audience; Methyl salicylate (MeSA) is a plant metabolite that induces plant defence resistance and an odorous volatile compound presenting green nuances. This volatile compound was shown to be present in wine samples, sometimes at concentrations above its olfactory detection threshold. MeSA is localized in grapes, particularly in the skins and stems, and is extracted during red wine vinification. It was detected at the highest concentrations in wines of several grape varieties, made from grapes affected by cryptogamic diseases, namely downy mildew caused by Plasmopara viticola, and black rot caused by Guignardia bidwellii. It has also been detected in wines from vines affected by Esca, a Grapevine Trunk Disease. MeSA can also be considered to be a chemical marker in grapes and wine indicative of the level of development of several vine cryptogamic diseases.

Phenolic compounds are important constituents of red wine involved in its sensory properties. Although wine tannins can come from microbial and oak sources, the main sources of polyphenol remains grape skins and seeds. In order to better understand the grape seed and skin tannins contribution to the final wine tannins content, an original approach of winemaking has been set up. Our protocol explains a simple method to determine the percentage of skin and seed tannins extracted in wine all along the winemaking. The advantages of this method are presented below:•This method allows us to describe tannins extraction kinetics in wine according to the berry part (skins and seeds).•Additionally, our protocol allows to specifically determine the percentage of tannins extraction according to the winemaking stages (alcoholic fermentation and post-fermentative maceration) and the berry part (skins and seeds).•To sum up, this method serves to enhance the comprehension of tannins extraction process in wine according to the berry part., Graphical abstract Image, graphical abstract

International audience; The formation of 3-methyl-2,4-nonanedione (MND) during red wine aging can contribute to the premature evolution of aroma, characterized by the loss of fresh fruit and development of dried fruit flavors. The identification of two new hydroxy ketones, 2-hydroxy-3-methylnonan-4-one (syn- and anti-ketol diastereoisomers) and 3-hydroxy-3-methyl-2,4-nonanedione (HMND), prompted the investigation of the precursors and pathways through which MND is produced and evolves. An HS-SPME-GC-MS method was optimized for their quantitation in numerous must and wine samples, providing insight into the evolution of MND, HMND, and ketols through alcoholic fermentation and wine aging. Alcoholic fermentation resulted in a significant decrease in MND and HMND and the simultaneous appearance of ketol diastereoisomers. The analysis of 167 dry red wines revealed significant increases in MND and anti-ketol contents through aging and a significant positive correlation between MND and anti-ketols. Additional experiments demonstrated that ketols are precursors to MND during red wine oxidation.

This work investigated the structural, biochemical, and molecular characteristics of grape skin cell wall during ripening, related to susceptibility to Botrytis cinerea. The comparative study between the two main grape cultivars in Champagne region, Pinot noir and Chardonnay, quantified: (1) the maturity and physical profile of grape skin, (2) the morphological characteristics, (3) soluble pectic polysaccharides located in grape skin cell walls, and (4) the gene expression of the two main degrading enzymes (VvPME1 and VvPG1) and PME activity. During the maturation period, the grape skins of the two cultivars appear different in their structure and composition. Chardonnay is characterized by higher relative humidity (RH) and level of VvPG1 expression, lower disease incidence and penetrometry values, and thicker cell walls than Pinot noir skins. Thus, the cell wall composition is sufficiently different between grape varieties from the same area to allow their discrimination and could be used to better manage the harvest date.

International audience; Context and purpose of the study: Like in other wine producing regions around the world, Bordeaux vineyards already experience the effects of climate change. Recent trends as well as model outputs for the future strongly support an increase of average and extreme temperatures. For the maturation period, this increase will by far exceed mean atmospheric temperature increase, as the ripening period will occur earlier in hotter climatic conditions. Therefore, a detrimental secondary metabolism response is expected in grape berries, and of particular concern are the impacts on phenolics and aromas and aroma precursors. The effects of high temperatures on secondary metabolism control have been partly characterized for phenolics, however mostly in artificial growing conditions, while little is known with respect to aromas. A better understanding of how high temperatures influence grape berry secondary metabolites could help vineyard growers to adapt to climate change and maintain wine quality. Material and methods: A two-year field study was carried out in 2015 and 2016 in a vineyard in Bordeaux, France. Two treatments, heated (H) and control (C), were applied to two varieties, Cabernet-Sauvignon and Sauvignon blanc, from fruit-set to maturity. Field heating was achieved by a very local greenhouse effect applied to the bottom of the rows, by enclosing most of the underlying soil surface by polycarbonate shields. As the training system was vertically trellised, the heated volume surrounded most of the bunches but did not disturb most of the leaves in the canopy. This simple and robust setup allowed an increase of berry temperature by about +1.5 degrees C in mean value, up to +5 degrees C at times during clear sky days. This moderate increase of temperature was indicative of the predicted future climatic conditions for the mid-21st century. Berry samples were collected at 4 time points from bunch closure to maturity for each cultivar and treatment. Primary and secondary metabolites were measured in whole berries or skins. Results and conclusions: With this moderate temperature increase, primary metabolite content in berries did not change significantly. In H samples, anthocyanins were reduced and tannins increased before veraison, and both decreased thereafter. H samples also exhibited lower concentrations of some amino acids, especially alanine, serine and phenylalanine. IBMP (2-methoxy-3-isobutylpyrazine) concentrations were also reduced in H samples of Cabernet-Sauvignon, in both seasons, especially at bunch closure stage, but the differences diminished at full maturity. For thiol 3-sulfanyl hexanol precursors, H samples again exhibited much lower concentrations for both varieties, with weak differences at early stages that increased at later stages (up to -70% decline at maturity in 2015 for Sauvignon blanc). These results demonstrate the potential negative impact of elevated temperature on polyphenols and aroma quality of grape berries. Significance and impact of the study: For viticulture to adapt to new climatic conditions, the negative impacts of high temperature on secondary metabolites and aromas, and therefore on wine quality, need to be contemplated. Thus, already established or new vineyard plantings must prepare and consider practices able to mitigate these impacts, for instance practices that increase bunch shading.

International audience; Phenolic compounds are among the most important quality factors of wines. They contribute to the organoleptic characteristics of wine such as color, astringency, and bitterness. Although tannins found in wine can come from microbial and oak sources, the main sources of polyphenols are grape skins and seeds. Since the 1960s, this subject has been widely studied by a large number of researchers covering different types of wine, climate conditions, growing practices, and grape varieties. As these works have been conducted under different conditions, the data collected can be conflicting. Moreover, even though the biosynthesis of the major proanthocyanidins units (+)-catechin and (-)-epicatechin is well-known, the mechanism of their polymerization remains unexplained. This is why the question remains: what factors influence the biosynthesis, the quantity, and the distribution of tannins in grape seeds and how can winemaking processes impact the extractability of seed tannins in wine?

International audience; The extraction of seed and skin tannins in wine has been investigated at three different grape maturity stages. For that, the tannins content and composition of seeds and skins at three different maturity stages were characterized. After that, an original approach of nanovinification was conducted. At each maturity stages, three winemaking modalities have been produced: (i) a control modality, (ii) a seed modality made of exclusively with seed and (iii) a skin modality made of exclusively with skins. The aim of this work is to describe and explain the seed tannins kinetics release in wine but also the impact of grape maturity on seed tannins extractability. For that, the evolution of seed and wine tannins content have been followed during the winemaking, from alcoholic fermentation to post-fermentative maceration.

The reproductive cycle of the grapevine (Vitis vinifera) extends over two consecutive years. Grapevine bud fruitfulness is determined by the number and the size of inflorescence primordia (IP) formed in latent buds during the first year. Many studies show that up to 60% of the seasonal variation in yield could be explained by bud fruitfulness. The regulation mechanism of the initiation and differentiation of IP remains little known, but this process seems to be modulated by several molecular regulation factors involved in flowering and embryo development. Some genes involved in flowering process and regulated by auxins (IAA) and abscisic acid (ABA) have also been recently identified in grapevine latent buds. Gene expression analysis of VvL1L, VvMFT1 and VvTFL1A in different types of fruitful buds (N0, N1 and N2) were realized during their development stages (88 to 159 days after budbreak) of the first year in 'Merlot' cultivar. VvL1L and VvTFL1A were up-regulated in early stages of bud development during the period of the initiation and differentiation of IP. The relative expression of VvMFT1 showed significant differences between the three types of buds (N0, N1 and N2) in relation with their development. IAA and ABA contents in latent buds at different stages during the first year were analyzed and correspond to gene expressions: VvTFL1A seems to be activated by IAA, while VvL1L and VvMFT1 might be regulated by IAA/ABA balance. These results may support the assumption of the involvement of VvL1L, VvMFT1 and VvTFL1A in the regulation of bud fruitfulness and their response to IAA/ABA balance.

International audience; Pulsed electric field (PEF) treatment is an emerging technology that is arousing increasing interest in vinification processes for its ability to enhance polyphenol extraction performance. The aim of this study was to investigate the effects of PEF treatment on grape skin histocytological structures and on the organization of skin cell wall polysaccharides and tannins, which, until now, have been little investigated. This study relates to the effects of two PEF treatments on harvested Cabernet Sauvignon berries: PEF1 (medium strength (4 kV/cm); short duration (1 ms)) and PEF2 (low intensity (0.7 kV/cm); longer duration (200 ms)). Histocytological observations and the study of levels of polysaccharidic fractions and total amounts of tannins allowed differentiation between the two treatments. Whereas PEF1 had little effect on the polyphenol structure and pectic fraction, PEF2 profoundly modified the organization of skin cell walls. Depending on the PEF parameters, cell wall structure was differently affected, providing variable performance in terms of polyphenol extraction and wine quality.

The treatment of Cabernet Sauvignon red grapes by pulsed electric fields (PEFs) is performed prior to vinification in order to enhance the extraction of polyphenols. PEF treatments of the longest duration and of the highest energy (E = 0.7 kV/cm, t PEF = 200 ms, W = 31 Wh/kg) changed the structure of grape skins and produced a wine that was richer in tannins (34 %), while treatment of the highest strength (E = 4 kV/cm, t PEF = 1 ms, W = 4 Wh/kg) altered the visual appearance of phenolic compounds in the skins and led to greater extraction of the anthocyanins (19 %). The PEF treatments caused the depolymerization of skin tannins, improving the diffusion of these decondensed tannins which are smaller. The PEF treatment of longest duration and of the highest energy had more impact on the parietal tannins and the cell walls of the skins while treatment of the highest strength modified more the vacuolar tannins. Changes in the operating parameters of the PEF treatment (E = 0.7 to 4 kV/cm, t PEF = 1 to 200 ms, W = 4 to 31 Wh/kg) did not affect alcohol content, total acidity nor volatile acidity in finished wines compared to the values of the control wine, but seemed to cause a slight increase in pH (1–2 %).

Esca is one of the major diseases affecting vineyards with direct impact on product yield; nevertheless, scientific studies concerning its impact on grape quality are scarce. As an attempt to better understand the mechanisms behind "Esca proper" development in grapes, this work focused on the identification of proteins whose expression is altered by the disease. 2-DEs were performed on protein extracts from grape skins at different stages of maturity for two consecutive vintages. Grapes were collected in 2009 and in 2010 from plants that did not present signs of infection by Esca proper since the 2004 vintage and from plants that presented cast leaf symptoms at least once since 2004. For the first time, 13 proteins were shown to be influenced by Esca proper during the ripening process. Extensive bioinformatics analysis allowed the grouping of proteins involved in (i) stress tolerance and defense response, (ii) oxidative phosphorylation, (iii) oxidation-reduction processes in mitochondria, and (iv) oxidation-reduction processes in chloroplasts. Of these 13 proteins, cysteine synthase is the only one implicated in a metabolic pathway of oenological interest. This study shows how foliar symptoms of Esca proper may impact stress-related pathways in grapes, which are characterized by modifications in the chain of oxidative phosphorylation and redox scavenging.

International audience; The grapevine (Vitis vinifera L.) is a widely cultivated species of major economic importance for wine production. The quality and quantity of grapes are criteria of prime importance to the wine industry, but they are highly variable from year to year. Unlike many perennial plants, cluster formation unfolds in two seasons: season 1 takes place in the bud until dormancy, and season 2 starts after budbreak in the following year. Season 1 corresponds to the initiation and differentiation of inflorescence primordia, controlled by many exogenous and endogenous factors, which explains up to 60% seasonal variation in yield. Season 2 consists of flowering and fruit development, which explains, respectively, 30% and 10% of seasonal variation in yield. It is therefore essential to understand the impact of these factors to better control the yield. This review aims to summarize past and present knowledge concerning the physiology of latent buds relating to their fruitfulness, and to assess the impact of environmental, hormonal, and regulation factors on the final yield. Avenues of further research to understand physiological, biochemical and molecular regulatory mechanisms of initiation and differentiation of clusters will be then proposed.; La vigne (Vitis vinifera L.) est une espèce cultivée représentant une importance économique incontournable par la production de vin. La qualité et la quantité des raisins sont des critères préoccupant la filière viti-vinicole, or elles sont très variables selon les millésimes. A la différence de nombreuses plantes pérennes, la formation des fruits se déroule sur deux cycles : une première phase dans le bourgeon jusqu’à la dormance et une seconde phase après le débourrement l’année suivante. La première phase correspond à celle de l’initiation et de la différentiation des primordia inflorescentiels, contrôlée par de nombreux facteurs exogènes et endogènes, et expliquerait jusqu’à 60 % de variation saisonnière du rendement. La deuxième phase consiste en la floraison et le développement des fruits, qui expliqueraient respectivement 30 et 10 % de la variation saisonnière du rendement. Il est donc primordial de comprendre l’impact de ces facteurs pour mieux réguler le rendement. Cette synthèse a pour objectif de faire le point sur les connaissances anciennes et récentes concernant la physiologie du bourgeon latent en rapport avec sa fertilité et d’évaluer l’impact des facteurs environnementaux, hormonaux et de régulation sur le rendement final. Des pistes de recherche afin de comprendre les mécanismes de régulation physiologique, biochimique et moléculaire de l’initiation et de la différentiation des grappes, seront par la suite proposées.

Background and Aims: The impact of grey mould (Botrytis cinerea (B. cinerea)) was quantified on chemical, phenolic and sensory qualities of grapes, derived musts and wines. Methods and Results: Analyses were carried out by using naturally or artificially infected grape berries at ripeness or overripeness. In grape seeds, chemical analyses revealed no major differences between healthy and rotten grapes. In grape skins of Botrytis-affected berries, concentrations of all the phenolic compounds (anthocyanins and proanthocyanidin monomers, dimers and trimer) decreased drastically. Mean degree of polymerization of the proanthocyanidin polymeric fraction was also affected in skins. Chemical analyses of musts and wines made with different percentages of rotten berries showed a moderate impact of the pathogen on their phenolic composition. Nevertheless, sensory analyses underlined a loss of wine sensory quality perceptible from a threshold as low as 5% of Botrytis-affected grapes onwards. Conclusion: Phenolic variations and the associated negative impact in grapes, derived musts and wines may be related to oxidation phenomena from B. cinerea. The main effects of severity/age of grey mould and the level of berry maturity are also discussed. Significance of the Study: B. cinerea drastically affects the phenolic and organoleptic properties of grape skins and derived wines. Therefore, prophylactic actions early in the vineyard, evaluation of the sanitary status of the harvested grapes and berry sorting are primordial even under low disease pressure.

Background and Aims: The impact of Esca, a cryptogamic disease affecting woody tissues of grapevines (trunk, branches and shoots), was investigated on phenolic and sensory qualities of Cabernet Sauvignon grapes, derived musts and wines. Methods and Results: Global phenolic analyses revealed no major difference between healthy and affected grapes whereas high-performance liquid chromatography analyses showed decreases in the skin concentrations of catechin and epicatechin and of anthocyanins for the two vintages assessed. Proanthocyanidin characteristics, and particularly mean degree of polymerisation, were strongly decreased in skin proanthocyanidin polymeric fractions. Chemical analyses of wines made with different percentages of fruit from Esca infected grapevines confirmed the moderate impact of Esca on phenolic composition. Nevertheless, sensory analyses revealed a loss of wine sensory quality perceptible with as little as 5% of affected fruit in wines. Conclusions: Phenolic variations in grapes and in derived wines may be related to changes to grapevine physiology caused by Esca fungi that impact on flavonoid metabolism. The chemical and sensory variations between wines can also be related to the delayed ripening delay of fruit from Esca infected grapevines. Significance of the Study: Esca moderately affected the phenolic composition of grapes and decreased the sensory quality of wines, suggesting a dramatic increase in the economic importance of Esca if no control methods are found.

Proanthocyanidins, or condensed tannins, are crucial polyphenolic compounds for grape and wine quality. Recently, significant advances were achieved in understanding the biosynthesis of their main subunits, (+)-catechin and (-)-epicatechin, produced by leucoanthocyanidin reductase (LAR) and anthocyanidin reductase (ANR), respectively. Expression and enzyme activity studies have been published but no data were previously available on their regulation by hormonal status. As abscisic acid (ABA) is involved in controlling anthocyanin biosynthesis, we examined the effects of the ABA signal on the proanthocyanidin pathway. The aim of this research was to determine whether ABA application on green grapes affected tannin content and composition, LAR and ANR activity, and the expression patterns of the genes encoding them (VvANR, VvLAR1, VvLAR2) and the associated transcription regulators (VvMYB5a, VvMYBPA1) in grape skin during ripening. Our results show that ABA affects tannin content and is involved in the tannin biosynthesis pathway in grape skin by decreasing LAR and ANR activity and repressing the expression of related genes a few days after application. This treatment reduced the tannin content of green grapes without modifying their composition but had a positive impact on tannin biosynthesis during veraison, as previously demonstrated for anthocyanins, suggesting that ANR and LAR were coregulated by ABA.

International audience; This work investigated the structural and biochemical changes during grape berry development which account potentially for the onset and increase in susceptibility to Botrytis cinerea. Using the cv. Sauvignon blanc, we quantified at seven developmental growth stages from herbaceous to over-mature berries: (1) fruit ontogenic resistance using three strains (II-transposa), (2) the morphological and maturity fruit characteristics and (3) preformed biochemical compounds located in the berry skin. From the mid-colour change stage onwards, susceptibility of unwounded fruit increased sigmoidally in both rot and sporulation severities at the berry surface. A principal component analysis identified a very close connection between fruit susceptibility and the level of fruit maturity. Berry susceptibility was significantly and positively correlated with the phenolic compounds in the skin cell walls and negatively correlated with the total tannin content in the skin and with water activity (Aw) at the fruit surface. On the berry, Aw decreased from 0.94 at bunch closure to 0.89 at berry maturity, with a relatively low value (0.90) at the stage of mid-colour change. Using artificial media, different Aw levels led to significant differences in mycelial growth (Aw ≤0.95 resulted in the lowest growth rate ≤0.34 mm day−1). Thus, besides the level of fruit maturity, both water activity on the fruit and the total tannin content in the skin may affect fungal growth and berry colonisation. The potential of these variables for use as indicators of grape berry susceptibility as well as associated mechanisms for the development of disease are discussed.

In grapevines, stomatal aperture decreases after a mid-morning peak during summer days. Afternoon stomatal closure increases in non-irrigated plants as water limitation progresses, which suggests the involvement of abscisic acid (ABA) in the control of stomatal aperture. The objective of this work was to study the seasonal and diurnal time-courses of CO2 assimilation rate, leaf conductance, leaf water potential (Y), and ABA concentration in xylem sap, leaves, flowers and berries in non-irrigated field-grown Tempranillo grapevines throughout reproductive development. Leaf Y decreased throughout fruit development because water availability decreased towards the end of the reproductive cycle. CO2 assimilation rate, leaf conductance and xylem ABA concentration also decreased during the course of the growing season. Combining all measurements xylem ABA was either not correlated, or only slightly correlated, with leaf water status (Y) and daily leaf conductance, respectively. This lack of relationship indicates that xylem ABA during fruit ripening had functions other than provision of a non-hydraulic signal. On a seasonal basis, xylem ABA concentration measured in non-irrigated grapevines was well related to berry ABA concentration, especially at the end of fruit development (veraison and harvest).

ata are provided on contrasting levels of different categories of polyamines and hydroxycinnamic acids during the course of disease development due to Botrytis cinerea on ripe grape berries that led to either noble rot or to grey mould. Noble rot development was mainly characterised by a greater accumulation of wall-bound polyamines at the beginning of infection, whereas the disease process that led to grey mould was marked by an increase of conjugated polyamines. During grey mould infection, coumaric acid was the predominant hydroxycinnamic acid present in both healthy and infected grape berries regardless of infection stage, whereas ferulic acid was present at lower concentrations than the other hydroxycinnamic acids identified. Moreover, changes in the relative proportions of each of the three hydroxycinnamic acids considered showed contrasting patterns according to the sort of rot which developed. The different roles of these compounds (hydroxycinnamic acids and polyamines) and their involvement in the interaction between Botrytis cinerea and grapevines is discussed. During development of grey mould, coumaric acid was the predominant hydroxycinnamic acid at the beginning and the end of infection, whereas caffeic acid predominated during mid stage.

The study was carried out in order to compare the development of berries growing on fruiting cuttings and on vines from the field, and to evaIuate this experimentaI system for grapevine physiology studies. The results show that the main developmental characteristics of the berries are close to those of berries from the vineyard. Fruit set is good and berry growth follows a double sigmoid curve. Final size is smaller than usual, even if seed number is higher for fruiting cuttings. The evolution of organics acids, sugar and mineral content is classical. At maturity, the berries from fruiting cuttings reach the same sugar content, but are less acidic, because the components of acid equilibrium are slightly modified. At flowering polyamine content is higher in the cluster of fruiting cutting but the variations aIong cluster development is the same than in the vineyard. Finally the application of exogenous putrescin on fruiting cuttings at flowering time improves fruit set, as in the field. Our data allow to conclude that fruiting cuttings behave mostly as bigger vines and could be a usefull tool for grapevine physiology studies.

International audience; Abscisic acid (ABA) has a well-known positive impact on grape ripening, especially color development, but its role in the initiation of anthocyanin synthesis remains unclear. To elucidate this point, ABA treatment was applied to a simple Vitis vinifera model, consisting of Cabernet Sauvignon cell suspensions that do not spontaneously produce anthocyanins under laboratory conditions. Endogenous ABA levels, the expression of some genes in the upstream part of the anthocyanin pathway, and anthocyanin content were determined. Exogenous ABA treatment sharply increased cell ABA content and induced both structural and regulatory genes involved in anthocyanin production. These changes were promptly detected, as early as 6 h after ABA treatment, whereas anthocyanin production was observed only after 4 days in culture. These results demonstrate that ABA promotes anthocyanin synthesis in grape cell culture.

International audience; The control of ripening of the non-climacteric grapevine fruit is still a matter of debate, but several lines of evidence point to an important role for the hormone abscisic acid (ABA). The effects of ABA treatments on Cabernet Sauvignon berries before and at véraison were studied using a 2-DE proteomic approach. Proteins from whole deseeded berries (before véraison) and berry flesh and skin (at véraison) treated with 0.76 mM ABA and collected 24 h after treatment were separated and analysed. A total of 60 protein spots showed significant variations between treated and control berries, and 40 proteins, mainly related to general metabolism and cell defence, were identified by LC MS/MS. Our results show that ABA acts mainly through the regulation of mostly the same proteins which are involved in the ripening process, and that several of these changes share common elements with the ABA-induced responses in vegetative tissues.

International audience; Proanthocyanidins, or condensed tannins, are crucial polyphenolic compounds for grape and wine quality. Recently, significant advances were achieved in understanding the biosynthesis of their main subunits: (+)-catechin and (−)-epicatechin, produced by catalysis of leucoanthocyanidin reductase (LAR) and anthocyanidin reductase (ANR), respectively. Expression studies had been published but no data were available on enzyme activity. In our work, we devised assays to measure LAR and ANR activity and determine their development throughout the growth of flowers, young berries, and skins of Vitis vinifera L. cv. Cabernet-Sauvignon. We also investigated the accumulation of compounds in these tissues and focused on the expression of both the structural genes and the transcription factors involved in regulating them: VvMYB5a and VvMYBPA1. Biosynthetic genes were expressed early and LAR and ANR were already active during flowering and at the beginning of berry growth, as well as during colour-change in skins. The profiles we determined correlated with total tannin, catechin, and epicatechin concentrations. The involvement of VvMYB5a and VvMYBPA1 was confirmed and specific expression patterns were also established for VvLAR transcripts.

International audience; Ripening of grape (Vitis vinifera L.) berry immediately precedes harvesting and the evolution of the skin tissue is important as it contains the key compounds for wine quality. Grape softening is thought to result from extensive cell wall modifications that occur during ripening. These modifications result from the activity of different cell wall-modifying enzymes. Two of the most significant pectin-degrading enzymes are pectin methylesterase (EC 3.1.1.11) and polygalacturonase (EC 3.2.1.15). In this work, the activities of both enzymes were monitored in skin tissue throughout berry development. Pectin methylesterase activity was present before the onset of veraison and increased during skin maturation. No polygalacturonase activity could be detected. The accumulation of mRNA encoding a pectin methylesterase and two polygalacturonase isoforms was examined using RT-PCR. Transcripts for pectin methylesterase were present in all stages analyzed with a maximal accumulation at the end of color change. Accumulation of VvPG1 transcript was closely correlated with berry softening, and expression of this gene was markedly increased during the color change. VvPG2 mRNA accumulation began before veraison and was low during skin ripening. A phylogenic analysis showed that this gene is classified in a different group than VvPG1. These findings suggest that both genes are associated with different mechanisms during skin development. VvPG1, in particular, is most likely to play a role in skin softening and VvPG2 in triggering the ripening process.

International audience; The characterization of proteins isolated from skin tissue is apparently an essential parameter for understanding grape ripening as this tissue contains the key compounds for wine quality. It has been particularly difficult to extract proteins from skins for analysis by two-dimensional electrophoresis gels and, therefore, a protocol for this purpose has been adapted. The focus was on the evolution of the proteome profile of grape skin during maturation. Proteome maps obtained at three stages of ripening were compared to assess the extent to which protein distribution differs in grape skin during ripening. The comparative analysis shows that numerous soluble skin proteins evolve during ripening and reveal specific distributions at different stages. Proteins involved in photosynthesis, carbohydrate metabolisms, and stress response are identified as being over-expressed at the beginning of colour-change. The end of colour-change is characterized by the over-expression of proteins involved in anthocyanin synthesis and, at harvest, the dominant proteins are involved in defence mechanisms. In particular, increases in the abundance of different chitinase and ß-1,3-glucanase isoforms were found as the berry ripens. This observation can be correlated with the increase of the activities of both of these enzymes during skin ripening. The differences observed in proteome maps clearly show that significant metabolic changes occur in grape skin during this crucial phase of ripening. This comparative analysis provides more detailed characterization of the fruit ripening process.

International audience; For two successive years, cell walls were isolated from the internal part of skin cells of Vitis vinifera L. cv. Cabernet Sauvignon grape berries grown in a vineyard. Procyanidin localization and composition were determined over the course of development. Tannins were mainly localized in the inner cell fraction, due to their biosynthesis and storage. Cell wall tannins always exhibited a higher mean degree of polymerization as compared to the internal cell fraction, which had a constant mDP. The mDP of cell wall tannins also tended to increase at the end of maturation. Our results suggest tannin polymerization near the cell wall but an aggregation in the vacuole during growth. The tannin composition was typical of skins, and small differences were noted between the two cell parts. Surprisingly, epigallocatechin-3-gallate was also detected, although in a very small amount. Epicatechin was present in significant proportions in both fractions, especially as an extension subunit, while epigallocatechin was likewise abundant, also as a terminal subunit. Last, procyanidin composition and organization seemed to be characteristic of the Cabernet Sauvignon variety.

Effects of Eutypa lata, the fungal agent of eutypiosis, on polyamine (putrescine (Put), spermidine (Spd), spermine (Spm), diaminopropane (Dap)) and tyramine (Tyr) contents in grapevine leaves were studied to determine the relationship between these components and disease development. The external symptoms of eutypiosis are small and necrotic leaves, which could be observed on only one arm or on two arms of the vine stock. This study was carried out on healthy leaves (S, borne by healthy vine stock), apparently healthy leaves (Aps, borne by one arm without symptoms, whereas the other arm was diseased), and moderately (M. mal.) and strongly (F. mal.) diseased leaves. We analysed free polyamines (free PAs, chlorhydric acid soluble fraction), conjugated polyamines (PAs-fpm, PAs covalently linked to molecules with low molecular mass like hydroxycinnamic acids), and bound polyamines (PAs-mm, covalently linked to macromolecules) in Vitis vinifera L. 'Cabernet sauvignon', a variety sensitive to eutypiosis. In diseased leaves, we noted a decrease of free PAs, PAs-fpm, free Tyr, and Tyr-fpm levels. By contrast, the Dap-fpm level was increased by the disease. Eutypiosis also induced an increase of Spm-mm and Dap-mm levels, whereas the Put-mm and Spd-mm levels were decreased. Free Put, free Spd, PAs-fpm, free Tyr, and Tyr-fpm levels were higher in apparently healthy leaves than in healthy leaves. The PAs-mm levels were slightly affected by eutypiosis in apparently healthy leaves. The specific role of each type of polyamines and tyramine is discussed with regard to the host-parasite relationship.Key words: polyamines, leaf, Eutypa lata, Vitis vinifera.

Cell walls were isolated from seeds of grape berries (Vitis vinifera L.), and proanthocyanidin composition was determined over the course of ripening for different levels of vine water status. During the ripening period the tannins from the cell walls were always more polymerized than those from the inner part of the cell. At maturity this difference becomes more significant compared to veraison, due to a significant increase in the mean degree of polymerization of the cell wall tannins. The tannin composition was typical of grape seed tannins and was quite similar in the two cell fractions studied, but the epicatechin gallate proportion was significantly higher in the cell wall fraction. There were no significant effects of water deficit on composition and polymerization of seed tannins. Keywords: Vitis vinifera L.; grape seeds; cell wall; tannins; water status

Aims: The objective was to better understand the mechanisms involved in grape ripening that brings about important changes in the physiology and chemistry of the fruit. So we focused on the involvement of two growth regulators: abscisic acid (ABA) and indol-acetic acid (IAA) in controlling grape berry ripening process.Methods and results: We described the evolution of the two plant growth regulators during the development of cv. Merlot grapes (Vitis vinifera L.). In order to better understand the role of ABA and IAA in the ripening control, these two growth regulators were applied on the grapes at the onset of veraison. The hormonal profile was established on treated berries and different physiological parameters were assayed to evaluate the effects of both applications. The partitioning of both plant growth regulators in nontreated berries showed that ABA and IAA accumulated at the onset of ripening. Moreover, it appeared that endogenous ABA decreased progressively in the flesh while accumulated in the skin from the beginning of the colour change to maturity. The hormonal treatments modified the hormonal profile and several physiological parameters: sugar, acidity, colour, and Botrytis sensibility.Conclusion: These findings suggest that both treatments have modified the ripening process. Exogenous ABA has induced advancement in grape ripening, while IAA application has delayed this process. These observations support the view that the grape ripening process may be influenced by the hormonal status.Significance and impact of study: This study gives new information about the ripening control of the non-climacteric fruits. In grape berries, it provides evidence of a possible co-involvement of ABA and IAA in controlling ripening process.

Grape berry ripening is characterised by numerous metabolic and physiological changes, placed under hormonal control. The phytohormone abscisic acid (ABA) is considered as a possible promoter of « véraison », since it accumulates at this time. In order to evaluate its impact, we assess the effect of ABA on several parameters of maturity: anthocyanins synthesis, phenylalanine ammonia-lyase activity, sugar contents, acidity and maturity index, both in untreated berry skin and in skin of berries submitted to applications of exogenous ABA. The maturation processes we focused on appear ABA-dependent, as they occured immediately after ABA accumulation in the skin. Treatment also shows a close relationship between ABA and the beginning of ripening. These results allow us to conclude that ABA content could modulate the triggering of « véraison ».

The effects of polyamines were examined for growth and polyamine contents in cultings, callus and primary adventitious roots of Vitis vinifera L. Variations in free, conjugated and wall-bound polyamines in cuttings were observed during rhizogenesis. The main polyamines in cuttings were conjugated polyamines while in callus and primary adventitious roots they were free polyamines. Exogenous polyamine addition did not modify the total number of roots per cutting but increased the mean size and number of long roots. Moreover, exogenous polyamines increased polyamine levels in callus and roots, particurlarly wall-bound and conjugated polyamines. The involvement of these classes of polyamines in morphogenic processes is discussed.