Aminoacids and flavonoids profiling in tempranillo berries can be modulated by the arbuscular mychorrhizal fungi

(1) Background: Vitis vinifera L. cv. Tempranillo is cultivated over the world for its wine of high quality. The association of Tempranillo with arbuscular mycorrhizal fungi (AMF) induced the accumulation of phenolics and carotenoids in leaves, affected the metabolism of abscisic acid (ABA) during berry ripening, and modulated some characteristics and quality aspects of grapes. The objective of this study was to elucidate if AMF influenced the profiles and the content of primary and secondary metabolites determinants for berry quality in Tempranillo. (2) Methods: Fruit-bearing cuttings inoculated with AMF or uninoculated were cultivated under controlled conditions. (3) Results: Mycorrhizal symbiosis modified the profile of metabolites in Tempranillo berries, especially those of the primary compounds. The levels of glucose and amino acids clearly increased in berries of mycorrhized Tempranillo grapevines, including those of the aromatic precursor amino acids. However, mycorrhizal inoculation barely influenced the total amount and the profiles of anthocyanins and flavonols in berries. (4) Conclusions: Mycorrhizal inoculation of Tempranillo grapevines may be an alternative to the exogenous application of nitrogen compounds in order to enhance the contents of amino acids in grapes, which may affect the aromatic characteristics of wines

VineyardFACE: Investigation of a moderate (+20 %) increase of ambient CO2 concentration on berry ripening dynamics and fruit composition of Cabernet-Sauvignon

Climate change and rising atmospheric carbon dioxide concentration are a concern for agriculture, including viticulture. Studies on elevated carbon dioxide have already been conducted on grapevines, mainly taking place in greenhouses using potted plants or using field grown vines under instant and higher CO2 enrichment, i.e., > 650 ppm. The VineyardFACE, located at Hochschule Geisenheim University, is an open field Free Air CO2 Enrichment (FACE) experimental set-up designed to study the effects of elevated carbon dioxide using adapted, field-grown vines (Vitis vinifera L. cv. Cabernet-Sauvignon). As the carbon dioxide fumigation started in 2014, the long-term effects of elevated carbon dioxide treatment can be investigated on berry ripening parameters and fruit metabolic composition. The present study investigates the effect on fruit composition under a moderate increase (+20 %; eCO2) of carbon dioxide concentration, as predicted for 2050 on Cabernet-Sauvignon. Berry growth, ripening dynamics and composition were determined and primary (sugars, organic acids, amino acids) and secondary metabolites (anthocyanins) were analysed. Compared to previous results of the early adaptive phase of the vines (Wohlfahrt et al., 2020), our results show little effects of eCO2 treatment on primary metabolites composition in berries. However, total anthocyanins concentration in berry skin was lower for eCO2 treatment in the hot and dry season of 2020, although the ratio between anthocyanins derivatives did not differ

Transcriptome Remodeling in Response to Leaf Removal and Exogenous Abscisic Acid in Berries of Grapevine (Vitis vinifera L.) Fruit Cuttings

Climate change is known to simultaneously increase berry sugars but decrease anthocyanins, leading to an imbalance between sugars and anthocyanins in grape berries. To restore the balance of sugars and anthocyanins, carbon limitation by leaf removal and exogenous abscisic acid (ABA) were separately or simultaneously applied to Vitis vinifera cv. Cabernet Sauvignon fruit cuttings to decipher their effects on berry quality with metabolite and whole-genome transcriptome analyses. Carbon limitation decreased the hexose concentration and fully blocked the accumulation of anthocyanins. However, exogenous ABA increased the anthocyanin concentration under both carbon limitation and sufficient conditions. Carbon limitation and exogenous ABA induced the profound remodeling of the whole-genome transcriptome and altered the anthocyanin concentration by regulating the transcription levels of genes involved in the anthocyanin biosynthesis pathways as well as in the genes involved in various types of hormone signaling. Moreover, two pertinent candidate genes were identified based on the co-expression network analysis between the berry metabolite and transcriptome results, including a transcriptional factor, ERF2, and a calcineurin B-like proteininteracting protein kinase gene, CIPK25. In summary, simultaneously modifying the carbon supply by leaf removal and spraying exogenous ABA could re-establish the balance between sugars and anthocyanins to improve the qualities of grape berries via whole-genome transcriptome remodeling

The effects of a moderate grape temperature increase on berry secondary metabolites

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°C in mean value, up to +5°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

Metabolite analysis reveals distinct spatio-temporal accumulation of anthocyanins in two teinturier variants of cv. ‘Gamay’ grapevines (Vitis vinifera L.)

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.Peer reviewe

The flavour of grape colour: anthocyanin content tunes aroma precursor composition by altering the berry microenvironment

Anthocyaninless (white) instead of black/red (coloured) fruits develop in grapevine cultivars without functional VviMYBA1 and VviMYBA2 genes, and this conditions the colour of wines that can be produced. To evaluate whether this genetic variation has additional consequences on fruit ripening and composition, we performed comparisons of microenvironment, transcriptomics, and metabolomics of developing grapes between near-isogenic white- and black-berried somatic variants of Garnacha and Tempranillo cultivars. Berry temperature was as much as 3.5 ºC lower in white- compared to black-berried Tempranillo. An RNA-seq study combined with targeted and untargeted metabolomics revealed that ripening fruits of white-berried variants were characterized by the up-regulation of photosynthesis-related and other light-responsive genes and by their higher accumulation of specific terpene aroma precursors, fatty acid-derived aldehyde volatiles, and phenylpropanoid precursor amino acids. MYBA1-MYBA2 function proved essential for flavonol trihydroxylation in black-berried somatic variants, which were also characterized by enhanced expression of pathogen defence genes in the berry skin and increased accumulation of C6-derived alcohol and ester volatiles and γ-aminobutyric acid. Collectively, our results indicate that anthocyanin depletion has side-effects on grape composition by altering the internal microenvironment of the berry and the partitioning of the phenylpropanoid pathway. Our findings show how fruit colour can condition other fruit features, such as flavour potential and stress homeostasis

Zebularine, a DNA Methylation Inhibitor, Activates Anthocyanin Accumulation in Grapevine Cells

Through its role in the regulation of gene expression, DNA methylation can participate in the control of specialized metabolite production. We have investigated the link between DNA methylation and anthocyanin accumulation in grapevine using the hypomethylating drug, zebularine and Gamay Teinturier cell suspensions. In this model, zebularine increased anthocyanin accumulation in the light, and induced its production in the dark. To unravel the underlying mechanisms, cell transcriptome, metabolic content, and DNA methylation were analyzed. The up-regulation of stress-related genes, as well as a decrease in cell viability, revealed that zebularine affected cell integrity. Concomitantly, the global DNA methylation level was only slightly decreased in the light and not modified in the dark. However, locus-specific analyses demonstrated a decrease in DNA methylation at a few selected loci, including a CACTA DNA transposon and a small region upstream from the UFGT gene, coding for the UDP glucose:flavonoid-3-O-glucosyltransferase, known to be critical for anthocyanin biosynthesis. Moreover, this decrease was correlated with an increase in UFGT expression and in anthocyanin content. In conclusion, our data suggest that UFGT expression could be regulated through DNA methylation in Gamay Teinturier, although the functional link between changes in DNA methylation and UFGT transcription still needs to be demonstrated

Effets de la nutrition azotée et du stress hydrique sur la maturation et la composition en anthocyanes des baies de Vitis vinifera L. au vignoble et en conditions contrôlées

*INRA CR Bordeaux, Unité d'Agronomie Diffusion du document : INRA CR Bordeaux, Unité d'Agronomie Diplôme : Dr. d'Universit

Effets de la nutrition azotée et du stress hydrique sur la maturation et la composition en anthocyanes des baies de Vitis vinifera L. au vignoble et en conditions contrôlées

Les incidences de la nutrition azotée et de l'alimentation hydrique de la vigne sur la maturation des raisins et sur la synthèse des anthocyanes dans les pellicules ont été étudiées sur des plants de Vitis vinifera L. cv. Merlot. Les expérimentations ont été conduites sur des plants cultivés au vignoble (variations simultanées des disponibilités en eau et en azote induites par un enherbement permanent) et en serre en hydroponique (boutures fructifères et plants en pots de deux ans). La maturation du raisin est favorisée par la limitation de la nutrition azotée et par la réduction de l'alimentation hydrique des vignes. Notre travail met également en évidence des interactions de la nutrition azotée et de l'alimentation hydrique avec l'ensemble de l'alimentation minérale de la vigne (en particulier P et K). Une alimentation hydrique réduite ou une nutrition limitée en azote stimulent la synthèse d'anthocyanes tandis qu'un niveau d'azote très élevé semble inhiber cette synthèse. Cet effet inhibiteur n'est pas observé sous contrainte hydrique. Les alimentations hydrique et azotée exercent donc probablement un contrôle sur l'activité d'une ou de plusieurs enzymes impliquées dans les premières étapes de la voie de biosynthèse des anthocyanes. Les niveaux de nutrition azotée et d'alimentation hydrique conduisent à des profils d'anthocyanes différents. La réduction de l'alimentation hydrique favorise la synthèse des dérivés de la malvidine. A l'inverse, une diminution de la nutrition azotée limite leur synthèse. Ces facteurs environnementaux semblent agir différemment sur les enzymes impliquées dans la synthèse des différentes formes d'anthocyanes. Les différents profils observés pourraient également être le fait de l'activation ou de la limitation du catabolisme des anthocyanes. Au vignoble, le niveau de réponse de la vigne (en particulier en termes de synthèse des anthocyanes) dépend essentiellement des conditions climatiques de l'année (millésime), du terroir et du mode d'entretien du sol.We studied the effect of nitrogen nutrition and water deficit on the ripening of berries and on anthocyanins biosynthesis in the berry skins of Merlot vines (Vitis vinifera L.) The experiments have been performed either on vines cultivated in vineyard (permanent grass cover allowed simultaneous variation of nitrogen and water supply) or under controlled environmental conditions (grapevine fruiting cuttings and two-year-old potted vines). Better berry ripening was observed under nitrogen or water limited supply. Our work showed interactions between nitrogen nutrition and water supply and the whole mineral nutrition of vines (especially P and K). Under reduced water and nitrogen nutrition, an increase of anthocyanins skin content was observed while a high nitrogen supply seemed to inhibit the anthocyanins synthesis. However, this latter effect was not observed under watering constraint. Our results suggest that water and nitrogen supply have a regulatory effect on the activity of the enzymes involved in the first steps of anthocyanins pathway. Different profiles of the anthocyanins species were obtained depending on the levels of nitrogen nutrition and watering supply. The synthesis of the malvidin derivatives was increased by a reduced water supply and was decreased by a limited nitrogen nutrition. The environmental factors seem to act differently on the esterification steps of anthocyanins biosynthesis. A regulation of the anthocyanins catabolism has to be considered too. In the vineyard, the level of anthocyanins biosynthesis depends essentially on the climatic conditions of the year (vintage), the "Terroir" effect and the cultural practices.BORDEAUX2-BU Santé (330632101) / SudocBORDEAUX1-BU Sciences-Talence (335222101) / SudocVILLENAVE D'ORNON-Bib. ISVV (335502201) / SudocSudocFranceF