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Developmental regulation of primary carbohydrate metabolism in grape berry (Vitis vinifera L.) cv.
Cabernet Sauvignon
Zhanwu Dai, Céline Léon, Regina Feil, John Lunn, Serge Delrot, Eric Gomes
To cite this version:
Zhanwu Dai, Céline Léon, Regina Feil, John Lunn, Serge Delrot, et al.. Developmental regulation of primary carbohydrate metabolism in grape berry (Vitis vinifera L.) cv. Cabernet Sauvignon. 5.
Journées Scientifiques du Réseau Français de Métabolomique et Fluxomique, May 2011, Paris, France.
1 p., 2011. �hal-02804263�
Similar metabolite concentration in vineyard and greenhouse
Most metabolites showed similar levels of concentration for berries grown in vineyardand greenhouse(Fig. 2), although the temperatures during the late berry development stages were quite different between the two growth conditions (Fig. 1). In contrast, the profile of sugar phosphate (including F6P, G1P, G6P, and T6P) showed different pattern during the late development stages between greenhouse and vineyard (Fig. 2). In vineyard, sugar phosphate gradually decreased with berry development, while they slightly increased after veraison in greenhouse. This difference might be resulted from the higher temperature in greenhouse than in vineyardduring the corresponding period.
Developmental regulation of primary carbohydrate metabolism in grape berry (Vitis vinifera L.) cv. Cabernet Sauvignon
1INRA, Univ. Bordeaux, ISVV, UMR 1287 Ecophysiologie et Génomique Fonctionnelle de la Vigne (EGFV), 210 Chemin de Leysotte, 33882 Villenave d'Ornon, France.
2Max Planck Institute of Molecular Plant Physiology, Am Mühlenberg 1, 14476 Potsdam‐Golm, Germany.
*Corresponding author. E‐mail: eric.gomes@bordeaux.inra.fr
Zhan Wu Dai
1, Céline Leon
1, Regina Feil
2, John Lunn
2, Serge Delrot
1, Eric Gomès
1*
Introduction
Carbohydrate metabolism in grape plays a central role in shaping final grape quality, not only because it produces the sugars that determine sweetness and alcohol content after fermentation, but also because it provides precursors for synthesis of organic acids, anthocyanins and aroma compounds etc. It is well known that carbohydrate metabolism is under developmental regulation, however, the underlying mechanisms are not yet clear. This experiment aims to identify metabolic coordination switches during grape development, to provide insights into the timing of developmental regulation of carbohydrate metabolism.
Mitochondrion
Oxaloacetate Citrate
Isocitrate Aconitate
2-ketoglutarate Succinyl-CoA
Succinate Fumarate Malate
Acetyl CoA Pyruvate
CO2
CO2
CO2
Erythrose-4P
3-Dehydroshikimate Pyruvate
Oxaloacetate PEP
Malate
2-PGA 3-PGA Glycerate-1,3-BP Glycerinaldehyde-3-P F1,6BP
F6P G6P
G1P
UDP-G
T6P
TRE Glucose
Fructose Sucrose
S6P ADPG
Shikimate
Materials and methods
Grape berries (cv. Cabernet Sauvignon) were sampled at 10 different developmental stages (from flowering to maturity), from either vineyard grown vines or fruit‐bearing cuttings grown in the greenhouse. The ten developmental stages include P1:10 days after flowering (DAF), P2:20DAF, P3:30DAF, P4:40DAF, and P5:veraison; and after veraison, berries were harvested according to their potential alcohol levels (%TVA), P6: 7%TVA, P7:8%TVA, P8:9%TVA, P9:10%TVA and P10: maturity. Temperatures were recorded for each experimental site and were used to calculate sum of temperature from flowering (oCday)(Fig.1). The concentrations of 27 metabolites from central carbon metabolism were measured using LC‐MS and enzymatic assays (Fig.2).
Figure 1. Daily temperature and sum of temperature (oCday) from flowering to maturity in vineyard and greenhouse. Greenhouse has similar temperature as in vineyard during the early development stages, but has higher temperature than those of vineyard during the final development stages (from 80 to 110 DAF).
Figure 2. Metabolite profiles of berries grown in vineyard or greenhouse. Metabolites are assigned to their metabolism pathways (sugar metabolism, glycolysis, and TCA cycle) and their evolutions in concentration (μnom g‐1FW for sucrose, glucose, and fructose; and nmol g‐1FW for the rest metabolites) are presented beside the corresponding metabolites. To facilitate comparing between the results of vineyard and greenhouse, the sum of temperature (oCday) was normalized to be 0 at veraison (indicated by the dashed line) for both conditions.
Vineyard Greenhouse
Glycolysis TCA cycle Sugar metabolism Veraison
Figure 3. Principle component analysis (PCA) of metabolite profiling during berry development. (A)The trajectories during development period for berries grown in vineyard (colored points) and greenhouse (black points). Arrows indicate the order of development, color for the berry skin color, point size for berry size.(B) correlation plots of metabolites for the first two PCs.
A B
Principle component analysis (PCA) readily discriminates the various stages of berry development, with similar trajectories for field grown and greenhouse samples. The first two principal components (PC1and PC2) explain about 86% of the total variance. PC1 separates the pre‐and post‐veraison stages, based largely on changes in sugars and glycolytic intermediates. PC2resolves the individual stages of development within the pre‐
and post‐veraison phases, related mainly to differences in TCA cycle intermediates, such as citrate, isocitrate and malate.
Figure 4. Heatmap and clustering of metabolistes during berry development for berries grown in vineyard. Each row represents a metabolite and each column represents a development stage. Values were centered and scaled in the row direction to form virtual color as presented in the color key.
The measured metabolites can be clustered into three main groups:
sugars and sugar‐phosphate metabolism, glycolysis and the TCA cycle. Sugars (including sucrose, fructose and glucose) stay at very low level before version and show dramatic increase from version.
Organic acids increase gradually with development, reach a peak at veraison, and thereafter decrease gradually. Interestingly, glycolytic intermediates and sugar phosphates declined during the early stages of development, but then remain relatively stable post‐
veraison.
Figure 5 Correlation matrix of all metabolites of berries grown in vineyard. Pearson correlation coefficient (positive or negative) between a pair of metabolites is represented by virtual color.
Metabolites within a given pathway were often positively correlated, however, negative correlations were observed between metabolites from different pathways. The main exception is the negative correlation between sugars and sugar phosphates.
Summary and overlook
Over all, it is clear that glycolytic and the TCA cycle intermediates are tightly coordinated with other metabolites within their respective pathways during development. In addition, there is a pronounced shift in metabolism around veraison, characterised by rapidly increasing sugar levels and decreasing organic acids. In contrast, glycolytic intermediates and sugar phosphates declined during the early stages of development, but then remain relatively stable post‐veraison. These results provide a rather detailed picture of the metabolite profiling of primary carbohydrate metabolism in grape berry. Integration with enzyme activity measurements and kinetic model analysis will be developed in the future.
