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Proteome changes in the skin of the grape cultivar Barbera among different stages of ripening.

Negri AS, Prinsi B, Rossoni M, Failla O, Scienza A, Cocucci M, Espen L - BMC Genomics (2008)

Bottom Line: These results give new insights to the skin proteome evolution during ripening, thus underlining some interesting traits of this tissue.Moreover, these data emphasize the relevance of this tissue as a physical barrier exerting an important part in berry protection.In fact, the level of many proteins involved in (a)biotic stress responses remarkably changed through the five stages taken into consideration, thus suggesting that their expression may be developmentally regulated.

View Article: PubMed Central - HTML - PubMed

Affiliation: Dipartimento di Produzione Vegetale, University of Milan, via Celoria 2, 20133 Milano, Italy. alfredo.negri@unimi.it

ABSTRACT

Background: Grape ripening represents the third phase of the double sigmoidal curve of berry development and is characterized by deep changes in the organoleptic characteristics. In this process, the skin plays a central role in the synthesis of many compounds of interest (e.g. anthocyanins and aroma volatiles) and represents a fundamental protective barrier against damage by physical injuries and pathogen attacks. In order to improve the knowledge on the role of this tissue during ripening, changes in the protein expression in the skin of the red cultivar Barbera at five different stages from véraison to full maturation were studied by performing a comparative 2-DE analysis.

Results: The proteomic analysis revealed that 80 spots were differentially expressed throughout berry ripening. Applying a two-way hierarchical clustering analysis to these variations, a clear difference between the first two samplings (up to 14 days after véraison) and the following three (from 28 to 49 days after véraison) emerged, thus suggesting that the most relevant changes in protein expression occurred in the first weeks of ripening. By means of LC-ESI-MS/MS analysis, 69 proteins were characterized. Many of these variations were related to proteins involved in responses to stress (38%), glycolysis and gluconeogenesis (13%), C-compounds and carbohydrate metabolism (13%) and amino acid metabolism (10%).

Conclusion: These results give new insights to the skin proteome evolution during ripening, thus underlining some interesting traits of this tissue. In this view, we observed the ripening-related induction of many enzymes involved in primary metabolism, including those of the last five steps of the glycolytic pathway, which had been described as down-regulated in previous studies performed on whole fruit. Moreover, these data emphasize the relevance of this tissue as a physical barrier exerting an important part in berry protection. In fact, the level of many proteins involved in (a)biotic stress responses remarkably changed through the five stages taken into consideration, thus suggesting that their expression may be developmentally regulated.

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Related in: MedlinePlus

2-DE maps of five stages through the ripening of Barbera. 2-DE maps of five different ripening stages from véraison until full ripeness of cultivar Barbera berry skins. The véraison stage (0 DAV) was considered as the moment when 50% of the berries started to change colour. Proteins (200 μg) were separated by IEF at pH 3–10, followed by 12.5% SDS PAGE and visualized by cCBB-staining.
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Figure 1: 2-DE maps of five stages through the ripening of Barbera. 2-DE maps of five different ripening stages from véraison until full ripeness of cultivar Barbera berry skins. The véraison stage (0 DAV) was considered as the moment when 50% of the berries started to change colour. Proteins (200 μg) were separated by IEF at pH 3–10, followed by 12.5% SDS PAGE and visualized by cCBB-staining.

Mentions: Proteins were extracted from the berry skin samples of cultivar Barbera previously washed in acetone through a protocol which made use of phenol followed by precipitation in ammonium acetate in methanol, which was previously indicated to be appropriate for this recalcitrant tissue [24]. 2-DE gels are shown in Fig. 1. The average number of detected spots was about 850 for each stage and did not vary significantly among the five different conditions. To ascertain the quantitative changes in the proteomic maps, their relative spot volumes (%Vol) were evaluated by software-assisted analysis. The ANOVA test (p < 0.01), coupled with a threshold of two-fold change in level, revealed 80 spots as being differentially expressed throughout berry ripening.


Proteome changes in the skin of the grape cultivar Barbera among different stages of ripening.

Negri AS, Prinsi B, Rossoni M, Failla O, Scienza A, Cocucci M, Espen L - BMC Genomics (2008)

2-DE maps of five stages through the ripening of Barbera. 2-DE maps of five different ripening stages from véraison until full ripeness of cultivar Barbera berry skins. The véraison stage (0 DAV) was considered as the moment when 50% of the berries started to change colour. Proteins (200 μg) were separated by IEF at pH 3–10, followed by 12.5% SDS PAGE and visualized by cCBB-staining.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2529320&req=5

Figure 1: 2-DE maps of five stages through the ripening of Barbera. 2-DE maps of five different ripening stages from véraison until full ripeness of cultivar Barbera berry skins. The véraison stage (0 DAV) was considered as the moment when 50% of the berries started to change colour. Proteins (200 μg) were separated by IEF at pH 3–10, followed by 12.5% SDS PAGE and visualized by cCBB-staining.
Mentions: Proteins were extracted from the berry skin samples of cultivar Barbera previously washed in acetone through a protocol which made use of phenol followed by precipitation in ammonium acetate in methanol, which was previously indicated to be appropriate for this recalcitrant tissue [24]. 2-DE gels are shown in Fig. 1. The average number of detected spots was about 850 for each stage and did not vary significantly among the five different conditions. To ascertain the quantitative changes in the proteomic maps, their relative spot volumes (%Vol) were evaluated by software-assisted analysis. The ANOVA test (p < 0.01), coupled with a threshold of two-fold change in level, revealed 80 spots as being differentially expressed throughout berry ripening.

Bottom Line: These results give new insights to the skin proteome evolution during ripening, thus underlining some interesting traits of this tissue.Moreover, these data emphasize the relevance of this tissue as a physical barrier exerting an important part in berry protection.In fact, the level of many proteins involved in (a)biotic stress responses remarkably changed through the five stages taken into consideration, thus suggesting that their expression may be developmentally regulated.

View Article: PubMed Central - HTML - PubMed

Affiliation: Dipartimento di Produzione Vegetale, University of Milan, via Celoria 2, 20133 Milano, Italy. alfredo.negri@unimi.it

ABSTRACT

Background: Grape ripening represents the third phase of the double sigmoidal curve of berry development and is characterized by deep changes in the organoleptic characteristics. In this process, the skin plays a central role in the synthesis of many compounds of interest (e.g. anthocyanins and aroma volatiles) and represents a fundamental protective barrier against damage by physical injuries and pathogen attacks. In order to improve the knowledge on the role of this tissue during ripening, changes in the protein expression in the skin of the red cultivar Barbera at five different stages from véraison to full maturation were studied by performing a comparative 2-DE analysis.

Results: The proteomic analysis revealed that 80 spots were differentially expressed throughout berry ripening. Applying a two-way hierarchical clustering analysis to these variations, a clear difference between the first two samplings (up to 14 days after véraison) and the following three (from 28 to 49 days after véraison) emerged, thus suggesting that the most relevant changes in protein expression occurred in the first weeks of ripening. By means of LC-ESI-MS/MS analysis, 69 proteins were characterized. Many of these variations were related to proteins involved in responses to stress (38%), glycolysis and gluconeogenesis (13%), C-compounds and carbohydrate metabolism (13%) and amino acid metabolism (10%).

Conclusion: These results give new insights to the skin proteome evolution during ripening, thus underlining some interesting traits of this tissue. In this view, we observed the ripening-related induction of many enzymes involved in primary metabolism, including those of the last five steps of the glycolytic pathway, which had been described as down-regulated in previous studies performed on whole fruit. Moreover, these data emphasize the relevance of this tissue as a physical barrier exerting an important part in berry protection. In fact, the level of many proteins involved in (a)biotic stress responses remarkably changed through the five stages taken into consideration, thus suggesting that their expression may be developmentally regulated.

Show MeSH
Related in: MedlinePlus