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Proteomic analysis of grape berry cell cultures reveals that developmentally regulated ripening related processes can be studied using cultured cells.

Sharathchandra RG, Stander C, Jacobson D, Ndimba B, Vivier MA - PLoS ONE (2011)

Bottom Line: Thirty two % of the identified proteins are currently annotated as hypothetical.The differential expression profile of the identified proteins, when compared with published literature on grape berry ripening, suggested common trends in terms of relative abundance in the different developmental stages between real berries and cell suspensions.The advantages of having suspension cultures that accurately mimic specific developmental stages are profound and could significantly contribute to the study of the intricate regulatory and signaling networks responsible for berry development and ripening.

View Article: PubMed Central - PubMed

Affiliation: Department of Viticulture and Oenology, Institute for Wine Biotechnology, Stellenbosch University, Stellenbosch, South Africa.

ABSTRACT

Background: This work describes a proteomics profiling method, optimized and applied to berry cell suspensions to evaluate organ-specific cultures as a platform to study grape berry ripening. Variations in berry ripening within a cluster(s) on a vine and in a vineyard are a major impediment towards complete understanding of the functional processes that control ripening, specifically when a characterized and homogenous sample is required. Berry cell suspensions could overcome some of these problems, but their suitability as a model system for berry development and ripening needs to be established first.

Methodology/principal findings: In this study we report on the proteomic evaluation of the cytosolic proteins obtained from synchronized cell suspension cultures that were established from callus lines originating from green, véraison and ripe Vitis vinifera berry explants. The proteins were separated using liquid phase IEF in a Microrotofor cell and SDS PAGE. This method proved superior to gel-based 2DE. Principal component analysis confirmed that biological and technical repeats grouped tightly and importantly, showed that the proteomes of berry cultures originating from the different growth/ripening stages were distinct. A total of twenty six common bands were selected after band matching between different growth stages and twenty two of these bands were positively identified. Thirty two % of the identified proteins are currently annotated as hypothetical. The differential expression profile of the identified proteins, when compared with published literature on grape berry ripening, suggested common trends in terms of relative abundance in the different developmental stages between real berries and cell suspensions.

Conclusions: The advantages of having suspension cultures that accurately mimic specific developmental stages are profound and could significantly contribute to the study of the intricate regulatory and signaling networks responsible for berry development and ripening.

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Related in: MedlinePlus

Functional categories of the identified proteins and their distribution indicated in percentage.
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pone-0014708-g005: Functional categories of the identified proteins and their distribution indicated in percentage.

Mentions: Among the 26 bands subjected to MALDI-TOF/MS analysis, 22 bands were identified. A protein was considered to be positively identified if a minimum of four peptides matched, showed ≥10% sequence coverage and a Molecular Weight Search (MOWSE) score of ≥70 (p<0.05). The detailed list of the identified proteins with their theoretical molecular weights, pIs, number of matched peptides and their respective expression profiles are reported in Table S1. The identified proteins could be categorized into different functional classes including metabolism, signaling, protein synthesis and defense (Figure 5).


Proteomic analysis of grape berry cell cultures reveals that developmentally regulated ripening related processes can be studied using cultured cells.

Sharathchandra RG, Stander C, Jacobson D, Ndimba B, Vivier MA - PLoS ONE (2011)

Functional categories of the identified proteins and their distribution indicated in percentage.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3040747&req=5

pone-0014708-g005: Functional categories of the identified proteins and their distribution indicated in percentage.
Mentions: Among the 26 bands subjected to MALDI-TOF/MS analysis, 22 bands were identified. A protein was considered to be positively identified if a minimum of four peptides matched, showed ≥10% sequence coverage and a Molecular Weight Search (MOWSE) score of ≥70 (p<0.05). The detailed list of the identified proteins with their theoretical molecular weights, pIs, number of matched peptides and their respective expression profiles are reported in Table S1. The identified proteins could be categorized into different functional classes including metabolism, signaling, protein synthesis and defense (Figure 5).

Bottom Line: Thirty two % of the identified proteins are currently annotated as hypothetical.The differential expression profile of the identified proteins, when compared with published literature on grape berry ripening, suggested common trends in terms of relative abundance in the different developmental stages between real berries and cell suspensions.The advantages of having suspension cultures that accurately mimic specific developmental stages are profound and could significantly contribute to the study of the intricate regulatory and signaling networks responsible for berry development and ripening.

View Article: PubMed Central - PubMed

Affiliation: Department of Viticulture and Oenology, Institute for Wine Biotechnology, Stellenbosch University, Stellenbosch, South Africa.

ABSTRACT

Background: This work describes a proteomics profiling method, optimized and applied to berry cell suspensions to evaluate organ-specific cultures as a platform to study grape berry ripening. Variations in berry ripening within a cluster(s) on a vine and in a vineyard are a major impediment towards complete understanding of the functional processes that control ripening, specifically when a characterized and homogenous sample is required. Berry cell suspensions could overcome some of these problems, but their suitability as a model system for berry development and ripening needs to be established first.

Methodology/principal findings: In this study we report on the proteomic evaluation of the cytosolic proteins obtained from synchronized cell suspension cultures that were established from callus lines originating from green, véraison and ripe Vitis vinifera berry explants. The proteins were separated using liquid phase IEF in a Microrotofor cell and SDS PAGE. This method proved superior to gel-based 2DE. Principal component analysis confirmed that biological and technical repeats grouped tightly and importantly, showed that the proteomes of berry cultures originating from the different growth/ripening stages were distinct. A total of twenty six common bands were selected after band matching between different growth stages and twenty two of these bands were positively identified. Thirty two % of the identified proteins are currently annotated as hypothetical. The differential expression profile of the identified proteins, when compared with published literature on grape berry ripening, suggested common trends in terms of relative abundance in the different developmental stages between real berries and cell suspensions.

Conclusions: The advantages of having suspension cultures that accurately mimic specific developmental stages are profound and could significantly contribute to the study of the intricate regulatory and signaling networks responsible for berry development and ripening.

Show MeSH
Related in: MedlinePlus