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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

Principal component analysis as performed on a matrix consisting of the objects as defined by the log and early stationary phase cultures derived from different growth stages and the variables defined as the average expressed values of each protein (band) detected in each of those cell cultures.Ripe, véraison and green refer to the different ripening stages of berry suspension cells (cultures I-III, respectively) used in the study. Log and early stationary refer to the growth phase of the cells sampled for the study.
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pone-0014708-g004: Principal component analysis as performed on a matrix consisting of the objects as defined by the log and early stationary phase cultures derived from different growth stages and the variables defined as the average expressed values of each protein (band) detected in each of those cell cultures.Ripe, véraison and green refer to the different ripening stages of berry suspension cells (cultures I-III, respectively) used in the study. Log and early stationary refer to the growth phase of the cells sampled for the study.

Mentions: Principal component analysis (PCA) was performed on a matrix consisting of the objects as defined by the early and late log phase cell cultures derived from each stage of berry development and the variables defined as the average expressed values of each protein (band) detected in each of those cell cultures. The early and late log phase matrices of stages I-III plot quite closely (Figure 4), confirming the homology in the samples per developmental stage. The PCA results show that the total proteome signal from the different stages, when considered as individual matrices, cluster far apart from each other, confirming that the berry cultures originating from berry explants in different developmental stages, maintained significant differences in the cultured state. Given the tight groupings in the two sampling points (log and early stationary phase), only the data from the log phase will be reported on in all further sections.


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)

Principal component analysis as performed on a matrix consisting of the objects as defined by the log and early stationary phase cultures derived from different growth stages and the variables defined as the average expressed values of each protein (band) detected in each of those cell cultures.Ripe, véraison and green refer to the different ripening stages of berry suspension cells (cultures I-III, respectively) used in the study. Log and early stationary refer to the growth phase of the cells sampled for the study.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0014708-g004: Principal component analysis as performed on a matrix consisting of the objects as defined by the log and early stationary phase cultures derived from different growth stages and the variables defined as the average expressed values of each protein (band) detected in each of those cell cultures.Ripe, véraison and green refer to the different ripening stages of berry suspension cells (cultures I-III, respectively) used in the study. Log and early stationary refer to the growth phase of the cells sampled for the study.
Mentions: Principal component analysis (PCA) was performed on a matrix consisting of the objects as defined by the early and late log phase cell cultures derived from each stage of berry development and the variables defined as the average expressed values of each protein (band) detected in each of those cell cultures. The early and late log phase matrices of stages I-III plot quite closely (Figure 4), confirming the homology in the samples per developmental stage. The PCA results show that the total proteome signal from the different stages, when considered as individual matrices, cluster far apart from each other, confirming that the berry cultures originating from berry explants in different developmental stages, maintained significant differences in the cultured state. Given the tight groupings in the two sampling points (log and early stationary phase), only the data from the log phase will be reported on in all further sections.

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