Vacuum Infiltration-Centrifugation Method for Apoplastic Protein Extraction in Grapevine.
The apoplastic fluid moving in the extracellular space external to the plasma membrane provides a means of delivering molecules and facilitates intercellular communications. However, the apoplastic fluid extraction from in planta systems remains challenging and this is particularly true for grapevine (Vitis vinifera L.), a worldwide-cultivated fruit plant. Here, we describe an optimized vacuum-infiltration-centrifugation method to extract soluble proteins from apoplastic fluid of grapevine leaves. This optimized method allows recovering of the grapevine apoplastic soluble proteins suitable for mono- and bi-dimensional gel electrophoresis for further proteomic analysis in order to elucidate their physiological functions.
Laboratoire Stress, Défenses et Reproduction des Plantes, Unité de Recherche Vignes et Vins de Champagne-EA 4707, SFR Condorcet - FR CNRS 3417, Université de Reims Champagne-Ardenne, Moulin de la Housse, BP 1039, 51687, Reims Cedex 2, France.
Bertrand
B
Delaunois
Laboratoire Stress, Défenses et Reproduction des Plantes, Unité de Recherche Vignes et Vins de Champagne-EA 4707, SFR Condorcet - FR CNRS 3417, Université de Reims Champagne-Ardenne, Moulin de la Housse, BP 1039, 51687, Reims Cedex 2, France.
Fabienne
F
Baillieul
Laboratoire Stress, Défenses et Reproduction des Plantes, Unité de Recherche Vignes et Vins de Champagne-EA 4707, SFR Condorcet - FR CNRS 3417, Université de Reims Champagne-Ardenne, Moulin de la Housse, BP 1039, 51687, Reims Cedex 2, France.
Christophe
C
Clément
Laboratoire Stress, Défenses et Reproduction des Plantes, Unité de Recherche Vignes et Vins de Champagne-EA 4707, SFR Condorcet - FR CNRS 3417, Université de Reims Champagne-Ardenne, Moulin de la Housse, BP 1039, 51687, Reims Cedex 2, France.
Philippe
P
Jeandet
Laboratoire Stress, Défenses et Reproduction des Plantes, Unité de Recherche Vignes et Vins de Champagne-EA 4707, SFR Condorcet - FR CNRS 3417, Université de Reims Champagne-Ardenne, Moulin de la Housse, BP 1039, 51687, Reims Cedex 2, France. sylvain.cordelier@univ-reims.fr.
Sylvain
S
Cordelier
D015153:Blotting, Western; D002498:Centrifugation; D005374:Filtration; D042541:Intracellular Space / Q000378:metabolism; D010940:Plant Proteins / Q000302:isolation & purification* / Q000378:metabolism*; D020543:Proteome; D014618:Vacuum; D027843:Vitis / Q000378:metabolism*
D010940:Plant Proteins; D020543:Proteome
Apoplastic fluid extraction; Apoplastic protein; Grapevine leaves; Secretome; VIC method; Vacuum-infiltration-centrifugation method; Vitis vinifera