title: Isolation of Chromoplasts and Suborganellar Compartments from Tomato and Bell Pepper Fruit.
abstract: Tomato is a model for fruit development and ripening. The isolation of intact plastids from this organism is therefore important for metabolic and proteomic analyses. Pepper, a species from the same family, is also of interest since it allows isolation of intact chromoplasts in large amounts. Here, we provide a detailed protocol for the isolation of tomato plastids at three fruit developmental stages, namely, nascent chromoplasts from the mature green stage, chromoplasts from an intermediate stage, and fully differentiated red chromoplasts. The method relies on sucrose density gradient centrifugations. It yields high purity organelles suitable for proteome analyses. Enzymatic and microscopy assays are summarized to assess purity and intactness. A method is also described for subfractionation of pepper chromoplast lipoprotein structures.
authors.affiliation: Génomique et Biotechnologie des Fruits, INRA, INP-ENSA Toulouse, Université de Toulouse, Avenue de l'Agrobiopole, Castanet-Tolosan, 31326, France.
authors.forename: Cristina
authors.initials: C
authors.lastname: Barsan
authors.affiliation: Laboratoire de Physiologie Cellulaire Végétale, CEA/CNRS/INRA/Université Grenoble-Alpes, Grenoble, France.
authors.forename: Marcel
authors.initials: M
authors.lastname: Kuntz
authors.affiliation: Génomique et Biotechnologie des Fruits, INRA, INP-ENSA Toulouse, Université de Toulouse, Avenue de l'Agrobiopole, Castanet-Tolosan, 31326, France. Jean-Claude.Pech@ensat.fr.
authors.forename: Jean-Claude
authors.initials: JC
authors.lastname: Pech
mesh_terms: D002212:Capsicum / Q000737:chemistry*; D002458:Cell Fractionation / Q000295:instrumentation / Q000379:methods*; D002499:Centrifugation, Density Gradient / Q000295:instrumentation / Q000379:methods; D003470:Culture Media / Q000737:chemistry; D057075:Enzyme Assays; D005638:Fruit / Q000737:chemistry; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) / Q000378:metabolism; D007425:Intracellular Membranes / Q000737:chemistry* / Q000648:ultrastructure; D008074:Lipoproteins / Q000737:chemistry / Q000302:isolation & purification; D018551:Solanum lycopersicum / Q000737:chemistry*; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification*; D018087:Plastids / Q000737:chemistry* / Q000648:ultrastructure; D013395:Sucrose / Q000737:chemistry
chemical_list: D003470:Culture Media; D008074:Lipoproteins; D010940:Plant Proteins; D013395:Sucrose; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating)
keywords: Capsicum annuum; Chromoplasts; Organelle purity; Solanum lycopersicum