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Plastids pp 123-135 | Cite as

Purification of Chloroplasts and Chloroplast Subfractions: Envelope, Thylakoids, and Stroma—From Spinach, Pea, and Arabidopsis thaliana

  • Maryse A. Block
  • Catherine Albrieux
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1829)

Abstract

Chloroplasts are specific organelles of plant cells dedicated to photosynthesis and delimited by a two-membrane chloroplast envelope. Their photosynthetic function is based on the development of an operational large internal membrane network, called the thylakoids, and on enzymatic processes present in the chloroplast matrix, called the stroma. Thylakoid membranes are clearly different from the chloroplast envelope and their biogenesis is dependent on biosynthetic and transport activities specific of the chloroplast envelope. Starting with the isolation of intact chloroplasts, the method presents the separation by differential centrifugation of the three main compartments of the chloroplast: the stroma, the thylakoids, and the chloroplast envelope. Three different protocols are provided, adapted for starting leaves of spinach, Arabidopsis, and pea.

Key words

Chloroplast Stroma Envelope Thylakoids Membrane Differential centrifugation Chloroplast fractionation 

Notes

Acknowledgments

This work was supported by the French National Research Agency (ANR-13-ADAP-0008 Reglisse and ANR-10-LABEX-04 GRAL Labex, Grenoble Alliance for Integrated Structural Cell Biology).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Laboratoire de Physiologie Cellulaire et Végétale, Centre National de la Recherche Scientifique, Institut National Recherche Agronomique, Commissariat à l’Energie Atomique et aux Energies Alternatives, CEA GrenobleUMR5168, Université Grenoble AlpesGrenobleFrance

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