Unit

UNIT 3.30 Purification of Intact Chloroplasts from Arabidopsis and Spinach Leaves by Isopycnic Centrifugation

  1. Daphné Seigneurin-Berny,
  2. Daniel Salvi,
  3. Jacques Joyard,
  4. Norbert Rolland

Published Online: 1 SEP 2008

DOI: 10.1002/0471143030.cb0330s40

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Seigneurin-Berny, D., Salvi, D., Joyard, J. and Rolland, N. 2008. Purification of Intact Chloroplasts from Arabidopsis and Spinach Leaves by Isopycnic Centrifugation. Current Protocols in Cell Biology. 40:3.30:3.30.1–3.30.14.

Author Information

  1. Laboratoire de Physiologie Cellulaire Végétale, CNRS and Université Joseph Fourier, Grenoble, France

Publication History

  1. Published Online: 1 SEP 2008
  2. Published Print: SEP 2008

Abstract

Chloroplasts are plant-specific organelles. They are the site of photosynthesis but also of many other essential metabolic pathways, such as syntheses of amino acids, vitamins, lipids, and pigments. This unit describes the isolation and purification of chloroplasts from Arabidopsis and spinach leaves. Differential centrifugation is first used to obtain a suspension enriched in chloroplasts (crude chloroplasts extract). In a second step, Percoll density gradient centrifugation is used to recover pure and intact chloroplasts. The Basic Protocol describes the purification of chloroplasts from Arabidopsis leaves. This small flowering plant is now widely used as a model organism in plant biology as it offers important advantages for basic research in genetics and molecular biology. The Alternate Protocol describes the purification of chloroplasts from spinach leaves. Spinach, easily available all through the year, remains a model of choice for the large-scale preparation of pure chloroplasts with a high degree of intactness. Curr. Protoc. Cell Biol. 40:3.30.1-3.30.14. © 2008 by John Wiley & Sons, Inc.

Keywords:

  • chloroplast;
  • Arabidopsis;
  • spinach;
  • purification;
  • leaves