Plant Physiology 75:670-674 (1984)
© 1984 American Society of Plant Biologists
Articles
Purification of Peroxisomes and Mitochondria from Spinach Leaf by Percoll Gradient Centrifugation 1
Jean-Paul Schwitzguebel and
Paul-André Siegenthaler
Laboratoire de Physiologie Végétale, Université de Neuchâtel, Chemin de Chantemerle 20, CH-2000 Neuchâtel, Switzerland
A procedure was developed to purify simultaneously peroxisomes and mitochondria from spinach (Spinacia oleracea L.) leaf under isoosmotic and low viscosity conditions. This method involved differential centrifugation and density gradient centrifugation on four layers of Percoll. Chlorophyll-free preparations of highly intact and active organelles were obtained and cross-contamination was negligible. Both organelles were stable for several hours, even if they remained in Percoll. Purified mitochondria were able to carry out the oxidation of different substrates with excellent respiratory control and ADP:O ratios. The method described in the present work was also suitable to purify mitochondria and peroxisomes from potato (Solanum tuberosum L.) tubers.
1 Supported by the Swiss National Science Foundation (Grant 3.661.0.80 to P. A. S.) and in part by the Emil-Barrel Foundation.
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