Open AccessPurification of Peroxisomes and Mitochondria from Spinach Leaf by Percoll Gradient Centrifugation
Author(s) -
JeanPaul Schwitzguébel,
PaulAndré Siegenthaler
Publication year - 1984
Publication title -
plant physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.554
H-Index - 312
eISSN - 1532-2548
pISSN - 0032-0889
DOI - 10.1104/pp.75.3.670
Subject(s) - percoll , spinach , spinacia , peroxisome , differential centrifugation , organelle , centrifugation , biology , photorespiration , mitochondrion , microbody , biochemistry , chromatography , chloroplast , chemistry , enzyme , gene
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.