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Preparation of leaf mitochondria from Arabidopsis thaliana
Author(s) -
Keech Olivier,
Dizengremel Pierre,
Gardeström Per
Publication year - 2005
Publication title -
physiologia plantarum
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.351
H-Index - 146
eISSN - 1399-3054
pISSN - 0031-9317
DOI - 10.1111/j.1399-3054.2005.00521.x
Subject(s) - percoll , arabidopsis thaliana , mitochondrion , organelle , biology , alternative oxidase , differential centrifugation , biochemistry , chloroplast , centrifugation , gene , mutant
Arabidopsis thaliana is, perhaps, the most important model species in modern plant biology. However, the isolation of organelles from leaves of this plant has been difficult. Here, we present two different protocols for the isolation of mitochondria, yielding either highly functional crude mitochondria or highly purified mitochondria. The crude mitochondria were well coupled with the substrates tested (malate + glutamate, glycine and NADH), exhibiting respiratory control ratios of 2.1–3.9. Purified mitochondria with very low levels of chlorophyll contamination were obtained by Percoll gradient centrifugation, yielding 1.2 mg of mitochondrial protein from 50 g of leaves.