Mitochondria pp 125-136 | Cite as

Isolation of Intact, Functional Mitochondria From the Model Plant Arabidopsis thaliana

  • Lee J. Sweetlove
  • Nicolas L. Taylor
  • Christopher J. Leaver
Part of the Methods in Molecular Biology™ book series (MIMB, volume 372)


The ability to isolate intact, functional mitochondria from plant tissues is a key technique in the study of the genome, proteome, and metabolic function of the plant mitochondrion. Traditionally, mitochondrial plant researchers have turned to specific plant systems and organs (such as potato tubers and pea shoots) from which mitochondria are readily isolated in large quantities. However, increasingly, research is focused on a small number of model species, and there is a need to adapt existing protocols to allow the isolation of mitochondria from these model species. Arguably, the most important of these is Arabidopsis thaliana, for which a formidable array of genetic resources is available. However, because of its relatively small size and the absence of large heterotrophic organs, Arabidopsis is a challenging plant from which to isolate mitochondria. Here, we present two methods for isolating mitochondria from Arabidopsis, either from heterotrophic cell suspension cultures or from hydroponic seedling cultures. We also present details of commonly used assays to assess the physical and functional integrity of the isolated organelles.

Key Words

Arabidopsis cell suspension culture leaf mitochondria isolation outer mitochondrial membrane integrity respiratory control ratio 


  1. 1.
    The Arabidopsis Genome Initiative. (2000) Analysis of the genome sequence of the flowering plant Arabidopsis thaliana. Nature 408, 796–815.CrossRefGoogle Scholar
  2. 2.
    Sessions, A., Burke, E., Presting, G., et al. (2002) A high-throughput Arabidopsis reverse genetics system. Plant Cell 14, 2985–2994.PubMedCrossRefGoogle Scholar
  3. 3.
    Millar, A. H. (2004) Location, location, location: surveying the intracellular real estate through proteomics in plants. Funct. Plant Biol. 31, 563–571.CrossRefGoogle Scholar
  4. 4.
    Gibon, Y., Blaesing, O. E., Hannemann, J., et al. (2004) A robot-based platform to measure multiple enzyme activities in Arabidopsis using a set of cycling assays: comparison of changes of enzyme activities and transcript levels during diurnal cycles and in prolonged darkness. Plant Cell 16, 3304–3325.PubMedCrossRefGoogle Scholar
  5. 5.
    Millar, A. H., Sweetlove, L. J., Giege, P., and Leaver, C. J. (2001) Analysis of the Arabidopsis mitochondrial proteome. Plant Physiol. 127, 1711–1727.PubMedCrossRefGoogle Scholar
  6. 6.
    Day, D. A., Neuberger, M., and Douce, R. (1985) Biochemical characterization of chlorophyll-free mitochondria from pea leaves. Austr. J. Plant Physiol. 12, 219–228.CrossRefGoogle Scholar
  7. 7.
    Millar, A. H. M., Liddell, A., and Leaver, C. J. (2001) Isolation and subfractionation of mitochondria from plants. Methods Cell Biol. 65, 53–74.PubMedCrossRefGoogle Scholar
  8. 8.
    May, M., and Leaver, C. (1993) Oxidative stimulation of glutathione synthesis in Arabidopsis thaliana suspension cultures. Plant Physiol. 103, 621–627.PubMedGoogle Scholar
  9. 9.
    Xiang, C., and Oliver, D. J. (1998) Glutathione metabolic genes coordinately respond to heavy metals and jasmonic acid in Arabidopsis. Plant Cell 10, 1539–1550.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2007

Authors and Affiliations

  • Lee J. Sweetlove
    • 1
  • Nicolas L. Taylor
    • 1
  • Christopher J. Leaver
    • 1
  1. 1.Department of Plant SciencesUniversity of OxfordOxfordUK

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