Subcellular Fractionation of Plant Tissues

Isolation of Chloroplasts and Mitochondria from Leaves
  • Alyson K. Tobin
Part of the Methods in Molecular Biology™ book series (MIMB, volume 59)


The successful isolation of intact, functional organelles from plant tissue is fraught with difficulties. Leaves are often covered in waxy cuticles, and frequently contain silica (as in grasses) and toxic components, such as phenolics, proteolytic enzymes, and high concentrations of acids and salts in the vacuole. In addition, all higher plants have one major barrier in common, the presence of a rigid, cellulose cell wall, which has to be broken in order to release the organelles. Mechanical isolation methods, i.e., where leaf material is macerated in a mechanical homogenizer, are likely to succeed only for a limited number of species, e.g., pea and spinach, where the leaves do not contain large amounts of tough, thickened tissue. Otherwise, the prolonged homogenization required to release significant numbers of organelles from tough leaves, e.g., of grasses, such as wheat or barley, results in most of them being broken and inactive. For this reason, the only viable method for obtaining good-quality organelles from species such as these is to isolate them from protoplasts. Although chloroplasts can be successfully isolated from protoplasts, the yield of mitochondria is so small that, unless very large-scale protoplast preparations are employed, the technique is generally inappropriate for mitochondrial isolation. This means that there are many plant species from whose leaves it has proven impossible to isolate good-quality mitochondria using existing techniques.


Oxygen Electrode Leaf Section Respiratory Control Ratio Chloroplast Intactness Mitochondrial Isolation 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


  1. 1.
    Lilley, R. McC., Fitzgerald, M. P., Rienits, K. G., and Walker, D. A. (1975) Criteria of intactness and the photosynthetic activity of spinach chloroplast preparations. New Phytol. 75, 1–10.CrossRefGoogle Scholar
  2. 2.
    Foster, J. G., Edwards, G. E., and Winter, K. (1982) Changes in levels of Phosphoenol pyruvate carboxylase with induction of Crassulacean acid metabolism in Mesembryanthemum crystallinum L Plant Cell Physiol. 23, 585–594.Google Scholar
  3. 3.
    Cooper, T. G. and Beevers, H. (1969) Mitochondria and glyoxysomes from castor bean endosperm Enzyme constituents and catalytic capacity. J. Biol. Chem. 244, 3507–3513.PubMedGoogle Scholar
  4. 4.
    Tolbert, N. E. (1981) Microbodies-peroxisomes and glyoxysomes. Annu. Rev. Plant Physiol. 26, 45–73.Google Scholar
  5. 5.
    Behrends, W., Rausch, U., Loffler, H. G., and Kindl, H (1982) Purification of glycolate oxidase from greening cucumber cotyledons. Planta 156, 566–571.CrossRefGoogle Scholar

Copyright information

© Humana Press Inc. 1996

Authors and Affiliations

  • Alyson K. Tobin
    • 1
  1. 1.Plant Sciences Laboratory, School of Biological and Medical SciencesUniversity of St. AndrewsUK

Personalised recommendations