Skip to main content
PMC home page
Plant Physiology logoLink to Plant Physiology
. 1985 Apr;77(4):1010–1012. doi: 10.1104/pp.77.4.1010

Isolation of Mitochondria from Soybean Leaves on Discontinuous Percoll Gradients 1

Terry C Hrubec 1,2, J Michael Robinson 1,2, Robert P Donaldson 1,2
PMCID: PMC1064649  PMID: 16664137

Abstract

A technique to isolate mitochondria from chamber-grown soybeans (Glycine max cv Williams) was developed. The mitochondria were isolated by centrifugation on discontinuous Percoll gradients which yielded a sharp band of mitochondria contaminated by only 4% of the total chlorophyll in the gradient. Contamination by peroxisomes was also slight. The isolated mitochondria oxidized malate plus glutamate, NADH, and malate with respiratory control. They also showed cyanide-insensitive, alternative pathway activity which was inhibited by salicylhydroxamic acid.

Full text

PDF
1010

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Albertsson P. A., Andersson B., Larsson C., Akerlund H. E. Phase partition--a method for purification and analysis of cell organelles and membrane vesicles. Methods Biochem Anal. 1982;28:115–150. doi: 10.1002/9780470110485.ch2. [DOI] [PubMed] [Google Scholar]
  2. Azcón-Bieto J., Lambers H., Day D. A. Effect of photosynthesis and carbohydrate status on respiratory rates and the involvement of the alternative pathway in leaf respiration. Plant Physiol. 1983 Jul;72(3):598–603. doi: 10.1104/pp.72.3.598. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bergman A., Gardeström P., Ericson I. Method to Obtain a Chlorophyll-free Preparation of Intact Mitochondria from Spinach Leaves. Plant Physiol. 1980 Sep;66(3):442–445. doi: 10.1104/pp.66.3.442. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  5. Gardeström P., Edwards G. E. Isolation of Mitochondria from Leaf Tissue of Panicum miliaceum, a NAD-Malic Enzyme Type C(4) Plant. Plant Physiol. 1983 Jan;71(1):24–29. doi: 10.1104/pp.71.1.24. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Jackson C., Dench J. E., Hall D. O., Moore A. L. Separation of mitochondria from contaminating subcellular structures utilizing silica sol gradient centrifugation. Plant Physiol. 1979 Jul;64(1):150–153. doi: 10.1104/pp.64.1.150. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Liang Z., Yu C., Huang A. H. Isolation of spinach leaf peroxisomes in 0.25 molar sucrose solution by percoll density gradient centrifugation. Plant Physiol. 1982 Oct;70(4):1210–1212. doi: 10.1104/pp.70.4.1210. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Nash D., Wiskich J. T. Properties of substantially chlorophyll-free pea leaf mitochondria prepared by sucrose density gradient separation. Plant Physiol. 1983 Mar;71(3):627–634. doi: 10.1104/pp.71.3.627. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Nishimura M., Douce R., Akazawa T. Isolation and characterization of metabolically competent mitochondria from spinach leaf protoplasts. Plant Physiol. 1982 Apr;69(4):916–920. doi: 10.1104/pp.69.4.916. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. SIMON E. W. The effect of digitonin on the cytochrome c oxidase activity of plant mitochondria. Biochem J. 1958 May;69(1):67–74. doi: 10.1042/bj0690067. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Walker G. H., Oliver D. J., Sarojini G. Simultaneous oxidation of glycine and malate by pea leaf mitochondria. Plant Physiol. 1982 Nov;70(5):1465–1469. doi: 10.1104/pp.70.5.1465. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Plant Physiology are provided here courtesy of Oxford University Press

RESOURCES