An Improved Method to Isolate Mitochondrial RNA from Green Plant Tissue

  • Fei Ye
  • Ralf Reski
Part of the Springer Protocols Handbooks book series (SPH)


In plant cells, mitochondrial RNA (mtRNA) constitutes about only 1% of the total RNA. From this, most RNAs are ribosomal RNAs. Thus, isolation of high purified mtRNA is necessary, not only for construction of a mitochondrial cDNA library, but also for the analysis of plant mitochondrial transcription. Several methods have been frequently used for isolation of plant mtRNA (1, 2, 3). However, these mtRNA preparations may be heavily contaminated by chloroplast RNA (cpRNA), especially when mtRNA is isolated from green leaves (1,4). It is believed that the cpRNA sticks to the mitochondrial membrane and therefore persists after gradient purification of mitochondria. Although micrococcal nuclease would be the enzyme to remove the non-mtRNA from mitochondrial membranes prior to lysis of mitochondria, treatments with micrococcal nuclease for the mtRNA isolation from green leaves have not been effective (4).


Potassium Acetate Micrococcal Nuclease Guanidinium Thiocyanate Sucrose Gradient Centrifugation Guanidine Thiocyanate 
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  1. 1.
    Stern, D. B. and Newton, K. J. (1986) Isolation of plant mitochondrial RNA. Methods Enzymol. 118, 488–496.PubMedCrossRefGoogle Scholar
  2. 2.
    Schuster, A. M. and Sisco, P. H. (1986) Isolation and characterization of single-stranded and double-stranded RNAs in mitochondria. Methods Enzymol. 118, 488–496.CrossRefGoogle Scholar
  3. 3.
    Schuster, W., Hiesel, R., Wissinger, B., Schobel, W., and Brennicke, A. (1988) Isolation and analysis of plant mitochondria and their genomes, in Plant Molecular Biology (Shaw, C. H., ed.), pp. 79–102, IRL Press, Oxford, UK, and Washington, DC.Google Scholar
  4. 4.
    Makaroff, C. A. and Palmer, J. D. (1987) Extensive mitochondrial specific transcription of the Brassica campestris mitochondrial genome. Nucleic Acids Res. 5, 5141–5156.CrossRefGoogle Scholar
  5. 5.
    Han, J. H., Stratowa, C., and Rutter, W. J. (1987) Isolation of full-length putative rat lysophospholipase cDNA using improved methods for mRNA isolation and cDNA cloning. Biochemistry 26, 1617–1625.PubMedCrossRefGoogle Scholar
  6. 6.
    Ye, F., Albaum, M., Markmann-Mulisch, U., and Abel, W. O. (1993) Improved method for the isolation of mitochondrial RNA from green leaves. BioTechniques 14, 184.PubMedGoogle Scholar
  7. 7.
    Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Lab. Press, Cold Spring Harbor, NY.Google Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2000

Authors and Affiliations

  • Fei Ye
    • 1
  • Ralf Reski
    • 2
    • 3
  1. 1.Department of BiologyMassachusetts Institute of TechnologyCambridge
  2. 2.Freiburg University Institute Biology IIFrieburg/BreisguaGermany
  3. 3.OR Institute for General BotanyHamburgGermany

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