Skip to main content
SpringerLink
Log in

Cytokinins

Extraction, Separation, and Analysis

  • Protocol
Plant Hormone Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 141))

Abstrect

Analysis of cytokinins in plant tissues has historically been laborious and time consuming because of the extremely low levels and diverse molecular structures of the cytokinins. The initial anticipation that cytokinins could be quantified by radioimmunoassay of crude plant extracts (1,2) was, unfortunately, not justified. Detailed analysis requires the sample to be well purified and the individual cytokinins to be separated, so that not only is interference in the assay avoided, but also problems associated with differential crossreactivity of individual cytokinins with the antibodies in the immunoassay (3). The (sometimes subtle) differences in the chemical properties of the cytokinin free bases, ribosides, nucleotides, and the O- and N-glucosides enables the ready separation and quantification of the majority of the cytokinins.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Weiler, E. W. (1979) Radioimmunoassay for the determination of free and conjugated abcisicacid.Planta 144, 255–263.

    Article  CAS  Google Scholar 

  2. Weiler, E. W. (1980) Radioimmunoassays for trans-zeatin and related cytokinins. Planta 149, 155–162.

    Article  CAS  Google Scholar 

  3. Hedden, P. (1993) Modern methods for the quantitative analysis of plant hormones. Annu. Rev. Plant Physiol. Plant Mol. Bol. 44, 107–129.

    Article  CAS  Google Scholar 

  4. Jameson, P. E. (1994) Cytokinin metabolism and compartmentation,in Cytokinins: Chemistry, Activity and Function (Mok, D. W. S. and Mok, M. C., eds.), CRC, Boca Raton, FL, pp. 113–128.

    Google Scholar 

  5. Tokota, T., Murofushi, N., and Takahashi, N. (1985) Extraction, purification,and identification, in Hormonal Regulation of Development I: Molecular Aspects of Plant Hormones (MacMillan, J., ed.),Springer-Verlag, Berlin, pp. 113–201.

    Google Scholar 

  6. Parker, C. W. and Letham, D. S. (1973) Regulators of cell division in plant tissues XVI. Metabolism of zeatin by radish cotyledons and hypocotyls. Planta 114, 199–218.

    Article  CAS  Google Scholar 

  7. Morris, J. W., Doumas, P., Morris, R. O., and Zaerr J. B. (1990) Cytokinins in vegetative and reproductive buds of Pseudotsuga menziesii. Plant Physiol. 93, 67–71.

    Article  PubMed  CAS  Google Scholar 

  8. Dietrich, J. T., Kaminek, M., Blevins, D. G., Reinbott, T. M., and Morris, R. O. (1995) Changes in cytokinins and cytokinin oxidase activity in developing maize kernels and the effects of exogenous cytokinin on kernel development. Plant Physiol. Biochem. 33, 327–336.

    CAS  Google Scholar 

  9. Bieleski, R. L. (1964) The problem of halting enzyme action when extracting plant tissues. Anal. Biochem. 9, 431–442.

    Article  PubMed  CAS  Google Scholar 

  10. Horgan, R. and Scott, I. M. (1987) Cytokinins, in Principles and Practices of Plant Hormone Analysis (Rivier, L. and Crozier, A., eds.),Academic, New York, pp. 303–365.

    Google Scholar 

  11. Summons, R. E., Palni, L. M. S., and Letham, D. S. (1983) Determination of intact zeatin nucleotide by direct chemical ionisation mass spectrometry. FEBS Lett. 151, 122–126.

    Article  CAS  Google Scholar 

  12. Meilan, R., Horgan, R., Heald, J. K., LaMotte, C. E., and Schultz, R. C. (1993) Identification of cytokinins in red pine seedlings. Plant Growth Regul. 13, 169–178.

    Article  CAS  Google Scholar 

  13. von Schwartzenberg, K., Bonnet-Masimbert, M., and Doumas, P. (1994) Isolation of two cytokinin metabolites from the rhizosphere of Norway spruce seedlings (Picea abies L. Karst.). Plant Growth Regul. 15, 117–124.

    Article  CAS  Google Scholar 

  14. Jameson, P. E., Letham, D. S., Zhang, R., Parker, C. W., and Badenoch-Jones, J.(1987) Cytokinin translocation and metabolism in Lupinspecies. I Zeatin riboside introduced into the xylem at the base of Lupinus angustifolius stems.Aust. J. Plant Physiol. 14, 695–718.

    Article  CAS  Google Scholar 

  15. Wang, J., Letham, D. S., Taverner, E., Badenoch-Jones, J., and Hocart, C. H. (1995) A procedure for quantification of cytokinins as free bases involving scintillation proximity immunoassay. Physiol.Plant. 95, 91–98.

    Article  CAS  Google Scholar 

  16. Palmer, M. V., Horgan, R., and Wareing, P. F. (1981) Cytokinin metabolism in Phaseolus vulgaris L.: Identification of endogenous cytokinins and metabolism of [8-14C] dihydrozeatin in stems of decapitated plants. Planta 153, 297–302.

    Article  CAS  Google Scholar 

  17. Muller, P. and Hilgenberg, W. (1986) Isomers of zeatin and zeatin riboside in clubroot tissue: evidence for trans-zeatin biosynthesis by Plasmodiophora brassicae. Physiol. Plant. 66, 245–250.

    Article  Google Scholar 

  18. Cappeiello, P. E. and King, G. J. (1990) Determination of zeatin and zeatin riboside in plant tissue by solid-phase extraction and ion-exchange chromatography. J. Chromatog. 504, 197–201.

    Article  Google Scholar 

  19. Biddington, N. L. and Thomas, T. H. (1976) Effect of pH on the elution of cytokinins from polyvinylpyrrolidone columns. J. Chromatog. 121, 107–109.

    Article  CAS  Google Scholar 

  20. Mousdale, D. M. A. and Knee, M. (1979) Poly-N-vinylpyrrolidone column chromatography of plant hormones with methanol as eluent.J. Chromatog. 177, 398–400.

    Article  CAS  Google Scholar 

  21. Parker, C. W. and Letham, D. S. (1974) Regulators of cell division in plant tissues XVII. Metabolism of zeatin in Zea mays seedlings.Planta 115, 337–344.

    Article  CAS  Google Scholar 

  22. Badenoch-Jones, J., Letham, D. S., Parker, C. W., and Rolfe, B. G. (1984) Quantitation of cytokinins in biological samples using antibodies against zeatin riboside. Plant Physiol. 75, 1117–1125.

    Article  PubMed  CAS  Google Scholar 

  23. Guinn, G. and Brummett, D. L. (1990) Solid phase extraction of cytokinins from aqueous solutions with C18 cartridges and their use in a rapid purification procedure. Plant Growth Regul. 9, 305–314.

    Article  CAS  Google Scholar 

  24. MacDonald, E. M. S. and Morris, R. O. (1985) Isolation of cytokinins by immunoaffinity chromatography and analysis by high-performance liquid chromatography-radioimmunoassay. Methods Enzymol. 110, 347–358.

    Article  CAS  Google Scholar 

  25. Morris, R. O., Zaerr, J. B., and Chapman, R. W. (1976) Trace enrichment of cytokinins from Douglas-fir xylem exudate. Planta 131, 271–274.

    Article  CAS  Google Scholar 

  26. Jameson, P. E. and Morris, R. O. (1989) Zeatin-like cytokinins in yeast: Detection by immunological methods. J. Plant Physiol. 135, 385–390.

    CAS  Google Scholar 

  27. Hocart, C. H., Letham, D. S., and Parker, C. W. (1990) Metabolism and translocation of exogenous zeatin riboside in germinating seeds and seedlings of Zea mays. J. Exp. Bot. 41, 1517–1524.

    Article  CAS  Google Scholar 

  28. Lewis, D. H., Burge, G. K., Schmierer, D. M., and Jameson, P. E.(1996) Cytokinins and fruit development in the kiwifruit (Actinidia deliciosa). I. Changes during fruit development. Physiol. Plant. 98, 179–186.

    Article  CAS  Google Scholar 

  29. Banowetz, G. M. (1994) Immunoanalysis of cytokinins, in Cytokinins: Chemistry, Activity and Function (Mok, D. W. S. and Mok, M. C., eds.), CRC, Boca Raton, FL, pp. 305–316.

    Google Scholar 

  30. Eason, J. R., Morris, R. O., and Jameson, P. E. (1996) The relationship between virulence and cytokinin production by Rhodococcus fascians (Tilford 1936) Goodfellow 1984. Plant Pathol. 45, 323–331.

    Article  CAS  Google Scholar 

  31. Pengelly, W. L. (1986) Validation of immunoassays, in Plant Growth Substances 1985 (Bopp, M., ed.), Springer-Verlag, Berlin, pp. 35–43.

    Google Scholar 

  32. Prinsen, E., Redig, P., Van Dongen, W., Esmans, E. L., and Van Onckelen, H. A. (1995) Quantitative analysis of cytokinins by electrospray tandem mass spectrometry. Rapid Commun. Mass Spectrom. 9, 948–953.

    Article  CAS  Google Scholar 

  33. MacDonald, E. M., Akiyoshi, D. E., and Morris, R. O. (1981) Combined high-performance liquid chromatography-radioimmunoassay for cytokinins. J. Chromatog. 214, 101–109.

    Article  CAS  Google Scholar 

  34. Trione, E. J., Krygier, B. B., Banowetz, G. M., and Kathrein, J. M. (1985) The development of monoclonal antibodies against the cytokinin zeatin riboside. J. Plant Growth Regul. 4, 101–109.

    Article  CAS  Google Scholar 

  35. Trione, E. J., Krygier, B. B., Kathrein, J. M., Banowetz, G. M., and Sayavedra-Soto, L. A. (1987) Monoclonal antibodies against the plant cytokinin isopentenyl adenosine. Physiol. Plant. 70, 467–472.

    Article  CAS  Google Scholar 

  36. Dawson, R. M. C., Elliott, D. C., Elliott, W. H., and Jones, K. M. (1986) Ion Exchange, gel filtration, and affinity chromatography media. In: Data for Biochemical Research, 3rd ed., Clarendon Press, Oxford, pp. 508–513.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institute of Molecular BioSciences, College of Sciences, Massey University, Palmerston North, New Zealand

    Paula E. Jameson

  2. The Horticulture and Food Research Institute of New Zealand, Palmerston North, New Zealand

    Huaibi Zhang

  3. New Zealand Institute for Coop and Food Research, Palmerston North, New Zealand

    David H. Lewis

Authors
  1. Paula E. Jameson
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Huaibi Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. David H. Lewis
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. School of Biological Sciences, University of Nottingham, UK

    Gregory A. Tucker  & Jeremy A. Roberts  & 

Rights and permissions

Reprints and permissions

Copyright information

© 2000 Humana Press Inc.

About this protocol

Cite this protocol

Jameson, P.E., Zhang, H., Lewis, D.H. (2000). Cytokinins. In: Tucker, G.A., Roberts, J.A. (eds) Plant Hormone Protocols. Methods in Molecular Biology™, vol 141. Humana Press. https://doi.org/10.1385/1-59259-067-5:101

Download citation

  • DOI: https://doi.org/10.1385/1-59259-067-5:101

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-577-5

  • Online ISBN: 978-1-59259-067-4

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation