, Volume 255, Issue 5, pp 1581–1594 | Cite as

Study of cytokinin transport from shoots to roots of wheat plants is informed by a novel method of differential localization of free cytokinin bases or their ribosylated forms by means of their specific fixation

  • Stanislav Yu. Veselov
  • Leila N. Timergalina
  • Guzel R. Akhiyarova
  • Guzel R. Kudoyarova
  • Alla V. Korobova
  • Igor Ivanov
  • Tatiana N. Arkhipova
  • Els Prinsen
New Methods in Cell Biology


The aim of the present report was to demonstrate how a novel approach for immunohistochemical localization of cytokinins in the leaf and particularly in the phloem may complement to the study of their long-distance transport. Different procedures of fixation were used to conjugate either cytokinin bases or their ribosides to proteins of cytoplasm to enable visualization and differential localization of these cytokinins in the leaf cells of wheat plants. In parallel to immunolocalization of cytokinins in the leaf cells, we immunoassayed distribution of free bases of cytokinins, their nucleotides and ribosides between roots and shoots of wheat plants as well as their presence in phloem sap after incubation of leaves in a solution supplemented with either trans-zeatin or isopentenyladenine. The obtained data show ribosylation of the zeatin applied to the leaves and its elevated level in the phloem sap supported by in vivo localization showing the presence of ribosylated forms of zeatin in leaf vessels. This suggests that conversion of zeatin to its riboside is important for the shoot-to-root transport of zeatin-type cytokinins in wheat. Exogenous isopentenyladenine was not modified, but diffused from the leaves as free base. These metabolic differences may not be universal and may depend on the plant species and age. Although the measurements of cytokinins in the phloem sap and root tissue is the most defining for determining cytokinin transport, study of immunolocalization of either free cytokinin bases or their ribosylated forms may be a valuable source of information for predicting their transport in the phloem and to the roots.


Triticum durum Zeatin Isopentenyladenosine Cytokinin ribosides Immunolocalization Phloem 



Carbonyl cyanide m- chlorophenylhydrazone


Equilibrium nucleoside transporters






Isopentenyl nucleotide


Phosphate buffer containing gelatin and Tween


Purine permease


Thin-layer chromatography




Zeatin riboside


Zeatin nucleotide



The work was partially supported by Russian Foundation for Basic Research (grant 15-04-04750).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.


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Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2018

Authors and Affiliations

  • Stanislav Yu. Veselov
    • 1
  • Leila N. Timergalina
    • 2
  • Guzel R. Akhiyarova
    • 2
  • Guzel R. Kudoyarova
    • 2
  • Alla V. Korobova
    • 2
  • Igor Ivanov
    • 2
  • Tatiana N. Arkhipova
    • 2
  • Els Prinsen
    • 3
  1. 1.Bashkir State UniversityUfaRussia
  2. 2.Ufa Institute of Biology, Ufa Research CentreRussian Academy of SciencesUfaRussia
  3. 3.Department of BiologyUniversity of AntwerpenAntwerpenBelgium

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