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Biological activity of cytokinins derived from Ortho- and Meta-Hydroxybenzyladenine

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Abstract

To determine the structure-activity relationships of natural aromatic cytokinins, the activity of 6-benzylaminopurine (BAP) and its hydroxylated derivatives was compared in three bioassays based on stimulation of tobacco callus growth, retention of chlorophyll in excised wheat leaves, and dark induction of betacyanin synthesis in Amaranthus cotyledons. The aromatic cytokinins 6-(2-hydroxybenzylamino)purine (ortho-topolin) and 6-(3-hydroxybenzylamino)purine (meta-topolin), their 9-ribosides and 9-glucosides, were synthesized by the condensation of 6-chloropurine and its 9-glycosides with the appropriate hydroxybenzylamine. The activity of free bases, 9-ribosides and 9-glucosides was compared with that of BAP, trans-zeatin and their 9-glycosides. Hydroxylation of the benzyl ring in the meta position increased the activity of BAP and its riboside in tobacco callus and chlorophyll retention bioassays, whereas ortho-hydroxylation decreased the activity. In contrast, in the Amaranthus bioassay meta-hydroxylation of BAP substantially decreased its activity. Ribosylation at position 9 had no significant effect on the activity of zeatin, BAP and both topolins. The activity of 9-glucosides of all cytokinins tested was near zero. The biological activity of meta-topolin and its riboside is comparable to that of the most active isoprenoid cytokinin, zeatin, in tobacco callus growth and senescence bioassays. The results establish the existence of a family of endogenous aromatic cytokinins centered around the highly active compound, meta-topolin. We also report here an improved chlorophyll retention bioassay based on incubation of 2.5 cm long detached wheat leaf segments in microtiter plate wells containing 150 µl of cytokinin solution. The consumption of cytokinin to be tested is 0.1 µmol per assay only. The amount as small as 1.5 pmol of substance can be estimated using this biotest.

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Authors and Affiliations

  1. Laboratory of Growth Regulators, Faculty of Natural Sciences of the Palacký, University & Institute of Experimental Botany ASCR, Šlechtitelů 11, 783 71, Olomouc, Czech Republic; telephone

    Jan Holub

  2. Isotope Laboratory, Institute of Experimental Botany ASCR, Vídeňská 1083, 142 20, Praha 4, Czech Republic

    Jan Hanuš

  3. Department of Plant Sciences, University of Cambridge, Downing Street, Cambridge, CB2 3EA, England

    David E. Hanke

  4. Laboratory of Growth Regulators, Faculty of Natural Sciences of the Palacký, University & Institute of Experimental Botany ASCR, Šlechtitelů 11, 783, 71 Olomouc, Czech Republic

    Miroslav Strnad

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  1. Jan Holub
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  2. Jan Hanuš
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  3. David E. Hanke
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  4. Miroslav Strnad
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Holub, J., Hanuš, J., Hanke, D.E. et al. Biological activity of cytokinins derived from Ortho- and Meta-Hydroxybenzyladenine. Plant Growth Regulation 26, 109–115 (1998). https://doi.org/10.1023/A:1006192619432

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