Abstract
Primary modes of action of cytokinins have been thought to involve stimulation of the electrogenic H+ pump and-or opening of plasmamembrane Ca2+ channels. In order to test these hypotheses, rapid changes in membrane transport in response to cytokinin application were studied in heterotrophic suspension-cultured callus of soybean (Glycine max (L.) Merr.) using electrophysiological techniques. Kinetin (N6-furfurylaminopurine; 2 μM) elicited membrane hyperpolarization of 13±1 mV. This effect occurred even at membrane poteintials more negative than the most negative ionic equilibrium potential, and therefore might have resulted either from stimulation of the electrogenic pump, or from closure of ionic channels. The former mechanism of action appears most likely because (i) kinetin-induced membrane hyperpolarization is not accompanied by a significant change in plasma-membrane resistivity and (ii) hyperpolarization is abolished by cyanide, which inhibits electrogenic pump activity by depletion of cellular ATP.
Electrogenic pumping is also activated by two other cytokinins: N6-(benzyl)adenine and trans-zeatin. However, it is unlikely that the hormonal effect on electrogenesis is directly related to transduction of the cytokinin signal, for the following reasons: (i) hormonally inactive, but chemically related compounds (cis-zeatin, adenine) also elicited membrane hyperpolarization; (ii) hormonally active, N9-substituted cytokinins failed to stimulate electrogenesis; (iii) the chemically unrelated cytokinin N,N′-diphenylurea also failed to stimulate electrogenesis.
The results imply that the kinetin effect on electrogenic pumping is related to adenine, or its metabolism, and not hormonal action. Adenine was absorbed by soybean cells, but not in sufficient quantities to have a significant effect on adeninenucleotide pools. It appears likely that the control of electrogenesis requires either the presence of a purine free base (i.e. no substituents at the N9 position) or phosphoribosylation of the free base. No evidence was found for cytokinin-induced Ca2+-channel opening, though it is argued that such an event might be physiologically relevant, yet undetectable with the methods employed. It is essential that future studies on cytokinin signal transduction — especially as they relate to membrane transport — take into account the possibility that metabolic effects unrelated to hormone action are dominant.
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Abbreviations
- bzl6Ade:
-
N6 (benzyl)adenine
- SRB:
-
Soybean Recording Buffer
- Vm :
-
membrane potential
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Parsons, A., Blackford, S. & Sanders, D. Kinetin-induced stimulation of electrogenic pumping in soybean suspension cultures is unrelated to signal transduction. Planta 178, 215–222 (1989). https://doi.org/10.1007/BF00393197
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DOI: https://doi.org/10.1007/BF00393197