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Nebularine Affects Plant Growth and Development but does not Interfere with Cytokinin Signaling

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Abstract

Nebularine is known for its high cytotoxicity in animals, whereas in plants it was originally believed to be an anticytokinin. In this study we show that in classical cytokinin bioassays, nebularine antagonized cytokinin function in senescence and callus biotests but not in the Amaranthus bioassay. Nebularine reversed the inhibitory effect of cytokinin on lateral root formation in Arabidopsis seedlings, and when applied alone caused increased lateral root formation and shortening of the main root. Systematic spraying of Arabidopsis plants with nebularine led to yellowing and formation of purple pigments, local drying, and withering, although younger plants showed a greater resilience. Comparison of nebularine cytotoxicity in plant and animal cells showed that the growth of tobacco BY-2 cells was inhibited with only about tenfold lower efficacy than mammalian cell lines. Most importantly, direct binding assay with Arabidopsis cytokinin receptors AHK3 and CRE1/AHK4 showed that nebularine did not compete for binding with the natural cytokinin trans-zeatin. Although nebularine reduced cytokinin-induced expression of the cytokinin reporter ARR5:GUS in planta, the same effect was observed for DR5:GUS, an auxin reporter gene. Taken together, the results indicate that the mode of action of nebularine does not involve cytokinin signaling and that the anticytokinin-like effect is rather a consequence of the inhibition of various processes as described for animal systems.

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Acknowledgments

The authors thank Tomáš Pospíšil for synthesis of nebularine and Dr. Vladimír Kryštof for performing the human cell viability assay and determing the competition of nebularine toward ATP on CDK2 and ABL. We are greatful to Prof. David Morris for helpful suggestions and critical reading of the manuscript. This work was supported in part by the grants MSM 6198959216 from the Ministry of Education, Youth and Sports Czech Republic, and 522/06/0022, 522/08/H003, and 522/07/P197 from the Czech Science Foundation.

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Authors and Affiliations

  1. Department of Biochemistry, Faculty of Science, Palacký University, Šlechtitelů 11, 78371, Olomouc, Czech Republic

    Hana Pospíšilová & Ivo Frébort

  2. Laboratory of Growth Regulators, Palacký University/Institute of Experimental Botany AS CR, Šlechtitelů 11, 78371, Olomouc, Czech Republic

    Jaroslav Nisler & Lukáš Spíchal

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  1. Hana Pospíšilová
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Correspondence to Hana Pospíšilová.

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Pospíšilová, H., Nisler, J., Spíchal, L. et al. Nebularine Affects Plant Growth and Development but does not Interfere with Cytokinin Signaling. J Plant Growth Regul 28, 321–330 (2009). https://doi.org/10.1007/s00344-009-9096-5

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  • DOI: https://doi.org/10.1007/s00344-009-9096-5

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