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Immunodetection and Identification of N6-(o-Hydroxybenzylamino)Purine as a Naturally Occurring Cytokinin in Populus × canadensis Moench cv Robusta Leaves

Immunodetection and Identification of N6-(o-Hydroxybenzylamino)Purine as a Naturally Occurring... Abstract A highly specific and sensitive enzyme-linked immunosorbent assay (ELISA) was developed for 9-β-d-ribofuranosyl-N6-(o-hydroxybenzylamino)purine [(oOH)[9R]BAP] and structurally related cytokinins. As little as 3 femtomoles of the compound could be detected by this method. Cross-reactivity studies demonstrated the specificity of four polyclonal antibodies for (oOH)[9R] BAP and its free base in preference to a range of natural cytokinins and other purines. After evaluating the method by internal standardization employing [2-3H](oOH)[9R]BAP of high specific radioactivity as recovery marker by dilution analyses and by immunohistograms, it was possible to apply ELISA to quantify (oOH)[9R]BAP in plant extracts. In addition to (oOH)[9R]BAP, an unknown cytokinin reacting with the same antibody was detected in high performance liquid chromatography-fractionated extracts of mature Populus × canadensis Moench cv Robusta. The structure of the new compound was determined by gas chromatography-mass spectrometry and finally confirmed by synthesis as N6-(o-hydroxybenzylamino)purine. 1 Research was supported by the Czechoslovak Academy of Sciences and by a grant from the Deutsche Forschungsgemeinschaft within the scope of SFB 137. This content is only available as a PDF. © 1992 American Society of Plant Biologists This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model) http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Plant Physiology Oxford University Press

Immunodetection and Identification of N6-(o-Hydroxybenzylamino)Purine as a Naturally Occurring Cytokinin in Populus × canadensis Moench cv Robusta Leaves

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References (23)

Publisher
Oxford University Press
Copyright
Copyright © 2021 American Society of Plant Biologists
ISSN
0032-0889
eISSN
1532-2548
DOI
10.1104/pp.99.1.74
Publisher site
See Article on Publisher Site

Abstract

Abstract A highly specific and sensitive enzyme-linked immunosorbent assay (ELISA) was developed for 9-β-d-ribofuranosyl-N6-(o-hydroxybenzylamino)purine [(oOH)[9R]BAP] and structurally related cytokinins. As little as 3 femtomoles of the compound could be detected by this method. Cross-reactivity studies demonstrated the specificity of four polyclonal antibodies for (oOH)[9R] BAP and its free base in preference to a range of natural cytokinins and other purines. After evaluating the method by internal standardization employing [2-3H](oOH)[9R]BAP of high specific radioactivity as recovery marker by dilution analyses and by immunohistograms, it was possible to apply ELISA to quantify (oOH)[9R]BAP in plant extracts. In addition to (oOH)[9R]BAP, an unknown cytokinin reacting with the same antibody was detected in high performance liquid chromatography-fractionated extracts of mature Populus × canadensis Moench cv Robusta. The structure of the new compound was determined by gas chromatography-mass spectrometry and finally confirmed by synthesis as N6-(o-hydroxybenzylamino)purine. 1 Research was supported by the Czechoslovak Academy of Sciences and by a grant from the Deutsche Forschungsgemeinschaft within the scope of SFB 137. This content is only available as a PDF. © 1992 American Society of Plant Biologists This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model)

Journal

Plant PhysiologyOxford University Press

Published: May 1, 1992

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