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

Miroslav Strnad | Zendy; Walter Peters | Zendy; E. Beck | Zendy; M. Kamínek | Zendy

Open Access

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

Author(s) -

Miroslav Strnad,

Walter Peters,

E. Beck,

M. Kamínek

Publication year - 1992

Publication title -

plant physiology

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 3.554

H-Index - 312

eISSN - 1532-2548

pISSN - 0032-0889

DOI - 10.1104/pp.99.1.74

Subject(s) - cytokinin , polyclonal antibodies , purine , purine metabolism , chemistry , chromatography , enzyme , biochemistry , biology , antibody , auxin , immunology , gene

A highly specific and sensitive enzyme-linked immunosorbent assay (ELISA) was developed for 9-beta-d-ribofuranosyl-N(6)-(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-(3)H](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 x canadensis Moench cv Robusta. The structure of the new compound was determined by gas chromatography-mass spectrometry and finally confirmed by synthesis as N(6)-(o-hydroxybenzylamino)purine.

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