Open AccessImmunodetection 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,
Erwin 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.