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Radioimmunoassays for trans-zeatin and related cytokinins

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Abstract

Radioimmunoassays for the quantitation of trans-zeatin and related cytokinins have been developed. Antisera produced against bovine serum albumin conjugates of trans-zeatinriboside have a high affinity (Ka=2.4.10-11 M) for zeatinriboside and for zeatin, but show a negligible cross reaction to isopentenyladenosine (0.1%) and cis-zeatinriboside (0.4%), only a slight cross reaction to dihydrozeatin (1.7%), and no cross reaction at all to other purines, such as adenosine and related, compounds, was observed. The assays are sensitive and measuring ranges extend from 0.06–30 pmol (0.02–10 ng) of zeatinriboside. This has been achieved by employing as tracers immunoreactive zeatin derivatives with high-specific activity, (tritiated zeatinriboside-dialcohol: 8.37.1011 Bq mmol-1 and zeatinribosyl-[125I]tyramine: ca. 1.9.1013 Bq mmol-1. The detection limit is 40 fmol (15pg) for the assay employing the tritiated tracer, and assay reproducibility is high (variation coefficients of triplicates less than 5%). Several hundred assays can be completed in one day, and, due to the high specificity of this assay, crude extracts may be used for analysis. The course of zeatin levels in developing fruits of Lycopersicon esculentum cv. Moneymaker is given.

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Abbreviations

GC:

gas chromatography

LC:

liquid chromatography

MS:

mass spectroscopy

RIA:

radioimmunoassay

TLC:

thin layer chromatography

UB:

unspecific binding

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Authors and Affiliations

  1. Lehrstuhl für Pflanzenphysiologie, Ruhr-Universität Bochum, Postfach 102148, D-4630, Bochum 10, Germany

    Elmar W. Weiler

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  1. Elmar W. Weiler
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Part 12 in the Series: “Use of Immunoassay in Plant Science”

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Weiler, E.W. Radioimmunoassays for trans-zeatin and related cytokinins. Planta 149, 155–162 (1980). https://doi.org/10.1007/BF00380877

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  • DOI: https://doi.org/10.1007/BF00380877

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