A Radioimmunoassay for Dihydrozeatin and Dihydrozeatin Riboside, and its Application to a Study of the in vitro Metabolism of Dihydrozeatin by Soybean Callus

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Summary

A radioimmunoassay was developed for the quantitation of DHZ and DHZR, using antibodies raised to a DHZR-BSA conjugate, and [3H]-DHZ as the tracer. The effective measuring range of the assay was 0.11–22.6 pmol for DHZ, and 0.071–14.2 pmol for DHZR. The antiserum showed sufficient cross-reaction with Z and ZR to necessitate their separation from the respective dihydro derivatives prior to RIA. This was adequately achieved by oxidation with KMnO4. Cross-reaction with kinetin, BA and HA was also unusually high, and this indicated the potential suitability of this assay system for their quantitation.

The in vitro metabolism of DHZ by soybean callus over an incubation period of 48 h was followed by RIA, and by monitoring the degradation of added [3H]-DHZ by HPLC. Both assay systems gave similar results, indicating a conversion of DHZ to DHZR.

Referencess (20)

  • D.J. Armstrong et al.

    Isolation of cytokinins from tRNA. Biochem.

    Biophys. Res. Commun.

    (1969)
  • Z. Cekan

    Assessment of reliability of steroid radioimmunoassays.

    J. Steroid Biochem.

    (1975)
  • B.A. Mcgaw et al.

    Dihydrozeatin metabolism in radish seedlings.

    Phytochemistry

    (1984)
  • J. Van Staden et al.

    6-(2,3,4,-trihydroxy-3-methylbutylamino)purine

    a cytokinin with biological activity., Phytochemistry

    (1982)
  • J. Badenoch-Jones et al.

    Quantitation of cytokinins in biological samples using antibodies against zeatin riboside.

    Plant Physiol.

    (1984)
  • R.L. Butcher

    Changes in gonadotropins and steroids associated with unilateral ovariectomy of the rat.

    Endocrinology

    (1977)
  • J.G. Cutting et al.

    Development of a radioimmunoassay for the cytokinin isopentenyladenosine.

    Crop Prod.

    (1983)
  • D. Ernst et al.

    Isolation and quantitation of isopentenyladenosine in the anise cell culture by single-ion monitoring radioimmunoassay and bioassay.

    Planta

    (1983)
  • C.E. Hansen et al.

    Concentration gradients of trans-zeatin in maize stem. Measurement by a specific enzyme immunoassay.

    Plant Physiol.

    (1984)
  • R. Horgan

    Modern methods for plant analysis

There are more references available in the full text version of this article.

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