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Plant Physiology 99:1090-1098 (1992)
© 1992 American Society of Plant Biologists

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Molecular Biology and Gene Regulation

A New Bioassay for Auxins and Cytokinins 1

Wout Boerjan, Chris Genetello, Marc Van Montagu and Dirk Inzé

Laboratorium voor Genetica, Universiteit Gent, B-9000 Gent, Belgium, Laboratoire Associé de l'Institut National de la Recherche Agronomique, Universiteit Gent, B-9000 Gent, Belgium

The authors have developed a sensitive bioassay that can be used to detect auxins as well as cytokinins. The bioassay is based on the expression in transformed tobacco (Nicotiana tabacum) mesophyll protoplasts of a chimeric gene, consisting of the upstream sequences of the Agrobacterium tumefaciens gene 5, coupled to the coding sequence of the beta-glucuronidase. The expression of this gene is induced by the presence of both auxin and cytokinin in the culture medium. Using this assay, indole-3-acetic acid was detected at 5 x 10–8 molar, whereas trans-zeatin could be detected at 5 x 10–11 molar. The assay can be performed in microtiter plates, allowing numerous samples to be analyzed simultaneously. Only 2.5 x 105 protoplasts are required for one individual assay in 250 microliters of culture medium and for qualitative results, the reaction is readily visualized by ultraviolet light.

1 This work was supported by grants from the "A.S.L.K.-Kankerfonds," the Services of the Prime Minister (I.U.A.P. 12OC0187), and the "Vlaams Actieprogramma Biotechnologie" (174KP490). D.I. is a Research Director of the Institut National de la Recherche Agronomique (Paris, France).




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T. A. Walsh, R. Neal, A. O. Merlo, M. Honma, G. R. Hicks, K. Wolff, W. Matsumura, and J. P. Davies
Mutations in an Auxin Receptor Homolog AFB5 and in SGT1b Confer Resistance to Synthetic Picolinate Auxins and Not to 2,4-Dichlorophenoxyacetic Acid or Indole-3-Acetic Acid in Arabidopsis
Plant Physiology, October 1, 2006; 142(2): 542 - 552.
[Abstract] [Full Text] [PDF]


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Copyright © 1992 by the American Society of Plant Biologists