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The effect of auxin on cytokinin levels and metabolism in transgenic tobacco tissue expressing an ipt gene

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Abstract

The ipt gene from the T-DNA of Agrobacterium tumefaciens was transferred to tobacco (Nicotiana tabacum L.) in order to study the control which auxin appears to exert over levels of cytokinin generated by expression of this gene. The transgenic tissues contained elevated levels of cytokinins, exhibited cytokinin and auxin autonomy and grew as shooty calli on hormone-free media. Addition of 1-naphthylacetic acid to this culture medium reduced the total level of cytokinins by 84% while 6-benzylaminopurine elevated the cytokinin level when added to media containing auxin. The cytokinins in the transgenic tissue were labelled with 3H and auxin was found to promote conversion of zeatin-type cytokinins to 3H-labelled adenine derivatives. When the very rapid metabolism of exogenous [3H]zeatin riboside was suppressed by a phenylurea derivative, a noncompetitive inhibitor of cytokinin oxidase, auxin promoted metabolism to adenine-type compounds. Since these results indicated that auxin promoted cytokinin oxidase activity in the transformed tissue, this enzyme was purified from the tobacco tissue cultures. Auxin did not increase the level of the enzyme per unit tissue protein, but did enhance the activity of the enzyme in vitro and promoted the activity of both glycosylated and non-glycosylated forms. This enhancement could contribute to the decrease in cytokinin level induced by auxin. Studies of cytokinin biosynthesis in the transgenic tissues indicated that trans-hydroxylation of isopentenyladenine-type cytokinins to yield zeatin-type cytokinins occurred principally at the nucleotide level.

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Abbreviations

Ade:

adenine

Ados:

adenosine

BA:

6-benzylaminopurine

C:

control

Con A:

concanavallin A

CP:

cellulose phosphate

IPT:

isopentenyl transferase

NAA:

1-naphthylacetic acid

NP:

normal phase

NPPU:

N-(3-nitrophenyl)-N′-phenylurea

RIA:

radioimmunoassay

RP:

reversed phase

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Authors and Affiliations

  1. Plant Cell Biology Group, Research School of Biological Sciences, Australian National University, PO Box 475, Canberra, Australia

    R. Zhang, X. Zhang, J. Wang & S. A. McKinney

  2. CSIRO Division of Plant Industry, PO Box 1600, 2601, Canberra, ACT, Australia

    R. Zhang & T. J. V. Higgins

  3. Cooperative Research Centre for Plant Science, Australian National University, PO Box 475, 2601, Canberra, ACT, Australia

    D. S. Letham

Authors
  1. R. Zhang
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  2. X. Zhang
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  3. J. Wang
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  4. D. S. Letham
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  5. S. A. McKinney
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  6. T. J. V. Higgins
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We wish to thank Dr. J. Zwar for supplying phenylurea derivitives.

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Zhang, R., Zhang, X., Wang, J. et al. The effect of auxin on cytokinin levels and metabolism in transgenic tobacco tissue expressing an ipt gene. Planta 196, 84–94 (1995). https://doi.org/10.1007/BF00193221

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  • DOI: https://doi.org/10.1007/BF00193221

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