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PLANT PHYSIOLOGY , Vol 106, Issue 4 1511-1520, Copyright © 1994 by American Society of Plant Biologists
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METABOLISM AND ENZYMOLOGY |
A Novel Metabolic Pathway for Indole-3-Acetic Acid in Apical Shoots of Populus tremula (L.) x Populus tremuloides (Michx.)
H. Tuominen, A. Ostin, G. Sandberg and B. Sundberg
Department of Forest Genetics and Plant Physiology, The Swedish University of Agricultural Sciences, 90183 Umea, Sweden
Metabolism of indole-3-acetic acid (IAA) in apical shoots of Populus
tremula (L.) x Populus tremuloides (Michx.) was investigated by feeding a
mixture of [12C]IAA, [13C6]IAA, and [1[prime]-14C]IAA through the base of
the excised stem. HPLC of methanolic plant extracts revealed eight major
radiolabeled metabolites after a 24-h incubation period. Comparison between
feeds with [5-3H]IAA and [1[prime]-14C]IAA showed that all detectable
metabolites were nondecarboxylative products. The purified radiolabeled
HPLC fractions were screened by frit-fast atom bombardment liquid
chromatography-mass spectrometry for compounds with characteristic fragment
pairs originating from the application with 12C and 13C isotopes. Samples
of interest were further characterized by gas chromatography-mass
spectrometry. Using this procedure, oxindole-3-acetic acid (OxIAA),
indole-3-acetyl-N-aspartic acid (IAAsp), oxindole-3-acetyl-N-aspartic acid
(OxIAAsp), and ring-hydroxylated oxindole-3-acetic acid were all identified
as IAA metabolites. Furthermore, a novel metabolic pathway from IAA via
IAAsp and OxIAAsp to OxIAA was established on the basis of refeeding
experiments with the different IAA metabolites.
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