In Vivo Measurement of Indole-3-acetic Acid Decarboxylation in Aging Coleus Petiole Sections

The concentration of indoleacetic acid (IAA) in plant tissues is regulated, in part, by its rate of decarboxylation. However, the commonly used in vitro assays for IAA oxidase may not accurately reflect total in vivo decarboxylation rates. A method for measuring in vivo decarboxylation was utilized in which (14)CO(2) is collected following uptake of [1-(14)C]IAA by excised tissue sections. After a 30-minute equilibration period, the evolution of (14)CO(2) was found to follow an approximately linear course with respect to both time and tissue weight.Decarboxylation rates were measured by this method in petiole sections of the Princeton clone of Coleus blumei Benth. Both the (14)CO(2) evolved per milligram tissue and the percent of [1-(14)C]IAA uptake decarboxylated were highest in sections from the youngest petioles tested, and declined in the older tissue. Thin layer chromatography of acetonitrile extracts from the [1-(14)C]IAA-treated petioles showed a decreasing amount of free IAA and an increase at the retardation factor of indoleacetylaspartate in the older sections. The decreased decarboxylation rates in the older petioles may be attributable to a generally lower metabolic rate and increased protection of the IAA by conjugation.