Affordable Access

Cytokinin oxidase/cytokinin dehydrogenase assay: optimized procedures and applications.

Authors
Type
Published Article
Journal
Analytical biochemistry
Publication Date
Volume
306
Issue
1
Pages
1–7
Identifiers
PMID: 12069407
Source
Medline
License
Unknown

Abstract

Spectrophotometric methods for determining the activity of cytokinin oxidase/cytokinin dehydrogenase (EC 1.5.99.12) were developed and optimized. A sensitive end-point method based on a combination of the electron acceptor 2,6-dichlorophenolindophenol and Schiff base formation of the reaction product with 4-aminophenol under acidic conditions can be applied to crude cell and tissue extracts. The assay was also adapted for other substrates than N6-(2-isopentenyl)adenine, such as zeatin and the aromatic cytokinins, although an enzyme which degrades the latter compounds has not yet been identified. The second novel method is an initial rate method based on the coupled redox reaction of phenazine methosulfate and 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide resulting in the formation of a formazan dye. This method can be used for kinetic studies with purified enzyme and is entirely substrate independent.

Statistics

Seen <100 times