Controlled indole-3-acetaldoxime production through ethanol-induced expression of CYP79B2

Michael Dalgaard Mikkelsen; Victoria L Fuller; Bjarne Gram Hansen; Majse Nafisi; Carl Erik Olsen; Henrik Bjørn Nielsen; Barbara Ann Halkier

doi:10.1007/s00425-009-0907-5

Controlled indole-3-acetaldoxime production through ethanol-induced expression of CYP79B2

Planta. 2009 May;229(6):1209-17. doi: 10.1007/s00425-009-0907-5. Epub 2009 Mar 5.

Authors

Michael Dalgaard Mikkelsen¹, Victoria L Fuller, Bjarne Gram Hansen, Majse Nafisi, Carl Erik Olsen, Henrik Bjørn Nielsen, Barbara Ann Halkier

Affiliation

¹ VKR Research Centre Pro-Active Plants, Department of Plant Biology and Biotechnology, University of Copenhagen, 40 Thorvaldsensvej, 1871, Frederiksberg C, Denmark.

PMID: 19263076
DOI: 10.1007/s00425-009-0907-5

Abstract

Indole-3-acetaldoxime (IAOx) is a key branching point between primary and secondary metabolism. IAOx serves as an intermediate in the biosynthesis of indole glucosinolates (I-GLSs), camalexin and the plant hormone indole-3-acetic acid (IAA). The cytochrome P450s CYP79B2 and CYP79B3 catalyze the conversion of tryptophan to IAOx. CYP83B1 channels IAOx into I-GLS biosynthesis, CYP71A13 channels IAOx into camalexin biosynthesis, whereas the IAOx-metabolizing enzyme in IAA biosynthesis is not known. In this report, we demonstrate controlled production of I-GLSs by introducing an ethanol (EtOH)-inducible CYP79B2 construct into double (cyp79b2 cyp79b3) or triple (cyp79b2 cyp79b3 cyp83b1) mutant lines. We show EtOH-dependent induction of camalexin and identify a number of candidate IAA homeostasis- or defense-related genes by clustered microarray analysis. The transgenic mutant lines are thus promising tools for elucidating the interplay between primary and secondary metabolism.

MeSH terms

Arabidopsis / genetics*
Arabidopsis / metabolism
Arabidopsis Proteins / genetics*
Arabidopsis Proteins / metabolism
Biosynthetic Pathways
Chromatography, Liquid
Cluster Analysis
Cytochrome P-450 Enzyme System / genetics*
Cytochrome P-450 Enzyme System / metabolism
Ethanol / pharmacology*
Gene Expression Profiling
Gene Expression Regulation, Enzymologic / drug effects*
Gene Expression Regulation, Plant / drug effects
Glucosinolates / chemistry
Glucosinolates / metabolism
Indoleacetic Acids / chemistry
Indoleacetic Acids / metabolism
Indoles / chemistry
Indoles / metabolism*
Mass Spectrometry
Molecular Structure
Mutation
Oligonucleotide Array Sequence Analysis / methods
Oximes / chemistry
Oximes / metabolism*
Plants, Genetically Modified
Thiazoles / chemistry
Thiazoles / metabolism

Substances

Arabidopsis Proteins
Glucosinolates
Indoleacetic Acids
Indoles
Oximes
Thiazoles
camalexin
indole-3-acetaldoxime
Ethanol
indoleacetic acid
Cytochrome P-450 Enzyme System
cytochrome P-450 79B2 protein, Arabidopsis
CYP83B1 protein, Arabidopsis
cytochrome P-450 CYP79B3, Sinapis alba