Arabidopsis Cytochrome P450 Monooxygenase 71A13 Catalyzes the Conversion of Indole-3-Acetaldoxime in Camalexin Synthesis

Author:

Nafisi Majse1,Goregaoker Sameer2,Botanga Christopher J.3,Glawischnig Erich4,Olsen Carl E.1,Halkier Barbara A.1,Glazebrook Jane3

Affiliation:

1. Plant Biochemistry Laboratory, Department of Plant Biology, Center for Molecular Plant Physiology, Faculty of Life Sciences, University of Copenhagen, 1871 Frederiksberg C, Denmark

2. Torrey Mesa Research Institute, San Diego, California 92114

3. Department of Plant Biology, Center for Microbial and Plant Genomics, University of Minnesota, Saint Paul, Minnesota 55108

4. Lehrstuhl für Genetik, Technische Universität München, 85350 Freising, Germany

Abstract

Abstract Camalexin (3-thiazol-2-yl-indole) is an indole alkaloid phytoalexin produced by Arabidopsis thaliana that is thought to be important for resistance to necrotrophic fungal pathogens, such as Alternaria brassicicola and Botrytis cinerea. It is produced from Trp, which is converted to indole acetaldoxime (IAOx) by the action of cytochrome P450 monooxygenases CYP79B2 and CYP79B3. The remaining biosynthetic steps are unknown except for the last step, which is conversion of dihydrocamalexic acid to camalexin by CYP71B15 (PAD3). This article reports characterization of CYP71A13. Plants carrying cyp71A13 mutations produce greatly reduced amounts of camalexin after infection by Pseudomonas syringae or A. brassicicola and are susceptible to A. brassicicola, as are pad3 and cyp79B2 cyp79B3 mutants. Expression levels of CYP71A13 and PAD3 are coregulated. CYP71A13 expressed in Escherichia coli converted IAOx to indole-3-acetonitrile (IAN). Expression of CYP79B2 and CYP71A13 in Nicotiana benthamiana resulted in conversion of Trp to IAN. Exogenously supplied IAN restored camalexin production in cyp71A13 mutant plants. Together, these results lead to the conclusion that CYP71A13 catalyzes the conversion of IAOx to IAN in camalexin synthesis and provide further support for the role of camalexin in resistance to A. brassicicola.

Publisher

Oxford University Press (OUP)

Subject

Cell Biology,Plant Science

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