First published online June 9, 2006; 10.1104/pp.106.082024
Plant Physiology 141:1248-1254 (2006)
© 2006 American Society of Plant Biologists
BIOCHEMICAL PROCESSES AND MACROMOLECULAR STRUCTURES
CYP71B15 (PAD3) Catalyzes the Final Step in Camalexin Biosynthesis1
Regina Schuhegger,
Majse Nafisi,
Madina Mansourova,
Bent Larsen Petersen,
Carl Erik Olsen,
Ale Svato,
Barbara Ann Halkier and
Erich Glawischnig*
Lehrstuhl für Genetik, Technische Universität München, D–85350 Freising, Germany (R.S., E.G.); Center for Molecular Plant Physiology, DK–1871 Frederiksberg C, Denmark (M.N., B.L.P., C.E.O., B.A.H.); and Max Planck Institute for Chemical Ecology, Beutenberg Campus, D–07745 Jena, Germany (M.M., A.S.)
Camalexin represents the main phytoalexin in Arabidopsis (Arabidopsis thaliana). The camalexin-deficient phytoalexin deficient 3 (pad3) mutant has been widely used to assess the biological role of camalexin, although the exact substrate of the cytochrome P450 enzyme 71B15 encoded by PAD3 remained elusive. 2-(Indol-3-yl)-4,5-dihydro-1,3-thiazole-4-carboxylic acid (dihydrocamalexic acid) was identified as likely intermediate in camalexin biosynthesis downstream of indole-3-acetaldoxime, as it accumulated in leaves of silver nitrate-induced pad3 mutant plants and it complemented the camalexin-deficient phenotype of a cyp79b2/cyp79b3 double-knockout mutant. Recombinant CYP71B15 heterologously expressed in yeast catalyzed the conversion of dihydrocamalexic acid to camalexin with preference of the (S)-enantiomer. Arabidopsis microsomes isolated from leaves of CYP71B15-overexpressing and induced wild-type plants were capable of the same reaction but not microsomes from induced leaves of pad3 mutants. In conclusion, CYP71B15 catalyzes the final step in camalexin biosynthesis.
1 This work was supported by the Deutsche Forschungsgemeinschaft (GL346/1) and the Max-Planck Gesellschaft.
The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Erich Glawischnig (egl{at}wzw.tum.de).
Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.106.082024.
* Corresponding author; e-mail egl{at}wzw.tum.de; fax 49–8161–71–5636.
Received April 13, 2006;
returned for revision May 18, 2006;
accepted May 24, 2006.
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