Plant Physiology Preview Published on August 8, 2002; 10.1104/pp.005439
Received March 13, 2002
Returned for revision April 25, 2002
Accepted May 24, 2002
Regulation of Transcript Levels of the Arabidopsis Cytochrome P450 Genes Involved in Brassinosteroid Biosynthesis
Simona Bancos , Takahito Nomura , Tatsuro Sato , Gergely Molnár , Gerard J. Bishop , Csaba Koncz , Takao Yokota , Ferenc Nagy , and Miklós Szekeres *
Institute of Plant Biology, Biological Research Center of the Hungarian Academy of Sciences, P.O. Box 521, H 6701 Szeged, Hungary (S.B., G.M., F.N., M.S.); Department of Biosciences, Teikyo University, Utsunomiya 320 8551, Japan (T.N., T.S., T.Y.); Institute of Biological Sciences, University of Wales, Aberystwyth SY23 3DD, United Kingdom (G.J.B.); and Max Planck-Institut für Züchtungsforschung, Carl von Linné-Weg 10, D 50829 Köln, Germany (C.K.)
* Corresponding author; email: szekeres{at}nucleus.szbk.u-szeged.hu.
Cytochrome P450 enzymes of the closely related CYP90 and CYP85 families catalyze essential oxidative reactions in the biosynthesis of brassinosteroid (BR) hormones. Arabidopsis CYP90B1/DWF4 and CYP90A1/CPD are responsible for respective C-22 and C-23 hydroxylation of the steroid side chain and CYP85A1 catalyzes C-6 oxidation of 6-deoxo intermediates, whereas the functions of CYP90C1/ROT3, CYP90D1, and CYP85A2 are still unknown. Semiquantitative reverse transcriptase-polymerase chain reaction analyses show that transcript levels of CYP85 and CYP90 genes are down-regulated by brassinolide, the end product of the BR biosynthesis pathway. Feedback control of the CYP90C1, CYP90D1, and CYP85A2 genes by brassinolide suggests that the corresponding enzymes might also participate in BR synthesis. CYP85 and CYP90 mRNAs show strong and transient accumulation during the 1st week of seedling development, as well as characteristic organ-specific distribution. Transcripts of CYP90A1 and CYP85A2 are preferentially represented in shoots and CYP90C1, CYP90D1, and CYP85A1 mRNAs are more abundant in roots, whereas CYP90B1 is ubiquitously expressed. Remarkably, the spatial pattern of CYP90A1 expression is maintained in the BR-insensitive cbb2 mutant, indicating the independence of organ-specific and BR-dependent regulation. Quantitative gas chromatography-mass spectrometry analysis of endogenous BRs in shoots and roots of Arabidopsis, pea (Pisum sativum), and tomato (Lycopersicon esculentum) reveal similar partitioning patterns of BR intermediates in these species. Inverse correlation between CYP90A1/CPD transcript levels and the amounts of the CYP90A1 substrate 6-deoxocathasterone in shoots and roots suggests that transcriptional regulation plays an important role in controlling BR biosynthesis.
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