First published online August 8, 2002; 10.1104/pp.005439
Plant Physiol, September 2002, Vol. 130, pp. 504-513
Regulation of Transcript Levels of the Arabidopsis Cytochrome
P450 Genes Involved in Brassinosteroid
Biosynthesis1
Simona
Banco ,
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.)
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.
1
This work was supported by the Hungarian
National Research Foundation (Országos Tudományos
Kutatás: Alap [OTKA], grant no. T 32432), by the Human
Frontiers Science Program (grant no. RG00162-2000 to G.J.B, C.K., and
T.Y.), by the Japan Society for the Promotion of Science (Grand-in-Aid
for Scientific Research no. 11460057 to T.Y and postdoctoral fellowship
to T.N.), by scientific exchange programs between the Deutsche
Forschungsgemeinschaft and the Hungarian Academy of Sciences (project
nos. 436-UNG-113/143 and D-132), by scientific exchange programs
between the Deutsches Zentrum für Luft und Raumfahrt e.V. and the
Hungarian Science and Technology Foundation (project nos. UNG-027-99
and D-7/99), and by the British Council (support to G.J.B.).
*
Corresponding author; e-mail
szekeres{at}nucleus.szbk.u-szeged.hu; fax 36-62-433434.
© 2002 American Society of Plant Physiologists
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