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The Arabidopsis dwarf1 Mutant Is Defective in the Conversion of 24-Methylenecholesterol to Campesterol in Brassinosteroid Biosynthesis1

Sunghwa Choe, Brian P. Dilkes, Brian D. Gregory, Amanda S. Ross, Heng Yuan, Takahiro Noguchi, Shozo Fujioka, Suguru Takatsuto, Atsushi Tanaka, Shigeo Yoshida, Frans E. Tax, and Kenneth A. Feldmann*

Department of Plant Sciences, University of Arizona, Tucson, Arizona 85721 (S.C., B.P.D., B.D.G., A.S.R., H.Y., A.T., F.E.T., K.A.F.); Institute of Physical and Chemical Research (RIKEN), Wako-shi, Saitama 351-0198, Japan (T.N., S.F., S.Y.); Department of Chemistry, Joetsu University of Education, Joetsu-shi, Niigata 943-8512, Japan (S.T.); and Department of Environment and Resources, Japan Atomic Energy Research Institute, 1233 Watanuki-machi, Takasaki-shi, Gunma 370-1292, Japan (A.T.)

Since the isolation and characterization of dwarf1-1 (dwf1-1) from a T-DNA insertion mutant population, phenotypically similar mutants, including deetiolated2 (det2), constitutive photomorphogenesis and dwarfism (cpd), brassinosteroid insensitive1 (bri1), and dwf4, have been reported to be defective in either the biosynthesis or the perception of brassinosteroids. We present further characterization of dwf1-1 and additional dwf1 alleles. Feeding tests with brassinosteroid-biosynthetic intermediates revealed that dwf1 can be rescued by 22alpha -hydroxycampesterol and downstream intermediates in the brassinosteroid pathway. Analysis of the endogenous levels of brassinosteroid intermediates showed that 24-methylenecholesterol in dwf1 accumulates to 12 times the level of the wild type, whereas the level of campesterol is greatly diminished, indicating that the defective step is in C-24 reduction. Furthermore, the deduced amino acid sequence of DWF1 shows significant similarity to a flavin adenine dinucleotide-binding domain conserved in various oxidoreductases, suggesting an enzymatic role for DWF1. In support of this, 7 of 10 dwf1 mutations directly affected the flavin adenine dinucleotide-binding domain. Our molecular characterization of dwf1 alleles, together with our biochemical data, suggest that the biosynthetic defect in dwf1 results in reduced synthesis of bioactive brassinosteroids, causing dwarfism.

1   This research was supported by the National Science Foundation (grant no. 9604439 to K.A.F.) and by a Grant-in-Aid for Scientific Research (B) from the Ministry of Education, Science, Sports, and Culture of Japan (grant no. 10460050 to S.F.).
*   Corresponding author; e-mail feldmann{at}ag.arizona.edu; fax 1-520-621-7186.

Plant Physiol. (1999) 119: 897-908
Copyright Clearance Center:   0032-0889/99/119//12
© 1999 American Society of Plant Physiologists




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