Open AccessArabidopsis det2 Is Defective in the Conversion of (24R)-24-Methylcholest-4-En-3-One to (24R)-24-Methyl-5α-Cholestan-3-One in Brassinosteroid Biosynthesis1
Author(s) -
Takahiro Noguchi,
Shozo Fujioka,
Suguru Takatsuto,
Akira Sakurai,
Shigeo Yoshida,
Jianming Li,
Joanne Chory
Publication year - 1999
Publication title -
plant physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.554
H-Index - 312
eISSN - 1532-2548
pISSN - 0032-0889
DOI - 10.1104/pp.120.3.833
Subject(s) - arabidopsis , campesterol , biosynthesis , catharanthus roseus , mutant , biochemistry , arabidopsis thaliana , biology , metabolic pathway , chemistry , stereochemistry , enzyme , gene , sterol , cholesterol
Previously, we have shown that the Arabidopsis det2 (deetiolated2) mutant is defective in the biosynthesis of brassinosteroids (BR) and that DET2 (a steroid 5alpha-reductase) acts early in the proposed BR biosynthetic pathway. In this paper we present further biochemical characterization of det2. We have undertaken metabolic experiments with 2H-labeled substrates of intermediates involved in the formation of campestanol from campesterol, and quantitative analysis of intermediates in Arabidopsis wild type and det2. The results of these studies indicate the early operating steps of BR biosynthesis as: campesterol --> 4-en-3beta-ol --> 4-en-3-one --> 3-one --> campestanol in Arabidopsis, with det2 deficient in the conversion of 4-en-3-one to 3-one. We have also detected these intermediates in the formation of campestanol from campesterol and their metabolic conversions using cultured cells of Catharanthus roseus. These studies confirmed the biosynthetic sequence of events from campesterol to campestanol as was found in Arabidopsis. As such, the originally proposed biosynthetic pathway should be modified.