Obtusifoliol 14α-demethylase (CYP51) antisense Arabidopsis shows slow growth and long life

被引:40
作者
Kushiro, M
Nakano, T
Sato, K
Yamagishi, K
Asami, T
Nakano, A
Takatsuto, S
Fujioka, S
Ebizuka, Y
Yoshida, S
机构
[1] Univ Tokyo, Grad Sch Pharmaceut Sci, Bunkyo Ku, Tokyo 1130033, Japan
[2] RIKEN, Plant Funct Lab, Wako, Saitama 3510198, Japan
[3] RIKEN, Mol Membrane Biol Lab, Wako, Saitama 3510198, Japan
[4] Joetsu Univ Educ, Dept Chem, Niigata 9438512, Japan
关键词
sterol; 14; alpha-demethylase; CYP51; obtusifoliol; brassionosteroid; transgenic; Arabidopsis;
D O I
10.1006/bbrc.2001.5122
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Obtusifoliol 14 alpha -demethylase is a plant orthologue of sterol 14 alpha -demethylase (CYP51) essential in sterol biosynthesis, We have prepared CYP51 antisense Arabidopsis in order to shed light on the sterol and steroid hormone biosynthesis in plants. Arabidopsis putative CYP51 cDNA AtCYP51 was obtained from Arabidopsis expressed sequence tag (EST) library and its function was examined in a yeast lanosterol 14 alpha -demethylase (Erg11) deficient mutant. A recombinant AtCYP51 protein fused with a yeast Erg11 signal-anchor peptide was able to complement the erg11 mutation, which confirmed AtCYP51 to be a functional sterol 14 alpha -demethylase, AtCYP51 was then used to generate transgenic Arabidopsis by transforming with pBI vector harboring AtCYP51 in the antisense direction under CaMV35S promoter. The resulting transgenic plants were decreased in accumulation of AtCYP51 mRNA and increased in the amount of endogenous obtusifoliol, They showed a semidwarf phenotype in the early growth stage and a longer life span than control plants. This newly found phenotype is different from previously characterized brassinosteroid (BR)-deficient campesterol biosynthesis mutants. (C) 2001 Academic Press.
引用
收藏
页码:98 / 104
页数:7
相关论文
共 25 条
[1]   DIFFERENT SUBSTRATE SPECIFICITIES OF LANOSTEROL 14-ALPHA-DEMETHYLASE (P-45014DM) OF SACCHAROMYCES-CEREVISIAE AND RAT-LIVER FOR 24-METHYLENE-24,25-DIHYDROLANOSTEROL AND 24,25-DIHYDROLANOSTEROL [J].
AOYAMA, Y ;
YOSHIDA, Y .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1991, 178 (03) :1064-1071
[2]   THE 4-BETA-METHYL GROUP OF SUBSTRATE DOES NOT AFFECT THE ACTIVITY OF LANOSTEROL 14-ALPHA-DEMETHYLASE (P-45014DM) OF YEAST - DIFFERENCE BETWEEN THE SUBSTRATE RECOGNITION BY YEAST AND PLANT STEROL 14-ALPHA-DEMETHYLASES [J].
AOYAMA, Y ;
YOSHIDA, Y .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1992, 183 (03) :1266-1272
[3]   Cloning and expression in Escherichia coli of the obtusifoliol 14 alpha-demethylase of Sorghum bicolor (L) Moench, a cytochrome P450 orthologous to the sterol 14 alpha-demethylases (CYP51) from fungi and mammals [J].
Bak, S ;
Kahn, RA ;
Olsen, CE ;
Halkier, BA .
PLANT JOURNAL, 1997, 11 (02) :191-201
[4]   Optimized expression and catalytic properties of a wheat obtusifoliol 14α-demethylase (CYP51) expressed in yeast -: Complementation of erg11Δ yeast mutants by plant CYP51 [J].
Cabello-Hurtado, F ;
Taton, M ;
Forthoffer, N ;
Kahn, R ;
Bak, S ;
Rahier, A ;
Werck-Reichhart, D .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1999, 262 (02) :435-446
[5]   Cloning and functional expression in yeast of a cDNA coding for an obtusifoliol 14 alpha-demethylase (CYP51) in wheat [J].
CabelloHurtado, F ;
Zimmerlin, A ;
Rahier, A ;
Taton, M ;
DeRose, R ;
Nedelkina, S ;
Batard, Y ;
Durst, F ;
Pallett, KE ;
WerckReichhart, D .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1997, 230 (02) :381-385
[6]   The Arabidopsis dwf7/ste1 mutant is defective in the Δ7 sterol C-5 desaturation step leading to brassinosteroid biosynthesis [J].
Choe, SW ;
Noguchi, T ;
Fujioka, S ;
Takatsuto, S ;
Tissier, CP ;
Gregory, BD ;
Ross, AS ;
Tanaka, A ;
Yoshida, S ;
Tax, FE ;
Feldmann, KA .
PLANT CELL, 1999, 11 (02) :207-221
[7]   Floral dip:: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana [J].
Clough, SJ ;
Bent, AF .
PLANT JOURNAL, 1998, 16 (06) :735-743
[8]   Molecular genetic studies confirm the role of brassinosteroids in plant growth and development [J].
Clouse, SD .
PLANT JOURNAL, 1996, 10 (01) :1-8
[9]   Brassinosteroids [J].
Fujioka, S ;
Sakurai, A .
NATURAL PRODUCT REPORTS, 1997, 14 (01) :1-10
[10]   MECHANISM OF ACTION OF POLYMERIC AURINTRICARBOXYLIC ACID, A POTENT INHIBITOR OF PROTEIN-NUCLEIC ACID INTERACTIONS [J].
GILBERTOGONZALEZ, R ;
HAXO, RS ;
SCHLEICH, T .
BIOCHEMISTRY, 1980, 19 (18) :4299-4303