CLONING OF HIGHER-PLANT OMEGA-3-FATTY-ACID DESATURASES

被引:228
作者
YADAV, NS
WIERZBICKI, A
AEGERTER, M
CASTER, CS
PEREZGRAU, L
KINNEY, AJ
HITZ, WD
BOOTH, JR
SCHWEIGER, B
STECCA, KL
ALLEN, SM
BLACKWELL, M
REITER, RS
CARLSON, TJ
RUSSELL, SH
FELDMANN, KA
PIERCE, J
BROWSE, J
机构
[1] DUPONT CO INC, CENT RES & DEV, EXPTL STN, WILMINGTON, DE 19880 USA
[2] WASHINGTON STATE UNIV, INST BIOL CHEM, PULLMAN, WA 99164 USA
关键词
D O I
10.1104/pp.103.2.467
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
Arabidopsis thaliana T-DNA transformants were screened for mutations affecting seed fatty acid composition. A mutant line was found with reduced levels of linolenic acid (18:3) due to a T-DNA insertion. Genomic DNA flanking the T-DNA insertion was used to obtain an Arabidopsis cDNA that encodes a polypeptide identified as a microsomal omega-3 fatty acid desaturase by its complementation of the mutation. Analysis of lipid content in transgenic tissues demonstrated that this enzyme is limiting for 18:3 production in Arabidopsis seeds and carrot hairy roots. This cDNA was used to isolate a related Arabidopsis cDNA, whose mRNA is accumulated to a much higher level in leaf tissue relative to root tissue. This related cDNA encodes a protein that is a homolog of the microsomal desaturase but has an N-terminal extension deduced to be a transit peptide, and its gene maps to a position consistent with that of the Arabidopsis fad D locus, which controls plastid omega-3 desaturation. These Arabidopsis cDNAs were used as hybridization probes to isolate cDNAs encoding homologous proteins from developing seeds of soybean and rapeseed. The high degree of sequence similarity between these sequences suggests that the omega-3 desaturases use a common enzyme mechanism.
引用
收藏
页码:467 / 476
页数:10
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