Plant Physiol. Journal of Pharmacology and Experimental Therapeutics
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PLANT PHYSIOLOGY , Vol 106, Issue 4 1453-1459, Copyright © 1994 by American Society of Plant Biologists

METABOLISM AND ENZYMOLOGY

Identification of a Gene that Complements an Arabidopsis Mutant Deficient in Chloroplast [omega]6 Desaturase Activity

D. L. Falcone, S. Gibson, B. Lemieux and C. Somerville
Carnegie Institution, Department of Plant Biology, 290 Panama Street, Stanford, California 94305 (D.L.F., C.S.)

Membrane lipids of the fad6 (formerly fadC) mutant of Arabidopsis, which is deficient in chloroplast [omega]6 desaturase activity, have increased levels of monounsaturated fatty acids and are deficient in trienoic fatty acids. A putative fad6 cDNA clone was isolated by probing a cDNA library with a degenerate oligonucleotide based on a conserved region within known [omega]3 desaturase genes. Expression of the cDNA in transgenic plants of a fad6 mutant restored normal levels of all fatty acids. When used as a hybridization probe, the cDNA identified a restriction fragment-length polymorphism that co-segregated with the fad6 mutation. Thus, on the basis of a genetic complementation test and genetic map position, the fad6 gene is encoded by the cDNA. The cDNA encoded a 418-amino acid polypeptide of 47,727 D that displayed a high degree of sequence similarity to a [delta]12 desaturase from the cyanobacterium Synechocystis. The fad6 gene exhibited less sequence homology to any known higher plant desaturase, including an endoplasmic reticulum-localized [omega]6 desaturase corresponding to the Arabidopsis fad2 gene.


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