A bifunctional oleate 12-hydroxylase: desaturase from Lesquerella fendleri.
LFAH12, an oleate 12-hydroxylase gene from Lesquerella fendleri (L.) was isolated on the basis of nucleotide sequence similarity to an oleate hydroxylase gene from Ricinus communis (L.). Transgenic Arabidopsis plants containing the Lesquerella gene under transcriptional control of the cauliflower mosaic virus 35S promoter accumulated ricinoleic, lesquerolic and densipolic acids in seeds, but not in leaves or roots. However, hydroxylase activity was detectable in crude extracts of vegetative tissues. The discrepancy between the presence of activity and the lack of hydroxy fatty acids suggests selective removal and breakdown of hydroxy fatty acids in vegetative organs. High levels of LFAH12 mRNA accumulation did not lead to correspondingly high levels of protein accumulation, suggesting that accumulation of the hydroxylase may be controlled post-transcriptionally. Expression of the L. fendleri gene in transgenic plants of a fad2 mutant of Arabidopsis, which is deficient in cytoplasmic oleate delta 12 desaturase activity, resulted in partial suppression of the mutant phenotype in roots. Thus, unlike the hydroxylase from R. communis, the L. fendleri enzyme has both hydroxylase and desaturase activities. Fusion of the 5' flanking region of the LFAH12 gene to the beta-glucuronidase coding sequence resulted in a high level of early seed-specific expression of beta-glucuronidase activity in transgenic Arabidopsis plants.
Broun
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Carnegie Institution of Washington, Department of Plant Biology, Stanford, CA 94305, USA.
Boddupalli
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Somerville
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C
D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics; D001483:Base Sequence; D019607:Brassicaceae / Q000201:enzymology* / Q000235:genetics*; D002367:Ricinus communis / Q000201:enzymology; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D044943:Fatty Acid Desaturases / Q000235:genetics* / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D006899:Mixed Function Oxygenases / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D044925:Oxidoreductases Acting on CH-CH Group Donors; D010641:Phenotype; D010940:Plant Proteins; D030821:Plants, Genetically Modified; D010947:Plants, Toxic; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D018749:RNA, Plant / Q000235:genetics / Q000378:metabolism; D014018:Tissue Distribution
D017931:DNA Primers; D010940:Plant Proteins; D012333:RNA, Messenger; D018749:RNA, Plant; D006899:Mixed Function Oxygenases; C094812:phosphatidylcholine 12-monooxygenase; D044943:Fatty Acid Desaturases; C021904:Phosphatidylcholine desaturase; D044925:Oxidoreductases Acting on CH-CH Group Donors