The production of unusual fatty acids in transgenic plants.

The ability to genetically engineer plants has facilitated the generation of oilseeds synthesizing non-native fatty acids. Two particular classes of fatty acids are considered in this review. First, so-called industrial fatty acids, which usually contain functional groups such as hydroxyl, epoxy, or acetylenic bonds, and second, very long chain polyunsaturated fatty acids normally found in fish oils and marine microorganisms. For industrial fatty acids, there has been limited progress toward obtaining high-level accumulation of these products in transgenic plants. For very long chain polyunsaturated fatty acids, although they have a much more complex biosynthesis, accumulation of some target fatty acids has been remarkably successful. In this review, we consider the probable factors responsible for these different outcomes, as well as the potential for further optimization of the transgenic production of unusual fatty acids in transgenic plants.

[1]  Qing Liu,et al.  Metabolic engineering of Arabidopsis to produce nutritionally important DHA in seed oil. , 2005, Functional plant biology : FPB.

[2]  Edgar B Cahoon,et al.  Fungal responsive fatty acid acetylenases occur widely in evolutionarily distant plant families. , 2003, The Plant journal : for cell and molecular biology.

[3]  A. Green From alpha to omega—producing essential fatty acids in plants , 2004, Nature Biotechnology.

[4]  S. Stymne,et al.  Acyl exchange between oleoyl-CoA and phosphatidylcholine in microsomes of developing soya bean cotyledons and its role in fatty acid desaturation , 1981, Lipids.

[5]  Sunita Chaudhary,et al.  Cloning of Δ12- and Δ6-desaturases from Mortierella alpina and recombinant production of γ-linolenic acid in Saccharomyces cerevisiae , 1999, Lipids.

[6]  H. Klee,et al.  Agrobacterium-Mediated Plant Transformation and its Further Applications to Plant Biology , 1987 .

[7]  J. Napier,et al.  Identification of Primula “front-end” desaturases with distinct n−6 or n−3 substrate preferences , 2006, Planta.

[8]  P. Calder n-3 Fatty acids and cardiovascular disease: evidence explained and mechanisms explored. , 2004, Clinical science.

[9]  S. Stymne,et al.  Evidence for cytochrome b5 as an electron donor in ricinoleic acid biosynthesis in microsomal preparations from developing castor bean (Ricinus communis L.). , 1992, The Biochemical journal.

[10]  Yoshikazu Tanaka,et al.  Production of γ-linolenic acid in Lotus japonicus and Vigna angularis by expression of the Δ6-fatty-acid desaturase gene isolated from Mortierella alpina , 2005 .

[11]  P. Strittmatter,et al.  Purification and properties of rat liver microsomal stearyl coenzyme A desaturase. , 1974, Proceedings of the National Academy of Sciences of the United States of America.

[12]  Qing Liu,et al.  Combined transgenic expression of Δ12-desaturase and Δ12-epoxygenase in high linoleic acid seeds leads to increased accumulation of vernolic acid. , 2006, Functional plant biology : FPB.

[13]  Alimuddin,et al.  Enhancement of EPA and DHA biosynthesis by over-expression of masu salmon Δ6-desaturase-like gene in zebrafish , 2005, Transgenic Research.

[14]  S. Boddupalli,et al.  A bifunctional oleate 12-hydroxylase: desaturase from Lesquerella fendleri. , 1998, The Plant journal : for cell and molecular biology.

[15]  R. Wait,et al.  Proteomic analysis of the endoplasmic reticulum from developing and germinating seed of castor (Ricinus communis) , 2002 .

[16]  D. Rice,et al.  Acyltransferases and their role in the biosynthesis of lipids : opportunities for new oils , 2001 .

[17]  L. Chuang,et al.  Identification of two novel microalgal enzymes involved in the conversion of the omega3-fatty acid, eicosapentaenoic acid, into docosahexaenoic acid. , 2004, The Biochemical journal.

[18]  X. Qiu,et al.  Isolation and characterization of a Δ5 FA desaturase from Pythium irregulare by heterologous expression in Saccharomyces cerevisiae and oilseed crops , 2002, Lipids.

[19]  P. Broun,et al.  Accumulation of Ricinoleic, Lesquerolic, and Densipolic Acids in Seeds of Transgenic Arabidopsis Plants That Express a Fatty Acyl Hydroxylase cDNA from Castor Bean , 1997, Plant physiology.

[20]  E. Cahoon,et al.  Industrial oils from transgenic plants. , 2003, Current opinion in plant biology.

[21]  Johnathan A. Napier,et al.  Progress toward the production of long-chain polyunsaturated fatty acids in transgenic plants , 2004, Lipids.

[22]  Chaofu Lu,et al.  A high-throughput screen for genes from castor that boost hydroxy fatty acid accumulation in seed oils of transgenic Arabidopsis. , 2006, The Plant journal : for cell and molecular biology.

[23]  E. Heinz First breakthroughs in sustainable production of “oceanic fatty acids” , 2006 .

[24]  J. Napier,et al.  The alternative pathway C20 Δ8‐desaturase from the non‐photosynthetic organism Acanthamoeba castellanii is an atypical cytochrome b 5‐fusion desaturase , 2006, FEBS letters.

[25]  T. Thomas,et al.  Expression of borage Δ6 desaturase in Saccharomyces cerevisiae and oilseed crops , 2002 .

[26]  S. Stymne,et al.  Phospholipid:diacylglycerol acyltransferase: an enzyme that catalyzes the acyl-CoA-independent formation of triacylglycerol in yeast and plants. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[27]  J. Napier,et al.  A growing family of cytochrome b5-domain fusion proteins , 1999 .

[28]  J. Browse,et al.  Mutants of Arabidopsis reveal many roles for membrane lipids. , 2002, Progress in lipid research.

[29]  J. Browse,et al.  Production of Polyunsaturated Fatty Acids by Polyketide Synthases in Both Prokaryotes and Eukaryotes , 2001, Science.

[30]  A. Kinney,et al.  The production of vegetable oils with novel properties: Using genomic tools to probe and manipulate plant fatty acid metabolism , 2005 .

[31]  M. Nishimura,et al.  A gene encoding a chloroplast omega-3 fatty acid desaturase complements alterations in fatty acid desaturation and chloroplast copy number of the fad7 mutant of Arabidopsis thaliana. , 1993, The Journal of biological chemistry.

[32]  L. Chuang,et al.  (Biochem. J., 384:357-366)Identification of two novel microalgal enzymes involved in the conversion of the ω3-fatty acid, eicosapentaenoic acid, into docosahexaenoic acid , 2004 .

[33]  R. Scarth,et al.  Modification of Brassica Oil Using Conventional and Transgenic Approaches , 2006 .

[34]  A. Kinney,et al.  Identification of an animal ω-3 fatty acid desaturase by heterologous expression in Arabidopsis , 1997 .

[35]  R. Mullen,et al.  Development and potential of genetically engineered oilseeds , 2005, Seed Science Research.

[36]  Fatty Acid Metabolism , 1988 .

[37]  D. Tocher,et al.  A vertebrate fatty acid desaturase with Δ5 and Δ6 activities , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[38]  P. Covello,et al.  High-Level Production of γ-Linolenic Acid in Brassica juncea Using a Δ6 Desaturase from Pythium irregulare , 2002, Plant Physiology.

[39]  John Shanklin,et al.  Desaturation and Hydroxylation , 2002, The Journal of Biological Chemistry.

[40]  T. Girke,et al.  Identification of a novel delta 6-acyl-group desaturase by targeted gene disruption in Physcomitrella patens. , 1998, The Plant journal : for cell and molecular biology.

[41]  P. Stumpf Plant lipid biosynthesis in 1959 and 1984. , 1984, Journal of lipid research.

[42]  S. Stymne,et al.  Electron-transport components of the 1-acyl-2-oleoyl-sn-glycero-3-phosphocholine delta 12-desaturase (delta 12-desaturase) in microsomal preparations from developing safflower (Carthamus tinctorius L.) cotyledons. , 1990, The Biochemical journal.

[43]  M. Iwabuchi,et al.  Δ12-Oleate Desaturase-related Enzymes Associated with Formation of Conjugated trans-Δ11, cis-Δ13 Double Bonds* , 2003, The Journal of Biological Chemistry.

[44]  T. Zank,et al.  The evolution of desaturases. , 2003, Prostaglandins, leukotrienes, and essential fatty acids.

[45]  A. Green,et al.  Transgenic expression of a Δ12-epoxygenase gene in Arabidopsis seeds inhibits accumulation of linoleic acid , 2001, Planta.

[46]  J. Browse,et al.  Arabidopsis mutants deficient in polyunsaturated fatty acid synthesis. Biochemical and genetic characterization of a plant oleoyl-phosphatidylcholine desaturase. , 1992, The Journal of biological chemistry.

[47]  Xiao Qiu,et al.  Stepwise engineering to produce high yields of very long-chain polyunsaturated fatty acids in plants , 2005, Nature Biotechnology.

[48]  J. Napier Plumbing the depths of PUFA biosynthesis: a novel polyketide synthase-like pathway from marine organisms. , 2002, Trends in plant science.

[49]  P. Stumpf Plants, fatty acids, compartments , 1981 .

[50]  S. Stymne,et al.  Plant Microsomal Phospholipid Acyl Hydrolases Have Selectivities for Uncommon Fatty Acids , 1995, Plant physiology.

[51]  A. Kinney,et al.  Formation of Conjugated Δ8,Δ10-Double Bonds by Δ12-Oleic-acid Desaturase-related Enzymes , 2001, The Journal of Biological Chemistry.

[52]  Yung-Sheng Huang,et al.  Enzymes for transgenic biosynthesis of long-chain polyunsaturated fatty acids. , 2004, Biochimie.

[53]  J. Napier,et al.  Eicosapentaenoic acid: biosynthetic routes and the potential for synthesis in transgenic plants. , 2004, Phytochemistry.

[54]  M. Bafor,et al.  Identification of non-heme diiron proteins that catalyze triple bond and epoxy group formation. , 1998, Science.

[55]  D. W. James,et al.  Mutants of Arabidopsis deficient in the synthesis of alpha-linolenate. Biochemical and genetic characterization of the endoplasmic reticulum linoleoyl desaturase. , 1993, The Journal of biological chemistry.

[56]  D. Taylor,et al.  Modification of seed oil content and acyl composition in the brassicaceae by expression of a yeast sn-2 acyltransferase gene. , 1997, The Plant cell.

[57]  J. Browse,et al.  A Mutation at the fad8 Locus of Arabidopsis Identifies a Second Chloroplast [omega]-3 Desaturase , 1994, Plant physiology.

[58]  B. Ruyter,et al.  Biotechnological approaches to modify rapeseed oil composition for applications in aquaculture , 2003 .

[59]  E. Heinz,et al.  Cloning and functional characterization of Phaeodactylum tricornutum front-end desaturases involved in eicosapentaenoic acid biosynthesis. , 2002, European journal of biochemistry.

[60]  E. Heinz,et al.  Relief for fish stocks: oceanic fatty acids in transgenic oilseeds. , 2005, Trends in plant science.

[61]  T. Shimmen,et al.  Purification and characterization of plant dynamin from tobacco BY-2 cells. , 2006, Plant & cell physiology.

[62]  Qing Liu,et al.  Metabolic engineering of new fatty acids in plants. , 2005, Current opinion in plant biology.

[63]  T. Starzl,et al.  Generation of cloned transgenic pigs rich in omega-3 fatty acids , 2006, Nature Biotechnology.

[64]  P. Shewry,et al.  Accumulation of Δ6-unsaturated fatty acids in transgenic tobacco plants expressing a Δ6-desaturase from Borago officinalis , 1999 .

[65]  P. Shewry,et al.  Expression of a borage desaturase cDNA containing an N-terminal cytochrome b5 domain results in the accumulation of high levels of delta6-desaturated fatty acids in transgenic tobacco. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[66]  P. Broun,et al.  Catalytic plasticity of fatty acid modification enzymes underlying chemical diversity of plant lipids. , 1998, Science.

[67]  A. Kinney,et al.  VARIATIONS IN THE BIOSYNTHESIS OF SEED-STORAGE LIPIDS. , 2001, Annual review of plant physiology and plant molecular biology.

[68]  A. Kinney,et al.  A novel omega3-fatty acid desaturase involved in the biosynthesis of eicosapentaenoic acid. , 2004, The Biochemical journal.

[69]  M. Truksa,et al.  Metabolic Engineering of Plants to Produce Very Long-chain Polyunsaturated Fatty Acids , 2006, Transgenic Research.

[70]  E. Heinz,et al.  In vivo desaturation of cis-delta 9-monounsaturated to cis-delta 9,12-diunsaturated alkenylether glycerolipids. , 1993, The Journal of biological chemistry.

[71]  J. Ohlrogge,et al.  Metabolic engineering of fatty acid biosynthesis in plants. , 2002, Metabolic engineering.

[72]  K. Riedel,et al.  Transgenic oilseeds as sustainable source of nutritionally relevant C20 and C22 polyunsaturated fatty acids , 2001 .

[73]  I. Hwang,et al.  Map-based cloning of a gene controlling omega-3 fatty acid desaturation in Arabidopsis. , 1992, Science.

[74]  A. Mañas-Fernández,et al.  Substrate specificity of acyl-Delta(6)-desaturases from Continental versus Macaronesian Echium species. , 2006, Phytochemistry.

[75]  B. Matthäus,et al.  A new database for seed oil fatty acids — the database SOFA , 2003 .

[76]  H.-G Opsahl-Ferstad,et al.  Erratum to “Biotechnological approaches to modify rapeseed oil composition for applications in aquaculture” , 2003 .

[77]  M. Mucenski,et al.  Identification and characterization of a lysophosphatidylcholine acyltransferase in alveolar type II cells. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[78]  J. Browse,et al.  A new class of Arabidopsis mutants with reduced hexadecatrienoic acid fatty acid levels. , 1998, Plant physiology.

[79]  J. Tomb,et al.  Identification of bifunctional Δ12/ω3 fatty acid desaturases for improving the ratio of ω3 to ω6 fatty acids in microbes and plants , 2006 .

[80]  P. Broun,et al.  An oleate 12-hydroxylase from Ricinus communis L. is a fatty acyl desaturase homolog. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[81]  P. Shewry,et al.  Heterologous reconstitution in yeast of the polyunsaturated fatty acid biosynthetic pathway. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[82]  P. Broun,et al.  Genetic engineering of plant lipids. , 1999, Annual review of nutrition.

[83]  G. Tucker Nutritional enhancement of plants. , 2003, Current opinion in biotechnology.

[84]  T. Mckeon,et al.  Biosynthesis of ricinoleate in castor oil. , 1999, Advances in experimental medicine and biology.

[85]  J. Widholm,et al.  Nucleotide Sequence of a Maize cDNA for a Class II, Acidic [beta]-1,3-Glucanase , 1994, Plant physiology.

[86]  S. Stymne,et al.  Evidence for the reversibility of the acyl-CoA:lysophosphatidylcholine acyltransferase in microsomal preparations from developing safflower (Carthamus tinctorius L.) cotyledons and rat liver. , 1984, The Biochemical journal.

[87]  H. Yamada,et al.  Production of polyunsaturated fatty acids by microorganisms. , 1992, Journal of nutritional science and vitaminology.

[88]  D. Murphy,et al.  Manipulation of plant oil composition for the production of valuable chemicals. Progress, problems, and prospects. , 1999, Advances in experimental medicine and biology.

[89]  D. Murphy Molecular breeding strategies for the modification of lipid composition , 2006, In Vitro Cellular & Developmental Biology - Plant.

[90]  X. Qiu Biosynthesis of docosahexaenoic acid (DHA, 22:6-4, 7,10,13,16,19): two distinct pathways. , 2003, Prostaglandins, leukotrienes, and essential fatty acids.

[91]  E. Heinz,et al.  Purification and PCR-based cDNA cloning of a plastidial n-6 desaturase , 1994, Plant Molecular Biology.

[92]  R. Reski,et al.  Cloning and functional characterisation of an enzyme involved in the elongation of Delta6-polyunsaturated fatty acids from the moss Physcomitrella patens. , 2002, The Plant journal : for cell and molecular biology.

[93]  E. Heinz,et al.  Isomeric sn-1-octadecenyl and sn-2-octadecenyl analogues of lysophosphatidylcholine as substrates for acylation and desaturation by plant microsomal membranes. , 1993, European journal of biochemistry.

[94]  K. Gable,et al.  Members of the Arabidopsis FAE1-like 3-Ketoacyl-CoA Synthase Gene Family Substitute for the Elop Proteins of Saccharomyces cerevisiae* , 2006, Journal of Biological Chemistry.

[95]  S. Stymne,et al.  The interconversion of diacylglycerol and phosphatidylcholine during triacylglycerol production in microsomal preparations of developing cotyledons of safflower (Carthamus tinctorius L.). , 1985, The Biochemical journal.

[96]  F. Garcia-Maroto,et al.  Cloning and molecular characterization of the Δ6-desaturase from two Echium plant species: Production of GLA by heterologous expression in yeast and tobacco , 2002, Lipids.

[97]  I. Feussner,et al.  Formation of conjugated Δ11Δ13‐double bonds by Δ12‐linoleic acid (1,4)‐acyl‐lipid‐desaturase in pomegranate seeds , 2002 .

[98]  A. Kinney,et al.  Biosynthetic origin of conjugated double bonds: production of fatty acid components of high-value drying oils in transgenic soybean embryos. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[99]  Johnathan A Napier,et al.  A Saccharomyces cerevisiae Gene Required for Heterologous Fatty Acid Elongase Activity Encodes a Microsomal β-Keto-reductase* , 2002, The Journal of Biological Chemistry.

[100]  J. Watts,et al.  Polyunsaturated fatty acid synthesis: what will they think of next? , 2002, Trends in biochemical sciences.

[101]  J. Browse,et al.  Cholinephosphotransferase and Diacylglycerol Acyltransferase (Substrate Specificities at a Key Branch Point in Seed Lipid Metabolism) , 1996, Plant physiology.

[102]  A. Green,et al.  Heterologous production of GLA and SDA by expression of an Echium plantagineum Δ6-desaturase gene , 2006 .

[103]  K. Feldmann,et al.  Arabidopsis FAD2 gene encodes the enzyme that is essential for polyunsaturated lipid synthesis. , 1994, The Plant cell.

[104]  S. Stymne,et al.  The biosynthesis of linoleate and α-linolenate in homogenates from developing soya bean cotyledons , 1980 .

[105]  A. Kinney,et al.  Co-expression of the borage Δ6 desaturase and the Arabidopsis Δ15 desaturase results in high accumulation of stearidonic acid in the seeds of transgenic soybean , 2006, Planta.

[106]  P. Covello,et al.  Identification and analysis of a gene from Calendula officinalis encoding a fatty acid conjugase. , 2001, Plant physiology.

[107]  J. Napier,et al.  Biosynthesis of Very-Long-Chain Polyunsaturated Fatty Acids in Transgenic Oilseeds: Constraints on Their Accumulationw⃞ , 2004, The Plant Cell Online.

[108]  A. Kinney Metabolic engineering in plants for human health and nutrition. , 2006, Current opinion in biotechnology.

[109]  X. Qiu,et al.  Identification of a (cid:1) 4 Fatty Acid Desaturase from Thraustochytrium sp. Involved in the Biosynthesis of Docosahexanoic Acid by Heterologous Expression in Saccharomyces cerevisiae and Brassica , 2001 .

[110]  John Shanklin,et al.  DESATURATION AND RELATED MODIFICATIONS OF FATTY ACIDS1. , 1998, Annual review of plant physiology and plant molecular biology.

[111]  J. Browse,et al.  Dissecting desaturation: plants prove advantageous. , 1996, Trends in cell biology.

[112]  D. Macherel,et al.  Site‐directed mutagenesis of histidine residues in the Δ12 acyl‐lipid desaturase of Synechocystis , 1995, FEBS letters.

[113]  S. Stymne,et al.  Remodelling of triacylglycerols in microsomal preparations from developing castor bean (Ricinus communis L.) endosperm , 1997, Planta.

[114]  J. Napier,et al.  Functional characterisation of two cytochrome b5-fusion desaturases from Anemone leveillei: the unexpected identification of a fatty acid Δ6-desaturase , 2003, Planta.

[115]  J. Shockey,et al.  Tung Tree DGAT1 and DGAT2 Have Nonredundant Functions in Triacylglycerol Biosynthesis and Are Localized to Different Subdomains of the Endoplasmic Reticulum[W] , 2006, The Plant Cell Online.

[116]  S. Stymne,et al.  Biosynthesis of gamma-linolenic acid in cotyledons and microsomal preparations of the developing seeds of common borage (Borago officinalis). , 1986, The Biochemical journal.

[117]  B. Fox,et al.  Eight histidine residues are catalytically essential in a membrane-associated iron enzyme, stearoyl-CoA desaturase, and are conserved in alkane hydroxylase and xylene monooxygenase. , 1994, Biochemistry.

[118]  J. Shanklin,et al.  Stearoyl-acyl-carrier-protein desaturase from higher plants is structurally unrelated to the animal and fungal homologs. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[119]  J. Ohlrogge,et al.  What limits production of unusual monoenoic fatty acids in transgenic plants? , 2002, Planta.

[120]  Kennedy Ep Biosynthesis of complex lipids. , 1961 .

[121]  D. Murphy Production of novel oils in plants. , 1999, Current opinion in biotechnology.

[122]  E. Cahoon,et al.  Transgenic Production of Epoxy Fatty Acids by Expression of a Cytochrome P450 Enzyme from Euphorbia lagascaeSeed , 2002, Plant Physiology.

[123]  J. Napier,et al.  Identification of a cDNA encoding a novel C18‐Δ9 polyunsaturated fatty acid‐specific elongating activity from the docosahexaenoic acid (DHA)‐producing microalga, Isochrysis galbana 1 , 2002, FEBS letters.

[124]  J. Napier,et al.  A Bifunctional Δ12,Δ15-Desaturase from Acanthamoeba castellanii Directs the Synthesis of Highly Unusual n-1 Series Unsaturated Fatty Acids* , 2006, Journal of Biological Chemistry.

[125]  Christine M. Williams,et al.  Long-chain n−3 PUFA: plant v. marine sources , 2006, Proceedings of the Nutrition Society.

[126]  J. Napier,et al.  Progress towards the production of very long-chain polyunsaturated fatty acid in transgenic plants: plant metabolic engineering comes of age , 2006 .

[127]  R. Mullen,et al.  Molecular Analysis of a Bifunctional Fatty Acid Conjugase/Desaturase from Tung. Implications for the Evolution of Plant Fatty Acid Diversity1 , 2002, Plant Physiology.

[128]  H. Sprecher,et al.  Elongation of long-chain fatty acids. , 2004, Progress in lipid research.

[129]  L. Kunst,et al.  All fatty acids are not equal: discrimination in plant membrane lipids. , 2000, Trends in plant science.

[130]  Grace Q Chen,et al.  Cloning and characterization of a cDNA encoding diacylglycerol acyltransferase from castor bean , 2004, Lipids.

[131]  T. Zank,et al.  Metabolic engineering of fatty acids for breeding of new oilseed crops: strategies, problems and first results. , 2003, Journal of plant physiology.

[132]  Petra Cirpus,et al.  Biosynthesis of docosahexaenoic acid in Euglena gracilis: biochemical and molecular evidence for the involvement of a Delta4-fatty acyl group desaturase. , 2003, Biochemistry.

[133]  Takao Shimizu,et al.  Cloning and Characterization of Mouse Lung-type Acyl-CoA:Lysophosphatidylcholine Acyltransferase 1 (LPCAT1) , 2006, Journal of Biological Chemistry.

[134]  J. Napier,et al.  The role of cytochrome b5 fusion desaturases in the synthesis of polyunsaturated fatty acids. , 2003, Prostaglandins, leukotrienes, and essential fatty acids.

[135]  D. W. James,et al.  A Mutant of Arabidopsis Deficient in the Elongation of Palmitic Acid , 1994, Plant physiology.

[136]  S. Cunnane Problems with essential fatty acids: time for a new paradigm? , 2003, Progress in lipid research.

[137]  J. Browse,et al.  Fluxes through the prokaryotic and eukaryotic pathways of lipid synthesis in the '16:3' plant Arabidopsis thaliana. , 1986, The Biochemical journal.

[138]  J. Napier,et al.  Production of very long chain polyunsaturated omega-3 and omega-6 fatty acids in plants , 2004, Nature Biotechnology.

[139]  S. Robert Production of Eicosapentaenoic and Docosahexaenoic Acid-Containing Oils in Transgenic Land Plants for Human and Aquaculture Nutrition , 2005, Marine Biotechnology.

[140]  A. Kinney,et al.  Production of γ-linolenic acid and stearidonic acid in seeds of marker-free transgenic soybean , 2004 .

[141]  S. Stymne,et al.  Plant microsomal phospholipases exhibit preference for phosphatidylcholine with oxygenated acyl groups , 1992 .

[142]  S. Stymne,et al.  The role of the acyl-CoA pool in the synthesis of polyunsaturated 18-carbon fatty acids and triacylglycerol production in the microsomes of developing safflower seeds. , 1983, Biochimica et biophysica acta.

[143]  J. Napier Transgenic plants as a source of fish oils: healthy, sustainable and GM , 2007 .

[144]  J. Ohlrogge,et al.  The genetics of plant lipids. , 1991, Biochimica et biophysica acta.

[145]  J. Napier,et al.  The production of very-long-chain PUFA biosynthesis in transgenic plants: towards a sustainable source of fish oils , 2005, The Proceedings of the Nutrition Society.

[146]  Edgar B Cahoon,et al.  Dimorphecolic Acid Is Synthesized by the Coordinate Activities of Two Divergent Δ12-Oleic Acid Desaturases* , 2004, Journal of Biological Chemistry.

[147]  T. Zank,et al.  Acyl Carriers Used as Substrates by the Desaturases and Elongases Involved in Very Long-chain Polyunsaturated Fatty Acids Biosynthesis Reconstituted in Yeast* , 2003, Journal of Biological Chemistry.

[148]  A. Kinney,et al.  Conjugated fatty acids accumulate to high levels in phospholipids of metabolically engineered soybean and Arabidopsis seeds. , 2006, Phytochemistry.

[149]  M. Bafor,et al.  Ricinoleic acid biosynthesis and triacylglycerol assembly in microsomal preparations from developing castor-bean (Ricinus communis) endosperm. , 1991, The Biochemical journal.

[150]  S. Stymne,et al.  Distribution of fatty acids in polar and neutral lipids during seed development in Arabidopsis thaliana genetically engineered to produce acetylenic, epoxy and hydroxy fatty acids , 2001 .

[151]  H. Moreau,et al.  In vivo characterization of the first acyl-CoA Delta6-desaturase from a member of the plant kingdom, the microalga Ostreococcus tauri. , 2005, The Biochemical journal.

[152]  T. Thomas,et al.  Expression of a cyanobacterial Δ6-desaturase gene results in γ-linolenic acid production in transgenic plants , 1996, Nature Biotechnology.

[153]  Yoshikazu Tanaka,et al.  Expression of Δ6, Δ5 desaturase and GLELO elongase genes from Mortierella alpina for production of arachidonic acid in soybean [Glycine max (L.) Merrill] seeds , 2006 .

[154]  T. Tonon,et al.  Fatty acid desaturases from the microalga Thalassiosira pseudonana , 2005, The FEBS journal.

[155]  Mark A. Smith,et al.  Heterologous expression of a fatty acid hydroxylase gene in developing seeds of Arabidopsis thaliana , 2003, Planta.