Washington State University

Bayer Cropscience Nv

John A Browse

Jennifer L Watts

James G Wallis

Jay M Shockey

Zhanguo Xin

Chaofu Lu

James P Spychalla

Peter Denolf

Judy Schnurr

Gregory B Tilton

Shuangyi Bai

Julie Jeannine Burgal

The amino acid and nucleic acid sequences of a &#x394;-desaturase enzyme and a &#x394;-desaturase enzyme are disclosed. The nucleic acid sequences can be used to design recombinant DNA constructs and vectors. These vectors can then be used to transform various organisms, including for example, plants and yeast. The transformed organisms will then produce polyunsaturated fatty acids. The amino acid sequences are useful for generating enzyme-specific antibodies that are useful for identifying the desaturases.

Desaturases and methods of using them for synthesis of polyunsaturated fatty acids

Aspects of the invention provide methods for differential regulation of fatty acid unsaturation in seed oil and membrane lipids of plants based on modulation of a previously unknown biosynthetic pathway involving a novel phosphatidylcholine:diacylglycerol cholinephosphotransferase (PDCT) that regulates phosphatidylcholine biosynthesis in developing oil seed plants. Specific aspects relate to inventive PDCT polypeptides including, for example, variants, deletions, muteins, fusion proteins, and orthologs thereof (collectively PDCT proteins), to nucleic acids encoding same, to plants comprising such PDCT sequences or proteins or devoid or depleted of such PDCT proteins or sequences, and to methods for generating plants having altered or no PDCT expression and/or activity, including but not limited to methods comprising mutagenesis, recombinant DNA, transgenics, etc.

Compositions and methods for differential regulation of fatty acid unsaturation in membrane lipids and seed oil

The present invention relates toplants comprising mutant phosphatidylcholine:diacylglycerol cholinephosphotransferase encoding (ROD1) genes, ROD1 nucleic acid sequences and proteins, as well as methods for generating and identifying said plants and alleles, which can be used to plants with increased levels of C18:1 in the seeds.

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Aspects of the invention provide methods for differential regulation of fatty acid unsaturation in seed oil and membrane lipids of plants based on modulation of a previously unknown biosynthetic pathway involving a novel phosphatidylcholine: diacylglycerol cholinephosphotransferase (PDCT) that regulates phosphatidylcholine biosynthesis in developing oil seed plants. Specific aspects relate to inventive PDCT polypeptides including, for example, variants, deletions, muteins, fusion proteins, and orthologs thereof (collectively PDCT proteins), to nucleic acids encoding same, to plants comprising such PDCT sequences or proteins or devoid or depleted of such PDCT proteins or sequences, and to methods for generating plants having altered or no PDCT expression and/or activity, including but not limited to methods comprising mutagenesis, recombinant DNA, transgenics, etc.

Particular aspects provide six novelcDNA clones, including cloned sequences of: DGAT (RcDGAT1 and RcDGAT2); RcLPAT; LACS (RcLACS4), and PDAT (RcPDAT1A and RcPDAT1B). Additional aspects provide methods for substantially enhanced accumulation of hydroxy fatty acid (HFA) in transgenic plant tissue (e.g., seeds), comprising expression of particular novel sequences. For example, expression of RcDGAT2 or RcPDAT1 in castor hydroxylase-expressinglines resulted in substantially enhanced accumulation of hydroxy fatty acid (HFA) (e.g., to over 30%; a 50-70% increase in HFA accumulation) relative to the hydroxylase-only expressing parental lines. Further aspects provide methods to increase at least one of total lipid content, percent seed germination, and seed weight in transgenic plants, comprising expression of RcDGAT2 in castor hydroxylase-expressing plant lines. Yet further aspects provide methods for expressing and accumulating hydroxyl fatty acid in yeast (e.g., TAG biosynthesis from diricinolein), comprising expression of RcDGAT2 RcDGAT2 coding sequences in yeast.

Enhancement of hydroxy fatty acid accumulation in oilseed plants

The present invention relates to genes encoding plant acyl-CoA synthetases and methods of their use. In particular, the present invention is related to plant acyl-coenzyme A synthetases. The present invention encompasses both native and recombinant wild-type forms of the enzymes, as well as mutant and variant forms, some of which possess altered characteristics relative to the wild-type enzyme. The present invention also relates to methods of using acyl-CoA synthetases, including altered expression in transgenic plants and expression in prokaryotes and cell culture systems.

Plant acyl-CoA synthetases

Recombinant expression of fat-1 gene ofin a wide variety of cells, including cells ofand, produces a polypeptide having &#x3c9;-3 desaturase activity.

&#x3c9;-3 fatty acid desaturase

The present invention provides compositions and methods related to acyl-coenzyme A thioesterases. In particular, the present invention is related to plant acyl-coenzyme A thioesterases. The present invention encompasses both native and recombinant wild-type forms of the enzymes, as well as mutant and variant forms, some of which possess altered characteristics relative to the wild-type enzyme. The present invention also relates to methods of using acyl-CoA thioesterases, including altered expression in transgenic plants and expression in prokaryotes and cell culture systems.

Acyl coenzyme A thioesterases

The amino acid and nucleic acid sequences of a &Dgr; -desaturase enzyme and a &Dgr; -desaturase enzyme are disclosed. The nucleic acid sequences can be used to design recombinant DNA constructs and vectors. These vectors can then be used to transform various organisms, including for example, plants and yeast. The transformed organisms will then produce polyunsaturated fatty acids. The amino acid sequences are useful for generating enzyme-specific antibodies that are useful for identifying the desaturases.

Recombinant expression of fat-1 gene of,in a wide variety of cells, including cells of,and,produces a polypeptide having &ohgr;-3 desaturase activity.

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Palouse, WA, United States of America

John A Browse