Delta-8 desaturase and its use in making polyunsaturated fatty acids

Abstract

Isolated nucleic acid fragments and recombinant constructs comprising such fragments encoding a delta-8 desaturase along with a method of making long chain polyunsaturated fatty acids (PUFAs) using this delta-8 desaturase in plants and oleaginous yeast.

1 . An isolated polynucleotide comprising:
 (a) a nucleotide sequence encoding a polypeptide having delta-8 desaturase activity, wherein the polypeptide has an amino acid sequence consisting essentially of SEQ ID NOs:2 or 113; or,   (b) a complement of the nucleotide sequence, wherein the complement and the nucleotide sequence consist of the same number of nucleotides and are 100% complementary.   
     
     
         2 - 38 . (canceled) 
     
     
         39 . Oil obtained from a seed obtained from a plant comprising a recombinant construct comprising an isolated polynucleotide comprising:
 (a) a nucleotide sequence encoding a polypeptide having delta-8 desaturase activity, wherein the polypeptide has an amino acid sequence consisting essentially of SEQ ID NOs:2; or,   (b) a complement of the nucleotide sequence, wherein the complement and the nucleotide sequence consist of the same number of nucleotides and are 100% complementary.   
     
     
         40 . Oil obtained from a seed obtained from a plant made by a method for producing a transformed plant comprising transforming a plant cell with an isolated polynucleotide comprising:
 (a) a nucleotide sequence encoding a polypeptide having delta-8 desaturase activity, wherein the polypeptide has an amino acid sequence consisting essentially of SEQ ID NOs:2; or,   (b) a complement of the nucleotide sequence, wherein the complement and the nucleotide sequence consist of the same number of nucleotides and are 100% complementary and   (c) regenerating a plant from the transformed plant cell.   
     
     
         41 . The oil of  claim 39 , wherein the recombinant construct is operably linked to at least one regulatory sequence. 
     
     
         42 . The oil of  claim 39 , wherein the plant is an oilseed plant. 
     
     
         43 . Oil obtained from a seed obtained from a plant made by a method for producing a transformed plant comprising:
 a) transforming a plant cell with an isolated polynucleotide comprising with:   (i) a nucleotide sequence encoding a polypeptide having delta-8 desaturase activity, wherein the polypeptide has an amino acid sequence consisting essentially of SEQ ID NOs:2; or,   (ii) a complement of the nucleotide sequence, wherein the complement and the nucleotide sequence consist of the same number of nucleotides and are 100% complementary,   and b) regenerating a plant from the transformed plant cell.   
     
     
         44 . The oil of  claim 43 , wherein the plant is a soybean plant. 
     
     
         45 . Oil obtained from a seed obtained from an oilseed plant comprising a recombinant construct comprising:
 a) a first recombinant DNA construct comprising an isolated polynucleotide encoding a delta-8 desaturase polypeptide, operably linked to at least one regulatory sequence; and   b) at least one additional recombinant DNA construct comprising an isolated polynucleotide, operably linked to at least one regulatory sequence, encoding a polypeptide selected from the group consisting of a delta-4, a delta-5, a delta-6, a delta-9, a delta-12, a delta-15, and a delta-17 desaturase, a delta-9 elongase, a C18 to C22 elongase and a C20 to C24 elongase.   
     
     
         46 . Oil obtained by a method for making long chain fatty acids in a plant cell comprising:
 a) transforming a cell with a recombinant construct comprising an isolated polynucleotide comprising:
 (i) a nucleotide sequence encoding a polypeptide having delta-8 desaturase activity, wherein the polypeptide has an amino acid sequence consisting essentially of SEQ ID NOs:2; or, 
 (ii) a complement of the nucleotide sequence, wherein the complement and the nucleotide sequence consist of the same number of nucleotides and are 100% complementary, 
   and b) selecting those transformed cells that make long chain polyunsaturated fatty acids.   
     
     
         47 . Oil obtained by a method for producing at least one polyunsaturated fatty acid in a soybean cell comprising:
 a) transforming a soybean cell with a recombinant DNA construct comprising an isolated polynucleotide encoding a delta-8 desaturase polypeptide, operably linked to at least one regulatory sequence and at least one additional recombinant DNA construct comprising an isolated polynucleotide, operably linked to at least one regulatory sequence, encoding a polypeptide selected from the group consisting of a delta-4, a delta-5, a delta-6, a delta-9, a delta-12, a delta-15, and a delta-17 desaturase, a delta-9 elongase, a C18 to C22 elongase and a C20 to C24 elongase;   b) regenerating a soybean plant from the transformed cell of step a); and   c) selecting those seeds obtained from the plants of step b), wherein the seeds have an altered level of polyunsaturated fatty acids when compared to the level in seeds obtained from a nontransformed soybean plant.