Delta-9 elongases and their use in making polyunsaturated fatty acids

Abstract

Isolated nucleic acid fragments and recombinant constructs comprising such fragments encoding delta-9 elongases along with a method of making long-chain polyunsaturated fatty acids (PUFAs) and using these delta-9 elongases in plants.

1 . An isolated polynucleotide comprising:
 (a) a nucleotide sequence encoding a polypeptide having delta-9 elongase activity, wherein the polypeptide has at least 80% amino acid identity, based on the Clustal V method of alignment, when compared to an amino acid sequence as set forth in SEQ ID NO:13 or SEQ ID NO:14;   (b) a nucleotide sequence encoding a polypeptide having delta-9 elongase activity, wherein the nucleotide sequence has at least 80% sequence identity, based on the BLASTN method of alignment, when compared to a nucleotide sequence as set forth in SEQ ID NO:11 or SEQ ID NO:12;   (c) a nucleotide sequence encoding a polypeptide having delta-9 elongase activity, wherein the nucleotide sequence hybridizes under stringent conditions to a nucleotide sequence as set forth in SEQ ID NO:11 or SEQ ID NO:12; or   (d) a complement of the nucleotide sequence of (a), (b) or (c), wherein the complement and the nucleotide sequence consist of the same number of nucleotides and are 100% complementary.   
     
     
         2 . The polypeptide of  claim 1 , wherein the amino acid sequence of the polypeptide has at least 85% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NO:13 or SEQ ID NO:14. 
     
     
         3 . The polypeptide of  claim 1 , wherein the amino acid sequence of the polypeptide has at least 90% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NO:13 or SEQ ID NO:14. 
     
     
         4 . The polypeptide of  claim 1 , wherein the amino acid sequence of the polypeptide has at least 95% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NO:13 or SEQ ID NO:14. 
     
     
         5 . The polynucleotide of  claim 1  wherein the nucleotide sequence comprises SEQ ID NO:11 or SEQ ID NO:12. 
     
     
         6 . The polynucleotide of  claim 1 , wherein the amino acid sequence of the polypeptide comprises
 (a) SEQ ID NO:13 or SEQ ID NO:14; or   (b) an amino acid sequence that differs from the amino acid sequences in (a) by at least one conservative amino acid substitution.   
     
     
         7 . The polynucleotide of  claim 1 , wherein the amino acid sequence of the polypeptide comprises SEQ ID NO:13 or SEQ ID NO:14. 
     
     
         8 . A recombinant DNA construct comprising the polynucleotide of  claim 1 ,  5 ,  6  or  7  operably linked to at least one regulatory sequence. 
     
     
         9 . A plant cell comprising in its genome the recombinant DNA construct of  claim 8 . 
     
     
         10 . A method for transforming a plant cell, comprising transforming a plant cell with the recombinant construct of  claim 8  and selecting those plant cells transformed with the recombinant construct of  claim 8 . 
     
     
         11 . A method for producing a transformed plant comprising transforming a plant cell with the polynucleotide of  claim 1 ,  5 ,  6  or  7  and regenerating a plant from the transformed plant cell. 
     
     
         12 . The method of  claim 11  wherein the plant is a soybean plant. 
     
     
         13 . A transgenic seed comprising in its genome the recombinant construct of  claim 8 . 
     
     
         14 . A transgenic seed obtained from the plant made by the method of  claim 11  or  12 . 
     
     
         15 . Oil or by-products obtained from the seed of  claim 13 . 
     
     
         16 . Oil or by-products obtained from the seed of  claim 14 . 
     
     
         17 . A method for making long-chain polyunsaturated fatty acids in a plant cell comprising:
 (a) transforming a plant cell with the recombinant construct of  claim 8 ; and   (b) selecting those transformed plant cells that make long-chain polyunsaturated fatty acids.   
     
     
         18 . A method for producing at least one polyunsaturated fatty acid in an oilseed plant cell comprising:
 (a) transforming an oilseed plant cell with a first recombinant DNA construct comprising an isolated polynucleotide encoding at least one delta-9 elongase 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 desaturase, a delta-5 desaturase, a delta-6 desaturase, a delta-8 desaturase, a delta-12 desaturase, a delta-15 desaturase, a delta-17 desaturase, a delta-9 desaturase, a delta-9 elongase, a C 14/16  elongase, a C 16/18  elongase, a C 18/20  elongase and a C 20/22  elongase;   (b) regenerating an oilseed plant from the transformed cell of step (a); and   (c) selecting those seeds obtained from the plants of step (b) having an altered level of polyunsaturated fatty acids when compared to the level in seeds obtained from a nontransformed oilseed plant.   
     
     
         19 . The method of  claim 18  wherein the oilseed plant is selected from the group consisting of soybean,  Brassica  species, sunflower, maize, cotton, flax, and safflower. 
     
     
         20 . An oilseed plant comprising in its genome the recombinant construct of  claim 8 . 
     
     
         21 . An oilseed plant comprising:
 (a) a first recombinant DNA construct comprising an isolated polynucleotide encoding at least one delta-9 elongase 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 desaturase, a delta-5 desaturase, a delta-6 desaturase, a delta-8 desaturase, a delta-12 desaturase, a delta-15 desaturase, a delta-17 desaturase, a delta-9 desaturase, a delta-9 elongase, a C 14/16  elongase, a C 16/18  elongase, a C 18/20  elongase and a C 20/22  elongase.   
     
     
         22 . The oilseed plant of  claim 20  or  21  wherein the oilseed plant is selected from the group consisting of soybean,  Brassica  species, sunflower, maize, cotton, flax and safflower. 
     
     
         23 . A transgenic seed obtained from the oilseed plant of  claim 20 . 
     
     
         24 . A transgenic seed obtained from the oilseed plant of  claim 21 . 
     
     
         25 . Oil or by-products obtained from the seed of  claim 23 . 
     
     
         26 . Oil or by-products obtained from the seed of  claim 24 . 
     
     
         27 . Oil obtained by the method of  claim 17  or  18 . 
     
     
         28 . Food or feed which incorporates the oil of  claim 25 . 
     
     
         29 . Food or feed which incorporates the oil of  claim 26 . 
     
     
         30 . Food or feed which incorporates the oil of  claim 27 . 
     
     
         31 . Food or feed comprising the seed of  claim 23 . 
     
     
         32 . Food or feed comprising the seed of  claim 24   
     
     
         33 . Food or feed comprising an ingredient derived from the processing of the seeds of  claim 23 . 
     
     
         34 . Food or feed comprising an ingredient derived from the processing of the seed of  claim 24 . 
     
     
         35 . The oil or by-products of  claim 25  wherein the by-product is lecithin. 
     
     
         36 . The oil or by-products of  claim 26  wherein the by-product is lecithin. 
     
     
         37 . Progeny plants obtained from the plant made by the method of  claim 11  or  12 . 
     
     
         38 . Progeny plants obtained from the oilseed plant of  claim 20  or  21 .