Production of dha and other lc pufas in plants

Abstract

The invention provides recombinant host organisms genetically modified with a polyunsaturated fatty acid (PUFA) synthase system and one or more accessory proteins that allow for and/or improve the production of PUFAs in the host organism. The present invention also relates to methods of making and using such organisms as well as products obtained from such organisms.

What is claimed is: 
     
         1 . A method of genetically modifying a soybean plant, descendant, cell, tissue, seed or part thereof, comprising modifying a soybean plant to include:
 (i) a nucleic acid molecule comprising a soybean codon-optimized coding sequence encoding an algal polyunsaturated fatty acid (PUFA) synthase that produces at least one PUFA; and   (ii) a nucleic acid molecule comprising a soybean codon-optimized coding sequence encoding algal phosphopantetheinyl transferase (PPTase).   
     
     
         2 . The method of  claim 1 , wherein the PUFA synthase and the PPTase enzymes are from  Schizochytrium  or  Thraustochytrium microalgae.    
     
     
         3 . The method of  claim 1 , wherein the PUFA synthase comprises an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8. 
     
     
         4 . The method of  claim 1 , wherein the PUFA synthase comprises the amino acid sequence of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8. 
     
     
         5 . The method of  claim 1 , wherein the PUFA synthase comprises the amino acid sequences of SEQ ID NOs:1-3 or SEQ ID NOs:6-8. 
     
     
         6 . The method of  claim 1 , wherein the PPTase comprises an amino acid sequence that is at least 80% identical to SEQ ID NO:5 or SEQ ID NO: 10. 
     
     
         7 . The method of  claim 1 , wherein the PPTase comprises the amino acid sequence of SEQ ID NO:5 or SEQ ID NO: 10. 
     
     
         8 . The method of  claim 3 , wherein the PPTase comprises an amino acid sequence that is at least 80% identical to SEQ ID NO:5 or SEQ ID NO: 10. 
     
     
         9 . The method of  claim 4 , wherein the PPTase comprises the amino acid sequence of SEQ ID NO:5 or SEQ ID NO: 10 
     
     
         10 . The method of  claim 1 , wherein the nucleic acid sequences of (i) and (ii) are contained in a single recombinant expression vector. 
     
     
         11 . The method of  claim 1 , wherein the nucleic acid sequence of (i) or (ii) is operably linked to a seed-specific promoter or a leaf-specific promoter. 
     
     
         12 . The method of  claim 1 , wherein the nucleic acid sequence of (i) or (ii) is operably linked to PvDlec2, LfKCS3, FAE 1, BoACP, BnaNapinC, ubiquitin or CsVMV promoter. 
     
     
         13 . The method of  claim 1 , further comprising modifying the soybean plant to include:
 (iii) a nucleic acid molecule comprising a nucleic acid sequence encoding an acyl-CoA synthetase (ACoAS).   
     
     
         14 . The method of  claim 13 , wherein the ACoAS comprises an amino acid sequence that is at least 80% identical to SEQ ID NO:4 or SEQ ID NO:9. 
     
     
         15 . The method of  claim 13 , wherein the ACoAS comprises the amino acid sequence of SEQ ID NO:4 or SEQ ID NO:9. 
     
     
         16 . The method of  claim 13 , wherein the nucleic acid sequence of (iii) is operably linked to a seed-specific promoter or a leaf-specific promoter. 
     
     
         17 . The method of  claim 13 , wherein the nucleic acid sequence of (iii) is operably linked to PvDlec2, LfKCS3, FAE 1, BoACP, BnaNapinC, ubiquitin or CsVMV promoter. 
     
     
         18 . The method of  claim 13 , wherein the nucleic acid sequence of (iii) encodes an acetyl CoA carboxylase (ACCase) or a type 2 diacylglycerol acyltransferase (DGAT2).