Oils, Lipids and Fatty Acids Produced in Transgenic Brassica Plant

Abstract

The invention relates to a method for producing eicosapentanoic acid, docosapentanoic acid and/or docohexanoic acid in transgenic plants. According to said method, the plant is provided with at least one nucleic acid sequence coding for a polypetide with a Δ6 desaturase activity, at least one nucleic acid sequence coding for a polypeptide with a Δ6 elongase activity, at least one nucleic acid sequence coding for a polypeptide with a Δ5 desaturase activity, and at least one nucleic acid sequence coding for a polypeptide with a Δ5 elongase activity, the nucleic acid sequence coding for a polypeptide with a Δ5 elongase activity being modified in relation to the nucleic acid sequence in the organism from which the sequence originates, such that it is adapted to the codon use in at least one type of plant. For the production of docosahexanoic acid, at least one nucleic acid sequence coding for a polypeptide with a Δ4 desaturase activity is also introduced into the plant.

1 . A process for producing eicosapentaenoic acid, docosapentaenoic acid and/or docosahexaenoic acid in a transgenic plant, comprising the provision in the plant of at least one nucleic acid sequence which codes for a polypeptide having a Δ6-desaturase activity; at least one nucleic acid sequence which codes for a polypeptide having a Δ6-elongase activity; at least one nucleic acid sequence which codes for a polypeptide having a Δ5-desaturase activity; and at least one nucleic acid sequence which codes for a polypeptide having a Δ5-elongase activity and optionally for a Δ4-desaturase,
 where the nucleic acid sequence which codes for a polypeptide having a Δ5-elongase activity is modified by comparison with the nucleic acid sequence in the organism from which the sequence is derived in that it is adapted to the codon usage in one or more plant species. 
 
     
     
         2 . The process as claimed in  claim 1 , where the nucleic acid sequence is adapted at least to the codon usage in oilseed rape, soybean and/or flax. 
     
     
         3 . The process as claimed in  claim 1 , where the nucleic acid sequence is adapted taking into account the natural frequency of individual codons. 
     
     
         4 . The process as claimed in  claim 1 , where the modified nucleic acid sequences corresponds to the nucleic acid sequence indicated in SEQ ID No. 64. 
     
     
         5 . The process as claimed in  claim 1 , where the nucleic acid sequences are expressed under the control of a seed-specific promoter. 
     
     
         6 . The process as claimed in  claim 5 , where the promoter is the USP, vicilin, napin, Glp, SBP, peroxireduxin, legumin, Fad3, conlinin or oleosin promoter. 
     
     
         7 . The process as claimed in  claim 6 , where the content of polyunsaturated C22 fatty acids in the seed oil is 5% by weight or more of the seed oil content. 
     
     
         8 . The process as claimed in  claim 1 , where additionally one or more nucleic acid sequences coding for a polypeptide having the activity of an ω3 desaturase and/or a Δ4-desaturase are provided in the plant. 
     
     
         9 . The process as claimed in  claim 8 , where the content of docosahexaenoic acid in the seed oil is 1% by weight or more of the seed oil content. 
     
     
         10 . The process as claimed in  claim 1 , where the eicosapentaenoic acid, docosapentaenoic acid and/or docosahexaenoic acid is present in the plant mainly bound as ester in phospholipids or triacylglycerides. 
     
     
         11 . The process as claimed in  claim 1 , where the plant is an oil-producing plant selected from the group consisting of  Brassica napus, Brassica juncea  and  Glycine max.    
     
     
         12 . The process as claimed in  claim 1 , further comprising the uptake of the eicosapentaenoic acid, docosapentaenoic acid and/or docosahexaenoic acid in the form of their oils, lipids or free fatty acids from the plant. 
     
     
         13 . An isolated nucleic acid molecule comprising a the nucleic acid sequence as shown in SEQ ID No. 64. 
     
     
         14 . A recombinant nucleic acid molecule comprising:
 a) one or more copies of at least one promoter which is active in plant cells,   b) at least one nucleic acid sequence which codes for a polypeptide having a Δ6-desaturase activity,   c) at least one nucleic acid sequence which codes for a polypeptide having a Δ5-desaturase activity,   d) at least one nucleic acid sequence which codes for a polypeptide having a Δ6-elongase activity,   e) at least one nucleic acid sequence which codes for a polypeptide having a Δ5-elongase activity and which is modified by comparison with the nucleic acid sequence in the organism from which the sequence is derived by being adapted to the codon usage in one or more plant species, and   f) one or more copies of at least one terminator sequence.   
     
     
         15 . The recombinant nucleic acid molecule as claimed in  claim 14 , where the modified nucleic acid sequence corresponds to the nucleic acid sequence indicated in SEQ ID No. 64. 
     
     
         16 . The recombinant nucleic acid molecule as claimed in  claim 14 , additionally comprising one or more nucleic acid sequences coding for a polypeptide having the activity of an ω3-desaturase and/or a Δ-4-desaturase. 
     
     
         17 . A transgenic plant comprising a recombinant nucleic acid molecule according to claim  22  or comprising the nucleic acid sequence indicated in SEQ ID No. 64. 
     
     
         18 . (canceled)