Plants with improved photosynthetic carbon fixation capacity

Abstract

Plants are provided with increased ribulose-1,5-bisphosphate (RuBP) regeneration capacity during the Calvin cycle through increased expression of sedoheptulose 1,7 bisphosphatase, in combination with reduced photo-respiratory losses through expression of glycolate catabolizing enzymes. Such plants have a greater growth rate and/or improved biomass and/or increased carbon fixation compared to untreated plants, or plants comprising only one of the features above.

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         2 . A method for improving the assimilation of carbon in a plant comprising the steps of:
 a. inserting into the genome of a plant a nucleic acid encoding an enzyme comprising sedoheptulose 1,7-bisphosphatase activity operably linked to a plant-expressible promoter, and optionally to a 3′ end region involved in transcription termination and/or polyadenylation; and   b. inserting into the genome of a plant one or more nucleic acids encoding one or more polypeptides having the enzymatic activity of glycolate dehydrogenase operably linked to a plant-expressible promoter, and optionally to a 3′ end region involved in transcription termination and/or polyadenylation, wherein said one or more polypeptides are located in chloroplasts of said plants.   
     
     
         3 . The method of  claim 2 , wherein said enzyme comprising sedoheptulose 1,7-bisphosphatase activity further comprises FBPase activity. 
     
     
         4 . The method of  claim 2 , wherein said enzyme comprising sedoheptulose 1,7-bisphosphatase activity is from a source heterologous with respect to the plant into which the nucleic acid is inserted, a plant species heterologous with respect to the plant into which the structural nucleic acid is inserted, from  Oryza sativa , or encoded by a nucleic acid wherein the codon usage has been adapted to the codon usage in wheat. 
     
     
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         11 . The method according to  claim 2 , wherein said promoter includes the promoter region of the ribulose-bisphosphate carboxylase small subunit gene of  Oryza sativa.    
     
     
         12 . The method according to  claim 11 , wherein said promoter comprises the complement of the nucleotide sequence of SEQ ID No. 12 from nucleotide position 2010 to 4759. 
     
     
         13 . The method according to  claim 2 , wherein said enzyme comprising sedoheptulose 1,7-bisphosphatase activity further comprises an amino acid sequence targeting said enzyme comprising sedoheptulose 1,7-bisphosphatase activity to the chloroplast or the chloroplast membrane or wherein said nucleic acid encoding an enzyme comprising sedoheptulose 1,7-bisphosphatase activity is integrated in the chloroplast genome of cells of said plant. 
     
     
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         16 . The method according to  claim 2 , wherein said one or more polypeptides having the enzymatic activity of a glycolate dehydrogenase are derived from the  E. coli  glc operon, a  Chlamydomonas  or  Synechocystis  glycolate dehydrogenase, or an  Arabidopsis  glycolate dehydrogenase. 
     
     
         17 . The method according to  claim 16 , wherein said one or more polypeptides comprise an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID No. 3, SEQ ID No. 4, and SEQ ID No. 5, SEQ ID No. 6, SEQ ID No. 7, SEQ ID NO. 8, or SEQ ID No. 9. 
     
     
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         19 . The method according to  claim 16 , wherein said polypeptides form a multi-subunit fusion protein. 
     
     
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         26 . The method according to  claim 16 , wherein said one or more polypeptides having glycolate dehydrogenase activity further comprise an amino acid sequence targeting said enzyme or subunits to the chloroplast or the chloroplast membrane or wherein said one or more nucleic acids encoding one or more polypeptides having glycolate dehydrogenase activity are integrated in the chloroplast genome of cells of said plant. 
     
     
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         29 . The method according to  claim 2 , wherein said promoter includes the promoter region of the ribulose-bisphosphate carboxylase small subunit gene of  Oryza sativa  or the promoter region of the cytosolic fructose 1,6 bisphosphatase gene of  Oryza sativa.    
     
     
         30 . The method according to  claim 29 , wherein said promoter comprises the complement of the nucleotide sequence of SEQ ID No. 12 from nucleotide position 2010 to 4759 or the nucleotide sequence of SEQ ID No. 11 from nucleotide position 75 to 1272. 
     
     
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         46 . A plant cell, a plant part, a plant or a seed comprising in its genome a nucleic acid encoding an enzyme comprising sedoheptulose 1,7-bisphosphatase activity operably linked to a plant-expressible promoter, and optionally to a 3′ end region involved in transcription termination and/or polyadenylation; and one or more nucleic acids encoding one or more polypeptides having the enzymatic activity of glycolate dehydrogenase operably linked to a plant-expressible promoter, and optionally to a 3′ end region involved in transcription termination and/or polyadenylation, wherein said one or more polypeptides are located in chloroplasts of said plants. 
     
     
         47 . The plant cell, a plant part, a plant or a seed of  claim 46 , wherein said enzyme comprising sedoheptulose 1,7-bisphosphate activity further comprises FBPase activity. 
     
     
         48 . The plant cell, a plant part, a plant or a seed of  claim 46 , wherein said enzyme comprising sedoheptulose 1,7-bisphosphatase activity is from a source heterologous with respect to the plant into which the nucleic acid is inserted, a plant species heterologous with respect to the plant into which the structural nucleic acid is inserted, from  Oryza sativa , or encoded by a nucleic acid wherein the codon usage has been adapted to the codon usage in wheat. 
     
     
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         55 . The plant cell, a plant part, a plant or a seed of  claim 46 , wherein said promoter includes the promoter region of the ribulose-bisphosphate carboxylase small subunit gene of  Oryza sativa.    
     
     
         56 . The plant cell, a plant part, a plant or a seed of  claim 55 , wherein said promoter comprises the complement of the nucleotide sequence of SEQ ID No. 12 from nucleotide position 2010 to 4759. 
     
     
         57 . The plant cell, a plant part, a plant or a seed according to  claim 46 , wherein said enzyme comprising sedoheptulose 1,7-bisphosphatase activity further comprises an amino acid sequence targeting said enzyme comprising sedoheptulose 1,7-bisphosphatase activity to the chloroplast or the chloroplast membrane or wherein nucleic acid encoding an enzyme comprising sedoheptulose 1,7-bisphosphatase activity is integrated in the chloroplast genome of cells of said plant. 
     
     
         58 . (canceled) 
     
     
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         60 . The plant cell, a plant part, a plant or a seed according to  claim 46 , wherein said one or more polypeptides having the enzymatic activity of a glycolate dehydrogenase are derived from the  E. coli  glc operon, a  Chlamydomonas  or  Synechocystis  glycolate dehydrogenase, or an  Arabidopsis  glycolate dehydrogenase. 
     
     
         61 . The plant cell, a plant part, a plant or a seed according to  claim 60 , wherein said one or more polypeptides comprise an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID No. 3, SEQ ID No 4, SEQ ID No. 5, SEQ ID No. 6, SEQ ID No. 7, SEQ ID NO. 8, or SEQ ID No. 9. 
     
     
         62 . (canceled) 
     
     
         63 . The plant cell, a plant part, a plant or a seed according to  claim 60 , wherein said polypeptides form a multi-subunit fusion protein. 
     
     
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         70 . The plant cell, a plant part, a plant or a seed according to  claim 60 , wherein said one or more polypeptides having glycolate dehydrogenase activity further comprise an amino acid sequence targeting said enzyme or subunits to the chloroplast or the chloroplast membrane or wherein said one or more nucleic acids encoding one or more polypeptides having glycolate dehydrogenase activity are integrated in the chloroplast genome of cells of said plant. 
     
     
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         73 . The plant cell, a plant part, a plant or a seed according to  claim 46 , wherein said promoter includes the promoter region of the ribulose-bisphosphate carboxylase small subunit gene of  Oryza sativa  or the promoter region of the cytosolic fructose 1,6 bisphosphatase gene of  Oryza sativa.    
     
     
         74 . The plant cell, a plant part, a plant or a seed according to  claim 73 , wherein said promoter comprises the complement of the nucleotide sequence of SEQ ID No. 12 from nucleotide position 2010 to 4759 or the nucleotide sequence of SEQ ID No. 11 from nucleotide position 75 to 1272. 
     
     
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         90 . A food product obtained from a plant or plant part or seed of  claim 46 . 
     
     
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