Plants with improved photosynthetic carbon fixation capacity
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.
1. 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, wherein said enzyme comprising sedoheptulose 1,7-bisphosphatase activity comprises an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID No. 2; and
b. inserting into the genome of said 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.
2. The method of claim 1 , wherein said enzyme comprising sedoheptulose 1,7-bisphosphatase activity further comprises FBPase activity or is from a source heterologous with respect to the plant into which the nucleic acid is inserted.
3. The method of claim 1 , wherein said enzyme comprising sedoheptulose 1,7-bisphosphatase activity comprises the amino acid sequence of SEQ ID No. 2.
4. The method of claim 3 , wherein said enzyme comprising sedoheptulose 1,7-bisphosphatase activity is encoded by a nucleic acid comprising the nucleotide sequence of SEQ ID No. 1.
5. The method according to claim 1 , wherein said promoter includes the promoter region of the ribulose-bisphosphate carboxylase small subunit gene of Oryza sativa.
6. The method according to claim 5 , wherein said promoter comprises the complement of the nucleotide sequence of SEQ ID No. 12 from nucleotide position 2010 to 4759.
7. The method according to claim 1 , 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.
8. The method according to claim 1 , wherein said one or more polypeptides having the enzymatic activity of a glycolate dehydrogenase are derived from the E. coli glc operon or wherein said one or more polypeptides having the enzymatic activity of a glycolate dehydrogenase is a Chlamydomonas or Synechocystis glycolate dehydrogenase or wherein said one or more polypeptides having the enzymatic activity of a glycolate dehydrogenase is an Arabidopsis glycolate dehydrogenase.
9. The method according to claim 8 , wherein said one or more polypeptides comprise an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID No. 3, SEQ ID No. 4 and SEQ ID No. 5 or to the amino acid sequence of SEQ ID No. 6, SEQ ID No. 7 or SEQ ID No. 8 or to the amino acid sequence of SEQ ID No. 9.
10. The method according to claim 8 , wherein said polypeptides form a multi-subunit fusion protein.
11. The method according to claim 1 , wherein said one or more polypeptides having glycolate dehydrogenase activity further comprise an amino acid sequence targeting said enzyme 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.
12. The method according to claim 1 , wherein said promoter includes the promoter region of the cytosolic fructose 1,6 bisphosphatase gene of Oryza sativa.
13. The method according to claim 12 , wherein said promoter comprises the nucleotide sequence of SEQ ID No. 11 from nucleotide position 75 to 1272.
14. The method according to claim 1 , wherein said plant is a monocotyledonous plant or a dicotyledonous plant or a Gymnosperm.
15. The method according to claim 14 , wherein said plant is selected from wheat, rice, maize, sorghum , millet, rye, oats, sugarcane, cotton, soybean or a Brassica plant.
16. 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 enzyme comprising sedoheptulose 1,7-bisphosphatase activity comprises an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID No. 2, and wherein said one or more polypeptides are located in chloroplasts of said plants.
17. The plant cell, a plant part, a plant or a seed of claim 16 , wherein said enzyme comprising sedoheptulose 1,7-bisphosphates activity further comprises FBPase activity or is from a source heterologous with respect to the plant into which the nucleic acid is inserted.
18. The plant cell, a plant part, a plant or a seed of claim 16 , wherein said enzyme comprising sedoheptulose 1,7-bisphosphatase activity comprises the amino acid sequence of SEQ ID No. 2.
19. The plant cell, a plant part, a plant or a seed of claim 18 , wherein said enzyme comprising sedoheptulose 1,7-bisphosphatase activity is encoded by a nucleic acid comprising the nucleotide sequence of SEQ ID No. 1.
20. The plant cell, a plant part, a plant or a seed of claim 16 , wherein said promoter includes the promoter region of the ribulose-bisphosphate carboxylase small subunit gene of Oryza sativa.
21. The plant cell, a plant part, a plant or a seed of claim 20 , wherein said promoter comprises the complement of the nucleotide sequence of SEQ ID No. 12 from nucleotide position 2010 to 4759.
22. The plant cell, a plant part, a plant or a seed according to claim 16 , 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.
23. The plant cell, a plant part, a plant or a seed according to claim 16 , wherein said one or more polypeptides having the enzymatic activity of a glycolate dehydrogenase are derived from the E. coli glc operon or wherein said one or more polypeptides having the enzymatic activity of a glycolate dehydrogenase is a Chlamydomonas or Synechocystis glycolate dehydrogenase or wherein said one or more polypeptides having the enzymatic activity of a glycolate dehydrogenase is an Arabidopsis glycolate dehydrogenase.
24. The plant cell, a plant part, a plant or a seed according to claim 23 , wherein said one or more polypeptides comprise an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID No. 3, SEQ ID No. 4 and SEQ ID No. 5 or to the amino acid sequence of SEQ ID No. 6, SEQ ID No. 7 or SEQ ID No. 8 or to the amino acid sequence of SEQ ID No. 9.
25. The plant cell, a plant part, a plant or a seed according claim 23 , wherein said polypeptides form a multi-subunit fusion protein.
26. The plant cell, a plant part, a plant or a seed according to claim 16 , wherein said one or more polypeptides having glycolate dehydrogenase activity further comprise an amino acid sequence targeting said enzyme 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.
27. The plant cell, a plant part, a plant or a seed according to claim 16 , 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.
28. The plant cell, a plant part, a plant or a seed according to claim 27 , 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.
29. The plant cell, a plant part, a plant or a seed according to claim 16 , wherein said plant is a monocotyledonous plant or a dicotyledonous plant or a Gymnosperm.
30. The plant cell, a plant part, a plant or a seed according to claim 29 , wherein said plant is selected from wheat, rice, maize, sorghum , millet, rye, oats, sugarcane, cotton, soybean or a Brassica plant.
31. A method for improving the assimilation of carbon in a plant or for increasing the growth rate or improving the biomass production of a plant, said method comprising the steps of inserting into the genome of a plant
a. 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, wherein said enzyme comprising sedoheptulose 1,7-bisphosphatase activity comprises an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID No. 2; and
b. 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.
32. The method of claim 1 , wherein said enzyme comprising sedoheptulose 1,7-bisphosphatase activity comprises an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID No. 2.
33. The method of claim 32 , said enzyme comprising sedoheptulose 1,7-bisphosphatase activity comprises the amino acid sequence of SEQ ID No. 2.