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1. (WO2010097623) TRANSGENIC PLANTS COMPRISING CONSTRUCTS ENCODING PHOSPHOENOLPYRUVATE CARBOXYKINASE AND/OR PYRUVATE ORTHO PHOS PHATE DIKINASE
Note: Text based on automatic Optical Character Recognition processes. Please use the PDF version for legal matters

Claims

1. A genetic construct comprising a senescence-specific promoter operably linked to at least one coding sequence, which encodes at least one polypeptide having phosphoenolpyruvate carboxykinase (PCK) activity and/or pyruvate orthophosphate dikinase (PPDK) activity.

2. A genetic construct comprising a promoter operably linked to at least one coding sequence, which encodes at least one polypeptide having phosphoenolpyruvate carboxykinase (PCK) activity and pyruvate orthophosphate dikinase (PPDK) activity.

3. A genetic construct according to claim 2, wherein the promoter is a senescence-specific promoter.

4. A genetic construct according to any preceding claim, wherein the promoter is isolated from a senescence-associated gene in Λrabidopsis.

5. A genetic construct according to any preceding claim, wherein the promoter is selected from a group consisting of: SAGl 2, SAGl 3, SAGlOl ,

SAG21 and SAGl 8, or a functional variant or fragment thereof.

6. A genetic construct according to any preceding claim, wherein the promoter is a SAGl 2 promoter, or a functional variant or fragment thereof.

7. A genetic construct according to any preceding claim, wherein the promoter comprises a nucleotide sequence substantially as set out in SEQ ID No.16, or a functional variant or fragment thereof.

8. A genetic construct according to claim 1, wherein the at least one coding sequence encodes (i) a phosphoenolpyruvate carboxykinase (PCK), or a functional variant or fragment thereof, and/or (ii) a pyruvate orthophosphate dikinase (PPDK), or a functional variant or fragment thereof.

9. A genetic construct according to any preceding claim, wherein the coding sequence, which encodes the polypeptide having PCK activity, is derived from Λrabidopsis.

10. A genetic construct according to any preceding claim, wherein the coding sequence, which encodes the polypeptide having PPDK activity, is derived from Λrabidopsis spp., Zea spp., Flaveria spp., or Cleome spp..

11. A genetic construct according to any preceding claim, wherein the coding sequence, which encodes the polypeptide having PCK activity, comprises a nucleic acid sequence substantially as set out in SEQ ID No: 17 or SEQ ID No.18, or a functional variant or fragment thereof.

12. A genetic construct according to any preceding claim, wherein the polypeptide having PCK activity comprises an amino acid sequence substantially as set out in SEQ ID No: 19, or a functional variant or fragment thereof.

13. A genetic construct according to any preceding claim, wherein the coding sequence, which encodes a polypeptide having PPDK activity, comprises a nucleic acid sequence substantially as set out in SEQ ID No:20, SEQ ID No:21, or SEQ ID No:22, or a functional variant or fragment thereof.

14. A genetic construct according to any preceding claim, wherein the polypeptide having PPDK activity comprises an amino acid sequence substantially as set out in SEQ ID No:23 or SEQ ID No.24, or a functional variant or fragment thereof.

15. A genetic construct according to claim 1 , wherein the construct encodes PCK or a functional variant or fragment thereof, and not PPDK.

16. A genetic construct according to claim 1 , wherein the construct encodes PPDK or a functional variant or fragment thereof, and not PCK.

17. A genetic construct according to any one of claims 1-14, wherein the construct encodes PCK or a functional variant or fragment thereof, and PPDK or a functional variant or fragment thereof.

18. A recombinant vector comprising the genetic construct according to any one of claims 1 to 17.

19. A recombinant vector substantially as illustrated in Figure 3.

20. A method of increasing the concentration of PCK and/or PPDK in a leaf of a test plant to above that of the corresponding concentration of PCK and/or

PPDK in a wild-type plant cultured under the same conditions, the method comprising altering plant metabolism in the test plant to achieve increased levels of PCK and/or PPDK in plant leaves after the initiation of leaf senescence.

21. A method of decreasing the concentration of nitrogen in the leaves of a test plant to below that of the corresponding concentration of nitrogen in a wild-type plant cultured under the same conditions, the method comprising altering plant metabolism in the test plant to achieve increased levels of PCK and/or PPDK in plant leaves after the initiation of leaf senescence.

22. A method of increasing the growth rate of a test plant compared to the corresponding growth rate of a wild-type plant cultured under the same conditions, the method comprising altering plant metabolism in the test plant to achieve increased levels of PCK and/or PPDK in plant leaves after the initiation of leaf senescence.

23. A method according to any one of claims 20-22, wherein the method comprises transforming a test plant cell with a genetic construct according to any one of claims 1-17, or a vector according to either claim 18 or claim 19.

24. A method according to any one of claims 20-23, wherein the method comprises increasing the concentration of PCK and PPDK in the plant leaves to above that of the corresponding concentration of PCK and PPDK in the wild-type plant cultured under the same conditions.

25. A cell comprising the genetic construct according to any one of claims 1 to 17, or the recombinant vector according to either claim 18 or claim 19.

26. A transgenic plant comprising the genetic construct according to any one of claims 1 to 17, or the recombinant vector according to either claim 18 or claim 19.

27. A transgenic plant according to claim 26, wherein the plant is from the Brassicaceae family.

28. A transgenic plant according to claim 26, wherein the plant is from the Poales family.

29. A transgenic plant according to claim 26, wherein the plant is from the Solanaceae family.

30. A plant propagation product obtainable from the transgenic plant according to any one of claims 26 to 29.

31. A plant propagation product according to claim 30, wherein the plant propagation product is a seed.

32. A method of producing a transgenic plant which remobilises nitrogen at a higher rate than a corresponding wild-type plant cultured under the same conditions, the method comprising the steps of:-i) transforming a plant cell with the genetic construct according to any one of claims 1-17, or the vector according to either claims 18 or 19; and ii) regenerating a plant from the transformed cell.

33. A method of producing a transgenic plant having a higher growth rate than a corresponding wild-type plant cultured under the same conditions, the method comprising the steps of:-i) transforming a plant cell with the genetic construct according to any one of claims 1-17, or the vector according to either claims 18 or 19; and ii) regenerating a plant from the transformed cell.

34. A harvested leaf containing a lower level of nitrogen than the corresponding level of nitrogen in a harvested leaf taken from a wild-type plant cultured under the same conditions, wherein the leaf is harvested from the transgenic plant according to any one of claims 26-29, or produced by the method according to either claim 32 or claim 33.

35. A smoking article comprising nitrogen-reduced tobacco obtained from a mutant tobacco plant, which mutant is capable of decreasing the concentration of nitrogen in senescent leaves.

36. A smoking article according to claim 35, wherein the smoking article is made from tobacco obtained from a mutant tobacco plant which has been transformed with the genetic construct according to any one of claims 1 to 17, or the vector according to either claim 18 or claim 19.