Sucrose phosphate synthase (SPS), its process for preparation its cDNA, and utilization of cDNA to modify the expression of SPS in plant cells

Abstract

Sucrose phosphate synthase (SPS), its process for preparation, its cDNA, and utilization of cDNA to modify the expression of SPS in the plant cells are provided.

We claim:  
     
       1. An isolated DNA encoding a sucrose phosphate synthase (SPS) derived from corn. 
     
     
       2. The DNA of  claim 1  comprising the SPS encoding region shown in FIG.  7 . 
     
     
       3. The DNA of  claim 1  comprising cDNA. 
     
     
       4. The DNA of  claim 1  comprising genomic DNA. 
     
     
       5. The DNA of  claim 1  as present in a recombinant construct, wherein said DNA encoding a sucrose phosphate synthase is operably linked to a second DNA which is not naturally linked to said DNA encoding a sucrose phosphate synthase. 
     
     
       6. A recombinant construct comprising, as operably linked components in the 5′ to 3′ direction of transcription, a transcription initiation region functional in a vegetal cell and a DNA encoding a sucrose phosphate synthase (SPS) derived from corn. 
     
     
       7. The construct of  claim 6  wherein said DNA encoding an SPS encodes a biologically active SPS. 
     
     
       8. The construct of  claim 7  wherein said DNA encoding an SPS is in a sense orientation as to said transcription initiation region. 
     
     
       9. The construct of  claim 8  further comprising a translation initiation region operably linked 3′ to said transcription initiation region and 5′ to said DNA encoding an SPS, wherein said translation initiation region is functional in a vegetal cell, and a transcription termination region functional in said vegetal cell 3′ to said DNA encoding an SPS. 
     
     
       10. The construct of  claim 9  wherein said transcription termination region is an SPS gene transcription termination region. 
     
     
       11. The construct of  claim 6  wherein said DNA encoding an SPS comprises the SPS encoding region shown in FIG.  7 . 
     
     
       12. The construct of  claim 6  wherein said transcription initiation region is tissue specific. 
     
     
       13. The construct of  claim 12  wherein said transcription initiation region is leaf specific. 
     
     
       14. A method of modifying the starch and sucrose levels in a tomato vegetal cell, said method comprising: 
       growing a tomato vegetal cell having integrated into its genome a construct comprising, as operably linked components in the 5′ to 3′ direction of transcription, a transcription initiation region functional in said tomato vegetal cell and a DNA encoding a sucrose phosphate synthase derived from corn, wherein said DNA encoding said sucrose phosphate synthase derived from corn is not naturally linked to said transcription initiation region, wherein said tomato vegetal cell is grown under conditions which permit said transcription initiation region to function, and wherein growing said tomato vegetal cell under said conditions permits said DNA encoding said sucrose phosphate synthase derived from corn to be expressed at a level which modifies the starch and sucrose levels in said tomato vegetal cell from a given ratio of starch to sucrose, as measured in control plant cells, to a different ratio of starch to sucrose.  
     
     
       15. The method of  claim 14  where said tomato vegetal cell is a leaf cell. 
     
     
       16. A tomato vegetal cell having integrated into its genome a recombinant construct of any one of claims  6 - 13 ,  60 - 61  and  64 - 65 . 
     
     
       17. A tomato plant comprising cells having integrated into its genome a recombinant construct of any one of claims  6 - 13 ,  60 - 61  and  64 - 65 . 
     
     
       18. A tomato vegetal cell having a modified ratio of starch to sucrose, wherein said cell is produced according to the method comprising growing a tomato vegetal cell having integrated into its genome a construct of any one of claims  6 - 13  under conditions which permit said transcription initiation region to function, and wherein growing said vegetal cell under said conditions permit said construct to be expressed at a level which modifies the starch and sucrose levels in said vegetal cell, 
       whereby the ratio of starch to sucrose level in said tomato vegetal cell is modified as compared to a given ratio of starch to sucrose measured in control plant cells.  
     
     
       19. A plant produced from a tomato vegetal cell of  claim 18 . 
     
     
       20. The method of  claim 14 , wherein said DNA encoding a sucrose phosphate synthase derived from corn is in a sense orientation as to said transcription initiation region. 
     
     
       21. The method of  claim 20  wherein said construct further comprises a translation initiation region functional in a tomato vegetal cell operably linked 3′ to said transcription initiation region and 5′ to said DNA encoding, said sucrose phosphate synthase derived from corn and a transcription termination region functional in said tomato vegetal cell operably linked 3′ to said DNA encoding said sucrose phosphate synthase derived from corn. 
     
     
       22. The method of  claim 14  wherein said transcription initiation region is tissue specific. 
     
     
       23. The method of  claim 22  wherein said transcription initiation region is leaf specific. 
     
     
       24. A tomato vegetal cell having modified levels of starch and sucrose, wherein said modified levels of starch and sucrose are produced according to the method of  claim 14 . 
     
     
       25. A method of increasing the yield of a tomato plant comprising: 
       growing a plant, wherein the genome of said plant comprises an integrated chimeric DNA construct capable of providing for expression of sucrose phosphate synthase derived from corn at a level sufficient to increase the amount of sucrose in tomato fruit by a factor of about 2 and decrease the amount of leaf starch by about 50% as compared to the amount of sucrose and starch measured in a control tomato plant, whereby an increase in plant yield is obtained.  
     
     
       26. An isolated DNA encoding a sucrose phosphate synthase wherein said DNA comprises at least about 10 nucleotides up to the full length of nucleotides represented by SEQ ID NO: 6. 
     
     
       27. The DNA sequence according to  claim 26 , wherein said DNA sequence encodes an amino acid sequence represented by a SEQ ID NO selected from the group consisting of SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4 and SEQ ID NO:5. 
     
     
       28. The DNA of  claim 26  comprising cDNA. 
     
     
       29. The DNA of  claim 26  comprising genomic DNA. 
     
     
       30. The isolated DNA encoding a sucrose phosphate synthase according to  claim 26  as present in a recombinant construct, wherein said DNA encoding a sucrose phosphate synthase is operably linked to a second DNA which is not naturally linked to said DNA encoding a sucrose phosphate synthase. 
     
     
       31. A recombinant construct comprising, as operably linked components in the 5′ to 3′ direction of transcription, a transcription initiation region functional in a tomato vegetal cell and said DNA encoding a sucrose phosphate synthase according to  claim 26 . 
     
     
       32. The construct of  claim 31  wherein said DNA encoding a sucrose phosphate synthase encodes a biologically active sucrose phosphate synthase. 
     
     
       33. The construct of  claim 32  wherein said DNA encoding a sucrose phosphate synthase is in a sense orientation as to said transcription initiation region. 
     
     
       34. The construct of  claim 33  further comprising a translation initiation region operably linked 3′ to said transcription initiation region and 5′ to said DNA encoding a sucrose phosphate synthase, wherein said translation initiation region is functional in a tomato vegetal cell, and a transcription termination region functional in said vegetal cell 3′ to said DNA encoding an SPS. 
     
     
       35. The construct of  claim 34  wherein said transcription termination region is a sucrose phosphate synthase gene transcription termination region. 
     
     
       36. The construct of  claim 31  wherein said transcription initiation region is tissue specific. 
     
     
       37. The construct of  claim 36  wherein said transcription initiation region is leaf specific. 
     
     
       38. A nucleic acid sequence encoding a peptide wherein said peptide has an amino acid sequence represented by a SEQ ID. NO: selected from the group consisting of SEQ ID. NO: 1, SEQ ID. NO: 2, SEQ ID. NO: 3, SEQ ID. NO: 4, and SEQ ID. NO: 5. 
     
     
       39. A tomato vegetal cell having integrated into its genome a recombinant construct according to  claim 30 . 
     
     
       40. A leaf cell having integrated into its genome a recombinant construct according to  claim 30 . 
     
     
       41. A tomato plant comprising cells according to claims  39  or  40 . 
     
     
       42. A tomato vegetal cell having a modified ratio of starch to sucrose, wherein said cell is produced according to the method comprising growing a tomato vegetal cell having integrated into its genome a construct according to  claim 30  under conditions which permit said transcription initiation region to function, and wherein growing said tomato vegetal cell under said conditions permit said construct to be expressed at a level which modifies the starch and sucrose levels in said tomato vegetal cell, 
       whereby the ratio of starch to sucrose level in said tomato vegetal cell is modified as compared to a given ratio of starch to sucrose measured in control plant cells.  
     
     
       43. A tomato plant produced from a tomato vegetal cell of  claim 42 . 
     
     
       44. A method of increasing the yield of a tomato plant sink tissue, said method comprising: 
       growing a tomato plant having integrated into its genome a construct comprising, as operably linked components in the 5′ to 3′ direction of transcription, a transcription initiation region functional in a tomato plant cell and a DNA encoding a sucrose phosphate synthase derived from corn, wherein said DNA encoding a sucrose phosphate synthase is not naturally linked to said transcription initiation region, and wherein said tomato plant cell is grown under conditions which permit said transcription initiation region to function,  
       whereby the amount of sucrose available to said tomato plant sink tissue is increased compared to the amount of sucrose measured in a control tomato plant sink tissue.  
     
     
       45. The method of  claim 44  wherein said transcription initiation region is tissue specific. 
     
     
       46. The method of  claim 45  wherein said transcription initiation region is functional in a fruit cell. 
     
     
       47. The method of  claim 45  wherein said transcription initiation region is functional in a leaf cell. 
     
     
       48. The method of  claim 47  wherein said transcription initiation region is a Rubisco small subunit promoter. 
     
     
       49. The method of  claim 44 , wherein increasing the amount of sucrose available in tomato plant sink tissue increases the amount of total solids per unit weight of said sink tissue compared to the amount of total solids per unit weight measured in a control tomato plant sink tissue. 
     
     
       50. The method of  claim 44  wherein said sink tissue is fruit tissue. 
     
     
       51. The method of  claim 40 , wherein said weight is dry weight. 
     
     
       52. A method of increasing the amount of soluble solids in a tomato plant sink tissue, said method comprising: 
       growing a tomato plant having integrated into its genome a construct comprising, as operably linked components in the 5′ to 3′ direction of transcription, a transcription initiation region functional in a tomato plant cell and a DNA encoding a sucrose phosphate synthase derived from corn, wherein said DNA encoding a sucrose phosphate synthase is not naturally linked to said transcription initiation region, and wherein said tomato plant cell is grown under conditions which will permit said transcription initiation region to function,  
       whereby the amount of soluble solids per unit weight of said tomato plant sink tissue is increased compared to the amount of solids measured in a control plant sink tissue; and  
       whereby starch is converted to sucrose in said tomato plant cell and whereby an increased level of sucrose is made available to said tomato plant sink tissue.  
     
     
       53. The method of  claim 52  wherein said transcription initiation region is tissue specific. 
     
     
       54. The method of  claim 53  wherein said transcription initiation region is functional in a tomato fruit cell. 
     
     
       55. The method of  claim 53  wherein said transcription initiation region is functional in a tomato leaf cell and wherein said sucrose is transported into said sink tissue. 
     
     
       56. The method of  claim 55  wherein said transcription initiation region is a Rubisco small subunit promoter. 
     
     
       57. The method of  claim 52  wherein said sink tissue is fruit tissue. 
     
     
       58. The method of  claim 52  wherein the amount of sucrose in said tomato plant sink tissue is increased compared to the amount of sucrose measured a control tomato plant sink tissue. 
     
     
       59. The method of  claim 52  wherein the amount of glucose and fructose in said sink tissue is increased compared to the amount of glucose and fructose measured in a control tomato plant sink tissue. 
     
     
       60. The construct of  claim 6  wherein said transcription initiation region is a cauliflower mosaic virus 35S promoter region. 
     
     
       61. The construct of  claim 13  wherein said transcription initiation region is a Rubisco small subunit promoter region. 
     
     
       62. The method of  claim 14  wherein said transcription limitation region is a cauliflower mosaic virus 35S promoter region. 
     
     
       63. The method of  claim 23  wherein said transcription limitation region is a Rubisco small subunit promoter region. 
     
     
       64. The construct of  claim 31  wherein said transcription initiation region is a cauliflower mosaic virus 35S promoter region. 
     
     
       65. The construct of  claim 37  wherein said transcription initiation region is a Rubisco small subunit promoter region. 
     
     
       66. The method of  claim 44  wherein said transcription initiation region is a cauliflower mosaic virus 35S promoter region. 
     
     
       67. The method of  claim 52  wherein said transcription initiation region is a cauliflower mosaic virus 35S promoter region. 
     
     
       68. A recombinant construct comprising, as operably linked components in the  5 ′ to  3 ′ direction of transcription, a transcription initiation region functional in a vegetal cell and the DNA encoding a sucrose phosphate synthase according to  claim 26 .  
     
     
       69. The recombinant construct according to  claim 68 , wherein said DNA encoding a sucrose phosphate synthase encodes a biologically active sucrose phosphate synthase.  
     
     
       70. The recombinant construct according to  claim 69 , wherein said DNA encoding a sucrose phosphate synthase is in a sense orientation as to said transcription initiation region.  
     
     
       71. The recombinant construct according to  claim 70 , further comprising a translation initiation region operably linked  3 ′ to said transcription initiation region and  5 ′ to said DNA encoding a sucrose phosphate synthase, and a transcription termination region functional in said vegetal cell  3 ′ to said DNA encoding a sucrose phosphate synthase.  
     
     
       72. The recombinant construct according to  claim 71 , wherein said transcription termination region is a sucrose phosphate synthase gene transcription termination region.  
     
     
       73. The recombinant construct according to  claim 68 , wherein said transcription initiation region is tissue specific.  
     
     
       74. The recombinant construct according to  claim 68 , wherein said transcription initiation region is leaf specific.  
     
     
       75. The recombinant construct according to  claim 68 , wherein said transcription initiation region is a cauliflower mosaic virus  35 S promoter region.  
     
     
       76. The recombinant construct according to  claim 74 , wherein said transcription initiation region is a Rubisco small subunit promoter region.