Glyphosate-tolerant 5-enolpyruvylshikimate-3-phosphate synthases

Abstract

Genes encoding Class II EPSPS enzymes are disclosed. The genes are useful in producing transformed bacteria and plants which are tolerant to glyphosate herbicide. Class II EPSPS genes share little homology with known, Class I EPSPS genes, and do not hybridize to probes from Class I EPSPS's. The Class II EPSPS enzymes are characterized by being more kinetically efficient than Class I EPSPS's in the presence of glyphosate. Plants transformed with Class II EPSPS genes are also disclosed as well as a method for selectively controlling weeds in a planted transgenic crop field.

1. An isolated DNA molecule which encodes an EPSPS enzyme having the sequence of SEQ ID NO:3. 
     
     
       2. A The DNA molecule of  claim 1  having the sequence of SEQ ID NO:2. 
     
     
       3. A The DNA molecule of  claim 1  having the sequence of SEQ ID NO:9. 
     
     
       4. A recombinant, double-stranded DNA molecule comprising in sequence:
 a) a promoter which functions in plant cells to cause the production of an RNA sequence;  
 b) a structural DNA sequence that causes the production of an RNA sequence which encodes a EPSPS enzyme having the sequence domains: 
 -R-X 1 -H-X 2 -E-(SEQ ID NO:37), in which 
 X 1  is G, S, T, C, Y, N, Q, D or E;  
 X 2  is S or T; and  
 
 -G-D-K-X 3 -(SEQ ID NO:38), in which 
 X 3  is S or T; and  
 
 -S-A-Q-X 4 -K-(SEQ ID NO:39), in which 
 X 4  is A, R, N, D, C, Q, E, G, H, I, L, K, M, F, P, S, T, W, Y or V; and  
 
 -N-X 5 -T-R-(SEQ ID NO:40), in which 
 X 5  is A, R, N, D, C, Q, E, G, H, I, L, K, M, F, P, S, T, W, Y or V,  
 
 provided that when X 1    is D, X   2    is T, X   3    is S, and X   4    is V, then X   5    is A, R, N, D, C, Q, E, G, H, I, L, K, M, F, S, T, W, Y, or V ; and  
 
 c) a 3′ non-translated region which functions in plant cells to cause the addition of a stretch of polyadenyl nucleotides to the 3′ end of the RNA sequence;  
 
       where the promoter is heterologous with respect to the structural DNA sequence and adapted to cause sufficient expression of the encoded EPSPS enzyme to enhance the glyphosate tolerance of a plant cell transformed with the DNA molecule. 
     
     
       5. A The DNA molecule of  claim 4  in which the structural DNA sequence encodes a fusion polypeptide comprising an amino-terminal chloroplast transit peptide and the EPSPS enzyme. 
     
     
       6. A The DNA molecule of  claim 4  in which X 1  is D or N; X 2  is S or T; X 3  is S or T; X 4  is V, I or L; and X 5  is P or Q, provided that when X 1    is D, X   2    is T, X   3    is S, and X   4    is V, then X   5    is Q . 
     
     
       7. A DNA molecule of  claim 6  in which the structural DNA sequence encodes an EPSPS enzyme selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:41 and SEQ ID NO:43. 
     
     
       8. A The DNA molecule of  claim 5  in which X 1  is D or N; X 2  is S or T; X 3  is S or T; X 4  is V, I or L; and X 5  is P or Q, provided that when X 1    is D, X   2    is T, X   3    is S, and X   4    is V, then X   5    is Q . 
     
     
       9. A DNA molecule of  claim 8  in which the structural DNA sequence encodes an EPSPS enzyme selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:41 and SEQ ID NO:43. 
     
     
       10. A The DNA molecule of claim  8   137  in which the EPSPS sequence is enzyme has the sequence set forth in SEQ ID NO:3. 
     
     
       11. A The DNA molecule of claim  10   4  in which the promoter is a plant DNA virus promoter. 
     
     
       12. A The DNA molecule of  claim 11  in which the promoter is selected from the group consisting of CaMV35S and FMV35S promoters. 
     
     
       13. A The DNA molecule of claim  10   5  in which the structural DNA sequence encodes a chloroplast transit peptide selected from the group consisting of SEQ ID NO:11 and SEQ ID NO:15. 
     
     
       14. A The DNA molecule of  claim 13  in which the 3′ non-translated region is selected from the group consisting of the NOS 3′ and the E9 3′ non-translated regions. 
     
     
       15. A method of producing genetically transformed plants which are tolerant toward glyphosate herbicide, comprising the steps of:
 a) inserting into the genome of a plant cell a recombinant, double-stranded DNA molecule comprising: 
 i) a promoter which functions in plant cells to cause the production of an RNA sequence,  
 ii) a structural DNA sequence that causes the production of an RNA sequence which encodes an EPSPS enzyme having the sequence domains: 
 -R-X 1 -H-X 2 -E-(SEQ ID NO:37), in which 
 X 1  is G, S, T, C, Y, N, Q, D or E;  
 X 2  is S or T; and  
 
 -G-D-K-X 3 -(SEQ ID NO:38), in which 
 X 3  is S or T; and  
 
 -S-A-Q-X 4 -K-(SEQ ID NO:39), in which 
 X 4  is A, R, N, D, C, Q, E, G, H, I, L, K, M, F, P, S, T, W, Y or V; and  
 
 -N-X 5 -T-R-(SEQ ID NO:40), in which 
 X 5  is A, R, N, D, C, Q, E, G, H, I, L, K, M, F, P, S, T, W, Y or V, provided that when X 1    is D, X   2    is T, X   3    is S, and X   4    is V, then X   5    is A, R, N, D, C, Q, E, G, H, I, L, K, M, F, S, T, W, Y or V ; and  
 
 
 iii) a 3′ non-translated DNA sequence which functions in plant cells to cause the addition of a stretch of polyadenyl nucleotides to the 3′ end of the RNA sequence;  
 
 
       where the promoter is heterologous with respect to the structural DNA sequence and adapted to cause sufficient expression of the polypeptide to enhance the glyphosate tolerance of a plant cell transformed with the DNA molecule;
 b) obtaining a transformed plant cell; and  
 c) regenerating from the transformed plant cell a genetically transformed plant which has increased tolerance to glyphosate herbicide.  
 
     
     
       16. A The method of  claim 15  in which X 1  is D or N; X 2  is S or T; X 3  is S or T; X 4  is V, I or L; and X 5  is P or Q, provided that when X 1    is D, X   2    is T, X   3    is S, and X   4    is V, then X   5    is Q . 
     
     
       17. A method of  claim 16  in which the structural DNA sequence encodes an EPSPS enzyme selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:41 and SEQ ID NO:43. 
     
     
       18. A The method of  claim 15  in which the structural DNA sequence encodes a fusion polypeptide comprising an amino-terminal chloroplast transit peptide and the EPSPS enzyme. 
     
     
       19. A The method of  claim 18  in which X 1  is D or N; X 2  is S or T; X 3  is S or T; X 4  is V, I or L; and X 5  is P or Q, provided that when X 1    is D, X   2    is T, X   3    is S, and X   4    is V, then X   5    is Q . 
     
     
       20. A method of  claim 19  in which the structural DNA sequence encodes an EPSPS enzyme selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:42 and SEQ ID NO:44. 
     
     
       21. A The method of claim  19   143  in which the EPSPS enzyme is that set forth in SEQ ID NO:3. 
     
     
       22. A The method of claim  21   15  in which the promoter is from a plant DNA virus. 
     
     
       23. A The method of  claim 22  in which the promoter is selected from the group consisting of CaMV35S and FMV35S promoters. 
     
     
       24. A glyphosate-tolerant plant cell comprising a the DNA molecule of claims  claim 4 ,  5  or  8 or  10  . 
     
     
       25. A The glyphosate-tolerant plant cell of  claim 24  in which the promoter is a plant DNA virus promoter. 
     
     
       26. A The glyphosate-tolerant plant cell of  claim 25  in which the promoter is selected from the group consisting of CaMV35S and FMV35S promoters. 
     
     
       27. A The glyphosate-tolerant plant cell of  claim 24  selected from the group consisting of corn, wheat, rice, barley, soybean, cotton, sugarbeet, oilseed rape, canola, flax, sunflower, potato, tobacco, tomato, alfalfa, poplar, pine, eukalyptus eucalyptus, apple, lettuce, peas, lentils, grape and turf grasses. 
     
     
       28. A glyphosate-tolerant plant comprising the plant cells cell of  claim 27 . 
     
     
       29. A The glyphosate-tolerant plant of  claim 28  in which the promoter is from a DNA plant virus promoter. 
     
     
       30. A The glyphosate-tolerant plant of  claim 29  in which the promoter is selected from the group consisting of CaMV35S and FMV35S promoters. 
     
     
       31. A The glyphosate-tolerant plant of  claim 30  selected from the group consisting of corn, wheat, rice, barley, soybean, cotton, sugarbeet, oilseed rape, canola, flax, sunflower, potato, tobacco, tomato, alfalfa, poplar, pine, eukalyptus eucalyptus, apple, lettuce, peas, lentils, grape and turf grasses. 
     
     
       32. A method for selectively controlling weeds in a field containing a crop having plant crop seeds or plants comprising the steps of:
 a) planting the crop seeds or plants which are glyphosate-tolerant as a result of a recombinant double-stranded DNA molecule being inserted into the crop seed or plant, the DNA molecule having: 
 i) a promoter which functions in plant cells to cause the production of an RNA sequence,  
 ii) a structural DNA sequence that causes the production of an RNA sequence which encodes an EPSPS enzyme having the sequence domains: 
 -R-X 1 -H-X 2 -E-(SEQ ID NO:37), in which 
 X 1  is G, S, T, C, Y, N, Q, D or E;  
 X 2  is S or T; and  
 
 -G-D-K-X 3 -(SEQ ID NO:38), in which 
 X 3  is S or T; and  
 
 -S-A-Q-X 4 -K-(SEQ ID NO:39), in which 
 X 4  is A, R, N, D, C, Q, E, G, H, I, L, K, M, F, P, S, T, W, Y or V; and  
 
 -N-X 5 -T-R-(SEQ ID NO:40), in which 
 X 5  is A, R, N, D, C, Q, E, G, H, I, L, K, M, F, P, S, T, W, Y or V, provided that when X 1    is D, X   2    is T, X   3    is S, and X   4    is V, then X   5    is A, R, N, D, C, Q, E, G, H, I, L, K, M, F, S, T, W, Y or V ; and  
 
 
 iii) a 3′ non-translated DNA sequence which functions in plant cells to cause the addition of a stretch of polyadenyl nucleotides to the 3′ end of the RNA sequence  
 
 
       where the promoter is heterologous with respect to the structural DNA sequence and adapted to cause sufficient expression of the EPSPS enzyme to enhance the glyphosate tolerance of the crop plant transformed with the DNA molecule; and
 b) applying to the crop and weeds in the field a sufficient amount of glyphosate herbicide to control the weeds without significantly affecting the crop.  
 
     
     
       33. A The method of  claim 32  in which X 1  is D or N; X 2  is S or T; X 3  is S or T; X 4  is V, I or L; and X 5  is P or Q, provided that when X 1    is D, X   2    is T, X   3    is S, and X   4    is V, then X   5    is Q . 
     
     
       34. A method of  claim 33  in which the structural DNA sequence encodes an EPSPS enzyme selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:42 and SEQ ID NO:44. 
     
     
       35. A The method of  claim 32  in which the structural DNA sequence encodes a fusion polypeptide comprising an amino-terminal chloroplast transit peptide and the EPSPS enzyme. 
     
     
       36. A The method of  claim 35  in which X 1  is D or N; X 2  is S or T; X 3  is S or T; X 4  is V, I or L; and X 5  is P or Q, provided that when X 1    is D, X   2    is T, X   3    is S, and X   4    is V, then X   5    is Q . 
     
     
       37. A method of  claim 36  in which the structural DNA sequence encodes an EPSPS enzyme selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:41 and SEQ ID NO:43. 
     
     
       38. A The method of claim  36   155  in which the DNA molecule encodes an EPSPS enzyme as set forth in SEQ ID NO:3. 
     
     
       39. A The method of claim  38   32  in which the DNA molecule further comprises a promoter selected from the group consisting of the CAMV35S and FMV35S promoters. 
     
     
       40. A The method of  claim 39  in which the crop plant is selected from the group consisting of corn, wheat, rice, barley, soybean, cotton, sugarbeet, oilseed rape, canola, flax, sunflower, potato, tobacco, tomato, alfalfa, poplar, pine, eukalyptus eucalyptus, apple, lettuce, peas, lentils, grape and turf grasses. 
     
     
       41. A The DNA molecule of  claim 5  in which the structural DNA sequence encodes a chloroplast transit peptide selected from the group consisting of SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15 and SEQ ID NO:17. 
     
     
       42. A The DNA molecule of  claim 41  in which the chloroplast transit peptide is encoded by a DNA sequence selected from the group consisting of SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14 and SEQ ID NO:16. 
     
     
       43. A The DNA molecule of  claim 5  in which the structural DNA sequence encodes a chloroplast transit peptide selected from the group consisting of SEQ ID NO:11 and SEQ ID NO:15. 
     
     
       44. A The DNA molecule of  claim 43  in which the chloroplast transit peptide is encoded by a DNA sequence selected from the group consisting of SEQ ID NO:10 and SEQ ID NO:14. 
     
     
       45. A The DNA molecule of  claim 41  in which the promoter is selected from the group consisting of CaMV 35S and FMV 35S promoters. 
     
     
       46. A The DNA molecule of  claim 42  in which the promoter is selected from the group consisting of CaMV 35S and FMV 35S promoters. 
     
     
       47. A The DNA molecule of  claim 43  in which the promoter is selected from the group consisting of CaMV 35S and FMV 35S promoters. 
     
     
       48. A The DNA molecule of  claim 44  in which the promoter is selected from the group consisting of CaMV 35S and FMV 35S promoters. 
     
     
       49. A The DNA molecule of  claim 45  in which the 3′ non-translated region is selected from the group consisting of the NOS 3′ and the E9 3′ non-translated regions. 
     
     
       50. A The DNA molecule of  claim 46  in which the 3′ non-translated region is selected from the group consisting of the NOS 3′ and the E9 3′ non-translated regions. 
     
     
       51. A The DNA molecule of  claim 47  in which the 3′ non-translated region is selected from the group consisting of the NOS 3′ and the E9 3′ non-translated regions. 
     
     
       52. A The DNA molecule of  claim 48  in which the 3′ non-translated region is selected from the group consisting of the NOS 3′ and the E9 3′ non-translated regions. 
     
     
       53. A DNA molecule of  claim 49  in which the structural DNA sequence encodes an EPSPS enzyme selected from the group consisting of SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:42 and SEQ ID NO:44. 
     
     
       54. A DNA molecule of  claim 50  in which the structural DNA sequence encodes an EPSPS enzyme selected from the group consisting of SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:42 and SEQ ID NO:44. 
     
     
       55. A DNA molecule of  claim 51  in which the structural DNA sequence encodes an EPSPS enzyme selected from the group consisting of SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:42 and SEQ ID NO:44. 
     
     
       56. A DNA molecule of  claim 52  in which the structural DNA sequence encodes an EPSPS enzyme selected from the group consisting of SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:42 and SEQ ID NO:44. 
     
     
       57. A The DNA molecule of claim  53   137  in which the structural DNA sequence contains an EPSPS encoding sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO: 4, and  SEQ ID NO:6, SEQ ID NO:41 and SEQ ID NO:43 . 
     
     
       58. A The DNA molecule of claim  54   137  in which the structural DNA sequence contains an EPSPS encoding sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:41 and SEQ ID NO: 43 as set forth in SEQ ID NO:9 . 
     
     
       59. A DNA molecule of  claim 55  in which the structural DNA sequence contains an EPSPS encoding sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:41 and SEQ ID NO:43. 
     
     
       60. A DNA molecule of  claim 56  in which the structural DNA sequence contains an EPSPS coding sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:41 and SEQ ID NO:43. 
     
     
       61. A DNA molecule of  claim 49  in which the structural DNA sequence encodes an EPSPS enzyme having the sequence of SEQ ID NO:3. 
     
     
       62. A DNA molecule of  claim 50  in which the structural DNA sequence encodes an EPSPS enzyme having the sequence of SEQ ID NO:3. 
     
     
       63. A DNA molecule of  claim 51  in which the structural DNA sequence encodes an EPSPS enzyme having the sequence of SEQ ID NO:3. 
     
     
       64. A DNA molecule of  claim 52  in which the structural DNA sequence encodes an EPSPS enzyme having the sequence of SEQ ID NO:3. 
     
     
       65. A DNA molecule of  claim 61  in which the structural DNA sequence contains an EPSPS encoding sequence selected from the group consisting of SEQ ID NO:2 and SEQ ID NO:9. 
     
     
       66. A DNA molecule of  claim 62  in which the structural DNA sequence contains an EPSPS encoding sequence selected from the group consisting of SEQ ID NO:2 and SEQ ID NO:9. 
     
     
       67. A DNA molecule of  claim 63  in which the structural DNA sequence contains an EPSPS encoding sequence selected from the group consisting of SEQ ID NO:2 and SEQ ID NO:9. 
     
     
       68. A DNA molecule of  claim 64  in which the structural DNA sequence contains an EPSPS encoding sequence selected from the group consisting of SEQ ID NO:2 and SEQ ID NO:9. 
     
     
       69. A The glyphosate-tolerant plant cell of claim  25   149  in which:
 (a) the promoter is selected from the group consisting of CaMV 35S and FMV 35S promoters;  
 (b) the structural DNA sequence encodes: 
 (i) a chloroplast transit peptide selected from the group consisting of SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15 and SEQ ID NO:17; and  
 (ii) an EPSPS enzyme selected from the group consisting of SEQ ID NO:3, SEQ ID NO: 5, and  SEQ ID NO: 7, SEQ ID NO: 42 and SEQ ID NO: 44 ; and    
 
 (c) the 3′ non-translated region is selected from the group consisting of the NOS 3′ and the E9 3′ non-translated regions.  
 
     
     
       70. A The glyphosate-tolerant plant cell of  claim 69  in which the structural DNA sequence comprises:
 (a) a chloroplast transit peptide encoding DNA sequence selected from the group consisting of SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14 and SEQ ID NO:16; and  
 (b) an EPSPS encoding sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, and SEQ ID NO: 6, SEQ ID NO: 41 and SEQ ID NO:43 .  
 
     
     
       71. A The glyphosate-tolerant plant cell of  claim 69  in which the structural DNA sequence comprises:
 (a) a chloroplast transist peptide encoding DNA sequence selected from the group consisting of SEQ ID NO:10 and SEQ ID NO:14; and  
 (b) a DNA sequence encoding an EPSPS enzyme having the sequence of SEQ ID NO:3.  
 
     
     
       72. A The glyphosate-tolerant plant cell of  claim 71  in which the structural DNA sequence comprises an EPSPS encoding sequence selected from the group consisting of SEQ ID NO:2 and as set forth in SEQ ID NO:9. 
     
     
       73. A The glyphosate-tolerant plant cell of  claim 71  selected from the group consisting of corn, wheat, rice, barley, soybean, cotton, sugarbeet, oilseed rape, canola, flax, sunflower, potato, tobacco, tomato, alfalfa, poplar, pine, eukalyptus eucalyptus, apple, lettuce, peas, lentils, grape and turf grasses. 
     
     
       74. A glyphosate-tolerant plant comprising a the DNA molecule of claims  5 ,  8  or  10   claim 131  in which:
 (a) the promoter is selected from the group consisting of CaMV 35S and FMV 35S promoters;  
 (b) the structural DNA sequence encodes: ;
 (i) a chloroplast transit peptide selected from the group consisting of SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15 and SEQ ID NO:17; and  
 (ii) an EPSPS enzyme selected from the group consisting of SEQ ID NO:3, SEQ ID NO: 5, and  SEQ ID NO: 7, SEQ ID NO: 42 and SEQ ID NO:44 ; and  
 
 (c) the 3′ non-translated region is selected from the group consisting of the NOS 3′ and the E9 3′ non-translated regions.  
 
     
     
       75. A The glyphosate-tolerant plant of  claim 74  in which the structural DNA sequence comprises:
 (a) a chloroplast transit peptide encoding DNA sequence selected from the group consisting of SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14 and SEQ ID NO:16; and  
 (b) an EPSPS encoding sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO: 4, and  SEQ ID NO: 6, SEQ ID NO: 41 and SEQ ID NO:43 .  
 
     
     
       76. A The glyphosate-tolerant plant of  claim 75  in which the structural DNA sequence comprises:
 (a) a chloroplast transit peptide encoding DNA sequence selected from the group consisting of SEQ ID NO:10 and SEQ ID NO:14; and  
 (b) a DNA sequence encoding an EPSPS enzyme having the sequence of SEQ ID NO:3.  
 
     
     
       77. A The glyphosate-tolerant plant of claim  76   74  in which the structural DNA sequence comprises an EPSPS encoding sequence selected from the group consisting of SEQ ID NO:2 and as set forth in SEQ ID NO:9. 
     
     
       78. A The glyphosate-tolerant plant of claim  77   74  selected from the group consisting of corn, wheat, rice, barley, soybean, cotton, sugarbeet, oilseed rape, canola, flax, sunflower, potato, tobacco, tomato, alfalfa, poplar, pine, eukalyptus eucalyptus, apple, lettuce, peas, lentils, grape and turf grasses. 
     
     
       79. A seed of a the glyphosate-tolerant plant of  claim 28 , wherein the seed comprises the recombinant DNA molecule. 
     
     
       80. A seed of a the glyphosate-tolerant plant of  claim 31 , wherein the seed comprises the recombinant DNA molecule. 
     
     
       81. A seed of a the glyphosate-tolerant plant of  claim 75 , wherein the seed comprises the recombinant DNA molecule. 
     
     
       82. A seed of a the glyphosate-tolerant plant of  claim 76 , wherein the seed comprises the recombinant DNA molecule. 
     
     
       83. A seed of a the glyphosate-tolerant plant of  claim 77 , wherein the seed comprises the recombinant DNA molecule. 
     
     
       84. A seed of a the glyphosate-tolerant plant of claim  78   129 , wherein the seed comprises the recombinant DNA molecule. 
     
     
       85. A seed of a the glyphosate-tolerant plant of claim  79   144 , wherein the seed comprises the recombinant DNA molecule. 
     
     
       86. A transgenic soybean plant which contains a heterologous gene which encodes an EPSPS enzyme having a K m  for phosphoenolpyruvate (PEP) between 1 and 150 μM and a K i (glyphosate)/K m (PEP) ratio between about 2 and 500, said plant exhibiting tolerance to N-phosphonomethylglycine herbicide at a rate of 1 lb/acre without significant yield reduction due to herbicide application. 
     
     
       87. Seed of a soybean plant of  claim 86 . 
     
     
       88. The DNA molecule of  claim 6  in which the structural DNA sequence contains an EPSPS encoding sequence selected from the group consisting of SEQ ID NO: 41  and SEQ ID NO: 43 . 
     
     
       89. The DNA molecule of  claim 8  in which the structural DNA sequence contains an EPSPS encoding sequence selected from the group consisting of SEQ ID NO: 41  and SEQ ID NO: 43 . 
     
     
       90. The method of  claim 16  in which the structural DNA sequence contains an EPSPS encoding sequence selected from the group consisting of SEQ ID NO: 41  and SEQ ID NO:  43 . 
     
     
       91. The method of  claim 19  in which the structural DNA sequence encodes an EPSPS enzyme having a sequence selected from the group consisting of SEQ ID NO: 42  and SEQ ID NO: 44 . 
     
     
       92. The method of  claim 33  in which the structural DNA sequence encodes an EPSPS enzyme having a sequence selected from the group consisting of SEQ ID NO: 42  and SEQ ID NO: 44 . 
     
     
       93. The method of  claim 36  in which the structural DNA sequence contains an EPSPS encoding sequence selected from the group consisting of SEQ ID NO: 41  and SEQ ID NO: 43 . 
     
     
       94. The DNA molecule of  claim 49  in which the structural DNA sequence encodes an EPSPS enzyme having a sequence selected from the group consisting of SEQ ID NO:  42  and SEQ ID NO:  44 . 
     
     
       95. The DNA molecule of  claim 50  in which the structural DNA sequence encodes an EPSPS enzyme having a sequence selected from the group consisting of SEQ ID NO:  42  and SEQ ID NO:  44 . 
     
     
       96. The DNA molecule of  claim 51  in which the structural DNA sequence encodes an EPSPS enzyme having a sequence selected from the group consisting of SEQ ID NO:  42  and SEQ ID NO:  44 . 
     
     
       97. The DNA molecule of  claim 52  in which the structural DNA sequence encodes an EPSPS enzyme having a sequence selected from the group consisting of SEQ ID NO:  42  and SEQ ID NO:  44 . 
     
     
       98. The glyphosate- tolerant plant cell of    claim 25    in which:      a )  the promoter is selected from the group consisting of CaMV  35 S and FMV  35 S promoters;        b )  the structural DNA sequence encodes:      i )  a chloroplast transit peptide selected from the group consisting of SEQ ID NO: 11 , SEQ ID NO: 13 , SEQ ID NO: 15  and SEQ ID NO: 17 ; and        ii )  an EPSPS enzyme selected from the group consisting of SEQ ID NO: 42  and SEQ ID NO: 44 ; and          c )  the  3 ′ non - translated region is selected from the group consisting of the NOS  3 ′ and the E 9   3 ′ non - translated regions.     
     
     
       99. The glyphosate- tolerant plant cell of    claim 26    in which the structural DNA sequence comprises:      a )  a chloroplast transit peptide encoding DNA sequence selected from the group consisting of SEQ ID NO: 10 , SEQ ID NO: 12 , SEQ ID NO: 14  and SEQ ID NO: 16 ; and        b )  an EPSPS encoding sequence selected from the group consisting of SEQ ID NO: 41  and SEQ ID NO: 43 .     
     
     
       100. The glyphosate- tolerant plant comprising the DNA molecule of    claim 4   ,  5  or  8  in which:      a )  the promoter is selected from the group consisting of CaMV  35 S and FMV  35 S promoters;        b )  the structural DNA sequence encodes:      i )  a chloroplast transit peptide selected from the group consisting of SEQ ID NO: 11 , SEQ ID NO: 13 , SEQ ID NO: 15  and SEQ ID NO: 17 ; and        ii )  an EPSPS enzyme selected from the group consisting of SEQ ID NO: 42  and SEQ ID NO: 44 ; and          c )  the  3 ′ non - translated region is selected from the group consisting of the NOS  3 ′ and the E 9   3 ′ non - translated regions.     
     
     
       101. The glyphosate- tolerant plant of    claim 28    in which the structural DNA sequence comprises:      a )  a chloroplast transit peptide encoding DNA sequence selected from the group consisting of SEQ ID NO: 10 , SEQ ID NO: 12 , SEQ ID NO: 14  and SEQ ID NO: 16 ; and        b )  an EPSPS encoding sequence selected from the group consisting of SEQ ID NO: 41  and SEQ ID NO: 43 .     
     
     
       102. An isolated DNA molecule that encodes a  5 - enolpyruvylshikimate -   3   - phosphate synthase  ( EPSPS )  enzyme having the sequence of SEQ ID NO: 70 .   
     
     
       103. A recombinant, double- stranded DNA molecule comprising in sequence:      a )  a promoter which functions in plant cells to cause the production of an RNA sequence;        b )  a structural DNA sequence that causes the production of an RNA sequence which encodes an EPSPS enzyme having the sequence of SEQ ID NO: 70 ; and        c )  a  3 ′ non - translated region that functions in plant cells to cause the addition of a stretch of polyadenyl nucleotides to the  3 ′ end of the RNA sequence;        where the promoter is heterologous with respect to the structural DNA sequence and adapted to cause sufficient expression of the encoded EPSPS enzyme to enhance the glyphosate tolerance of a plant cell transformed with the DNA molecule.     
     
     
       104. The DNA molecule of  claim 103 , wherein the structural DNA sequence further causes the production of an RNA sequence that encodes an amino- terminal chloroplast transit peptide that is fused to the EPSPS enzyme.   
     
     
       105. The DNA molecule of  claim 104 , wherein the chloroplast transit peptide has the sequence of SEQ ID NO: 11  or SEQ ID NO: 15 . 
     
     
       106. The DNA molecule of  claim 103 , wherein the promoter is a plant DNA virus promoter. 
     
     
       107. The DNA molecule of  claim 106 , wherein the promoter is a CaMV 35 S promoter or an FMV 35 S promoter. 
     
     
       108. The DNA molecule of  claim 103 , wherein the  3 ′ non- translated region is a NOS  3 ′ or an E 9   3 ′ non - translated region.   
     
     
       109. A method of producing a genetically transformed plant which is tolerant toward glyphosate herbicide, comprising the steps of:
   a )  inserting into the genome of a plant cell a recombinant double - stranded DNA molecule comprising:      i )  a promoter that functions in plant cells to cause the production of an RNS sequence;        ii )  a structural DNA sequence that causes the production of an RNS sequence which encodes an EPSPS enzyme having the sequence of SEQ ID NO:  70 ; and        iii )  a  3 ′ non - translated DNA sequence that functions in plant cells to cause the addition of a stretch of polyadenyl nucleotides to the  3′ end of the RNS sequence;        wherein the promoter is heterologous with respect to the structural DNA sequence and adapted to cause sufficient expression of the polypeptide to enhance the glyphosate tolerance of a plant cell transformed with the DNA molecule;        b )  obtaining a transformed plant cell; and        c )  regenerating from the transformed plant cell a genetically transformed plant which has increased tolerance to glyphosate herbicide.     
     
     
       110. The method of  claim 109 , wherein the structural DNA further causes the production of an RNA sequence that encodes an amino- terminal chloroplast transit peptide that is fused to the EPSPS enzyme.   
     
     
       111. The method of  claim 110 , wherein the chloroplast transit peptide has the sequence of SEQ ID NO: 11  or SEQ ID NO: 15 . 
     
     
       112. The method of  claim 109 , in which the promoter is a plant DNA virus promoter. 
     
     
       113. The method of  claim 112 , in which the promoter is a CaMV 35 S promoter or an FMV 35 S promoter. 
     
     
       114. The method of  claim 109 , wherein the  3 ′ non- translated DNA sequence is a NOS  3 ′ or an E 9   3 ′ non - translated sequence.   
     
     
       115. A glyphosate- tolerant plant cell comprising a DNA sequence encoding an EPSPS enzyme having the sequence of SEQ ID NO:  70 .   
     
     
       116. A glyphosate- tolerant plant comprising a DNA sequence encoding an EPSPS enzyme having the sequence of SEQ ID NO:  70 .   
     
     
       117. The plant of  claim 116 , wherein the plant is corn, wheat, rice, barley, soybean, cotton, sugarbeet, oilseed rape, canola, flax, sunflower, potato, tobacco, tomato, alfalfa, poplar, pine, eucalyptus, apple, lettuce, peas, lentils, grape or turf grasses. 
     
     
       118. The plant of  claim 117 , wherein the plant is corn. 
     
     
       119. The plant of  claim 117 , wherein the plant is soybean. 
     
     
       120. The plant of  claim 117 , wherein the plant is canola. 
     
     
       121. The plant of  claim 117 , wherein the plant is cotton. 
     
     
       122. A seed of the plant of  claim 116 , wherein the seed comprises the DNA sequence encoding an EPSPS enzyme having the sequence of SEQ ID NO:  70 . 
     
     
       123. The seed of  claim 122 , wherein the seed is corn, wheat, rice, barley, soybean, cotton, sugarbeet, oilseed rape, canola, flax, sunflower, potato, tobacco, tomato, alfalfa, poplar, pine, eucalyptus, apple, lettuce, peas, lentils, grape or turf grass seed. 
     
     
       124. The seed of  claim 123 , wherein the seed is corn seed. 
     
     
       125. The seed of  claim 123 , wherein the seed is soybean seed. 
     
     
       126. The seed of  claim 123 , wherein the seed is canola seed. 
     
     
       127. The seed of  claim 123 , wherein the seed is cotton seed. 
     
     
       128. A glyphosate tolerant plant cell comprising the recombinant DNA molecule of  claim 103 . 
     
     
       129. A plant comprising the glyphosate tolerant plant cell of  claim 128 . 
     
     
       130. A method for selectively controlling weeds in a field containing a crop having planted crop seeds or plants comprising the steps of:
   a )  planting the crop seeds or plants which are glyphosate - tolerant as a result of a recombinant double - stranded DNA molecule being inserted into the crop seed or plant, the DNA molecule having:      i )  a promoter which functions in plant cells to cause the production of an RNA sequence,        ii )  a structural DNA sequence that causes the production of an RNA sequence which encodes an EPSPS enzyme having the sequence of SEQ ID NO: 70 ; and        iii )  a  3 ′ non - translated DNA sequence which functions in plant cells to cause the addition of a stretch of polyadenyl nucleotides to the  3 ′ end of the RNA sequence,          where the promoter is heterologous with respect to the structural DNA sequence and adapted to cause sufficient expression of the EPSPS enzyme to enhance the glyphosate tolerance of the crop plant transformed with the DNA molecule; and        b )  applying to the crop and weeds in the field a sufficient amount of glyphosate herbicide to control the weeds without significantly affecting the crop.     
     
     
       131. A recombinant, double- stranded DNA molecule comprising in sequence:      a )  a promoter which functions in plant cells to cause the production of an RNA sequence;        b )  a structural DNA sequence that causes the production of an RNA sequence which encodes an EPSPS enzyme having the sequence of SEQ ID NO: 3 , SEQ ID NO: 5  or SEQ ID NO:  7 ;        c )  a  3 ′ non - translated region which functions in plant cells to cause the addition of a stretch of polyadenyl nucleotides to the  3 ′ end of the RNA sequence;        where the promoter is heterologous with respect to the structural DNA sequence and adapted to cause sufficient expression of the encoded EPSPS enzyme to enhance the glyphosate tolerance of a plant cell transformed with the DNA molecule.     
     
     
       132. The DNA molecule of  claim 131  in which the structural DNA sequence encodes a fusion polypeptide comprising an amino- terminal chloroplast transit peptide and the EPSPS enzyme.   
     
     
       133. The DNA molecule of  claim 131  in which the promoter is a plant DNA virus promoter. 
     
     
       134. The DNA molecule of  claim 133  in which the promoter is selected from the group consisting of CaMV 35 S and FMV 35 S promoters. 
     
     
       135. The DNA molecule of  claim 132  in which the structural DNA sequence encodes a chloroplast transit peptide selected from the group consisting of SEQ ID NO:  11  and SEQ ID NO:  15 . 
     
     
       136. The DNA molecule of  claim 131  in which the  3 ′ non- translated region is selected from the group consisting of the NOS  3 ′ and the E 9   3 ′ non - translated regions.   
     
     
       137. A method of producing genetically transformed plants which are tolerant toward glyphosate herbicide, comprising the steps of:
   a )  inserting into the genome of a plant cell a recombinant, double - stranded DNA molecule comprising:      i )  a promoter which functions in plant cells to cause the production of an RNA sequence,        ii )  a structural DNA sequence that causes the production of an RNA sequence which encodes an EPSPS enzyme having the sequence of SEQ ID NO: 3 , SEQ ID NO: 5  or SEQ ID NO: 7 ; and        iii )  a  3 ′ non - translated DNA sequence which functions in plant cells to cause the addition of a stretch of polyadenyl nucleotides to the  3 ′ end of the RNA sequence;          where the promoter is heterologous with respect to the structural DNA sequence and adapted to cause sufficient expression of the polypeptide to enhance the glyphosate tolerance of a plant cell transformed with the DNA molecule;        b )  obtaining a transformed plant cell; and        c )  regenerating from the transformed plant cell a genetically transformed plant which has increased tolerance to glyphosate herbicide.     
     
     
       138. The method of  claim 137  in which the structural DNA sequence encodes a fusion polypeptide comprising an amino- terminal chloroplast transit peptide and the EPSPS enzyme.   
     
     
       139. The method of  claim 130 , wherein the chloroplast transit peptide has the sequence of SEQ ID NO:  11  or SEQ ID NO:  15 . 
     
     
       140. The method of  claim 137  in which the promoter is a plant DNA virus. 
     
     
       141. The method of  claim 140  in which the promoter is a CaMV 35 S promoter or a FMV 35 S promoter. 
     
     
       142. The method of  claim 137 , wherein the  3 ′ non- translated DNA sequence is a NOS  3 ′ or an e 9   3 ′ non - translated sequence.   
     
     
       143. A glyphosate- tolerant plant cell comprising the DNA molecule of    claim 131   .   
     
     
       144. A plant comprising the glyphosate- tolerant plant cell of    claim 143   .   
     
     
       145. A glyphosate- tolerant plant cell comprising an EPSPS enzyme having the sequence of SEQ ID NO: 3 , SEQ ID NO: 5  or SEQ ID NO: 7 .   
     
     
       146. A glyphosate- tolerant plant comprising an EPSPS enzyme having the sequence of SEQ ID NO:  3 , SEQ ID NO:  5  or SEQ ID NO:  7 .   
     
     
       147. The glyphosate- tolerant plant cell of    claim 143    or    145    selected from the group consisting of corn, wheat, rice, barley, soybean, cotton, sugarbeet, oilseed rape, canola, flax, sunflower, potato, tobacco, tomato, alfalfa, poplar, pine, eucalyptus, apple, lettuce, peas, lentils, grape, and turf grasses.   
     
     
       148. The glyphosate- tolerant plant of    claim 144    or    146    selected from the group consisting of corn, wheat, rice, barley, soybean, cotton, sugarbeet, oilseed rape, canola, flax, sunflower, potato, tobacco, tomato, alfalfa, poplar, pine, eucalyptus, apple, lettuce, peas, lentils, grapes, and turf grasses.   
     
     
       149. A method for selectively controlling weeds in a field containing a crop having planted crop seeds or plants comprising the steps of:
   a )  planting the crop seeds or plants which are glyphosate - tolerant as a result of a recombinant double - stranded DNA molecule being inserted into the crop seed or plant, the DNA molecule having:      i )  a promoter which functions in plant cells to cause the production of an RNA sequence,        ii )  a structural DNA sequence that causes the production of an RNA sequence which encodes an EPSPS enzyme having the sequence of SEQ ID NO: 3 , SEQ ID NO: 5 , or SEQ ID NO: 7 ; and        iii )  a  3 ′ non - translated DNA sequence which functions in plant cells to cause the addition of a stretch of polyadenyl nucleotides to the  3 ′ end of the RNA sequence,          wherein the promoter is heterologous with respect to the structural DNA sequence and adapted to cause sufficient expression of the EPSPS enzyme to enhance the glyphosate tolerance of the crop plant transformed with the DNA molecule; and        b )  applying to the crop and weeds in the field a sufficient amount of glyphosate herbicide to control the weeds without significantly affecting the crop.