Glyphosate-tolerant 5-enolpyruvylshikimate-3-phosphate synthases

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.sub.1 -H-X.sub.2 -E-(SEQ ID NO:37), in which
• X.sub.1 is G, S, T, C, Y, N, Q, D or E;
• X.sub.2 is S or T;
• and
• -G-D-K-X.sub.3 -(SEQ ID NO:38), in which
• X.sub.3 is S or T;
• and
• -S-A-Q-X.sub.4 -K-(SEQ ID NO:39), in which
• X.sub.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.sub.5 -T-R-(SEQ ID NO:40), in which
• X.sub.5 is A, R, N, D, C, Q, E, G, H, I, L, K, M, F, P, 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.

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.sub.1 H-X.sub.2 -E-(SEQ ID NO:37), in which
• X.sub.1 is G, S, T, C, Y, N, Q, D or E;
• X.sub.2 is S or T;
• and
• -G-D-K-X.sub.3 -(SEQ ID NO:38), in which
• X.sub.3 is S or T;
• and
• -S-A-Q-X.sub.4 -K-(SEQ ID NO:39), in which
• X.sub.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.sub.5 -T-R-(SEQ ID NO:40), in which
• X.sub.5 is A, R, N, D, C, Q, E, G, H, I, L, K, M, F, P, 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.

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 domains:
• -R-X.sub.1 -H-X.sub.2 -E-(SEQ ID NO:37), in which
• X.sub.1 is G, S, T, C, Y, N, Q, D or E;
• X.sub.2 is S or T;
• and
• -G-D-K-X.sub.3 -(SEQ ID NO:38), in which
• X.sub.3 is S or T;
• and
• -S-A-Q-X.sub.4 -K-(SEQ ID NO:39), in which
• X.sub.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.sub.5 -T-R-(SEQ ID NO:40), in which
• X.sub.5 is A, R, N, D, C, Q, E, G, H, I, L, K, M, F, P, 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.