Mutated 5-enolpyruvylshikimate-3-phosphate synthase, gene coding for said protein and transformed plants containing said gene

• US 6,566,587 B1
• Filed: 01/20/1998
• Issued: 05/20/2003
• Est. Priority Date: 07/19/1995
• Status: Expired due to Term

• Current Assignee: Bayer SAS (Bayer AG)
• Original Assignee: Bayer Cropscience SA (Bayer AG)
• Inventors: Freyssinet, Georges, Lebrun, Michel, Degryse, Eric, Sailland, Alain
• Primary Examiner(s): Nelson, Amy J.
• Assistant Examiner(s): Kruse, David H
• Application Number:
  US08/945,144
• Time in Patent Office: 1,946 Days
• Field of Search: 536/23.2, 536/23.6, 47/581, 435/410, 435/413, 435/419, 435/430.1, 435/440, 800/300, 800/320.1
• US Class Current: 800/300
• CPC Class Codes: C12N 15/8275 Glyphosate
• CPC Class Codes: C12N 9/1092 3-Phosphoshikimate 1-carboxyvinyltransferase (2.5.1.19), i.e. 5-enolpyruvylshikimate-3-phosphate synthase

Abstract

The present invention relates to DNA molecules of plant origin encoding a modified 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) enzyme wherein the a first EPSPS coding sequence that normally encodes a threonine residue of a mature EPSPS sequence is modified to encode isoleucine of the mature EPSPS sequence, and a second EPSPS coding sequence that normally encodes a proline residue of a mature EPSPS sequence is modified to encode serine of the mature EPSPS sequence, wherein the first and second residues are respectively located at relatively positions 102 and 106 of a mature EPSPS sequence encoded by said DNA molecule; and the production of a transgenic plant resistant or tolerant to a herbicide of the phosphonomethylglycine family, e.g., glyphosate. The mutated enzyme, which substantially maintains the catalytic activity of the wild-type enzyme, allows for increased tolerance or tolerance of the plant to a herbicide of the phosphonomethylglycine family, and allows for the substantially normal growth or development of the plant, its organs, tissues or cells.

Claims

1. A modified plant DNA molecule encoding a modified EPSPS enzyme wherein said enzyme comprises:

a first amino substitution from threonine to isoleucine at the threonine which corresponds to position 102 of the amino acid sequence of SEQ ID NO;

3; and

a second amino acid substitution from proline to serine at the proline which corresponds to position 106 of the amino acid sequence of SEQ ID NO;

3.

2. The modified plant DNA molecule as claimed in claim 1 wherein the modified plant DNA molecule is of maize origin.

4. A vector comprising the following components, which are operably associated in the direction of transcription:

(a) a promoter functional in a plant cell; and

(b) the modified plant DNA molecule of claim 1.

5. The vector of claim 4 further comprising a nucleic acid encoding a chloroplast transit peptide operably associated with, and in the order of transcription between, the promoter functional in a plant cell and the modified plant DNA molecule.

3. An isolated polynucleotide molecule having the sequence of SEQ ID NO:

4.

6. A transformed plant cell comprising a vector comprising the following components, which are operably associated in the direction of transcription:

(a) a promoter functional in a plant cell;

(b) a nucleic acid encoding a chloroplast transit peptide;

(c) a modified DNA molecule of maize origin encoding a modified EPSPS enzyme wherein said enzyme comprises;

a first amino acid substitution from threonine to isoleucine at the threonine which corresponds to position 102 of the amino acid sequence of SEQ ID NO;

3; and

a second amino acid substitution from proline to serine at the proline which corresponds to position 106 of the amino acid sequence of SEQ ID NO;

3; and

(d) an untranslated transcription termination signal region.

7. The plant cell of claim 6 which is a monocot plant cell with increased tolerance to glyphosate herbicides relative to a non-transformed plant cell.

8. The plant cell of claim 6 which is a dicot plant cell with increased tolerance to glyphosate herbicide relative to a non-transformed plant cell.

9. A transgenic plant comprising a vector comprising the following components, which are operably associated in the direction of transcription:

(a) a promoter functional in a plant cell;

(b) a nucleic acid encoding a chloroplast transit peptide;

(c) a modified DNA molecule of maize origin encoding a modified EPSPS enzyme wherein said enzyme comprises;

a first amino acid substitution from threonine to isoleucine at the threonine which corresponds to position 102 of the amino acid sequence of SEQ ID NO;

3; and

a second amino acid substitution from proline to serine at the proline which corresponds to position 106 of the amino acid sequence of SEQ ID NO;

3; and

(d) an untranslated transcription termination signal region.

10. The transgenic plant of claim 9 which is a monocot with increased tolerance to glyphosate herbicides relative to a non-transgenic plant.

11. The transgenic plant of claim 9 which is a dicot with increased tolerance to glyphosate herbicides relative to a non-transgenic plant.

12. A method for selectively controlling plants which method comprises the steps of:

a) planting crop seeds or plants which have increased glyphosate tolerance as a result of a chimeric gene being inserted into said crop seeds or plants, said chimeric gene having

(i) a promoter region functional in a plant cell; and

(ii) a modified plant DNA molecule encoding a modified EPSAPS enzyme wherein said enzyme comprises;

a first amino acid substitution from threonine to isoleucine at the threonine which corresponds to position 102 of the amino acid sequence of SEQ ID NO;

3; and

a second amino acid substitution from proline to serine at the proline which corresponds to position 106 of the amino acid sequence of SEQ ID NO;

3; and

(iii) an untranslated transcription termination signal region; and

b) applying to said crop seeds or plants a sufficient amount of glyphosate to control untransformed plants without significantly affecting said crop seeds or plants that comprise the chimeric gene.

13. A plant transformed with a modified DNA molecule encoding a plant EPSPS enzyme having a first amino acid substitution from threonine to isoleucine at the threonine which corresponds to position 102 of the amino acid sequence of SEQ ID NO:

3; and

a second amino acid substitution from proline to serine at the proline which corresponds to position 106 of the amino acid sequence of SEQ ID NO;

3.