Modified DMO enzyme and methods of its use

• US 7,884,262 B2
• Filed: 06/05/2007
• Issued: 02/08/2011
• Est. Priority Date: 06/06/2006
• Status: Active Grant

• Current Assignee: Monsanto Technology LLC (Bayer AG), Board of Regents of the University of Nebraska
• Original Assignee: Monsanto Technology LLC (Bayer AG)
• Inventors: Feng, Paul C. C., Clemente, Thomas E., Flasinski, Stanislaw, Dumitru, Razvan, Weeks, Donald P.
• Primary Examiner(s): O Hara; Eileen B
• Application Number:
  US11/758,657
• Publication Number:
  US 20080015110A1
• Time in Patent Office: 1,344 Days
• Field of Search: None
• US Class Current: 800/278
• CPC Class Codes: C12N 15/8274 for herbicide resistance
• CPC Class Codes: C12N 9/0069 acting on single donors with incorporation of molecular oxygen, i.e. oxygenases (1.13)

Abstract

The invention provides a modified variant of dicamba monooxygenase (DMO). The invention relates to the unexpected finding that cells expressing this DMO exhibit high levels of tolerance to the herbicide dicamba. Compositions comprising DMO-encoding nucleic acids and methods of use are provided.

Claims

1. A nucleic acid molecule selected from the group consisting of:

a) a nucleic acid molecule encoding the polypeptide of SEQ ID NO;

1;

b) a nucleic acid molecule comprising the sequence of SEQ ID NO;

2; and

c) a nucleic acid molecule encoding a polypeptide with at least 90% sequence identity to the polypeptide of SEQ ID NO;

1, wherein the polypeptide has dicamba monooxygenase activity and comprises cysteine at a position corresponding to amino acid 112 of SEQ ID NO;

1.

2. The nucleic acid molecule of claim 1, wherein the nucleic acid molecule encodes the dicamba monooxygenase encoded by plasmid pKLP36-TEV-TP-DMOc (ATCC Accession No. PTA-7357).

3. A DNA construct comprising the nucleic acid molecule of claim 1 operably linked to a promoter.

4. The construct of claim 3, wherein the promoter is functional in a plant cell.

5. The construct of claim 3, wherein the nucleic acid molecule is operably linked to a chloroplast transit peptide.

6. A method of producing a dicamba tolerant plant comprising introducing into the plant the construct of claim 3.

7. The method of claim 6, comprising introducing said construct into said plant by stably transforming a starting plant cell and regenerating the cell into said dicamba tolerant plant.

8. The method of claim 6, wherein the dicamba tolerant plant is produced by crossing a parent plant with itself or a second plant, wherein the parent plant and/or the second plant comprises the transformation construct and the dicamba tolerant plant inherits the transformation construct from said parent plant and/or the second plant.

9. A plant cell transformed with the nucleic acid molecule of claim 1.

10. The cell of claim 9, wherein the plant cell is a dicotyledonous plant cell.

11. The cell of claim 10, wherein the dicotyledonous plant cell is a soybean, cotton, maize or rapeseed plant cell.

12. The cell of claim 9, wherein the plant cell is a monocotyledonous plant cell.

13. A plant tissue culture comprising the cell of claim 9.

14. A transgenic plant transformed with the nucleic acid molecule of claim 1.

15. The transgenic plant of claim 14, wherein the plant is a dicotyledonous plant.

16. The transgenic plant of claim 14, wherein the plant is a monocotyledonous plant.

17. The transgenic plant of claim 14, wherein the plant is a soybean, cotton, maize or rapeseed plant.

18. A method of controlling weed growth in a crop growing environment comprising a plant of claim 14 or a seed thereof, the method comprising applying to the crop growing environment an amount of dicamba herbicide effective to control weed growth.

19. The method of claim 18, wherein the dicamba herbicide is applied over the top to the crop growing environment.

20. The method of claim 18, wherein the amount of dicamba herbicide does not damage said plant or seed thereof and damages a plant of the same genotype as the plant lacking the nucleic acid.

22. A method of producing food, feed or an industrial product comprising:

a) obtaining the plant of claim 14 or a part thereof; and

b) preparing the food, feed or industrial product from the plant or part thereof.

23. The method of claim 22, wherein the food or feed is oil, meal, grain, starch, flour, or protein.

24. The method of claim 22, wherein the industrial product is biofuel, fiber, industrial chemicals, a pharmaceutical or nutraceutical.

21. A polypeptide sequence comprising an amino acid sequence with at least 90% identity to SEQ ID NO:

1, wherein the polypeptide has dicamba monooxygenase activity and comprises cysteine at a position corresponding to amino acid 112 of SEQ ID NO;

1.