Herbicide-tolerant plants through bypassing metabolic pathway

• US 20080163392A1
• Filed: 11/06/2007
• Published: 07/03/2008
• Est. Priority Date: 10/30/2000
• Status: Active Grant

• Current Assignee: Bayer Cropscience SA (Bayer AG)
• Original Assignee: Bayer Cropscience SA (Bayer AG)
• Inventors: Rolland, Anne, Zink, Olivier, Freyssinet, Georges, Sailland, Alain, Paget, Eric
• Granted Patent: US 8,124,846 B2
• Time in Patent Office: Days
• US Class Current: 800/278
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C12N 15/8274 for herbicide resistance
• CPC Class Codes: C12N 9/0008 acting on the aldehyde or oxo group of donors (1.2)
• CPC Class Codes: C12N 9/0071 acting on paired donors with incorporation of molecular oxygen (1.14)
• CPC Class Codes: C12N 9/0073 with NADH or NADPH as one donor, and incorporation of one atom of oxygen 1.14.13
• CPC Class Codes: C12Y 114/13018 4-Hydroxyphenylacetate 1-monooxygenase (1.14.13.18)

Abstract

The invention concerns a novel method for making herbicide-tolerant plants, in particular to HPPD inhibiting herbicides, the nucleic acid sequences coding for enzymes capable of being used in said method, expression cassettes containing them and transgenic plants comprising at least one of said expression cassettes.

Claims

1. A method for making plants tolerant to a herbicide, characterized in that at least one enzyme making it possible to bypass the metabolic pathway inhibited by said herbicide is expressed in said plant, wherein said at least one enzyme includes HPAH or HPAC.

2. The method as claimed in claim 1, characterized in that the herbicide is an HPPD inhibitor.

3. The method as claimed in claim 2, characterized in that an HPP oxidase and/or an HPAH and/or an HPAC is expressed in the plant.

4. A polypeptide, characterized in that it has HPP oxidase activity.

5. The polypeptide as claimed in claim 4, characterized in that it is insensitive to HPPD inhibitors.

6. The polypeptide as claimed in claim 4, characterized in that it comprises the amino acid sequence of SEQ ID No. 2, the fragments thereof and the sequences homologous thereto.

7. The polypeptide as claimed in claim 6, characterized in that it comprises an amino acid sequence selected from the group comprising SEQ ID No. 4 and SEQ ID No. 6, the fragments thereof and the sequences homologous thereto.

16. (canceled)

29. A method for making plants tolerant to an herbicide which is an HPPD inhibitor, comprising the step of growing plants expressing at least one enzyme insensitive to said herbicide that allows conversion of para-hydroxyphenylpyruvate (HPP) to 4-hydroxyphenylacetic acid (4-HPA) and at least one enzyme insensitive to said herbicide that allows conversion of 4-HPA to homogentisate as a result of nucleic acid sequences encoding said enzymes being inserted into said plants.

30. The method as claimed in claim 29 wherein at least one of HPP oxidase, an HPAH, and HPAC is expressed in said plant.

31. A polypeptide characterized in that it has HPAH activity or HPAC activity.

8. The polypeptide of claim 31 characterized in that it has HPAH activity.

9. The polypeptide as claimed in claim 8, characterized in that it is insensitive to HPPD inhibitors.

10. The polypeptide as claimed in either of claim 8, characterized in that it comprises the amino acid sequence of SEQ ID No. 8, the fragments thereof and the sequences homologous thereto.

11. The polypeptide of claim 31 characterized in that it has HPAC activity.

12. The polypeptide as claimed in claim 11, characterized in that it is insensitive to HPPD inhibitors.

13. The polypeptide as claimed in claim 11, characterized in that it comprises the amino acid sequence of SEQ ID No. 10, the fragments thereof and the sequences homologous thereto.

14. The polypeptide as claimed in claim 13, characterized in that it comprises an amino acid sequence selected from the group comprising SEQ ID No. 12 and SEQ ID No. 14, the fragments thereof and the sequences homologous thereof.

15. A nucleic acid sequence, characterized in that it encodes a polypeptide as claimed in claim 31.

17. The nucleic acid sequence of claim 15, wherein said nucleic acid sequence encodes an HPAH, characterized in that it comprises a nucleic acid sequence selected from the group comprising the coding sequences of SEQ ID No. 7 or SEQ ID No. 17, the sequences homologous thereto, the fragments thereof and the sequences capable of selectively hybridizing to said SEQ ID No. 7 or 17.

18. The nucleic acid sequence of claim 15, wherein said nucleic acid sequence encodes an HPAC, characterized in that it comprises a nucleic acid sequence selected from the group comprising the coding sequences of SEQ ID No. 9, SEQ ID No. 11, SEQ ID No. 13 or SEQ ID No. 19, the sequences homologous thereto, the fragments thereof and the sequences capable of selectively hybridizing to said SEQ ID No. 9, 11, 13 or 19.

19. An expression cassette comprising a coding sequence, characterized in that the coding sequence comprises a nucleic acid sequence as claimed in claim 15.

20. A cloning and/or expression vector, characterized in that it comprises at least one expression cassette as claimed in claim 19.

21. A transformed plant cell, characterized in that it comprises at least one expression cassette as claimed in claim 19.

22. A transformed plant, characterized in that it comprises at least one expression cassette as claimed in claim 19.

23. A seed of a transformed plant as claimed in claim 22, characterized in that it comprises at least one expression cassette comprising a coding sequence, wherein said coding sequence encodes a polypeptide that has HPAH or HPAC activity.

24. A method of transforming plants, characterized in that at least one expression cassette as claimed in claim 19 is introduced into their genome.

25. A method for selective weeding of plants, in particular of crops, using an HPPD inhibitor, characterized in that this herbicide is applied to transformed plants as claimed in claim 22.

26. A method of weed killing in an area of a field comprising seeds as claimed in claim 23 or transformed plants, wherein said transformed plants comprise at least one expression cassette comprising a coding sequence encoding a polypeptide having HPAH or HPAC activity, characterized in that it comprises the application, in said area of the field, off a dose, which is toxic for said weeds, of an HPPD-inhibiting herbicide, without however substantially affecting the seeds or transformed plants according to the invention.

27. A method of growing transformed plants, wherein said transformed plants comprise at least one expression cassette comprising a coding sequence encoding a polypeptide having HPAH or HPAC activity, characterized in that it comprises sowing the seeds as claimed in claim 23 in an area of a field suitable for growing said plants, applying to said area of said field a dose, which is toxic for the weeds, of a herbicide having HPPD as the target, in the event of weeds being present, without substantially affecting said seeds or said transformed plants, then harvesting the plants grown when they reach the desired maturity and, optionally, separating the seeds from the harvested plants.

28. The method as claimed in claim 25, characterized in that the herbicide is applied in pre-emergence and/or in post-emergence.