Transgenic or non-transgenic plants with mutated protoporphyrinogen oxidase having increased tolerance to herbicides

Patent metadata

Field	Value
Publication number	US-10982227-B2
Application number	US-201816124992-A
Country	US
Kind code	B2
Filing date	Sep 7, 2018
Priority date	Aug 12, 2013
Publication date	Apr 20, 2021
Grant date	Apr 20, 2021

Abstract

The present invention refers to a method for controlling undesired vegetation at a plant cultivation site, the method comprising the steps of providing, at said site, a plant that comprises at least one nucleic acid comprising a nucleotide sequence encoding a wild-type or a mutated protoporphyrinogen oxidase (PPO) which is resistant or tolerant to a PPO-inhibiting herbicide by applying to said site an effective amount of said herbicide. The invention further refers to plants comprising wild-type or mutated PPO enzymes, and methods of obtaining such plants.

First claim

The invention claimed is:

1. A method for controlling undesired vegetation at a plant cultivation site, the method comprising the steps of:
a) providing, at said site, a plant that comprises at least one nucleic acid comprising a nucleotide sequence encoding a mutated protoporphyrinogen oxidase (PPO) which is resistant or tolerant to a PPO inhibiting herbicide and
b) applying to said site an effective amount of said herbicide,

wherein the mutated PPO comprises a variant of SEQ ID NO: 2 having at least 75% identity to SEQ ID NO: 2, in which the amino acid at or corresponding to position 128 of SEQ ID NO:2 is Leu, Ala, Val, Ile, Met, Tyr, Gly, Asn, Cys, Phe, Ser, Thr, Gln, or His, and the amino acid at or corresponding to position 420 of SEQ ID NO:2 is Val.

2. The method according to claim 1 , wherein the plant comprises at least one additional heterologous nucleic acid comprising a nucleotide sequence encoding a herbicide tolerance enzyme.

3. The method according to claim 1 wherein the PPO inhibiting herbicide is applied in conjunction with one or more additional herbicides.

4. A mutated PPO polypeptide comprising a variant of SEQ ID NO: 2 having at least 75% identity to SEQ ID NO: 2, in which the amino acid at or corresponding to position 128 of SEQ ID NO:2 is Leu, Ala, Val, Ile, Met, Tyr, Gly, Asn, Cys, Phe, Ser, Thr, Gln, or His, and the amino acid at or corresponding to position 420 of SEQ ID NO:2 is Val, wherein said mutated PPO polypeptide confers increased resistance or tolerance to a PPO inhibiting herbicide when expressed in a plant as compared to a wild type plant.

5. A transgenic plant cell transformed by and expressing a nucleic acid encoding a mutated PPO polypeptide as defined in claim 4 , wherein expression of the nucleic acid in the plant cell results in increased resistance or tolerance to a PPO inhibiting herbicide as compared to a wild type variety of the plant cell.

6. A transgenic plant comprising a plant cell of claim 5 , wherein expression of the mutated PPO polypeptide encoding nucleic acid in the plant results in the plant's increased resistance to PPO inhibiting herbicide as compared to a wild type plant.

7. A plant cell mutagenized to obtain a plant cell which expresses a nucleic acid encoding a mutated PPO polypeptide of claim 4 .

8. A plant that expresses a mutagenized or recombinant mutated PPO polypeptide of claim 4 , and wherein said mutated PPO confers upon the plant increased herbicide tolerance as compared to the corresponding wild-type variety of the plant when expressed therein.

9. A method for growing a plant of claim 8 while controlling weeds in the vicinity of said plant, said method comprising the steps of:
a) growing said plant; and
b) applying a herbicide composition comprising a PPO-inhibiting herbicide to the plant and weeds, wherein the herbicide normally inhibits protoporphyrinogen oxidase, at a level of the herbicide that would inhibit the growth of a corresponding wild-type plant.

10. A seed produced by a plant of claim 8 , wherein the seed is true breeding for an increased resistance to a PPO inhibiting herbicide as compared to a wild type variety of the seed.

11. A method of producing a transgenic plant cell with an increased resistance to a PPO inhibiting herbicide as compared to a wild type variety of the plant cell comprising, transforming the plant cell with an expression cassette comprising a nucleic acid encoding a mutated PPO polypeptide as defined in claim 4 .

12. A method of producing a transgenic plant comprising, (a) transforming a plant cell with an expression cassette comprising a nucleic acid encoding a mutated PPO polypeptide as defined in claim 4 , and (b) generating a plant with an increased resistance to PPO inhibiting herbicide from the plant cell.

13. The method of claim 11 , wherein the expression cassette further comprises a transcription initiation regulatory region and a translation initiation regulatory region that are functional in the plant.

14. An expression cassette comprising a nucleic acid encoding a mutated PPO polypeptide as defined in claim 4 , a transcription initiation regulatory region and a translation initiation regulatory region that are functional in the plant, and a chloroplast-targeting sequence comprising a nucleotide sequence that encodes a chloroplast transit peptide.

15. The expression cassette of claim 14 , wherein the targeting sequence comprises a nucleotide sequence that encodes a transit peptide comprising the amino acid sequence of SEQ ID NO: 49, 50, 51, 52, or 53.

16. The method of claim 1 , wherein the mutated PPO comprises a variant of SEQ ID NO: 2 having at least 85% identity to SEQ ID NO: 2, in which the amino acid at or corresponding to position 128 of SEQ ID NO: 2 is Leu, Ala, Val, Ile, Met, Tyr, Gly, Asn, Cys, Phe, Ser, Thr, Gln, or His, and the amino acid at or corresponding to position 420 of SEQ ID NO: 2 is Val.

17. The method of claim 1 , wherein the mutated PPO comprises a variant of SEQ ID NO: 2 having at least 90% identity to SEQ ID NO: 2, in which the amino acid at or corresponding to position 128 of SEQ ID NO: 2 is Leu, Ala, Val, Ile, Met, Tyr, Gly, Asn, Cys, Phe, Ser, Thr, Gln, or His, and the amino acid at or corresponding to position 420 of SEQ ID NO: 2 is Val.

18. The method of claim 1 , wherein the mutated PPO comprises a variant of SEQ ID NO: 2 having at least 95% identity to SEQ ID NO: 2, in which the amino acid at or corresponding to position 128 of SEQ ID NO: 2 is Leu, Ala, Val, Ile, Met, Tyr, Gly, Asn, Cys, Phe, Ser, Thr, Gln, or His, and the amino acid at or corresponding to position 420 of SEQ ID NO: 2 is Val.

19. The mutated PPO polypeptide of claim 4 , wherein the mutated PPO polypeptide comprises a variant of SEQ ID NO: 2 having at least 85% identity to SEQ ID NO: 2, in which the amino acid at or corresponding to position 128 of SEQ ID NO: 2 is Leu, Ala, Val, Ile, Met, Tyr, Gly, Asn, Cys, Phe, Ser, Thr, Gln, or His, and the amino acid at or corresponding to position 420 of SEQ ID NO: 2 is Val.

20. The mutated PPO polypeptide of claim 4 , wherein the mutated PPO polypeptide comprises a variant of SEQ ID NO: 2 having at least 90% identity to SEQ ID NO: 2, in which the amino acid at or corresponding to position 128 of SEQ ID NO: 2 is Leu, Ala, Val, Ile, Met, Tyr, Gly, Asn, Cys, Phe, Ser, Thr, Gln, or His, and the amino acid at or corresponding to position 420 of SEQ ID NO: 2 is Val.

21. The mutated PPO polypeptide of claim 4 , wherein the mutated PPO polypeptide comprises a variant of SEQ ID NO: 2 having at least 95% identity to SEQ ID NO: 2, in which the amino acid at or corresponding to position 128 of SEQ ID NO: 2 is Leu, Ala, Val, Ile, Met, Tyr, Gly, Asn, Cys, Phe, Ser, Thr, Gln, or His, and the amino acid at or corresponding to position 420 of SEQ ID NO: 2 is Val.

22. The transgenic plant cell of claim 5 , wherein the mutated PPO polypeptide comprises a variant of SEQ ID NO: 2 having at least 85% identity to SEQ ID NO: 2, in which the amino acid at or corresponding to position 128 of SEQ ID NO: 2 is Leu, Ala, Val, Ile, Met, Tyr, Gly, Asn, Cys, Phe, Ser, Thr, Gln, or His, and the amino acid at or corresponding to position 420 of SEQ ID NO: 2 is Val.

23. The transgenic plant cell of claim 5 , wherein the mutated PPO polypeptide comprises a variant of SEQ ID NO: 2 having at least 90% identity to SEQ ID NO: 2, in which the amino acid at or corresponding to position 128 of SEQ ID NO: 2 is Leu, Ala, Val, Ile, Met, Tyr, Gly, Asn, Cys,

Assignees

• Basf Agro Bv

Inventors

• Aponte Raphael
• Tresch Stefan
• Witschel Matthias
• Lerchl Jens
• Massa Dario
• Seiser Tobias
• Mietzner Thomas
• Paulik Jill Marie
• Brommer Chad

Classifications

• C12N15/82 Primary

  for plant cells {, e.g. plant artificial chromosomes (PACs)} · CPC title

• A01H5/00

  Angiosperms, i.e. flowering plants, characterised by their plant parts; Angiosperms characterised otherwise than by their botanic taxonomy · CPC title

• A01N63/00

  Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates (containing compounds of determined constitution A01N27/00 - A01N59/00; unicellular algae A01N65/03) · CPC title

• C12Y103/03004

  Protoporphyrinogen oxidase (1.3.3.4) · CPC title

• C12N9/001

  acting on the CH-CH group of donors (1.3) · CPC title