HPPD variants and methods of use

Patent metadata

Field	Value
Publication number	US-12305181-B2
Application number	US-201916537801-A
Country	US
Kind code	B2
Filing date	Aug 12, 2019
Priority date	Mar 11, 2014
Publication date	May 20, 2025
Grant date	May 20, 2025

Abstract

In the present invention, HPPD enzymes and plants containing them showing a full tolerance against several classes of HPPD-inhibitors are described.
A set of HPPD enzymes have been designed which have either no or only a significantly reduced affinity to HPPD inhibitors and, at the same time, the rate of dissociation of the HPPD inhibitors of the enzyme is increased to such an extent that the HPPD inhibitors no longer act as slow-binding or slow, tight-binding inhibitors but, instead of this, have become fully reversible inhibitors.
In particular, isolated polynucleotides encoding HPPD inhibitor tolerance polypeptides are provided. Additionally, amino acid sequences corresponding to the polynucleotides are encompassed.

First claim

The invention claimed is:

1. A recombinant nucleic acid molecule encoding a 4-hydroxyphenylpyruvate dioxygenase (HPPD) protein comprising an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 1, wherein said amino acid sequence comprises a proline at the amino acid position corresponding to amino acid position 335 of SEQ ID NO:1 and a tyrosine at the amino acid position corresponding to amino acid position 336 of SEQ ID NO: 1, and wherein said HPPD protein is tolerant to an HPPD inhibitor herbicide,
wherein the amino acid sequence further comprises
(i) an alanine at the amino acid position corresponding to amino acid position 339 of SEQ ID NO:1; or
(ii) a glutamic acid, or glutamine at the amino acid position corresponding to amino acid position 340 of SEQ ID NO:1.

2. The recombinant nucleic acid molecule of claim 1 , wherein said HPPD amino acid sequence further comprises:
(i) a proline or leucine at the amino acid position corresponding to amino acid position 215 of SEQ ID NO:1.

3. The recombinant nucleic acid molecule of claim 1 , wherein its nucleotide sequence is a synthetic sequence that has been designed for expression in a plant.

4. The recombinant nucleic acid molecule of claim 1 , wherein its nucleotide sequence is operably linked to a promoter capable of directing expression of the nucleotide sequence in a plant cell.

5. A host cell comprising the recombinant nucleic acid molecule of claim 1 .

6. The host cell of claim 5 that is a bacterial host cell.

7. The host cell of claim 5 that is a plant cell.

8. A transgenic plant comprising the recombinant nucleic acid molecule of claim 1 .

9. The plant of claim 8 , wherein said plant is selected from the group consisting of maize, sorghum, wheat, sunflower, tomato, crucifers, peppers, potato, cotton, rice, soybean, sugar beet, sugarcane, tobacco, barley, and oilseed rape.

10. A transgenic seed comprising the recombinant nucleic acid molecule of claim 1 .

11. A method of controlling weeds in a field comprising planting the plant of claim 8 in a field and applying to said field an effective concentration of an HPPD inhibitor herbicide.

12. The method of claim 11 , wherein said HPPD inhibitor herbicide is selected from the group consisting of N-(1,2,5-oxadiazol-3-yl)benzamides; N-(tetrazol-4-yl)- or N-(triazol-3-yl)-arylcarboxamides, N-(1,3,4-oxadiazol-2-yl)benzamides, N-(tetrazol-5-yl)- or N-(triazol-3-yl)arylcarboxamides, pyridazinone derivatives, substituted 1,2,5-oxadiazoles, oxoprazin derivatives, triketones, isoxazoles, and pyrazolinates.

13. The method of claim 12 , wherein said HPPD inhibitor herbicide is selected from the group consisting of 2-chloro-3-ethoxy-4-(methylsulfonyl)-N-(1-methyl-1H-tetrazol-5-yl)benzamide and 2-Chloro-3-(methoxymethyl)-4-(methylsulfonyl)-N-(1-methyl-1H-tetrazol-5-yl)benzamide, 2-methyl-N-(5-methyl-1,3,4-oxadiazol-2-yl)-3-(methylsulfonyl)-4-(trifluoromethyl)benzamide, 2-chloro-3-ethoxy-4-(methylsulfonyl)-N-(1-methyl-1H-tetrazol-5-yl)benzamide, 4-(difluoromethyl)-2-methoxy-3-(methylsulfonyl)-N-(1-methyl-1H-tetrazol-5-yl)benzamide, 2-chloro-3-(methylsulfanyl)-N-(1-methyl-1H-tetrazol-5-yl)-4-(trifluoromethyl)benzamide, 2-(methoxymethyl)-3-(methylsulfinyl)-N-(1-methyl-1H-tetrazol-5-yl)-4-(trifluoromethyl)benzamide, tembotrione, sulcotrione, mesotrione, isoxaflutole, pyrasulfotole, and topramezone.

Assignees

• BASF Agricultural Solutions Seed US LLC
• Basf Agricultural Solutions Us Llc

Inventors

• Poree Fabien
• Laber Bernd
• Lange Gudrun
• Dubald Manuel
• Armstrong Roxanne

Classifications

• C12Y113/11001

  Catechol 1,2-dioxygenase (1.13.11.1) · CPC title

• A01N43/713

  having rings with four or more nitrogen atoms as the only ring hetero atoms · CPC title

• C12N9/0069 Primary

  acting on single donors with incorporation of molecular oxygen, i.e. oxygenases (1.13) · CPC title

• C12Y113/11027

  4-Hydroxyphenylpyruvate dioxygenase (1.13.11.27) · CPC title

• C12N15/8274 Primary

  for herbicide resistance · CPC title