# US 11,441,154 B2 · Plants having increased tolerance to herbicides

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- Number: US 11,441,154 B2
- Document ID: US11441154B2
- Status: Granted
- Kind: B2
- Application number: 16/129,379
- Filed: 2018-09-12
- Published: 2019-02-28
- Granted: 2022-09-13
- Earliest claimed: 2012-06-19
- Assignees: BASF Agro B.V.
- Applicants: BASF Agro B.V.
- Inventors: Jens Lerchl; Stefan Tresch; Dario Massa; Tobias Seiser; Matthias Witschel; Raphael Aponte; Jill Marie Paulik; Chad Brommer
- Web page: https://patentgrep.com/patent/US11441154B2

## CPC

- C12N 15/8274: {for herbicide resistance}
- C12N 9/001: {acting on the CH-CH group of donors (1.3)}
- C12Y 103/03004: Protoporphyrinogen oxidase (1.3.3.4)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12Y: ENZYMES

## 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.

## Claims

### Claim 1

An isolated nucleic acid encoding a mutated protoporphyrinogen oxidase (mut-PPO), wherein the encoded mut-PPO is a variant of SEQ ID NO: 2 having at least 75% identity to SEQ ID NO: 2 and comprising an amino acid substitution at a position corresponding to position Leu397 of SEQ ID NO:2 and an amino acid substitution at a position corresponding to position Phe420 of SEQ ID NO:2, wherein the amino acid at the position corresponding to position Leu397 of SEQ ID NO:2 is substituted with aspartate, glutamate, or glutamine, and wherein the amino acid at the position corresponding to position Phe420 of SEQ ID NO:2 is substituted with valine or methionine.

### Claim 2 (depends on 1)

A plant or plant cell comprising the nucleic acid encoding a mut-PPO polypeptide of claim 1, wherein expression of the nucleic acid in the plant or plant cell results in increased resistance or tolerance to a PPO inhibiting herbicide as compared to a wild type variety of the plant or plant cell.

### Claim 3 (depends on 2)

A seed produced by the plant of claim 2, 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, and wherein the seed comprises the nucleic acid.

### Claim 4 (depends on 1)

The isolated nucleic acid of claim 1, wherein the encoded mut-PPO is a variant of SEQ ID NO: 2 having at least 85% identity to SEQ ID NO: 2 and comprising an amino acid substitution at a position corresponding to position Leu397 of SEQ ID NO:2 and an amino acid substitution at a position corresponding to position Phe420 of SEQ ID NO:2, wherein the amino acid at the position corresponding to position Leu397 of SEQ ID NO:2 is substituted with aspartate, glutamate, or glutamine, and wherein the amino acid at the position corresponding to position Phe420 of SEQ ID NO:2 is substituted with valine or methionine.

### Claim 5 (depends on 1)

The isolated nucleic acid of claim 1, wherein the encoded mut-PPO is a variant of SEQ ID NO: 2 having at least 90% identity to SEQ ID NO: 2 and comprising an amino acid substitution at a position corresponding to position Leu397 of SEQ ID NO:2 and an amino acid substitution at a position corresponding to position Phe420 of SEQ ID NO:2, wherein the amino acid at the position corresponding to position Leu397 of SEQ ID NO:2 is substituted with aspartate, glutamate, or glutamine, and wherein the amino acid at the position corresponding to position Phe420 of SEQ ID NO:2 is substituted with valine or methionine.

### Claim 6 (depends on 1)

The isolated nucleic acid of claim 1, wherein the encoded mut-PPO is a variant of SEQ ID NO: 2 having at least 95% identity to SEQ ID NO: 2 and comprising an amino acid substitution at a position corresponding to position Leu397 of SEQ ID NO:2 and an amino acid substitution at a position corresponding to position Phe420 of SEQ ID NO:2, wherein the amino acid at the position corresponding to position Leu397 of SEQ ID NO:2 is substituted with aspartate, glutamate, or glutamine, and wherein the amino acid at the position corresponding to position Phe420 of SEQ ID NO:2 is substituted with valine or methionine.

### Claim 7 (depends on 1)

The isolated nucleic acid of claim 1, wherein the encoded mut-PPO is a variant of SEQ ID NO: 2 having at least 98% identity to SEQ ID NO: 2 and comprising an amino acid substitution at a position corresponding to position Leu397 of SEQ ID NO:2 and an amino acid substitution at a position corresponding to position Phe420 of SEQ ID NO:2, wherein the amino acid at the position corresponding to position Leu397 of SEQ ID NO:2 is substituted with aspartate, glutamate, or glutamine, and wherein the amino acid at the position corresponding to position Phe420 of SEQ ID NO:2 is substituted with valine or methionine.

### Claim 8 (depends on 2)

The plant or plant cell of claim 2, wherein the encoded mut-PPO is a variant of SEQ ID NO: 2 having at least 85% identity to SEQ ID NO: 2 and comprising an amino acid substitution at a position corresponding to position Leu397 of SEQ ID NO:2 and an amino acid substitution at a position corresponding to position Phe420 of SEQ ID NO:2, wherein the amino acid at the position corresponding to position Leu397 of SEQ ID NO:2 is substituted with aspartate, glutamate, or glutamine, and wherein the amino acid at the position corresponding to position Phe420 of SEQ ID NO:2 is substituted with valine or methionine.

### Claim 9 (depends on 2)

The plant or plant cell of claim 2, wherein the encoded mut-PPO is a variant of SEQ ID NO: 2 having at least 90% identity to SEQ ID NO: 2 and comprising an amino acid substitution at a position corresponding to position Leu397 of SEQ ID NO:2 and an amino acid substitution at a position corresponding to position Phe420 of SEQ ID NO:2, wherein the amino acid at the position corresponding to position Leu397 of SEQ ID NO:2 is substituted with aspartate, glutamate, or glutamine, and wherein the amino acid at the position corresponding to position Phe420 of SEQ ID NO:2 is substituted with valine or methionine.

### Claim 10 (depends on 2)

The plant or plant cell of claim 2, wherein the encoded mut-PPO is a variant of SEQ ID NO: 2 having at least 95% identity to SEQ ID NO: 2 and comprising an amino acid substitution at a position corresponding to position Leu397 of SEQ ID NO:2 and an amino acid substitution at a position corresponding to position Phe420 of SEQ ID NO:2, wherein the amino acid at the position corresponding to position Leu397 of SEQ ID NO:2 is substituted with aspartate, glutamate, or glutamine, and wherein the amino acid at the position corresponding to position Phe420 of SEQ ID NO:2 is substituted with valine or methionine.

### Claim 11 (depends on 2)

The plant or plant cell of claim 2, wherein the encoded mut-PPO is a variant of SEQ ID NO: 2 having at least 98% identity to SEQ ID NO: 2 and comprising an amino acid substitution at a position corresponding to position Leu397 of SEQ ID NO:2 and an amino acid substitution at a position corresponding to position Phe420 of SEQ ID NO:2, wherein the amino acid at the position corresponding to position Leu397 of SEQ ID NO:2 is substituted with aspartate, glutamate, or glutamine, and wherein the amino acid at the position corresponding to position Phe420 of SEQ ID NO:2 is substituted with valine or methionine.