# US 10,308,953 B2 · Plants having increased tolerance to herbicides > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,308,953 B2 - Document ID: US10308953B2 - Status: Granted - Kind: B2 - Application number: 15/105,270 - Filed: 2014-12-17 - Published: 2016-12-29 - Granted: 2019-06-04 - Earliest claimed: 2013-12-18 - Assignees: BASF Agro B.V. - Applicants: BASF Agro B.V. - Inventors: Raphael Aponte; Stefan Tresch; Matthias Witschel; Jens Lerchl; Dario Massa; Tobias Seiser; Thomas Mietzner; Jill Marie Paulik; Chad Brommer - Web page: https://patentgrep.com/patent/US10308953B2 ## CPC - C12N 15/8274: {for herbicide resistance} - A01N 37/10: Aromatic or araliphatic carboxylic acids, or thio analogues thereof; Derivatives thereof - A01N 43/54: 1,3-Diazines; Hydrogenated 1,3-diazines - A01N 43/653: 1,2,4-Triazoles; Hydrogenated 1,2,4-triazoles - A01N 43/84: six-membered rings with one nitrogen atom and either one oxygen atom or one sulfur atom in positions 1,4 - C12N 9/001: {acting on the CH-CH group of donors (1.3)} - C12N 9/1059: {Cellulose synthases (2.4.1.12; 2.4.1.29)} - 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) - A01N: PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF (preservation of food or foodstuff A23); BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES (preparations for medical, dental or toiletry purposes which kill or prevent the growth or proliferation of unwanted organisms A61K); PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS - 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 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 (mutated-PPO) which is resistant or tolerant to a “PPO inhibiting herbicide”, and which comprises the amino acid sequence of SEQ ID NO: 11 with a double amino acid substitution selected from the group consisting of R98A/F390V; L367E/F390V; L367Q/F390V; L367E/F390M; and L367Q/F390M; and b) applying to said site an effective amount of said herbicide. ### Claim 2 (depends on 1) 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. ### Claim 3 (depends on 1) The method according to claim 1 wherein the PPO inhibiting herbicide is applied in conjunction with one or more additional herbicides. ### Claim 4 A transgenic plant cell comprising a nucleic acid encoding a mutated-PPO polypeptide, 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, and wherein the encoded mutated-PPO comprises the amino acid sequence of SEQ ID NO: 11 with a double amino acid substitution selected from the group consisting of R98A/F390V; L367E/F390V; L367Q/F390V; L367E/F390M; and L367Q/F390M. ### Claim 5 (depends on 4) A transgenic plant comprising a plant cell as defined in claim 4, 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. ### Claim 6 A plant that expresses a mutagenized or recombinant mutated-PPO polypeptide comprising the amino acid sequence of SEQ ID NO: 11 with a double amino acid substitution selected from the group consisting of R98A/F390V; L367E/F390V; L367Q/F390V; L367E/F390M; and L367Q/F390M, 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. ### Claim 7 (depends on 4) A seed produced by a transgenic plant comprising a plant cell as defined in claim 4, 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. ### Claim 8 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 comprising the amino acid sequence of SEQ ID NO: 11 with a double amino acid substitution selected from the group consisting of R98A/F390V; L367E/F390V; L367Q/F390V; L367E/F390M; and L367Q/F390M. ### Claim 9 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 comprising the amino acid sequence of SEQ ID NO: 11 with a double amino acid substitution selected from the group consisting of R98A/F390V; L367E/F390V; L367Q/F390V; L367E/F390M; and L367Q/F390M, and (b) generating a plant with an increased resistance to PPO inhibiting herbicide from the plant cell. ### Claim 10 (depends on 9) The method of claim 9, wherein the expression cassette further comprises a transcription initiation regulatory region and a translation initiation regulatory region that are functional in the plant.