# US 12,599,073 B2 · Wheat CENH3 alleles

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- Number: US 12,599,073 B2
- Document ID: US12599073B2
- Status: Granted
- Kind: B2
- Application number: 18/426,902
- Filed: 2024-01-30
- Published: 2024-10-10
- Granted: 2026-04-14
- Earliest claimed: 2018-10-12
- Assignees: Syngenta Crop Protection AG
- Applicants: Syngenta Crop Protection AG
- Inventors: Jian Lv; Kun Yu; Juan Wei; Chunxia Liu; Hongju Zhou; Timothy Kelliher
- Web page: https://patentgrep.com/patent/US12599073B2

## CPC

- A01H 6/4678: {Triticum sp. [wheat]}
- C12N 15/63: Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N 15/8213: {Targeted insertion of genes into the plant genome by homologous recombination}
- A01H: NEW PLANTS OR {NON-TRANSGENIC} PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The present invention relates to wheat plants comprising a mutation causing an alteration of the amino acid sequence in centromere histone H3 (CENH3), which have the biological activity of a haploid inducer. Further, the present invention provides methods of generating the wheat plants of the present invention and haploid and doubled haploid wheat plants obtainable by crossing the wheat plants of the present invention with wildtype wheat plants.

## Claims

### Claim 1

A method of generating a haploid-inducing wheat plant, the method comprising: a. obtaining at least a wheat plant cell comprising an A genome, a B genome, and a D genome; b. mutating the B genome to obtain a homozygous knock-out mutation in a CENH3α gene; c. mutating the D genome to obtain a homozygous knock-out mutation in a CENH3α gene; d. mutating the A genome to obtain a knock-down mutation at a 5′ splice site of an intron in a CENH3α gene; and e. generating a wheat plant therefrom; whereby the wheat plant generated from step e. produces haploid progeny when crossed with a wildtype wheat plant.

### Claim 2 (depends on 1)

The method of claim 1, wherein the knock-down mutation in the CENH3α gene is a restored frame shift mutation.

### Claim 3 (depends on 2)

The method of claim 2, wherein the restored frame shift mutation is selected from the group consisting of SEQ ID NO: 56, a nucleic acid sequence 98% identical to SEQ ID NO: 56, SEQ ID NO: 63, a nucleic acid sequence 98% identical to SEQ ID NO: 63, or a combination thereof.