# US 10,306,848 B2 · Generation of haploid plants and improved plant breeding

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- Number: US 10,306,848 B2
- Document ID: US10306848B2
- Status: Granted
- Kind: B2
- Application number: 14/939,181
- Filed: 2015-11-12
- Published: 2016-03-03
- Granted: 2019-06-04
- Earliest claimed: 2009-10-06
- Assignees: The Regents of the University of California
- Applicants: The Regents of the University of California
- Inventors: Simon Chan; Ravi Maruthachalam
- Web page: https://patentgrep.com/patent/US10306848B2

## CPC

- C12N 15/8241: {Phenotypically and genetically modified plants via recombinant DNA technology}
- C07K 14/415: from plants
- C12N 15/8261: {with agronomic (input) traits, e.g. crop yield}
- C12N 15/8287: {for fertility modification, e.g. apomixis}
- Y02A 40/146: Genetically Modified [GMO] plants, e.g. transgenic plants
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE

## Abstract

Methods and compositions for generating haploid organisms are described.

## Claims

### Claim 1

A plant comprising a mutated CENH3 gene, wherein the mutated CENH3 gene encodes a mutant CENH3 polypeptide having a CENH3 tail-domain and a CENH3 histone-fold domain, wherein the mutant CENH3 polypeptide is identical to a wildtype CENH3 polypeptide but for 1-7 amino acid substitutions that occur in the CENH3 histone-fold domain, wherein the plant comprising the mutated CENH3 gene, when crossed with a wildtype diploid plant, generates haploid progeny.

### Claim 2 (depends on 1)

The plant of claim 1, wherein the plant comprises one copy of an allele of the mutated CENH3 gene.

### Claim 3 (depends on 1)

The plant of claim 1, wherein the plant comprises two copies of an allele of the mutated CENH3 gene.

### Claim 4 (depends on 1)

The plant of claim 1, wherein the mutant CENH3 polypeptide is at least 95% identical to any of SEQ ID NOs:49-94.

### Claim 5 (depends on 1)

The plant of claim 1, wherein the wildtype CENH3 polypeptide comprises any of SEQ ID NOs: 49-94.

### Claim 6 (depends on 1)

A method of generating an F1 progeny plant having half the ploidy of a parent plant expressing an endogenous wildtype CENH3 protein, the method comprising, crossing the parent plant to the plant of claim 1; and selecting F1 progeny generated from the crossing step having half the ploidy of the parent plant.

### Claim 7 (depends on 6)

The method of claim 6, wherein the parent plant is the pollen parent.

### Claim 8 (depends on 6)

The method of claim 6, wherein the parent plant is the ovule parent.

### Claim 9 (depends on 6)

The method of claim 6, further comprises converting at least one selected haploid plant into a doubled haploid plant.

### Claim 10 (depends on 6)

The method of claim 6, wherein the wildtype CENH3 polypeptide comprises any of SEQ ID NOs: 49-94.

### Claim 11 (depends on 6)

The method of claim 6, wherein the mutant CENH3 polypeptide is at least 95% identical to any of SEQ ID NOs: 49-94.