# US 12,108,719 B2 · Methods for creating doubled haploid plants

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 12,108,719 B2
- Document ID: US12108719B2
- Status: Granted
- Kind: B2
- Application number: 17/218,954
- Filed: 2021-03-31
- Published: 2021-07-22
- Granted: 2024-10-08
- Earliest claimed: 2015-07-16
- Assignees: Monsanto Technology LLC
- Applicants: Monsanto Technology LLC
- Inventors: Fenggao Dong; Michael Graham; Huachun Larue; Daniel Ovadya
- Web page: https://patentgrep.com/patent/US12108719B2

## CPC

- A01H 3/04: by treatment with chemicals
- A01H 1/06: Processes for producing mutations, e.g. treatment with chemicals or with radiation (specific mutations prepared by genetic engineering on plant cell or plant tissues C12N15/00 {; process for producing transgenic plants C12N15/82})
- A01H 5/10: Seeds
- A01H 6/4684: {Zea mays [maize]}
- A01N 43/84: six-membered rings with one nitrogen atom and either one oxygen atom or one sulfur atom in positions 1,4
- A01H 1/08: Methods for producing changes in chromosome number
- A01H 6/4678: {Triticum sp. [wheat]}
- A01H: NEW PLANTS OR {NON-TRANSGENIC} PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- 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

## Abstract

Provided are methods for increasing the efficiency of creating doubled haploid plants by increasing the number of chances of forming a double haploid seed through treatment of a monocot plant with a plant growth regulator. In certain embodiments, maize plants are produced that comprise multiple co-dominant ears. Also provided are plants comprising the potential to generate increased numbers of doubled haploid offspring.

## Claims

### Claim 1

A method of producing co-dominant ears on a haploid maize plant, the method comprising contacting the haploid maize plant with a plant growth regulator, wherein the haploid maize plant produces at least two co-dominant ears, wherein each of the at least two co-dominant ears are receptive to pollination during pollen release from the haploid maize plant,
wherein plant growth regulator is paclobutrazol.

### Claim 2

A method of improving the number of DH1 seeds harvested from a DH0 maize plant, the method comprising contacting the DH0 maize plant with a plant growth regulator at developmental stage V2, V3, V4, V5, V6, V7, V8, V9, V10, V11, or V12* and contacting the DH0 maize plant with a chromosome doubling agent at any stage of its life cycle, to produce a DH0 maize plant that produces at least one DH1 maize seed and at least two co-dominant ears,
wherein each of the at least two co-dominant ears are receptive to pollination during pollen release from the DH0 maize plant,
wherein plant growth regulator is paclobutrazol.

### Claim 3 (depends on 2)

The method of claim 2, wherein the total number of DH1 maize seeds produced by the DH0 maize plant with at least two co-dominant ears is greater than the number of DH1 maize seeds produced by control DH0 maize plants that exhibit a dominant ear.

### Claim 4 (depends on 2)

The method of claim 2, wherein the DH0 maize plant produces a first co-dominant ear and a second co-dominant ear and the second co-dominant ear produces more DH1 maize seeds than the first co-dominant ear.

### Claim 5 (depends on 2)

The method of claim 2, wherein the DHo maize plant produces a first co-dominant ear, a second co-dominant ear, and a third co-dominant ear, and the third co-dominant ear produces more DH1 maize seeds than the first co-dominant ear.

### Claim 6 (depends on 2)

The method of claim 2, further comprising genotyping the DH0 maize plant prior to contacting the DH0 maize plant with the plant growth regulator or the chromosome doubling agent.

### Claim 7 (depends on 2)

The method of claim 2, further comprising obtaining DH1 maize seeds from the DH0 maize plant.

### Claim 8 (depends on 7)

The method of claim 7, further comprising genotyping the DH1 maize seeds obtained from the DH0 maize plant or genotyping a plant grown from the DH1 maize seeds.

### Claim 9 (depends on 8)

The method of claim 8, further comprising growing a DH1 maize seed selected based on the genotyping.

### Claim 10 (depends on 2)

The method of claim 2, wherein the method results in a DE4 doubling efficiency of at least about 15%, results in a DE4 doubling efficiency of at least about 15%, results in a DE30 doubling efficiency of at least about 15%, and/or results in a DE50 doubling efficiency of at least about 15%.

### Claim 11 (depends on 1)

The method of claim 1, wherein the plant is contacted with the plant growth regulator by drenching, gassing, injecting, or spraying.

### Claim 12 (depends on 2)

The method of claim 2, wherein the DH0 maize plant is contacted with the chromosome doubling agent before it is contacted with the plant growth regulator.

### Claim 13 (depends on 2)

The method of claim 2, wherein the DH0 maize plant is contacted with the chromosome doubling agent after it is contacted with the plant growth regulator.

### Claim 14 (depends on 2)

The method of claim 2, wherein the DH0 maize plant is contacted with the chromosome doubling agent within 1 minute, 5 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 6 hours, 12 hours, or 24 hours before or after contact with the plant growth regulator.

### Claim 15 (depends on 14)

The method of claim 14, wherein the chromosome doubling agent and the plant growth regulator are contacted with the DH0 maize plant at the same time.

### Claim 16 (depends on 1)

The method of claim 1, wherein the maize plant is contacted with the plant growth regulator at developmental stage V4, VS, or V6.

### Claim 17 (depends on 1)

The method of claim 1, wherein the maize plant is contacted with the plant growth regulator at developmental stage V6, V7, V8, V9, or V10.

### Claim 18 (depends on 2)

The method of claim 2, wherein three or more, four or more, or five or more co-dominant ears are produced.