# US 11,304,392 B2 · Haploid induction compositions and methods for use therefor

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- Number: US 11,304,392 B2
- Document ID: US11304392B2
- Status: Granted
- Kind: B2
- Application number: 16/562,504
- Filed: 2019-09-06
- Published: 2020-02-27
- Granted: 2022-04-19
- Earliest claimed: 2013-03-15
- Assignees: Syngenta Participations AG
- Applicants: Syngenta Participations AG
- Inventors: Timothy Kelliher; Satya Chintamanani; Brent Delzer; Michael L Nuccio; Robert Arthur Dietrich; Suresh Babu Kadaru; Todd Lee Warner; William Paul Bullock
- Web page: https://patentgrep.com/patent/US11304392B2

## CPC

- A01H 1/08: Methods for producing changes in chromosome number
- A01H 1/045: {using molecular markers}
- C12N 9/18: Carboxylic ester hydrolases {(3.1.1)}
- C12N 15/8218: {Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS]}
- C12N 15/8247: {involving modified lipid metabolism, e.g. seed oil composition}
- C12N 15/8261: {with agronomic (input) traits, e.g. crop yield}
- C12N 15/8287: {for fertility modification, e.g. apomixis}
- C12Q 1/6895: for plants, fungi or algae
- C12Y 301/01002: Arylesterase (3.1.1.2)
- C12N 2310/14: interfering nucleic acids [NA]
- C12N 2310/20: involving clustered regularly interspaced short palindromic repeats [CRISPR]
- Y02A 40/146: Genetically Modified [GMO] plants, e.g. transgenic plants
- 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)
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Y: ENZYMES
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE

## Abstract

Provided here are methods of using a mutated patatin-like phospholipase IIα (“pPLAIIα,” renamed here MATRILINEAL) to induce haploid induction in plants, cloning a pPLAIIα to induce haploid induction in plants, and genetically engineering a plant to contain a mutated pPLAIIα. Also provided are methods of applying topical and spray chemicals, lipids, and RNAi molecules to plants during pollination in order to induce haploid production. Further provided are methods of chemically treating plants during pollination to induce haploids while also reducing embryo abortion and increasing seed set.

## Claims

### Claim 1

A plant containing a human-induced, non-transgenic knockout mutation within a gene encoding a patatin-like phospholipase, wherein the patatin-like phospholipase is selected from the group consisting of SEQ ID NO: 69, SEQ ID NO: 136, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 141, SEQ ID NO: 142, SEQ ID NO:143, and SEQ ID NO: 144.

### Claim 2 (depends on 1)

The plant of claim 1, wherein the human-induced, non-transgenic knockout mutation is an insertion of at least one nucleotide, a deletion of at least one nucleotide, or a substitution of at least one nucleotide, which creates a premature stop codon in the patatin-like phospholipase gene.

### Claim 3 (depends on 1)

A seed, a pollen grain, a plant part, or the progeny of the plant of claim 1; wherein the seed, the pollen grain, the plant part, or the progeny comprises the mutation.

### Claim 4 (depends on 2)

The plant of claim 2, wherein the plant is selected from the group consisting of maize (Zea mays), sorghum (Sorghum bicolor), millet (Setaria italica), barley (Hordeum vulgare), stiff brome (Brachypodium distachyon), rice (Oryza sativa), wheat (Triticum aestivum), banana (Musa acuminata), palm (Elaeis guineensis), oats (Avena sativa), sunflower (Helianthus annuus), sugar beet (Beta vulgaris), Arabidopsis thaliana, and soybean (Glycine max).

### Claim 5 (depends on 4)

The plant of claim 4, wherein the plant is a maize plant.

### Claim 6 (depends on 2)

The plant of claim 2, wherein the patatin-like phosphase gene comprises SEQ ID NO: 68 or SEQ ID NO: 70.

### Claim 7 (depends on 4)

The plant of claim 4, wherein the plant is a rice plant.

### Claim 8 (depends on 7)

The plant of claim 7, wherein the patatin-like phosphase gene comprises SEQ ID NO: 84.