# US 12,178,174 B2 · Rice grain with reduced ROS1a activity

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- Number: US 12,178,174 B2
- Document ID: US12178174B2
- Status: Granted
- Kind: B2
- Application number: 17/842,998
- Filed: 2022-06-17
- Published: 2023-01-19
- Granted: 2024-12-31
- Earliest claimed: 2015-11-18
- Assignees: Commonwealth Scientific and Industrial Research Organisation; Institute of Botany, Chinese Academy of Sciences
- Applicants: Commonwealth Scientific and Industrial Research Organisation; Institute of Botany, Chinese Academy of Sciences
- Inventors: Ronald Chun Wai Yu; Crispin Alexander Howitt; Philip John Larkin; Chun-Ming Liu; Xiao-Ba Wu; Jinxin Liu
- Web page: https://patentgrep.com/patent/US12178174B2

## CPC

- A01H 6/4636: {Oryza sp. [rice]}
- 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 1/102: {involving modified carbohydrate or sugar alcohol metabolism, e.g. starch biosynthesis}
- A01H 1/12: {Processes for modifying agronomic input traits, e.g. crop yield}
- A01H 1/121: {Plant growth habits}
- A01H 5/10: Seeds
- C12N 9/2497: {hydrolysing N- glycosyl compounds (3.2.2)}
- Y02P 60/87: Re-use of by-products of food processing for fodder production
- 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)
- Y02P: CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS

## Abstract

The present invention relates to rice grain with thickened aleurone. Also provided is a rice plant comprising at least one genetic variation which reduces the activity of at least one ROS1a gene in the plant. Grain of the invention, or aleurone therefrom, has improved nutritional properties, and hence is particularly useful for human and animal feed products.

## Claims

### Claim 1

A method of producing a rice plant which produces grain comprising an aleurone that is thickened compared to aleurone from a corresponding wild-type grain, the method comprising the steps of
i) introducing into one or more rice cells a mutation of an endogenous ROS1a gene such that the mutated ROS1a gene encodes a ROS1a polypeptide whose amino acid sequence is different compared to the amino acid sequence of the corresponding wild-type ROS1a polypeptide,
ii) obtaining one or more rice plants from the one or more cells obtained from step i), the one or more rice plants comprising the mutations of the endogenous ROS1a gene introduced in step i), and
iii) detecting the mutation of the endogenous ROS1a gene and measuring the aleurone of the one or more plants of step ii), and selecting a plant of step ii) which produces grain comprising the mutation of the endogenous ROS1a gene and an aleurone that is thickened compared to aleurone from a corresponding wild-type grain thereby producing the rice plant.

### Claim 2

A method of selecting a rice plant which produces a rice grain comprising an aleurone that is thickened compared to aleurone from a corresponding wild-type grain, the method comprising the steps of
i) screening a population of rice plants or grain, each of which were obtained from a mutagenic treatment of progenitor rice cells, grain or plants for grain with aleruone that is thickened compared to aleurone from a corresponding wild-type grain and the presence of a mutation in a ROS1a gene, and
ii) selecting from the population of step (i) a rice plant which comprises said mutation and which produces grain with an aleurone that is thickened compared to aleurone from a corresponding wild-type grain, thereby selecting the rice plant.

### Claim 3 (depends on 1)

The method of claim 1, further comprising harvesting grain from the plant selected in step iii).

### Claim 4 (depends on 3)

The method of claim 3, further comprising producing one or more generations of progeny plants from the harvested grain.

### Claim 5 (depends on 4)

The method of claim 4, further comprising harvesting grain from the one or more generations of progeny plants.

### Claim 6 (depends on 4)

The method of claim 4, wherein the one or more generations of progeny plants are homozygous for the mutation of the endogenous ROS1a gene.

### Claim 7 (depends on 5)

The method of claim 5, wherein the grain harvested from the one or more generations of progeny plants are homozygous for the mutation of the endogenous ROS1a gene.

### Claim 8 (depends on 5)

The method of claim 5, further comprising processing the grain to produce flour, bran, wholemeal, malt starch, or oil.

### Claim 9 (depends on 5)

The method of claim 5, further comprising processing the grain or bran, flour, wholemeal, malt, starch or oil from the grain, to produce a food or beverage ingredient.

### Claim 10 (depends on 5)

The method of claim 5, further comprising mixing the grain or bran, flour, wholemeal, malt, starch or oil from the grain, with a food or beverage ingredient.

### Claim 11 (depends on 4)

The method of claim 4, wherein producing one or more generations of progeny plants from the harvested grain comprises backcrossing with a recurrent parent plant.

### Claim 12 (depends on 2)

The method of claim 2, further comprising harvesting grain from the plant selected in step ii).

### Claim 13 (depends on 12)

The method of claim 12, further comprising producing one or more generations of progeny plants from the harvested grain.

### Claim 14 (depends on 13)

The method of claim 13, further comprising harvesting grain from the one or more generations of progeny plants.

### Claim 15 (depends on 13)

The method of claim 13, wherein the one or more generations of progeny plants are homozygous for the mutation of the endogenous ROS1a gene.

### Claim 16 (depends on 14)

The method of claim 14, wherein the grain harvested from the one or more generations of progeny plants are homozygous for the mutation of the endogenous ROS1a gene.

### Claim 17 (depends on 14)

The method of claim 14, further comprising processing the grain to produce flour, bran, wholemeal, malt starch, or oil.

### Claim 18 (depends on 14)

The method of claim 14, further comprising processing the grain or bran, flour, wholemeal, malt, starch or oil from the grain, to produce a food or beverage ingredient.

### Claim 19 (depends on 14)

The method of claim 14, further comprising mixing the grain or bran, flour, wholemeal, malt, starch or oil from the grain, with a food or beverage ingredient.

### Claim 20 (depends on 13)

The method of claim 13, wherein producing one or more generations of progeny plants from the harvested grain comprises backcrossing with a recurrent parent plant.