# US 7,476,779 B2 · Generation of plants with altered oil content

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- Number: US 7,476,779 B2
- Document ID: US7476779B2
- Status: Granted
- Kind: B2
- Application number: 10/508,442
- Filed: 2003-03-19
- Published: 2005-11-17
- Granted: 2009-01-13
- Earliest claimed: 2002-03-20
- Assignees: Agrinomics LLC
- Inventors: Jonathan Lightner; Jeremy E. Coate; Stephanie K. Clendennen; Nancy Anne Federspiel; Debra K. Schuster
- Web page: https://patentgrep.com/patent/US7476779B2

## CPC

- A23D 9/00: Other edible oils or fats, e.g. shortenings or cooking oils (preservation of other edible oils A23B20/30)
- C12N 9/88: Lyases (4.)
- C12N 15/8247: {involving modified lipid metabolism, e.g. seed oil composition}
- A23D: EDIBLE OILS OR FATS, e.g. MARGARINES, SHORTENINGS OR COOKING OILS (preservation thereof A23B20/00; production, refinement or preservation of animal or vegetable fats or oils C11B, C11C)
- 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 is directed to plants that display an altered oil content phenotype due to altered expression of an ICL nucleic acid. The invention is further directed to methods of generating plants with an altered oil content phenotype.

## Claims

### Claim 1

A method of producing an altered oil content phenotype in a plant, said method comprising:
a) introducing into progenitor cells of the plant a plant transformation vector comprising a CsVMV promoter and a nucleotide sequence that encodes a sequence that encodes an isocitrate lysase (ICL) polypeptide,
b) growing the transformed progenitor cells to produce a transgenic plant, wherein said polynucleotide sequence is expressed, and
c) identifying said transgenic plant with an altered oil content phenotype by measuring the oil content in said transgenic plant and comparing it to that of the oil content measured in a control plant.

### Claim 2 (depends on 1)

The method of claim 1 wherein the ICL polypeptide has at least 50% sequence identity to the amino acid sequence presented as SEQ ID NO:2, and wherein the sequence comprises an isocitrate lyase domain.

### Claim 3 (depends on 1)

The method of claim 1 wherein the ICL polypeptide has at least 80% sequence identity to the amino acid sequence presented as SEQ ID NO:2.

### Claim 4 (depends on 1)

The method of claim 1 wherein the ICL polypeptide has at least 90% sequence identity to the amino acid sequence presented as SEQ ID NO:2.

### Claim 5 (depends on 1)

The method of claim 1 wherein the ICL polypeptide has the amino acid sequence presented as SEQ ID NO:2.

### Claim 6 (depends on 1)

The method of claim 1 wherein an ICL polypeptide is over-expressed in the transgenic plant, and wherein the altered oil content phenotype is a high oil phenotype.

### Claim 7 (depends on 6)

A plant obtained by a method of claim 6, wherein the ICL polypeptide has greater than 95% sequence identity to the amino acid sequence presented as SEQ ID NO:2.

### Claim 8 (depends on 7)

The plant of claim 7, which is selected from the group consisting of rapeseed, soy, corn, sunflower, cotton, cocoa, safflower, oil palm, coconut palm, flax, castor and peanut.

### Claim 9 (depends on 7)

A plant part obtained from a plant according to claim 7.

### Claim 10 (depends on 9)

The plant part of claim 9, which is a seed.

### Claim 11

A transgenic plant comprising a chimeric DNA construct comprising a CsVMV promoter and a DNA encoding an isocitrate lysase (ICL) polypeptide, wherein the ICL polypeptide has greater than 95% sequence identity to the amino acid sequence presented as SEQ ID NO:2, whereby the transgenic plant has an increased level of ICL relative to a non-transgenic plant and wherein oil from the plant has increased content compared to a plant lacking the chimeric DNA construct.

### Claim 12

A method of improving the oil content produced from a plant, said method comprising:
a) introducing into a plant a chimeric DNA construct comprising a plant specific transcription initiation region, a CsVMV promoter and a DNA encoding an isocitrate lysase (ICL) which, when introduced into cells of said plant increases ICL activity in an amount sufficient to increase the quantity of oil produced by the plant compared to a plant lacking the chimeric DNA construct
b) identifying said plant with an increased oil content by measuring the oil content in said plant and comparing it to that of the oil content measured in the plant lacking the chimeric DNA construct, thereby providing a method of improving the oil content produced from a plant.

### Claim 13 (depends on 12)

The method according to claim 12, wherein the DNA encoding ICL is from a plant selected from the group consisting of rapeseed, soy, corn, sunflower, cotton, cocoa, safflower, oil palm, flax, castor and peanut.