# US 7,612,253 B2 · Methods of modulating cytokinin related processes in a plant using B3 domain proteins

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- Number: US 7,612,253 B2
- Document ID: US7612253B2
- Status: Granted
- Kind: B2
- Application number: 11/588,092
- Filed: 2006-10-25
- Published: 2007-03-01
- Granted: 2009-11-03
- Earliest claimed: 2002-06-21
- Assignees: The Regents of the University of California
- Inventors: John Harada; Sandra Stone; Julie Pelletier
- Web page: https://patentgrep.com/patent/US7612253B2

## CPC

- C07K 14/415: from plants
- C12N 15/8295: {Cytokinins}
- 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)
- 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 plant genetic engineering. In particular, it relates to methods of modulating cytokinin related processes in a plant and selecting a plant having a phenotype associated with an altered cytokinin-related process.

## Claims

### Claim 1

A method of delaying senescence, reducing stature, increasing the number of stems, or increasing biomass in a plant, the method comprising: (i) introducing into the plant a construct comprising a plant promoter operably linked to a heterologous polynucleotide, the heterologous polynucleotide encoding a B3 domain protein comprising an amino acid sequence with at least 95% identity to SEQ ID NO:15, and (ii) selecting a plant exhibiting, relative to a plant lacking the construct, a phenotype selected from the group consisting of delayed senescence, shorter stature, increased number of stems, and increased biomass.

### Claim 2 (depends on 1)

The method of claim 1, wherein the B3 domain protein comprises SEQ ID NO:15.

### Claim 3 (depends on 1)

The method of claim 1, wherein the plant promoter is a constitutive promoter.

### Claim 4 (depends on 1)

The method of claim 1, wherein the plant promoter is a tissue specific promoter.

### Claim 5 (depends on 4)

The method of claim 4, wherein the plant promoter is a floral specific promoter.

### Claim 6 (depends on 4)

The method of claim 4, wherein the plant promoter preferentially directs expression in ovules, pistils, anthers, fruits, seed coats, vascular tissues, provascular tissues, or apical meristems.

### Claim 7 (depends on 1)

The method of claim 1, wherein the plant promoter is a senescence inducible promoter.

### Claim 8 (depends on 1)

The method of claim 1, wherein the selecting step comprises selecting a plant exhibiting delayed senescence relative to a plant without the construct.

### Claim 9 (depends on 8)

The method of claim 8, wherein the selecting step comprises selecting a plant with delayed senescence in a vegetative plant structure.

### Claim 10 (depends on 9)

The method of claim 9, wherein the vegetative structure is a leaf, stem or root.

### Claim 11 (depends on 8)

The method of claim 8, wherein the selecting step comprises selecting a plant with delayed senescence in a reproductive plant structure.

### Claim 12 (depends on 11)

The method of claim 11, wherein the reproductive structure is a seed, embryo, ovule, flower, pistil, anther or fruit.

### Claim 13 (depends on 1)

The method of claim 1, wherein the selecting step comprises selecting a plant with increased biomass relative to a plant without the construct.

### Claim 14 (depends on 8)

The method of claim 8, wherein the selecting step comprises selecting a plant with delayed ovule senescence relative to a plant without the construct.

### Claim 15 (depends on 1)

The method of claim 1, wherein the selecting step comprises selecting a plant with shorter stature relative to a plant without the construct.

### Claim 16 (depends on 1)

The method of claim 1, wherein the construct is introduced by a sexual cross.

### Claim 17 (depends on 1)

The method of claim 1, wherein the B3 domain protein comprises SEQ ID NO:12.

### Claim 18 (depends on 1)

The method of claim 1, wherein the B3 domain protein comprises SEQ ID NO:14.