# US 7,611,898 B2 · Agrobacterium transformation of stolons

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- Number: US 7,611,898 B2
- Document ID: US7611898B2
- Status: Granted
- Kind: B2
- Application number: 11/431,142
- Filed: 2006-05-09
- Published: 2007-02-22
- Granted: 2009-11-03
- Earliest claimed: 2005-05-09
- Assignees: The Samuel Roberts Noble Foundation
- Inventors: Zengyu Wang; Yaxin Ge
- Web page: https://patentgrep.com/patent/US7611898B2

## CPC

- A01H 4/008: {Methods for regeneration to complete plants}
- C12N 15/8205: {Agrobacterium mediated transformation}
- 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)

## Abstract

The invention provides methods for transforming monocotyledonous plants with Agrobacterium, for example, using stolons as a target tissue. The invention allows creation of transgenic plants without the need for callus as a target tissue for transformation, thus providing a rapid method for the production of transgenic plants. The ability to directly regenerate transgenic plants from stolons significantly reduces the time, labor and other complications required to produce transgenic plants. Plants produced by such methods are also provided by the invention.

## Claims

### Claim 1

A method of transforming a stolon-producing turfgrass plant comprising contacting a stolon of the plant with an Agrobacterium comprising a recombinant DNA and selecting a transformed tissue comprising the recombinant DNA, wherein the stolon is contacted with the Agrobacterium without first forming a callus culture of cells from the stolon.

### Claim 2 (depends on 1)

The method of claim 1, wherein the stolon is excised from the plant prior to contacting the stolon with the Agrobacterium.

### Claim 3 (depends on 1)

The method of claim 1, wherein the turfgrass plant is a bermudagrass, a creeping bentgrass or a zoysiagrass.

### Claim 4 (depends on 1)

The method of claim 1, wherein the turfgrass plant is sterile.

### Claim 5 (depends on 1)

The method of claim 1, wherein the turfgrass plant is a triploid sterile hybrid turfgrass cultivar.

### Claim 6 (depends on 1)

The method of claim 1, wherein the recombinant DNA comprises an expression cassette comprising a promoter active in cells of said plant operably linked to a heterologous coding sequence.

### Claim 7 (depends on 6)

The method of claim 6, wherein said coding sequence encodes a polypeptide, antisense construct or siRNA construct.

### Claim 8 (depends on 7)

The method of claim 7, wherein the coding sequence encodes a polypeptide, wherein the polypeptide comprises a herbicide resistance polypeptide, an insect resistance polypeptide, a disease resistance polypeptide, a selectable marker polypeptide or a screenable marker polypeptide.

### Claim 9 (depends on 8)

The method of claim 8, wherein the polypeptide comprises a selectable marker that confers resistance to a selective agent.

### Claim 10 (depends on 9)

The method of claim 9, further comprising contacting the stolon with the selective agent.

### Claim 11 (depends on 1)

The method of claim 1, wherein the recombinant DNA is a binary vector.

### Claim 12 (depends on 1)

The method of claim 1, further comprising culturing said stolon on growth media to obtain a transgenic plant.

### Claim 13 (depends on 12)

The method of claim 12, wherein the transgenic plant is formed without producing a callus.

### Claim 14 (depends on 12)

The method of claim 12, further comprising planting said plant in soil.

### Claim 15 (depends on 14)

The method of claim 14, further comprising growing said plant to sexual maturity and obtaining a transgenic seed therefrom.

### Claim 16 (depends on 6)

The method of claim 6, wherein said promoter is a constitutive promoter, inducible promoter or tissue specific promoter.