Rice

Summary

  • United States and Australian patents were granted to Japan Tobacco on methods for the transformation of Indica rice. Patents are pending in other jurisdictions. The patent specification discloses the use of immature embryo cells of Indica rice for transformation with Agrobacterium spp. The claims granted recite wide ranges for typical components of media used in the selection medium. Note that other varieties of rice, such as Japonica rice, are not covered by the claims as granted (Update July 2003).
  • In 2001, the National Science Council of R.O.C. in Taiwan was granted a United States patent directed to a method for the transformation of immature rice embryos with Agrobacterium. The invention is not limited to any particular rice variety. A particular feature of the transformation method is that the rice embryos and Agrobacterium are co-cultivated with a dicot suspension culture. In one of the preferred embodiments, the dicot suspension culture is made of potato cells. According to the inventors, such culture is rich in phenolic compounds, which induce the vir genes of the Ti plasmid. Thus, the phenolic compounds of the culture assist the transformation process.
  • Paradigm Genetics filed a PCT application disclosing the transformation of a rice panicle with Agrobacterium using vacuum infiltration. This method is carried out in planta (the transformed plant part is still attached to the whole plant) and avoids in vitro regeneration steps.
  • The Indian company Avestha Gengraine Technologies has filed a PCT application disclosing a method of transformation of Indica rice using excised shoot tip tissue as a target for Agrobacterium.

Rice – Specific Patent Information – part 1

Patent Number Title, Independent Claims and Summary of Claims Assignee
US 6329571

  • Earliest priority – 22 October 1996
  • Filed – 22 October 1997
  • Granted – 11 December 2001
  • Expected expiry – 21 October 2017
Title – Method for Transforming Indica Rice

Claim 1A method for transforming rice comprising transforming immature embryo cells of Indica rice by Agrobacterium method and selecting transformed cells, characterized in that a medium containing

  • 2000 to 4000 mg/l of KNO3,
  • 60 to 200 mg/l of MgSO4,
  • 200 to 600 mg/l of KH2PO4,
  • 100 to 450 mg/l of CaCl2,
  • 200 to 600 mg/l of (NH4)2SO4,
  • 1 to 7 mg/l of H3BO3,
  • 2 to 20 mg/l of MnSO4,
  • 20 to 50 mg/l of EDTA or a salt thereof,
  • 3 to 8 mg/l of Fe,
  • 50 to 200 mg/l of myoinositol,
  • 0.5 to 10 mg/l of 2,4-dichlorophenoxyacetic acid,
  • 0.01 to 5 mg/l of a cytokinin,
  • 5000 to 80,000 mg/l of a sugar, and
  • a gelling agent,which medium has a pH of 4.5 to 6.5,

is used as a medium for selecting said transformed cells.

A method for transformation of Indica rice using immature embryo cells as target tissue for Agrobacterium. Ranges for the media components are disclosed in the independent claim.

Japan Tobacco

AU 736027 B2

  • Earliest priority – 22 October 1996
  • Filed – 22 October 1997
  • Granted – 26 July 2001
  • Expected expiry – 21 October 2017
Title – Method for Transforming Indica Rice

Claim 1A method for transforming rice comprising transforming immature embryo cells of Indica rice by Agrobacterium method and selecting transformed cells, characterized in that a medium containing:

  • 2000 to 4000 mg/l of KNO3,
  • 60 to 200 mg/l of MgSO4,
  • 200 to 600 mg/l of KH2PO4,
  • 100 to 450 mg/l of CaCl2,
  • 200 to 600 mg/l of (NH4)2SO4,
  • 1 to 7 mg/l of H3BO3,
  • 2 to 20 mg/l of MnSO4,
  • 20 to 50 mg/l of EDTA or a salt thereof,
  • 3 to 8 mg/l of Fe,
  • 50 to 200 mg/l of myoinositol,
  • 0.5 to 10 mg/l of 2,4-dichlorophenoxyacetic acid,
  • 0.01 to 5 mg/1 of a cytokinin,
  • 5000 to 80,000 mg/l of a sugar, and
  • a gelling agent, which medium has a pH of 4.5 to 6.5,

is used as a medium for selecting said transformed cells.

Transformation of immature embryo cells of Indica rice by Agrobacterium. The composition of the medium used to select the transformed cells is disclosed.

EP 897013 A1

  • Earliest priority – 22 October 1996
  • Filed – 27 October 1997
  • Application deemed withdrawn – 18 December 2003
Title – Method for Transforming Indica Rice

Claim 1A method for transforming rice comprising transforming immature embryo cells of Indica rice by Agrobacterium method and selecting transformed cells, characterized in that a medium containing:

  • 2000 to 4000 mg/l of KNO3,
  • 60 to 200 mg/l of MgSO4,
  • 200 to 600 mg/l of KH2PO4,
  • 100 to 450 mg/l of CaCl2,
  • 200 to 600 mg/l of (NH4)2SO4,
  • 1 to 7 mg/l of H3BO3,
  • 2 to 20 mg/l of MnSO4,
  • 20 to 50 mg/l of EDTA or a salt thereof,
  • 3 to 8 mg/l of Fe,
  • 50 to 200 mg/l of myoinositol,
  • 0.5 to 10 mg/l of 2,4-dichlorophenoxyacetic acid,
  • 0.01 to 5 mg/1 of a cytokinin,
  • 5000 to 80,000 mg/l of a sugar, and
  • a gelling agent, which medium has a pH of 4.5 to 6.5,

is used as a medium for selecting said transformed cells.

Remarks

  1. National phase entry of WO 1998/17813 in Canada (CA 2240454) has lapsed according to CIPO.
  2. National phase entry of WO 1998/17813 in China (CN 1206435) is deemed withdrawn according to CNPO.
  3. National phase entry of WO 1998/17813 in Japan (JP H10-117776) has been rejected by the JPO.

Note: Patent information on this page was last updated on 8 February 2006.

Rice – Specific Patent Information – part 2

Patent Number Title, Independent Claims and Summary of Claims Assignee
US 6215051

  • Earliest priority – 4 November 1992
  • Filed – 4 May 1998
  • Granted – 10 April 2001
  • Expected expiry – 3 May 2018
Title – Aarobacterium-mediated method for transforming rice
(note that Agrobacterium has the incorrect spelling, as shown above, on the patent)

Claim 1A method for the production of a transgenic plant of rice crop comprising the steps:

A) infecting an immature embryo of rice crop with the genus Agrobacterium for transformation;
B) co-culturing the infected embryo with a dicot suspension culture during the step of transformation;
C) allowing the transformed embryo in step (B) to grow into a callus in a selective medium comprising a sufficient amount of a plant growth hormone for the growth of rice crop; and
D) allowing the cultured callus to regenerate root and shoot in a regeneration medium comprising a pre-determined amount of nutrients for the growth of rice crop.

Claim 8A method for the production of a transgenic rice plant comprising the steps of:

A) transforming an immature rice embryo with a gene encoding a desired gene product by culturing the embryo in a dicot suspension culture with bacteria from the genus Agrobacterium, said bacteria comprising said gene;
B) growing the transformed embryo from step (A) into a callus in a selective medium comprising a rice plant growth hormone; and
C) regenerating root and shoot from said cultured callus in a regeneration medium comprising nutrients for the growth of rice crop.

Method for the production of transgenic rice plants by co-culturing an immature rice embryo and Agrobacterium with a dicot suspension culture. The transformed embryo grows into a callus that in turn regenerates roots and shoots.

National Science Council of R.O.C.

Remarks

Related Japanese applications and United States patents are not directed to transformation of rice with Agrobacterium. They refer to a gene expression system with a promoter region from the alpha amylase genes.
WO 2001/12828 A1

  • Earliest priority – 18 August 1999
  • Filed – 17 August 2000
  • OPI – 22 February 2001
Title – Methods and Apparatus for transformation of Monocotyledenous plants using Agrobacterium in combination with vacuum filtration

Claim 1 – See below *
Claim 20An in planta method of transforming a rice plant comprising: A) contacting at least one panicle of the rice plant with a solution or suspension comprising at least one Agrobacterium clone; and
B) subjecting the rice plant to a vacuum effective to cause entry of the Agrobacterium clone into at least one flower of the panicle.
Claim 38An in planta method of producing a transgenic rice plant, comprising: A) contacting at least one panicle of a first rice plant with a solution comprising at least one Agrobacterium clone wherein the Agrobacterium clone comprises at least one heterologous gene;
B) subjecting the first rice plant to a vacuum effective to cause entry of the Agrobacterium clone into at least one flower of the panicle;
C) cultivating the first rice plant to maturity; and
D) collecting seeds of the first rice plant expressing the heterologous gene.
Claim 40 – See below *

An in planta method for transforming rice panicles by contacting the panicle with Agrobacterium in a suspension containing a heterologous gene and subjecting the plant to vacuum so Agrobacterium enters the plant part. After the transformation the plant is cultivated into maturity and seeds express the heterologous gene.

* Independent claims 1 and 40 recite use of vacuum infiltration to transform monocotyledonous plants, and are introduced in the General Monocot Transformation Methods section.

Paradigm Genetics

Remarks

National phase entry of WO 01/12828 in Australia (AU 67807/00 A) has lapsed.
WO 02/057407 A2

  • Earliest priority – 17 January 2001
  • Filed – 14 January 2002
  • OPI – 25 July 2002
Title – Novel Method for Transgenic Plants by Transformation and Regeneration of Indica Rice Plant Shoot Tips

Claim 1A novel method of transforming excised shoot tip tissue of the Indica rice cultivars by using the Agrobacterium method.

A method of transformation of Indica rice using excised shoot tip tissue as a target for Agrobacterium.

Avestha Gengraine Technologies Pty. Ltd.

Remarks

  1. related application in India IN 2001CH00047
  2. National phase entry of WO 02/57407 in Europe (EP 1444339) is still pending.

Note: Patent information on this page was last updated on 8 February 2006.