Pollen Transformation

Pollen Transformation – patent granted to USDA

The invention is a method for the genetic transformation of any plant by using pollen as starting material for transformation with Agrobacterium. A culture medium useful for pollen germination and pollen tube growth in presence of Agrobacterium is also claimed.

Patents granted to the United States Department of Agriculture (USDA)

Specific Patent Information

Patent Number Title, Independent Claims and Summary of Claims Assignee
AU733080 B2

  • Earliest priority – 15 July 1997
  • Filed – 14 July 1998
  • Granted – 3 May 2001
  • Expected expiry – 14 July 2018
Title – Pollen-based transformation system using solid media

Claim 1
A method for producing a transgenic plant comprising:

a. obtaining pollen from a first plant,
b. applying a lawn of Acrrobacteria to a solid pollen culture medium, the Agrobacteria comprising at least one heterologous gene sequence capable of being transferred to a plant cell,
c. applying the pollen to the solid medium,
d. allowing the pollen to germinate and grow on the medium, thereby producing transgenic pollen,
e. applying the transgenic pollen to the stigma of a second plant capable of being fertilized by the pollen of the first plant, thereby fertilizing the second plant,
f. obtaining transgenic seed from the second plant,
g. germinating the transgenic seed to obtain a transgenic plant.

Claim 8
A medium for pollen germination and pollen tube growth comprising agarose, sucrose, KNO3, MnSO4, H3BO3, MgSO4, and gibberellic acid.

The Australian patent 733 080 claims the same method for transforming pollen of a plant with Agrobacterium as the United States patent. However, the Australian patent claims in addition a specific medium for pollen germination and pollen tube growth.

United States Department of Agriculture (USDA)

EP 996328 B1

  • Earliest priority – 15 July 1997
  • Filed – 14 July 1998
  • Granted – 5 March 2003
  • Expected expiry – 14 July 2018
Title – Pollen-based transformation system using solid media

Claim 1
A method for producing a transgenic plant comprising:

a. obtaining pollen from a first plant,
b. applying a lawn of Agrobacteria to a solid pollen culture medium, the Agrobacteria comprising at least one heterologous gene sequence capable of being transferred to a plant cell,
c. applying the pollen to the solid medium,
d. allowing the pollen to germinate and grow on the medium, thereby producing transgenic pollen,
e. applying the transgenic pollen to the stigma of a second plant capable of being fertilized by the pollen of the first plant, thereby fertilizing the second plant,
f. obtaining transgenic seed from the second plant,
g. germinating the transgenic seed to obtain a transgenic plant.

The European patent 996328 claims the same method for transforming pollen of a plant with Agrobacterium as the United States patent.

Designated contracting States at the time of grant are: Austria, Belgium, Switzerland, Cyprus, Germany, Denmark, Spain, Finland, France, United Kingdom, Greece, Ireland, Italy, Liechtenstein, Luxembourg, Monaco, Netherlands, Portugal, Sweden

US 5929300

  • Earliest priority – 15 July 1997
  • Filed – 15 July 1997
  • Granted – 27 July 1999
  • Expected expiry – 15 July 2017
Title – Pollen-based transformation system using solid media

Claim 1
A method for producing a transgenic plant comprising:

a. obtaining pollen from a first plant,

b. applying a lawn of Agrobacteria to a solid pollen culture medium, the Agrobacteria comprising at least one heterologous gene sequence capable of being transferred to a plant cell,

c. applying the pollen to the solid medium,

d. allowing the pollen to germinate and grow on the medium, thereby producing transgenic pollen,

e. applying the transgenic pollen to the stigma of a second plant capable of being fertilized by the pollen of the first plant, thereby fertilizing the second plant,

f. obtaining transgenic seed from the second plant,

g. germinating the transgenic seed to obtain a transgenic plant.

Delta and Pine Land Co., acquired exclusive licensing rights to the pollen-transformation system developed by the USDA in the United States. (Source: Ag Biotech InfoNet, January 26, 2001).

The United States patent 5 929 300 claims:

  • a method for producing a transgenic plant by transforming pollen of a plant with Agrobacterium carrying a foreign gene and then fertilizing a second plant with the transgenic pollen to obtain transgenic seed. The transgenic plant is obtained by germination of the transgenic seeds. The Agrobacterium must be applied as a lawn to a solid pollen culture medium and the pollen must also be applied to the solid culture medium.  Thus, it appears that other methods of applying the Agrobacterium (e.g., by pipetting drops onto the medium) might not fall within the scope of the claims.
WO 1999/03326

  • Earliest priority – 15 July 1997
  • Filed – 14 July 1998
  • OPI – 28 January 1999
Title – Pollen-based transformation system using solid media

Claim 1
A method for producing a transgenic plant comprising:

a. obtaining pollen from a first plant,
b. applying a lawn of Acrrobacteria [sic] to a solid pollen culture medium, the Agrobacteria comprising at least one heterologous gene sequence capable of being transferred to a plant cell,
c. applying the pollen to the solid medium,
d. allowing the pollen to germinate and grow on the medium, thereby producing transgenic pollen,
e. applying the transgenic pollen to the stigma of a second plant capable of being fertilized by the pollen of the first plant, thereby fertilizing the second plant,
f. obtaining transgenic seed from the second plant,
g. germinating the transgenic seed to obtain a transgenic plant.

Claim 8
A medium for pollen germination and pollen tube growth comprising agarose, sucrose, KNO3, MnSO4, H3BO3, MgSO4, and gibberellic acid.
Remarks
  1. National phase entry of WO 1999/03326 in Canada (CA 2296501) is pending.
  2. National phse entry of WO 1999/03326 in China (CN 1098029 C) has been published as granted on 8 January 2003.
  3. Related patent documents also filed in Brazil (BR 9811791), The Hong Kong Special Administrative Region of the People’s Republic of China (HK 1026118), Taiwan (TW 577922) and South Africa (ZA 9806240).

Note: Patent information on this page was last updated on 21 February 2006.

Patent application filed by the United States Department of Agriculture (USDA)

The invention disclosed in the European application is the same as in the United States patent and the recently granted Australian patent AU-B-733 080 (former Australian application AU 84005/98 A1).

Bibliographic data

EP 996 328 A1
Title Pollen-based transformation system using solid media
Application No. & Filing Date
EP 934497
July 14, 1998
Publication Date May 3, 2000
Language English
Remarks Applications also filed in Brazil (BR 9811791), China (CN 1263434), and South Africa (ZA 9806240).
To view or download the patent document as a PDF file, click on EP 996 328.

Summary of the invention

The independent claims as filed in the EP application 996 328 A1 are similar to the granted claims of its related United States and Australian patents. As in the Australian patent, the EP application also contains a filed claim referring to a medium for pollen germination and pollen tube growth.

EP Patent application filed by the United States Department of Agriculture (USDA)

Claims in plain English

DisclaimerTHE FOLLOWING CLAIMS ARE MEANT ONLY TO ASSIST READING OF THE CLAIMS AND ARE NOT MEANT AS A LEGAL INTERPRETATION OF THE CLAIM SCOPE.
EP 996 328 A1
Claim 1A method for producing a transgenic plant by:

A) applying Agrobacteria having a foreign gene to a pollen culture on a solid medium;
B) allowing germination and growth of the transgenic pollen on the medium;
C) fertilizing a second plant with the transgenic pollen;
D) obtaining seed; and
E) germinating the seed to obtain a transgenic plant.

Claim 8A medium for pollen germination and pollen tube growth having agarose, sucrose, KNO3, MnSO4, H3BO3, MgSO4, and gibberellic acid.


Actual pending claims

EP 996 328 A1
Claim 1*A method for producing a transgenic plant comprising:

A) obtaining pollen from a first plant;
B) applying a lawn of Agrobacteria to a solid pollen culture medium, the Agrobacteria comprising at least one heterologous gene sequence capable of being transferred to a plant cell;
C) applying the pollen to the solid medium;
D) allowing the pollen to germinate and grow on the medium, thereby producing transgenic pollen;
E) applying the transgenic pollen to the stigma of a second plant capable of being fertilized by the pollen of the first plant, thereby fertilizing the second plant;
F) obtaining transgenic seed from the second plant;
G) germinating the transgenic seed to obtain a transgenic plant.

Claim 8**

A medium for pollen germination and pollen tube growth comprising agarose, sucrose, KNO3, MnSO4 , H3BO3, MgSO4, and gibberellic acid.

Note: The Australian application AU 84005/98 A1 was granted. See Patents granted to the USDA for more information on this new patent.

* Identical to claim 1 of the granted United States and Australian patents.

**Identical to claim 8 of the granted Australian patent.