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Patent application filed by the University of Guelph (Canada)

The present disclosure is directed to transformation of any explant from monocot plants using Agrobacterium. According to the applicants, an advantage of the disclosed method is that vacuum infiltration of the tissue in the presence of Agrobacterium and a phenolic compound such as acetosyringone yields better transformation efficiency than by wounding. According to the inventors, the method is also simpler than previously described methods. Because the known patent applications have lapsed, the disclosed methods may be in the public domain. If you’d like to use it and are uncertain that it is in the public domain in your jurisdiction, it may be best to contact the assignee, for which a recent contact address disclosed on the website is plorenz@uoguelph.ca.

Specific PCT Application Information

PCT Number Title, Independent Claims and Summary of Claims Assignee
WO 00/58484 A2

  • Earliest priority – 26 March 1999
  • Filed – 26 September 2001
  • OPI – 5 October 2000
  • Expected expiry – N/A
 Title – Transformation of monocotyledoneous plants using Agrobacterium

Claim 1A method for the transformation of a monocot plant comprising,
i) exposing explant tissue of said monocot plant to an Agrobacterium strain under vacuum in the presence of a phenolic compound, said Agrobacterium strain comprising a heterologous gene of interest within a vector;
ii) removing said Agrobacterium from said explant tissue;
iii) adding an antibiotic against said Agrobacterium; and
iv) selecting explant tissue for occurrence of said heterologous gene of interest.
Claim 10A method for the transformation of a monocot plant comprising,
i) placing explant tissue of said monocot plant into media comprising a suspension of Agrobacterium to obtain a mixture, said Agrobacterium strain comprising a heterologous gene of interest within a vector;
ii) maintaining said mixture under vacuum in the presence of acetosyringone;
iii) releasing said vacuum and further incubating said explant tissue in the presence of said Agrobacterium;
iv) transferring said explant tissue to fresh media comprising acetosyringone and incubating said explant tissue in the dark
v) washing said explant tissue with an antibiotic against said Agrobacterium,
vi) transferring said explant tissue to fresh media and allowing said explant tissue to differentiate, thereby producing differentiated calli;
vii) placing said differentiated calli onto media containing a selection agent, and maintaining said differentiated calli in the light; and
viii) obtaining calli that grow in the presence of the selection agent.
Claim 17A method for the transformation of a monocot plant comprising,
i) placing explant tissue of said monocot plant into media comprising a phenolic compound, and a suspension of Agrobacterium to obtain a mixture, said Agrobacterium strain comprising a heterologous gene of interest within a vector;
ii) washing said explant tissue with an antibiotic against said Agrobacterium and transferring said explant tissue to fresh media comprising acetosyringone and incubating said explant tissue in the dark;
iii) transferring said explant tissue to fresh media and allowing said explant tissue to differentiate, thereby producing differentiated calli;
iv) placing said differentiated calli to media containing a selection agent, and maintaining said differentiated calli in the light; and
v) obtaining calli that grow in the presence of the selection agent.

The present PCT application recites:

  • method of transforming monocot plant
  • by vacuum infiltration of a plant tissue
  • with Agrobacterium having a heterologous gene of interest on a vector
  • in the presence of a phenoloic compound, e.g. acetosyringone.

University of Guelph

(Canada)
Remarks Related applications in Australia (AU 112261/00), Canada (CA 2368841), and US (US 2002/112261) have lapsed or been withdrawn.

Note: Patent information on this page was last updated on 5 February 2006.