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Sonication of plant tissue

Granted patent and patent application filed by The Ohio State Research Foundation

The inventors describe a method called sonication-assisted Agrobacterium-mediated transformation. It consists of subjecting the target tissue to ultrasound while immersed in an Agrobacterium suspension. The enhanced transformation rates probably result from micro-wounding both on the surface of and deep within the target tissue caused by the energy released in the process. High intensity ultrasound results in cell lysis, but sublethal doses cause temporary suppression of mRNA and protein synthesis as well as moderate rupture of the cell wall. The wounding caused by lower energy ultrasonic frequency may aid in the production of signal phenolics and enhance the accessibility of putative cell-wall binding factors to the bacterium.

The disclosure describes a method for transforming a plant sample with Agrobacterium by sonicating the plant in the presence of Agrobacterium. Preferably, the process does not take longer than 60 seconds and not less than 0.1 seconds.

Specific Patent Information

Patent Number Title, Independent Claims and Summary of Claims Assignee
US 5693512 A

  • Earliest priority – 1 March 1996
  • Filed – 1 March 1996
  • Granted – 2 December 1997
  • Expected expiry – 28 February 2016
 Title – Mehod for transforming plant tissue by sonication

Claim 1
A method for transforming a plant sample comprising the following steps:a. providing a non-tumor inducing vector containing nucleic acid to be transferred to the plant sample, wherein the vector is a non-tumor inducing Agrobacterium;
b. combining the plant sample with said vector;
c. sonicating the plant sample; wherein the vector is combined with the plant sample before, during, or after sonication;
d. lastly growing the plant sample and selecting for the transformed plant sample.

The United States patent claims

  • a method for transforming a plant where the plant sample is combined with a non-oncogenic Agrobacterium and sonicated. The sonication is applied before, during or after the plant sample is combined with Agrobacterium.
 The Ohio State Research Foundation
EP 904362 B1

  • Earliest priority – 1 March 1996
  • Filed – 28 February 1997
  • Granted – 21 January 2004
  • Expected expiry – 27 February 2017
 Title – Method for transforming plant tissue

Claim 1
A method for transforming a plant sample comprising the following steps:a. providing a non-tumor inducing vector containing nucleic acid to be transferred to the plant sample, wherein the vector is a non-tumor inducing Agrobacterium;
b. combining the plant sample with said vector;
c. sonicating the plant sample, wherein the vector is combined with the plant sample before, during or after sonication;
d. lastly growing the plant sample.

Designated contracting States at the time of grant are: Belgium, Switzerland (patent lapsed as reported by INPADOC), France, United Kingdom, Italy, Liechtenstein (patent lapsed as reported by EPO), Netherlands, Portugal, Sweden (patent lapsed as reported by INPADOC)

The difference in claim 1 between EP 904362 and US 5693512 is the last step, where US 5693512 includes selecting for the transformed plant sample, which is not included in that of WO 904362.
WO 1997/32016 A1

  • Earliest priority – 1 March 1996
  • Filed – 28 February 1997
  • OPI – 4 September 1997
 Title – Method for transforming plant tissue

Claim 1
A method for transforming a plant sample comprising the following steps:a. providing a non-tumor inducing vector containing nucleic acid to be transferred to the plant sample;
b. combining the plant sample with said vector;
c. sonicating the plant sample;
d. lastly growing the plant sample.

PCT application WO 1997/32016 recites a method of transforming a plant sample which includes sonicating the plant sample (which, according to the specification ‘includes whole plants as well as all the parts and portions thereof…‘).

According to the specification, the term ‘vector’ in claim 1 ‘is a non-tumor inducing bacteria or strain of bacteria…’.

Sonication of the plant sample can take place before, during, or after introducing the vector to the plant sample as follows – ‘The vector is combined with the plant sample before, during or after sonication…‘

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