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Conifers

Summary

Botanical aspects

Conifers is a major group within the Gymnosperms plants (plants with naked seeds that appear in a cone). They usually have needle-shaped or scale-like leaves, and nearly all are evergreen. The trees have a conical shape and have cones for pollen and seed production.

Within Conifers, Pinus is the largest genus with about 120 species. It is also the most widespread genus of trees in the Northern Hemisphere. The natural distribution of pines ranges from arctic and subarctic regions of Eurasia and North America to subtropical and tropical (usually montane) regions of Central America and Asia. Pines are also extensively planted in temperate regions of the Southern Hemisphere. Many pines are fast growing species tolerant of poor soils and relatively arid conditions, making them popular in reforestation. Due to their occurrence in some very cold and dry environments where disease and stand-destroying disturbance are rare, pines are collectively the most long-lived of conifers. Ages of over 1000 years are common for some pine species.

Important pine products include wood, turpentine, and edible seeds. The wood of several Pinus species (e.g. P. elliottii) is widely used for construction. Synthetic products derived from turpentine (e.g. terpin, anethole, camphor, and dl-menthol) are used in pharmaceutical preparations (e.g. expectorant in humans), in perfumery, in the elaboration of cigarettes, cosmetics, toilet products, and to impart flavor in different products.

IP aspects

There are a couple of patents directed to transformation of conifers by microprojectile bombardment, but very few related to transformation through Agrobacterium.

A United States patent on transformation of pine tissue with Agrobacterium was granted to North Carolina State University in 1989. The patent claims a method for the transformation of a differentiated tissue of pine with A. tumefaciens. The strain of A. tumefaciens used for transformation can be selected from: a strain capable of causing crown gall, a strain with a co-integrated Ti-plasmid and a strain with a binary vector system.

Recently, Genesis Research and Development Corporation has filed a United States patent application directed to transformation of nodal stem segments of a plant from the genus Pinus with Agrobacterium.

Conifers – Specific Patent Information

Patent Number Title, Independent Claims and Summary of Claims Assignee
US 4886937

  • Earliest priority – 20 May 1985
  • Filed – 22 June 1988
  • Granted – 12 December 1989
  • Expected expiry – 20 May 2005
 Title – Method for Transforming Pine

Claim 1
A method for transforming pine, comprising inoculating differentiated pine tissue with a strain of Agrobacterium tumefaciens selected from the group consisting of (i) a strain containing a Ti plasmid which is capable of causing crown gall in said pine, (ii) a strain containing a plasmid comprising the vir region of said Ti plasmid and TDNA borders, and (iii) a strain containing a first plasmid comprising the vir region of said Ti plasmid and a second plasmid comprising TDNA borders.
Claim 5

A transformed pine comprising differentiated pine tissue, said differentiated pine tissue comprising cells transformed by a strain of Agrobacterium tumefaciens selected from the group consisting of (i) a strain containing a Ti plasmid which is capable of causing crown gall in said pine, (ii) a strain containing a plasmid comprising the vir region of said Ti plasmid and TDNA borders, and (iii) a strain containing a first plasmid comprising the vir region of said Ti plasmid and a second plasmid comprising TDNA borders.

A method for transforming a differentiated pine tissue with a strain of A. tumefaciens selected from: a strain capable of causing crown gall in pines, a strain with a Ti-plasmid having both the vir genes and the T-borders and a strain having two plasmids, one with the vir genes and the other with the T-borders. A transformed pine attained with the mentioned method is also claimed.

North Carolina State University
US 2002/16981 A1

  • Earliest priority – 15 September 1998
  • Filed – 20 March 2001
  • Abandoned – 6 October 2003 (failure to respond to an office action)
 Title – Methods for Producing Genetically Modified Plants, Plant Materials and Plant Products Produced Thereby

Claim 1
A method for producing genetically modified plant material of the Eucalyptus or Pinus species, comprising:

(a) culturing nodal stem segments of a target plant selected from the Eucalyptus and Pinus species;

(b) transforming the stem segments with a genetic construct by incubating the nodal stem segments with an Agrobacterium culture transformed with the genetic construct;

(c) promoting regeneration of adventitious shoot buds from the transformed stem segments;

(d) selecting transformed adventitious shoot buds; and

(e) regenerating transformed plant material from the transformed adventitious shoot buds.

A method for producing genetically modified material of Pinus by incubating nodal stem segments with an Agrobacterium culture transformed with a genetic construct. Transformed plant material is regenerated from transformed adventitious shoot buds. The invention also includes transformation of Eucalyptus species. This part is discussed under Particular dicots – Eucalyptus.

Genesis Research and Development Corporation

 Remarks
  1. US 2002/0016981 was a continuation-in-part of now granted US 6255559, which is the priority document of WO 2000/15813. Claim 1 of WO 2000/15813 does not limit the type of plant to be transformed.
  2.  Related applications also filed in Brazil (BR 9913740 A), Indonesia (ID 29949) and South Africa (ZA 200101818). Granted United States patent US 6 255 559 is also related to this application, but Agrobacterium -mediated transformation of Pinus is not the subject matter of the invention.
  3. National phase entry of WO 2000/15813 in Australia (AU 61270/99), Canada (CA 2341781), China (CN 1326510), Europe (EP 1114169), New Zealand (NZ 510474) are deemed lapsed/withdrawn.
WO 2002/031112 A2

  • Earliest priority – 10 October 2000
  • Filed – 10 October 2001
  • OPI – 18 April 2002
 Title – Enhanced transformation and regeneration of transformed embryogenic pine tissue

Claim 1
A method for regenerating transgenic plants of pine of the genus Pinus subgenus Pinus which comprises:

(A) subjecting pine cells to Agrobacterium infection for Agrobacterium transformation;
(B) minimizing damage to cells subsequent to Agrobacterium infection;
(C) rapidly selecting transformed cells;
(D) culturing said transformed cells to produce transgenic somatic embryos; and
(E) germinating said transgenic somatic embryos to produce transgenic plants.
Claim 25
A method for regenerating transgenic plants of pine of the genus Pinus subgenus Pinus which comprises:

(A) subjecting pine cells to Agrobacterium infection for Agrobacterium transformation;
(B) eradicating Agrobacterium;
(C) minimizing damage to cells during and subsequent to Agrobacterium eradication;
(D) rapidly selecting transformed cells;
(E) culturing said transformed cells to produce transgenic somatic embryos; and
(F) germinating said transgenic somatic embryos to produce transgenic plants.
Claim 39
A method for minimizing damage to transformed cells of pine of the genus Pinus subgenus Pinus following infection by Agrobacteriumfor Agrobacterium transformation  which comprises:

(a) washing transformed cells in a liquid wash medium;
(b) plating said cells on a support membrane;
(c) resuspending said cells in a liquid wash medium; and
(d) recovering washed, transformed cells with minimal physical damage.
Claim 46
A method for pine cell tissue culture which comprises culturing pine cells on a support membrane placed over a gel medium.
Claim 52
A method for selecting transformed cells of pine of the genus Pinus subgenus Pinus which comprises:

(A) culturing cells which have been subjected to transformation on a support membrane placed over a gel medium;
(B) contacting said cells with a selection agent; and
(C) selecting transformed cells.
Claim 58
A method for eradicating Agrobacterium from cells of pine of the genus Pinus subgenus Pinus which comprises:

(A) culturing cells on a support membrane over a layer containing an eradicant, said layer positioned in or over a gel medium; and
(B) recovering cells from which said Agrobacterium contaminant has been eradicated.

This patent document was retrieved on the BIOS Patent Lens using the terms “((conifer or pine or pinus) in the title or abstract) and (transform* in the title or abstract)”.

Claims in this application recite a method of Agrobacterium-mediated transformation of pine cells, where physical damage of cells after  Agrobacterium infection is kept to a minimum (e.g. by use of vessels or support membranes).

Westvaco Corporation
Remarks
  1. National phase entry of WO 2002/031112 in Australia (AU 1308102 A), Canada (CA 2424313 A1), and United States (US 2002100083 A1) are pending.
  2. Related patent applications also filed in Brazil (BR 0114563 A) and South Africa (ZA 200303570 A).
  3. National phase entry of WO 2002/031112 in New Zealand (NZ 525632) has been published as granted on 24 September 2004.

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