Conifers - Specific Patent Information

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.

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 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 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.

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.

A method for minimizing damage to transformed cells of pine of the genus Pinus subgenus Pinus following infection by Agrobacterium for 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.

A method for pine cell tissue culture which comprises culturing pine cells on a support membrane placed over a gel medium.

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.

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.