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US 2007/0192894 A1Published application
Transformation and regeneration of Allium plants
AssigneeNew Zealand Institute for Crop & Food Research Limited
InventorsCarolyn Lister, Colin Eady
Life of this patent
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Abstract
The invention relates to a novel transformation method for plants of the genus Allium, in particular to onion plants. Plants transformed by the method are also provided. The method preferably involves an Agrobacterium tumefaciens-mediated transformation, more preferably involving immature embryos as the explant source and employing a binary vector.
Claims16
1-15. (canceled)
A method of transforming an Allium plant selected from the group consisting of Allium porrum, Allium sativum and Allium fistulosum comprising the following steps:
- (a) transforming embryo cells of the Allium plant with DNA sequences via a vector or direct gene transfer to produce transformed plant material, wherein transformation is achieved by:
- (i) wounding the embryo cells and transferring the embryo cells into a suspension of Agrobacterium,
- (ii) transferring the embryo cells from step (i) to a culture medium;
- (iii) co-cultivating the embryo cells and the Agrobacterium attached to the embryo cells for a period of 1-12 days from the end of step (ii);
- (b) selecting the transformed plant material derived from step (a), by transferring the embryo cells to a selection medium containing the appropriate selection agents to kill the agrobacteria and preferentially grow the transgenic embryo cells to produce transformed plant material;
- (c) culturing the transformed plant material from (b) to produce secondary embryos and regenerating transformed plant material from the secondary embryos;
- and
- (d) obtaining a transformed Allium plant from the secondary embryo;
- wherein the method of transforming is carried out without a passage through a callus phase.
6 more elements
A method of transforming an Allium plant selected from the group consisting of Allium porrum, Allium sativum and Allium fistulosum comprising the following steps:
- (a) transforming embryo cells of the Allium plant with DNA sequences via a vector or direct gene transfer to produce transformed plant material, wherein transformation is achieved by:
- (i) wounding the embryo cells and transferring the embryo cells into a suspension of Agrobacterium containing a plasmid with a functional T-DNA region that is capable of transfer to plant cells,
- (ii) transferring the embryo cells from step (i) to a culture medium;
- (iii) co-cultivating the embryo cells and the Agrobacterium attached to the embryo cells for a period of 1-12 days from the end of step (ii);
- (b) selecting the transformed plant material derived from step (a), by transferring the embryo cells to a selection medium containing the appropriate selection agents to kill the agrobacteria and preferentially grow the transgenic embryo cells to produce transformed plant material;
- (c) culturing the transformed plant material from (b) to produce secondary embryos and regenerating transformed plant material from the secondary embryos;
- and
- (d) obtaining a transformed Allium plant from the secondary embryo;
- wherein the method of transforming is carried out without a passage through a callus phase.
6 more elements
A method of transforming an Allium plant selected from the group consisting of Allium cepa, Allium porrum, Allium sativum and Allium fistulosum comprising the following steps:
- (a) transforming embryo cells of the Allium plant with DNA sequences via a vector or direct gene transfer to produce transformed plant material, wherein transformation is achieved by:
- (i) wounding the embryo cells and transferring the embryo cells into a suspension of Agrobacterium,
- (ii) transferring the embryo cells from step (i) to a culture medium;
- (iii) co-cultivating the embryo cells and the Agrobacterium attached to the embryo cells for a period of 1-12 days from the end of step (ii);
- (b) identifying the transformed plant material derived from step (a), by detecting the expression of a reporter gene;
- (c) culturing the transformed plant material from (b) to produce secondary embryos and regenerating transformed plant material from the secondary embryos;
- and
- (d) obtaining a transformed Allium plant from the secondary embryo;
- wherein the method of transforming is carried out without a passage through a callus phase.
6 more elements
A method of transforming an Allium genus plant selected from the group consisting of Allium porrum, Allium sativum and Allium fistulosum comprising the following steps:
- (a) transforming regeneration competent cells of the Allium plant with DNA sequences via a vector or direct gene transfer to produce transformed plant material, wherein transformation is achieved by:
- (i) wounding the regeneration competent cells and transferring the cells into a suspension of Agrobacterium,
- (ii) transferring the cells from step (i) to a culture medium;
- (iii) co-cultivating the cells and the Agrobacterium attached to the embryo cells for a period of 1-12 days from the end of step (ii);
- (b) selecting the transformed plant material derived from step (a), by transferring the embryo cells to a selection medium containing the appropriate selection agents to kill the agrobacteria and preferentially grow the transgenic cells to produce transformed plant material;
- (c) culturing the transformed plant material from (b) to produce secondary embryos or meristematic cultures and regenerating shoot cultures from this material;
- and
- (d) obtaining a transformed Allium plant from the shoot cultures.
5 more elements
A method of transforming an Allium genus plant selected from the group consisting of Allium porrum, Allium sativum and Allium fistulosum comprising the following steps:
- (a) transforming regeneration competent cells of the Allium plant with DNA sequences via a vector or direct gene transfer to produce transformed plant material, wherein transformation is achieved by:
- (i) wounding the embryo cells and transferring the embryo cells into a suspension of Agrobacterium containing a plasmid with a functional T-DNA region that is capable of transfer to plant cells,
- (ii) transferring the embryo cells from step (i) to a culture medium;
- (iii) co-cultivating the embryo cells and the Agrobacterium attached to the embryo cells for a period of 1-12 days from the end of step (ii);
- (b) selecting the transformed plant material derived from step (a), by transferring the embryo cells to a selection medium containing the appropriate selection agents to kill the agrobacteria and preferentially grow the transgenic embryo cells to produce transformed plant material;
- (c) culturing the transformed plant material from (b) to produce secondary embryos and regenerating transformed plant material from the secondary embryos;
- and
- (d) obtaining a transformed Allium plant from the secondary embryo;
- wherein the method of transforming is carried out without a passage through a callus phase.
6 more elements
Dependencies are inferred from the claim text.