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US 2003/0154517 A1Published application
Efficiency agrobacterium-mediated wheat transformation method
AssigneeMonsanto Technology LLC
InventorsHelen Allison, Xiaorong Feng, Joyce E. Fry, Tianci Hu, Fengming Lu, Maxim Radionenko
Life of this patent
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Abstract
The present invention relates to an improved transformation and regeneration system for wheat. In particular, the invention relates to enhancements in the glyphosate selection system. The transformation method is efficient and reliable for production of fertile plants with improved agronomic qualities.
Claims31
A method for transforming a monocotyledenous plant using an Agrobacterium-mediated process, comprising:
- (a) preculturing at least one immature embryo from a monocotyledenous plant in a first medium containing increased MS salts from between about 1.5 times standard to about 3 times standard and a concentration of picloram from between about 2.5 mg/L to about 5 mg/L for a period of time sufficient to form a precultured embryo;
- (b) contacting the precultured embryo with Agrobacterium capable of transferring at least one gene construct thereto;
- (c) co-cultivating the precultured embryo with Agrobacterium;
- and
- (d) regenerating plants expressing the gene construct.
2 more elements
A method for transforming a monocotyledonous plant using an Agrobacterium-mediated process, comprising:
- (a) preculturing at least one immature embryo from a monocotyledonous plant in a first medium containing a concentration of glyphosate that is not sufficient to kill plant cells for a period of time sufficient to form a precultured embryo;
- (b) contacting the precultured embryo with Agrobacterium capable of transferring at least one gene construct to the embryo comprising a gene that confers resistance to a glyphosate-containing herbicide;
- (c) co-cultivating the precultured embryo with Agrobacterium;
- (d) culturing the precultured embryo in a second medium containing a concentration of glyphosate that is not sufficient to kill plant cells;
- (e) culturing the embryo in a third medium containing a selective amount of glyphosate to select for embryos expressing the gene that confers resistance to a glyphosate-containing herbicide;
- and
- (e) regenerating plants expressing the gene construct.
4 more elements
A method for transforming a monocotyledonous plant using an Agrobacterium-mediated process, comprising:
- (a) preculturing at least one immature embryo from a monocotyledonous plant for a period of time sufficient to form a precultured embryo;
- (b) contacting the precultured embryo with Agrobacterium capable of transferring at least one gene construct to the embryo comprising a gene that confers resistance to a glyphosate-containing herbicide;
- (c) co-cultivating the precultured embryo with Agrobacterium;
- (d) culturing the precultured embryo in a medium containing aromatic amino acids and a selective amount of glyphosate to select for embryos expressing the gene;
- and
- (e) regenerating plants expressing the gene construct.
3 more elements
A method for transforming a monocotyledonous plant using an Agrobacterium-mediated process, comprising:
- (a) preculturing at least one immature embryo from a monocotyledonous plant for a period of time sufficient to form a precultured embryo;
- (b) contacting the precultured embryo with Agrobacterium capable of transferring at least one gene construct to the embryo;
- (c) co-cultivating the embryo with Agrobacterium;
- (d) culturing the embryo in a first medium containing a selective agent to select for embryos expressing the gene;
- and
- (e) regenerating plants expressing the gene construct in a second medium containing copper.
3 more elements
A method for transforming a monocotyledonous plant using an Agrobacterium-mediated process, comprising:
- (a) preculturing at least one immature embryo from a monocotyledonous plant in a first medium containing increased MS salts, increased picloram, and a concentration of glyphosate insufficient to kill plant cells for a period of time sufficient to form a precultured embryo;
- (b) contacting the precultured embryo with Agrobacterium capable of transferring at least one gene construct to the embryo comprising a gene that confers glyphosate resistance;
- (c) co-cultivating the embryo with Agrobacterium;
- (d) culturing the embryo in a second medium containing a concentration of glyphosate insufficient to kill plant cells;
- (e) culturing the embryo in a third medium containing aromatic amino acids and a selective amount of glyphosate to select for embryos expressing the gene;
- and
- (f) regenerating plants expressing the gene in a fourth medium containing copper.
4 more elements
A method for transforming a wheat plant using an Agrobacterium-mediated process, comprising:
- (a) preculturing at least one immature embryo from a monocotyledonous plant in a first medium containing doubled MS salts, picloram at 4 mg/L, and glyphosate at 0.02 mM for a period of time sufficient to form a precultured embryo;
- (b) contacting the precultured embryo with Agrobacterium capable of transferring at least one gene construct to the embryo comprising a gene that confers glyphosate resistance;
- (c) co-cultivating the embryo with Agrobacterium;
- (d) culturing the embryo in a second medium containing glyphosate at 0.02 mM;
- (e) culturing the embryo in a third medium containing aromatic amino acids at a concentration of 0.1 μM and a selective amount of glyphosate to select for embryos expressing the gene;
- and
- (f) regenerating plants expressing the gene in a fourth medium containing copper at a concentration from about 2 μM to 20 μM.
4 more elements
Dependencies are inferred from the claim text.