Genetic engineering of cotton plants and lines
AssigneeAgracetus, Inc.
InventorPaul F. Umbeck
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Abstract
A method is disclosed to achieve genetic transformation of cotton plants and lines. Immature cotton tissues are genetically transformed in vitro, by Agrobacterium-mediated genetic transformation. The resultant cotton tissues are subjected to a selection agent or agents to screen for transformants. The transformed cultures are then induced to commence somatic embryogenesis. One possible regime for regenerating such somatic embryos into whole cotton plants is disclosed.
Claims16
A method of of introducing genes into cotton plants and plant lines comprising the steps of:
- exposing hypocotyl tissue of immature cotton plants to a culture of transformation competent non-oncogenic Agrobacterium tumefaciens harboring a Ti plasmid having a T-DNA region including both a foreign chimeric gene and a selection agent resistance gene, both genes including appropriate regulatory sequences so as to be expressed in the cells of cotton plants;
- culturing the exposed tissue in the presence of a selection agent for which the resistance gene encodes for resistance so as to select for plant cells transformed with the T-DNA region;
- inducing somatic embryo formation in the exposed tissue in culture;
- and
- regenerating the somatic embryos into whole cotton plants.
The method of clim 6 wherein two antibiotics and two antibiotic resistance genes are used.
A method for introducing genes into cotton plants and plant lines, comprising the following steps in sequence;
- (a) surface sterilizing cotton seeds;
- (b) allowing said cotton seeds to germinate thus forming immature cotton plants, said immature cotton plants including hypocotyl tissue;
- (c) exposing said hypocotyl tissue to a culture of transformation competent non-oncogenic Agrobacterium tumefaciens harboring a Ti plasmid having a T-DNA region including both a foreign chimeric gene and a selection agent resistance gene;
- (d) culturing said hypocotyl tissue on a medium containing at least one antibiotic toxic to said Agrobacterium tumefaciens but not toxic to cotton cells;
- (e) culturing said tissue of step (d) in the presence of a selection agent for which the resistance gene encodes for resistance so as to select for plant cells transformed with the T-DNA region;
- (f) inducing somatic embryo formation in the exposed tissue in culture;
- and
- (g) regenerating the somatic embryos into whole cotton plants.
Dependencies are inferred from the claim text.