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US 8,541,653 B2Granted
Transformation of soybean
AssigneeBASF Plant Science GmbH
InventorsPaula Olhoft, Leslie Grist, Libby Bernal, Sara Price, Diana Arias, Haiping Hong
Life of this patent
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Abstract
The present invention relates to improved methods for the incorporation of DNA into the genome of a soybean (Glycine max) plant utilizing meristematic cells of primary or higher leaf nodes as target tissue by means of Agrobacterium-mediated transformation and subsequent regeneration of the transformed cells into a whole plant.
Claims18
A method for producing a transgenic soybean plant comprising the steps of:
- (a) providing an axillary meristematic tissue of a primary or higher leaf node of a soybean seedling,
- (b) co-cultivating said axillary meristematic tissue with a co-cultivation medium comprising an Agrobacterium comprising a transgenic T-DNA, said transgenic T-DNA comprising
- (i) at least one plant expression cassette for an agronomically valuable trait, and
- (ii) optionally one or more selectable marker genes,
- (c) transferring said co-cultivated axillary meristematic tissue on a shoot induction medium comprising
- (i) at least one plant growth factor in a concentration suitable to induce de novo shoot induction from said axillary meristematic tissue, and
- (ii) optionally one or more selection compounds which in combination with the selectable marker gene of (b) allow for identification, or selection, or identification and selection of a plant cell, tissue or plant comprising said selectable marker gene, and
- (iii) optionally one or more antibiotics suitable to inhibit Agrobacterium growth,
- and cultivating said co-cultivated axillary meristematic tissue until shoots are induced and developed therefrom and isolating said shoots, and
- (d) transferring said isolated shoots to a rooting medium and cultivating said shoots on said rooting medium until said shoots have formed roots, and further regenerating the so derived plantlets into mature plants, which comprise inserted into their genome a T-DNA comprising
- (i) said at least one plant expression cassette for an agronomically valuable trait, and
- (ii) optionally said at least one selectable marker gene,
- wherein the method further comprises wounding said axillary meristematic tissue prior to, during or immediately after co-cultivation of step (b), and wherein at least one of the co-cultivation medium of step (b) and the shoot induction medium of step (c) comprises at least one thiol compound.
10 more elements
Dependencies are inferred from the claim text.