Methods for plant transformation using spectinomycin selection
The present invention relates to methods and compositions for transforming soybean, corn, cotton, or canola explants using spectinomycin as a selective agent for transformation of the explants. The method may further comprise treatment of the explants with cytokinin during the transformation and regeneration process.
1. A method for producing a transgenic canola plant containing at least two heterologous nucleic acid sequences, comprising:
(a) providing a canola explant comprising a first heterologous nucleic acid sequence that confers resistance to an herbicide or that confers resistance to kanamycin and related aminoglycosides;
(b) transforming the explant to comprise a second heterologous nucleic acid sequence comprising a selectable marker gene conferring spectinomycin resistance, wherein the selectable marker gene comprises an aadA gene fused to a sequence encoding a chloroplast transit peptide; and
(c) regenerating an explant from step (b) that exhibits spectinomycin resistance into a transgenic canola plant containing at least the first and second heterologous nucleic acid sequences.
2. The method of claim 1 , wherein the explant comprises an embryonic meristem.
3. The method of claim 1 , wherein the first heterologous nucleic acid sequence confers resistance to glyphosate, bialaphos, phosphinothricin, Basta, glufosinate, 2,4-D, kanamycin and related aminoglycosides, hygromycin, an acetyl-coA carboxylase inhibitor, an oxygen radical generator, or dicamba.
4. The method of claim 1 , wherein the transgenic plant arises from transformation of a meristem that results in transformation of germline tissue.