Organogenic transformation and regeneration
This invention provides methods of transforming organogenic plant cells and regenerating plants from transformed cells.
1. A method of generating a plant comprising transformed plant cells, the method comprising:
culturing at least one non-apical meristemic cell to produce at least one shoot;
culturing at least one leaf base explant from the shoot to produce one or more organogenic cells;
introducing at least one nucleic acid segment into the organogenic cells to produce one or more transformed organogenic cells, wherein the nucleic acid segment is introduced into the organogenic cells using Agrobacterium -mediated delivery; and
generating at least one plant from the transformed organogenic cells without going through an undifferentiated intermediate.
2. The method of claim 1 , wherein the non-apical meristemic cell comprises a pineapple cell.
3. The method of claim 1 ,
wherein the nucleic acid segment comprises at least one sense nucleic acid segment that corresponds to at least a portion of at least one endogenous gene;
wherein the nucleic acid segment comprises at least one sense nucleic acid segment that corresponds to at least a portion of at least one exogenous gene;
wherein the nucleic acid segment comprises at least one antisense nucleic acid segment that corresponds to at least a portion of at least one endogenous gene;
wherein the nucleic acid segment encodes at least one polypeptide transcription factor; or,
wherein the nucleic acid segment encodes at least one promoter and/or at least one enhancer, which nucleic acid segment homologously recombines with at least one promoter and/or at least one enhancer of at least one endogenous gene.
4. The method of claim 1 , wherein the nucleic acid segment encodes a polypeptide.
5. The method of claim 4 , wherein the polypeptide is heterologous to the organogenic cells.
6. The method of claim 4 , wherein the polypeptide is homologous to at least one endogenous polypeptide of the organogenic cells.
7. The method of claim 4 , wherein the polypeptide comprises at least one carotenoid biosynthetic polypeptide that is selected from the group consisting of: an isomerase, an isopentenyl diphosphate isomerase, a geranylgeranyl pyrophosphate synthase, a phytoene synthase, a phytoene desaturase, a ζ-carotene desaturase, a lycopene β-cyclase, a lycopene ε-cyclase, a β-carotene hydroxylase, and an ε-hydroxylase.