Pollen-based transformation system using solid media

Multicellular living organisms and unmodified parts thereof and – Method of introducing a polynucleotide molecule into or...

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435414, 435415, 435419, 435469, 800294, 800312, 800314, 8003173, 800260, C12N 504, C12N 1584, C12N 1582, A01H 400

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059293004

ABSTRACT:
A medium is disclosed that supports pollen germination and pollen tube growth in the presence of Agrobacterium, the medium comprising agarose, sucrose, NO.sub.3, MnSO.sub.4, H.sub.3 BO.sub.3, MgSO.sub.4 and gibberellic acid. A method is disclosed for the genetic transformation of plants and lines by a pollen-based Agrobacterium-mediated transformation. The method comprises the steps of obtaining pollen from a first plant, applying a lawn of Agrobacteria to a solid pollen culture medium, the Agrobacterium comprising at least one heterologous gene sequence capable of being transferred to a plant cell, applying the pollen to the solid medium, allowing the pollen to germinate and grow on the medium, whereby the Agrobacterium attaches to the pollen tube or mediates transfer of the heterologous gene sequence to the germinating pollen to obtain transgenic pollen, applying the transgenic pollen to the stigma of a second plant capable of being fertilized by the pollen of the first plant, thereby fertilizing the second plant, obtaining transgenic seed from the second plant and germinating the transgenic seed to obtain a transgenic plant. The method is particularly suited for use with so-called "dry stigma" pollen.

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