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OwnerHeqiang Huo, Zhaoyuan Lianinventors
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Disclosed is an innovative method for genetic transformation through injection of Agrobacterium containing plasmid DNA expressing PLETHORAs (PLT5) or/and WOUNDINDUCED DEDIFFERENTIATION 1 (WIND1) in multiple plant species including Snapdragon and tomato. In addition, this plasmid has also been tested that it can significantly improve the genetic transformation method is not genotype dependent. Genetic transformation can be achieved by promoting de-novo regeneration of shoots in adult plants, which will of significant impact on some perennial plant species such as citrus and blue berry etc. The transgenic de-novo shoot regenerated from adult plants can by-pass the juvenile phase of plants regenerated through the tissue culture method. Expression of PLT5, WIND1, or both can also greatly improve plant regeneration and transgenic transformation through in-vitro tissue culture in recalcitrant plant species.
A vector comprising, as operably linked components, an optional first expression cassette which comprises a nucleic acid sequence encoding at least one selectable marker, an optional second expression cassette which comprises a cloning site, a third expression cassette which comprises at least one DR DNA sequence that encodes PLT, WUS1, WIND1 and/or BBM-WUS, and an optional fourth expression cassette comprising a gene of interest.
A method of transforming a plant to express at least one heterologous nucleic acid sequence comprising delivering to the plant at least one agrobacterium comprising the vector of any of claim 1-10 or 27, the vector engineered to comprise at least one transgene, wherein the delivering results in introduction of the at least one transgene into at least one plant cell of the plant such that the at least one transgene is stably integrated into a genome of said at least one plant cell, or optionally, wherein delivering to the plant at least one agrobacterium comprising the vector of any of claim 1-10 or 27 comprises delivering two or more agrobacteria, wherein an agrobacterium of the two or more agrobacteria comprises a different vector comprising a different DR DNA sequence relative to another agrobacterium of the two or more agrobacteria.
A method of promoting de novo shoot regeneration in a plant, comprising delivering to the plant at least one agrobacterium comprising the vector of any of claim 1-10 or 27, the vector engineered to comprise at least one transgene, wherein the delivering results in introduction of the at least one transgene into at least one plant cell of the plant, wherein the expression of the at least one DR DNA sequence increases de novo shoot regeneration in the plant compared to natural plant cells lacking the at least one DR DNA sequence, or optionally, wherein delivering to the plant at least one agrobacterium comprising the vector of any of claim 1-10 or 27 comprises delivering two or more agrobacteria, wherein an agrobacterium of the two or more agrobacteria comprises a different vector comprising a different DR DNA sequence relative to another agrobacterium of the two or more agrobacteria.
A plant cell of a recalcitrant plant stably transformed with a recombinant DNA construct comprising at least one DR DNA sequence that encodes PLT5 or WIND1, and, optionally, a gene of interest.
Dependencies are inferred from the claim text.