Genetically transformed plants
AssigneeMonsanto Technology LLC
InventorsStephen G. Rogers, Robert B. Horsch, Robert T. Fraley
Life of this patent
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Abstract
This invention relates to genetically transformed, non-tumorous plant cells. A modified Ti plasmid is created which contains a left T-DNA border, one or more desired genes, and a right T-DNA border. This region does not contain tumorigenic or phytohormone-altering genes. The Ti plasmid is inserted into plant cells, where the T-DNA region is transferred into the plant genome. The transformed plant cells may be regenerated into morphologically normal plants which will pass the desired gene(s) to their descendants.
Claims24
A method for genetically transforming dicotyledonous plant cells which comprises contacting dicotyledonous plant cells, which are susceptible to genetic transformation by Agrobacterium cells, with Agrobacterium cells containing a chimeric Ti plasmid comprising a T-DNA region which region comprises in sequence,
- (i) a first Agrobacterium T-DNA border sequence,
- (ii) a chimeric gene which functions in said plant cells comprising a 5′ promoter region from a gene which exists naturally in a plant cell or naturally enters a plant cell, a structural coding sequence encoding a polypeptide expressible in said plant cells and a 3′ non-translated region encoding a polyadenylation signal, said promoter and 3′ non-translated region being operably linked to said structural coding sequence;
- and
- (iii) a second Agrobacterium T-DNA border sequence,
- said T-DNA border sequences enabling the transfer and incorporation of T-DNA into the genome of said plant cell and said T-DNA region including the T-DNA border sequences containing no plant tumorigenic genes.
A method for genetically transforming dicotyledonous plant cells which comprises contacting dicotyledonous plant cells, which are susceptible to genetic transformation by Agrobacterium cells, with Agrobacterium cells containing a chimeric Ti plasmid comprising a T-DNA region which region comprises in sequence,
- (i) a first Agrobacterium T-DNA border sequence,
- (ii) a chimeric gene which functions in said plant cells comprising a 5′ promoter region from a gene which exists naturally in a plant cell or naturally enters a plant cell, a structural coding sequence encoding a polypeptide expressible in said plant cells and a 3′ non-translated region encoding a polyadenylation signal, said promoter and 3′ non-translated region being operably linked to said structural coding sequence;
- and
- (iii) a second Agrobacterium T-DNA border sequence,
- said T-DNA border sequences enabling the transfer and incorporation of T-DNA into the genome of said plant cell and said T-DNA region including the T-DNA border sequences containing no plant tumorigenic genes,
- wherein the chimeric plasmid is prepared by in vivo recombination between a plant tumor inducing plasmid and a pMON128 or pMON129 plasmid containing a T-DNA border and a chimeric gene which functions as a selectable marker in said transformed plant cells.
A method for genetically transforming dicotyledonous plant cells which comprises contacting dicotyledonous plant cells, which are susceptible to genetic transformation by Agrobacterium cells, with Agrobacterium cells containing a chimeric Ti plasmid comprising a T-DNA region which region comprises in sequence,
- (i) a first Agrobacterium T-DNA border sequence,
- (ii) a chimeric gene which functions in said plant cells comprising a 5′ promoter region from a gene which exists naturally in a plant cell or naturally enters a plant cell, a structural coding sequence encoding a polypeptide expressible in said plant cells and a 3′ non-translated region encoding a polyadenylation signal, said promoter and 3′ non-translated region being operably linked to said structural coding sequence;
- and
- (iii) a second Agrobacterium T-DNA border sequence,
- said T-DNA border sequences enabling the transfer and incorporation of T-DNA into the genome of said plant cell and said T-DNA region including the T-DNA border sequences containing no plant tumorigenic genes,
- wherein the chimeric plasmid is prepared by in vivo recombination between a disarmed plant tumor inducing plasmid and a pMON128 or pMON129 plasmid containing T-DNA border and a chimeric gene which functions as a selectable marker in said transformed plant cells.
A method for producing genetically transformed dicotyledonous plant cells which comprises,
- (a) contacting dicotyledonous plant cells with Agrobacterium cells comprising a T-DNA region in which the T-DNA region comprises,
- (i) a first T-DNA border sequence,
- (ii) a chimeric gene which functions as a selectable marker in said transformed plant cells, and
- (iii) a second T-DNA border sequence, said T-DNA border sequences enabling the transfer and incorporation of T-DNA into the genome of said plant cells and said T-DNA region including the T-DNA border sequences contains no tumorigenic genes, and
- (b) obtaining therefrom non-tumorigenic transformed plant cells.
A method for producing genetically transformed dicotyledonous plant cells comprising,
- (a) contacting dicotyledonous plant cells with Agrobacterium cells containing a T-DNA region to effect transfer and incorporation of said T-DNA region into said plant cells,
- wherein the T-DNA region comprises, (i) a first T-DNA border sequence, (ii) a chimeric gene encoding a polypeptide and (iii) a second T-DNA border sequence, and wherein the T-DNA region does not comprise tumorigenic genes, and
- (b) obtaining therefrom non-tumorigenic transformed plant cells.
A method for producing genetically transformed dicotyledonous plant cells comprising transferring and incorporating a chimeric gene that functions in dicotyledonous plant cells into the genome of the dicotyledonous plant cells by contacting the dicotyledonous plant cells with Agrobacterium cells harboring a chimeric plasmid comprising a T-DNA region containing the chimeric gene but not containing plant tumorigenic genes.
Dependencies are inferred from the claim text.