Novel methods and constructs for plant transformation
AssigneeSakata Seed Corporation
InventorsSteven F Fabijanski, Laurian Robert, Johann Schernthaner, Tong Wu
Life of this patent
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Abstract
The present invention relates to novel methods for the transformation of crop species, novel methods for the selection and identification of transformed plant cells and novel methods for recovery of regenerated whole plants. The method also relates to the development of plants with novel traits and plants that contain novel recombinant DNA constructs.
Claims64
A method for preparing a transformed plant, comprising:
- (a) introducing into plant cells a construct comprising at least one Agrobacterium oncogene to obtain transformed cells comprising said construct, the expression of said at least one Agrobacterium oncogene in said transformed cells enabling said cells to grow under conditions insufficient for the growth of untransformed cells, whereby transformed cells are selected;
- and
- (b) negating the effect of said at least one Agrobacterium oncogene in said transformed cells and regenerating morphologically normal transformed plants from said transformed cells.
A DNA vector comprising, in the 5′-3′ direction, a DNA sequence homologous to a left portion of the right border region of the T-DNA region of a Ti-plasmid, a recombinase recognition site, and a region of DNA homologous to a right portion of the right border region of the T-DNA region of a Ti-plasmid.
A DNA vector comprising, in the 5′-3′ direction, a DNA sequence homologous to a left portion of the left border region of the T-DNA region of a Ti-plasmid, a recombinase recognition site, and a region of DNA homologous to a right portion of the left border region of the T-DNA region of a Ti-plasmid.
A modified Ti-plasmid comprising a recombinase recognition site 3′ to the right T-DNA border.
A modified Ti-plasmid comprising a recombinase recognition site 5′ to the left T-DNA border.
A modified Ti-plasmid comprising a recombinase recognition site 3′ to the right T-DNA border and a recombinase recognition site 5′ to the left T-DNA border.
Modified Ti-plasmid pTi-C58 CIMB.
Modified Ti-plasmid pTi-C58 TIMB.
Modified Ti-plasmid pTi-C58 RBC-1.
DNA vector pRBC-1.
DNA vector pLBC-1.
DNA vector pLBC-2.
A construct for transformation of plant cells comprising, in the 5′ to 3′ direction, a first recombinase recognition site, at least one oncogene, and a second recombinase recognition site.
A construct comprising, in the 5′ to 3′ direction, a first transposase recognition site, at least one oncogene, and a second transposase recognition site.
A construct comprising at least one oncogene and a DNA binding protein coding sequence which encodes a DNA binding protein.
A method for modifying a Ti-plasmid to contain recombinase or transposase recognition sites comprising:
- (a) introducing a first DNA vector into Agrobacterium cells comprising a Ti-plasmid under conditions sufficient for a homologous recombination event to occur between said DNA vector and said Ti-plasmid, said first DNA vector comprising, in the 5′-3′ direction, a DNA sequence homologous to a left portion of the right border region of the T-DNA region of said Ti-plasmid, a recombinase or transposase recognition site, and a region of DNA homologous to a right portion of the right border region of the T-DNA region of said Ti-plasmid, to obtain Agrobacterium cells comprising a modified Ti-plasmid with a recombinase or transposase recognition site within the T-DNA region and in proximity to the right T-DNA border sequence;
- and,
- (b) introducing a second DNA vector into said Agrobacterium cells from step (a) under conditions sufficient for a homologous recombination event to occur between said second DNA vector and said modified Ti-plasmid, said second DNA vector comprising, in the 5′-3′ direction, a DNA sequence homologous to a left portion of the left border region of the T-DNA region of said modified Ti-plasmid, a recombinase or transposase recognition site, and a region of DNA homologous to a right portion of the left border region of the T-DNA region of said modified Ti-plasmid, to obtain Agrobacterium cells comprising a further-modified Ti-plasmid, said further-modified Ti-plasmid comprising a recombinase or transposase recognition site within the T-DNA region of and in proximity to the left T-DNA border sequence of said further-modified Ti-plasmid and a recombinase or transposase recognition site within the T-DNA region of and in proximity to the right T-DNA border sequence of said further-modified Ti-plasmid.
Dependencies are inferred from the claim text.