Methods and compositions for recombination a gene-deficient strains of
View Patent ↗The present disclosure provides novel compositions and methods for the production and use of Agrobacterium tumefaciens strains (for example LBA4404) that are deficient in RecA activity relative to the parent strain. Combinations with other gene-deficient-strains of Agrobacterium tumefaciens are also disclosed. Specifically, two exemplary s recA minus strains, UIA777 where chloramphenicol resistant gene disrupting the recA gene and UIA770 where kanamycin resistant gene disrupting the recA gene are provided.
1. A modified strain of Agrobacterium tumefaciens , wherein said modified strain is A. tumefaciens LBA4404 recA minus strain UIA777 or UIA770 as deposited with ATCC, and assigned Accession numbers PTA-123889 and PTA-123888, respectively.
2. The modified strain of claim 1 , wherein the strain comprises a Ti plasmid.
3. A method of producing a transgenic plant, comprising:
a. contacting targeted plant cells with a modified strain of Agrobacterium tumefaciens of claim 1 ;
b. selecting and screening plant cells comprising DNA from said Agrobacterium strain integrated into genome of the targeted plant cells; and
c. regenerating whole transgenic plants from plant cells selected/screened in step (b).
4. The method of claim 3 , wherein the selecting step is carried out using a selectable marker comprising an antibiotic resistance gene selected from a chloramphenicol resistance gene, a kanamycin resistance gene, a spectinomycin resistance gene, a gentamycin resistance, or combinations thereof.
5. The method of claim 3 , wherein said transgenic plant is selected from the group consisting of maize, soybean, rye, barley, cotton, and canola.
6. A composition comprising the modified Agrobacterium tumefaciens strain of claim 1 , wherein the strain is further modified to comprise a binary vector for transformation of plants.
7. The composition of claim 6 wherein said binary vector comprises a gene encoding a product regulating an agronomic trait selected from the group consisting of an insecticidal resistance trait, herbicide tolerance trait, nitrogen use efficiency trait, water use efficiency trait, nutritional quality trait, DNA binding trait, selectable marker trait, or any combination thereof.