Maize - Specific Patent Information - part 1

A method of producing transformed corn, said method comprising:

A) making a wound in a corn seedling with newly emerging radicle and stem, said wound being made in an area of the seedling containing rapidly dividing cells, wherein said area extends from the base of the scutellar node to slightly beyond the coleoptile node; and
B) inoculating the wound with vir⁺ A. tumefaciens.

1. A transformed Gramineae.

A fertile transgenic Zea mays plant comprising stably incorporated exogenous DNA.

A transformed corn plant having a transformed gene, the plant produced by an Agrobacterium-mediated transformation or direct gene transfer transformation.

A transformed corn plant produced by an Agrobacterium or direct cell transfer whereby the plant or other differentiated organs or tissues provide an expression of exogenous DNA in the corn product.

A transformed corn plant produced by a regenerated protoplast or single cell cultures.

A transformed corn plant produced by a method equivalent to an Agrobacterium tumefaciens-mediated transformation.

A method for transforming maize using Agrobacterium comprising the steps of:

A) contacting at least one immature embryo from a maize plant with Agrobacterium capable of transferring at least one gene to the embryo;
B) co-cultivating the embryo with Agrobacterium;
C) culturing the embryo in a medium comprising N6 salts, an antibiotic at concentrations capable of inhibiting the growth of Agrobacterium, and a selective agent to select for embryos expressing the gene; and
D) regenerating plants expressing the gene.

A method for transforming maize using Agrobacterium comprising the steps of:

A) contacting at least one immature embryo from a maize plant with Agrobacterium capable of transferring at least one gene to the embryo in a medium comprising N6 salts;
B) co-cultivating the embryo with Agrobacterium in a medium comprising N6 salts;
C) culturing the embryo in a medium comprising N6 salts, an antibiotic at concentrations capable of inhibiting the growth of Agrobacterium, and a selective agent to select for embryos expressing the gene; and
D) regenerating plants expressing the gene in a medium comprising MS salts.

A method for transforming maize using Agrobacterium comprising the steps of:

A) contacting at least one immature embryo from a maize plant with Agrobacterium capable of transferring at least one gene to the embryo in a medium comprising N6 or MS salts;
B) co-cultivating the embryo with Agrobacterium in a medium comprising MS salts;
C) culturing the embryo in a medium comprising N6 salts, an antibiotic at concentrations capable of inhibiting the growth of Agrobacterium, and a selective agent to select for embryos expressing the gene; and
D) regenerating plants expressing the gene in a medium comprising MS salts.

A method for optimizing the production of transgenic maize plants of a first genotype using Agrobacterium-mediated transformation comprising the steps of:

A) isolating immature embryos from maize;
B) separating the embryos into treatment groups;
C) incubating each treatment group separately in a medium comprising N6 or MS salts and in a suspension of Agrobacterium at concentrations ranging from about 1×10⁸ cfu/ml to about 1×10¹⁰ cfu/ml;
D) co-cultivating the embryos with Agrobacterium on a solid medium;
E) culturing the embryos in a medium comprising N6 salts, an antibiotic at concentrations capable of inhibiting the growth of Agrobacterium, and a selective agent to select for embryos transformed by Agrobacterium;
F) identifying the treatment group with the highest transformation frequency; and
G) using the concentration of Agrobacterium generating the highest transformation frequency to transform other embryos from the first genotype.

A method for transforming maize using Agrobacterium comprising the steps of:

A) contacting at least one immature embryo from a maize plant with Agrobacterium capable of transferring at least one gene to the embryo;
B) co-cultivating the embryo with Agrobacterium;
C) culturing the embryo in a medium containing salts other than MS salts, an antibiotic at concentrations capable of inhibiting the growth of Agrobacterium, and a selective agent to select for embryos expressing the gene; and
D) regenerating plants expressing the gene.

A method for transforming maize using Agrobacterium comprising the steps of:

A) contacting at least one immature embryo from a maize plant with Agrobacterium capable of transferring at least one gene to the embryo;
B) co-cultivating the embryo with Agrobacterium;
C) culturing the embryo in a medium comprising N6 salts, an antibiotic at concentrations capable of inhibiting the growth of Agrobacterium, and a selective agent to select for embryos expressing the gene; and
D) regenerating plants expressing the gene.

A method for transforming maize using Agrobacterium comprising the steps of:

A) contacting tissue from a maize plant with Agrobacterium capable of transferring at least one gene to the tissue in a non-LS salt medium;
B) co-cultivating the tissue with Agrobacterium in a non-LS salt medium;
C) culturing the tissue in a non-LS salt medium and a selective agent to select for tissue expressing the gene; and
D) regenerating, in a non-LS salt medium, plants expressing the gene.