Cacao

A method of inducing Agrobacterium -mediated transformation of cacao, which method comprises:

A) co-culturing a cacao floral tissue explant or cell with a non-tumorigenic Agrobacterium to produce a transformed embryogenic callus, wherein the cacao tissue explant or cell is cultured on a primary callus growth medium, said primary callus growth medium comprising DKW basal salts, a carbon source, and at least two growth regulators;
B) culturing the embryogenic callus produced in step (A) on a secondary callus growth medium having the property of inducing homeostatic growth and bipolar callus development, wherein the secondary callus growth medium is comprised of a low salt WPM basal medium, at least one growth regulator and a carbon source; and
C) culturing the callus produced in step (B) on a hormone-free embryo development medium having the property of inducing embryo differentiation, wherein the embryo development medium is comprised of a DKW basal medium and a carbon source.

A method of inducing Agrobacterium-mediated transformation of cacao comprising the steps of:

(a) coculturing a cacao tissue explant or cell with a non-tumorigenic Agrobacterium to produce a transformed embryogenic callus;
(b) culturing the embryogenic callus produced in step (a) on an embryo development medium, said medium having the property of inducing embryo differentiation to obtain mature transformed somatic embryos.

Claim 16
A method of regenerating transformed cacao plantlets comprising the steps of:
(a) providing a transformed cacao somatic embryo;
(b) germinating said somatic embryo on a primary embryo conversion medium; and
(c) regenerating a transformed cacao plantlet on a secondary embryo conversion medium from a germinated embryo produced in step (b).

Claim 24
A method of regenerating transformed cacao plantlets comprising the steps of:
(a) providing a transformed cacao somatic embryo;
(b) culturing said somatic embryo on a plant regeneration medium.