Methods for plant regeneration, transformation and production of insect resistant transgenic okra

A method of regeneration of okra plant, wherein said method comprising:

• a. obtaining the explant from either seeds, seedlings or whole plant, wherein the explants are selected from a group consisting of cotyledon with petiole, embryo, immature embryo, anther, and root;
• b. culturing the explants on a growth medium containing cytokinin and auxin alone or in combination to obtain multiple shoot buds or callus;
• c. culturing the shoot buds on a rooting medium to obtain rooted plantlets;
• and
• d. transferring the rooted plantlets in soil, to obtain regenerated plants.

A method for transforming okra plant, plant cells and tissues, wherein the said method comprising:

• a. surface sterilizing the seeds, and imbibing the seeds in water
• b. germinating the seeds of step (a) in a tissue culture medium to obtain seedlings,
• c. obtaining the explants from step (b),
• d. co-cultivating the explants of step (c) in a medium containing recombinant Agrobacterium strain,
• e. culturing the explants of step (d) on a tissue culture medium to select transformed plant cells and tissues,
• f. culturing the transformed plant cells and tissues of step (e) on a suitable tissue culture medium to obtain shoot buds, and
• g. culturing the shoot buds of step (f) in a rooting medium to obtain rooted transformed plants.

A method for producing transformed okra plant, plant cells and tissues, wherein said method comprising:

• a. surface sterilization of seeds and imbibing the seeds in water,
• b. germinating the seeds of step (a) in a tissue culture medium to obtain seedlings or explants,
• c. obtaining the explants from step (b),
• d. bombarding the explants of step (c) with tungsten or gold particles coated with DNA sequence of interest,
• e. culturing the explants of step (d) in a tissue culture medium to select the transformed plant cells and tissues,
• f. culturing the transformed plant cells and tissues of step (e) in a tissue culture medium to obtain shoot buds and/or callus, and
• g. culturing the shoot buds and/or callus of step (f) in a rooting medium for rooting to obtain transformed plants,