Abstract

Protoplasts isolated enzymatically from cotyledons of wild Glycine species were cultured in agarose-solidified droplets of Kao protoplast culture medium. Incubation of droplets in liquid Kao medium mixed with a B5-based formulation resulted in protoplast-derived colonies. Subculture of tissues with liquid SC2 medium containing B5 salts and vitamins, 3% sucrose, 1.1 mg 1 −1 BAP and 0.005 mg 1 −1 IBA, followed by transfer to agar solidified SC2 medium, induced shoots in two accessions, G. clandestina G1231 and G. canescens G1171. Shoots elongated on agar B5-based medium with 0.2 mg 1 −1 BAP and 0.005 mg 1 −1 IBA(SC6), and rooted on hormone-free, half-strength semi-solid B5 medium.

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