Induction of Somatic Embryogenesis and Efficient Plant Regeneration in Trifolium alexandrinum L.

Calli were induced from root, hypocotyl and cotyledon explants of two berseem cultivars BL⁻¹ and Wardan. For cultivar Wardan MS medium supplemented with 2,4-D (2 mg1⁻¹) and kin (0.5 mg 1⁻¹) was found to be the most appropriate and hypocotyl gave maximum response while for cultivar BL-1, MS culture medium supplemented with 2,4-D (2 mg 1⁻¹), NAA (2 mg 1⁻¹) and Kin (1 mg V⁻¹) was found to be the most suitable for root explants with 100 percent callus induction. Calli were subcultured on six different differentiation media for induction of somatic embryogenesis. Green somatic structures developed after 6–8 weeks of subculturing. Cotyledon derived calli differentiated better as compared to hypocotyl derived calli. Maximum differentiation (53.33%) was observed on MS+Kin (2 mg V) medium from cotyledon derived calli of variety BL-1. These differentiated green structures were then transferred to regeneration media. Maximum shoot regeneration was achieved on MS medium supplemented with kin (2 mg 1⁻¹) and NAA (0.5 mg 1⁻¹).