Plant regeneration from protoplasts derived from cell suspension of adventive somatic embryos in sugarcane (Saccharum spp. hybrid cv. CoL-54 and cv. CP-43/33)

Adventive somatic embryos were initiated from the cut edges of juvenile leaf explants of two cultivars of sugarcane (Saccharum spp. hybrid cv. CoL-54 and cv. CP-43/33). This response was achieved using MS medium containing 9 μmol (2 mg l-1) 2,4-D and 500 mg l-1 CH under either continuous or 16-h photoperiod. Regeneration from somatic embryos was achieved under either continuous or 16-h photoperiod on MS basal medium in 5–6 weeks. Using adventive somatic embryos of 20–25 days of age as an explant source, homogeneous cell suspension cultures were initiated in both AA and MS media supplemented with 2 mg l-1 2,4-D and 500 mg l-1 CH. Protoplasts were isolated from homogeneous cell suspension cultures, an average yield being 2.5×107 ml-1 for both the cultivars. The best division efficiency (1.5 and 0.80%) and microcalluses for cv. CoL-54 and cv. CP-43/33, respectively were achieved using modified KPR medium under dark conditions in 6–8 weeks. Microcalluses were proliferated and plant regeneration was achieved from protocalluses.