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 lea f 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 mu mol (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 adve ntive somatic embryos of 20-25 days of age as an explant source, homogeneou s cell suspension cultures were initiated in both AA and MS media supplemen ted with 2 mg l(-1) 2,4-D and 500 mg l(-1) CH. Protoplasts were isolated fr om homogeneous cell suspension cultures, an average yield being 2.5 x 10(7) 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 achi eved using modified KPR medium under dark conditions in 6-8 weeks. Microcal luses were proliferated and plant regeneration was achieved from protocallu ses.