Somatic embryogenesis of (Saccharum officinarum var Hawaii) and isozyme comparison of regenerated plant by isoelectricfocusing

Abstract

Callus induction was established from the young leaves of sugarcane (Saccharum officinarum var. Hawaii) on modified Murashige and Skoog (MS) medium supplemented with various concentrations of 2,4-D and 10% coconut water. The medium supplemented with 3 mg/l 2,4-D was the optimum on callus induction for the explants derived from 1.5-4 cm above the youngest node. While the medium supplemented with 3 mg/l 2,4-D and 10% coconut water produced green pin headed structures on the explant derived from 5-6 cm above the node. The calli were proliferated and maintained on modified MS medium supplemented with 0.5 mg/l 2,4-D. Embryogenic induction was achieved on modified MS medium with or without 0.5 mg/l 2,4-D and 10% coconut water. The embryogenic calli were transferred to modified MS medium containing 1.0% or 1.5% saccharosee for embrygenic development. In order to promote shoot elongation and pre-embryogenic callus recycling, the embryoid cluster was transferred to modified MS medium supplemented with 3 mg/l 2,4-D and 10% coconut water. Then the plantlets were transferred to modified MS medium supplemented with 1 mg/l IAA for for root development. The plantlets were transferred to pots with vermiculite and watered with complete fertilized for a month before establishing in soil. The regenerated plants could be derived in 3 groups by morphological comparisons. From isozyme isoelectricfocusing, there are 2 samples of regenerated plants showed two bands missing in esterase zymogram as compared to the donors.