High frequency plant regeneration from sugarcane callus

Abstract

Sub-apical slices and leaf-roll explants of sugarcane (Saccharum hybrids) cv. B.O. 91 formed callus on Murashige and Skoog’s (MS) basal medium supplemented with 5 mg/1 2,4-dichlorophenoxyacetic acid (2,4-D). Callus was maintained on 3 mg/1 2,4-D augmented medium. Concentrations of 2,4-D resulted in differences in growth rate and morphogenetic patterns. Rapid regeneration via somatic embryogenesis was obtained on 1 mg/1 kinetin. 0.5-2.0 cm tall shoots were obtained within 4 weeks on Kn containing medium, however, the root growth remained suppressed. The resulting shoots were rooted on 1/2 strength basal medium (sucrose @ 20 g/l) supplemented with 0.2 mg/l α-naphthalene acetic acid (NAA). Within 5 weeks healthy plantlets (12.2 ± 1.8 cm) bearing 14.3 ± 7.3 roots were obtained on rooting medium. These were successfully established in soil following a 3 week hardening phase. The procedure holds promise for genetic manipulation via cellular/molecular breeding in this cultivar.