Plant Regeneration Through Somatic Embryogenesis in Sugarcane

Abstract

An efficient protocol for plant regeneration through somatic embryogenesis was standardized using three cultivars of sugarcane (CoJ64, CoJ83 & CoJ86). Callus cultures were established by culturing the spindle segments on Murashige and Skoog medium supplemented with 2,4-d (4.0 mg/L) and kinetin (0.5 mg/L). The embryogenic calli were transferred to different media containing various concentrations of sugars (sucrose 3, 6 % and maltose 3, 6 %), proline (560 mg/L), activated charcoal (2.0 g/L), ABA (2.0 mg/L), cefotaxime (250 ppm) and agar (1.0 and 1.6 %) for optimization of somatic embryogenesis. Embryogenic calli were characterized by their creamy white, nodular and friable appearance. Putative embryogenic calli were further confirmed through histological studies. For shoot regeneration, the embryogenic callus was transferred to MS medium supplemented with BAP (0.5 mg/L) and kinetin (0.5 mg/L). Roots were induced from the regenerated shoots by transferring to MS medium supplemented with NAA (5.0 mg/L) and high level of sucrose (70 g/L). Somatic embryogenesis and subsequent plant regeneration was found genotype dependent. Hardening of the rooted plantlets was done before transplanting to field. The hardened plantlets exhibited good survival ranging from 85 to 90 %.