Establishment of efficient tissue culture systems for genetic transformation in sugarcane

Leaf roll explants (1.0–1.5 cm long) prepared from spindles of a commercial sugarcane variety CoJ 83 were cultured on different media to achieve regeneration through two modes viz., somatic embryogenesis (callus phase) and direct shoot regeneration. Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxyacetic acid- 2,4–0 (4 mg l⁻¹ kinetin (0.5 mg l⁻¹) and maltose (30 g l⁻¹) promoted highest callus induction (95.24%) from the cultured segments. Shoot regeneration (76.25%) from callus cultures was attained on MS medium supplemented with 6-benzylamlnopurine (0.5 mg l⁻¹. Shoot elongation and rooting of shoots was obtained on half strength MS medium. Highest frequency of direct shoot regeneration (85.98%) occurred on half strength MS medium supplemented with naphthaleneacetic acidNAA (5 mg l⁻¹ kinetin (0.5 mg l⁻¹) and sucrose (30 g l⁻¹). Shoot growth was achieved on half strength MS medium supplemented with NAA (3 mg l⁻¹), kinetin (0.5 mg l⁻¹) and sucrose (30 g l⁻¹). Subsequently, rooting of shoots was achieved on half strength MS medium supplemented with NAA (5 mg l⁻¹) and sucrose (70 g l⁻¹). Thus, two efficient tissue culture systems have been established in sugarcane, which can be used for genetic transformation.