Micropropagation of Ocimum kilimandscharicum Guerke (Labiatae)

An efficient plant regeneration protocol has been developed from nodal explants of Ocimum kilimandscharicum Guerke, a medicinally important herbaceous plant species belonging to the family Lamiaceae. Axillary shoot bud proliferation was initiated from nodal explants cultured on MS medium supplemented with various concentrations of 6-benzyladenine (BA) (0.5-3.0 mg/l), kinetin (KN) (0.5-3.0 mg/l) and 2-isoPentenyladenine (2-iP) (0.5-3.0 mg/l). The maximum number of shoots (6.09±0.05), with average length 3.83±0.11 cm, was achieved with medium containing 1.0 mg/l BA. Shoot culture was established by repeated subculturing of the original nodal explants on shoot multiplication medium after each harvest of newly formed shoots. In this way, 20-30 shoots were obtained from a single nodal explant after 5 months. Rooting of shoots was achieved on half-strength MS medium supplemented with 1.5 mg/1 Indole-3-butyric acid (IBA) and 2% sucrose. Well-developed plantlets transferred to plastic pots containing soil and vermiculite (1:1) showed 81.13% survival. The genetic fidelity of in-vitro-raised field-grown plants to the donor plant was ascertained from random amplified polymorphic DNA (RAPD) markers. This protocol can be used for commercial propagation and for future genetic improvement studies.