Studies on in Vitro Propagation of Ocimum Tenuiflorum and Testing of Clonal Fidelity of Micro Plantlets

Abstract

The present study was an attempt to develop the regeneration protocol from different explants of Ocimum tenuiflorum. The effect of media combinations on shoot multiplication, callusing, regeneration and rooting of microshoots of Ocimum tenuiflorum cultured on Murashige and Skoog (MS) medium were investigated. The best shoot proliferation in terms of shoot number and shoot quality was obtained on medium supplemented with 2.5 μM 6-benzylaminopurine (BA) and 5μM Naphthaleneacetic acid (NAA). Shoot regeneration was observed after 5-6 weeks of subcultured on MS medium supplemented with 5.0 μM BA and 2.5 μM NAA. The rooting was induced in microshoots by pulse treatment of 50 μM indole-3-butyric acid (IBA) for 36 hours. These shoots rooted on basal MS medium after 6 days of pulse treatment of IBA. The rooted plants were successfully acclimatized and transferred to the pots with about than 70 % survival. The testing of clonal fidelity of these plants using Random Amplified Polymorphic (RAPD) and Inter Simple Specific Repeats (ISSR) markers showed not a single polymorphic band. Thus these plants seem to be genetically uniform.