Direct shoot regeneration from leaf explants of Spilanthes acmella

Multiple shoots of Spilanthes acmella were induced from nodal buds of in vivo and in vitro seedlings on Murashige and Skoog (MS) medium containing 1.0 mg/cubic dm 6-benzyladenine (BA) and 0.1 mg/cubic dm alpha-naphthalene-acetic acid (NAA). Adventitious shoots were successfully regenerated from the leaf explants derived from the above mentioned multiple shoots. The efficiency of shoot regeneration was tested in the MS medium containing BA, kinetin, or 2-isopentenyl adenine in combination with NAA, indole-3-acetic acid (IAA), or indole-3-butyric acid (IBA) and gibberellic acid. Maximum number of shoots per explant (20+/-0.47) was recorded with 3.0 mg/cubic dm BA and 1.0 mg/cubic dm IAA. An anatomical study confirmed shoot regeneration via direct organogenesis. About 95% of the in vitro shoots developed roots after transfer to half strength MS medium containing 1.0 mg/cubic dm IBA. 95% of the plantlets were successfully acclimatized and established in soil. The transplanted plantlets showed normal flowering without any morphological variation.