Callus induction and plant regeneration in Cardiospermum halicacabum Linn. an important medicinal plant

Cardiospermum halicacabum Linn. is an important medicinal twining herb belonging to the family sapindaceae. A method for rapid micropropagation of C. helicacabum through plant regeneration from leaf and nodal explant derived calli has been developed. The nodal and leaf segments were cultured on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxy acetic acid (2,4-D; 0.5-9 mu M) for callus induction. Callus production was highest at 5 mu M 2,4-D where 96 and 90% of cultured leaf and nodal cuttings produced callus, respectively. The viable calli were maintained at reduced concentration of 2,4-D (2 mu M). These calli were transferred to MS medium supplemented with various concentrations of 6-benzyladenine (BA; 2-10 mu M) or kinetin (2-10 mu M) alone or in combination with indole 3-acetic acid (IAA; 0.2-1.0 mu M) for shoot regeneration. The addition of low concentrations of IAA into BA or kinetin containing medium significantly increased the frequency of shoot regeneration in both nodal cuttings and leaf-derived calli. The highest number of adventitious shoots (28 per callus) formed at 8 mu M Kin and 0.5 mu M IAA. For rooting of the shoots, half-strength MS medium supplemented with different concentrations of indole 3-acetic acid, indole 3-butyric acid (LBA) and (alpha)-naphthalene acetic acid (NAA) 1-5 mu M was tried. The optimal result was observed on half-strength MS medium supplemented with 2.5 mu M IBA, on which 91% of the regenerated shoots developed roots with an average of 4.2 roots per shoot within 45 days. The in vitro raised plantlets were acclimatized and transferred to soil with 90% success. This in vitro propagation protocol should be useful for conservation as well as mass propagation of this medicinal plant. (c) 2006 Elsevier B.V. All rights reserved.