Assessment of Culture Medium Components and Growth Regulators for in Vitro Seedlings Development and Regeneration of Blepharis sindica

Blepharis sindica is a rare medicinal plant of family Acanthaceae. Characterization, conservation and mass propagation of this rare species are essential for its sustainable use. In vitro culture requirement for growth and development of seedlings and micropropagation are analyzed in present investigation. Dry seeds were surface sterilized with 0.1% (w/v) mercuric chloride (HgCl2) for 2 to 3 min and inoculated on various concentration of sucrose and salts, NH4NO3 or KNO3 on agar (0.8%) gelled semi-solid MS medium for development of seedlings. The cultures were maintained in growth chamber at 28 ± 2°C, 35–45 µmol m−2s−1 spectral flux photon for 12 h photoperiod and 60% relative humidity. Half-strength NH4NO3 of MS medium containing 6% (w/v) sucrose was found to be suitable for growth and development of seedlings. Cotyledonary nodes of seedlings were used as explants for induction of axillary shoots. Two to four axillary shoots of 2.17 ± 0.15 cm length were induced from each cotyledonary nodes on MS medium containing half-strength NH4NO3, 6% (w/v) sucrose and 0.1 µM BAP. In vitro multiplied shoots were rooted suitably on 1/4 strength MS medium containing 0.1 µM IBA and 100 mgL−1 activated charcoal. Above 90% of the plantlets were hardened in the greenhouse.