Cassia sophera Linn. is an important medicinal plant belonging to family Fabaceae. It is extensively used in Homeopathy and is well known for its medicinal properties. The present study describes a simple, efficient and reproducible regeneration system for in vitro propagation of C. sophera through cotyledonary node (CN) explant excised from 21 d old axenic seedlings. Explants were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations of thidiazuron (TDZ). Multiple shoots were produced on all the concentrations of TDZ; however 2.5 μM concentration proved to be optimal for the production of maximum number of shoots. To avoid adverse effects of prolonged exposure to TDZ in long term establishment, the cultures were transferred to TDZ free MS medium fortified with various concentrations of 6- benzyl aminopurine (BA) for multiplication, proliferation and elongation of induced shoots. Emergence of new shoot buds and multiplication continued up to second subculture passage and maximum number (14.9 ± 1.4) of shoots obtained on MS + BA (1.0 μM). Best rooting response was observed on half strength MS containing Indole-3-butyric acid (IBA) (1.0 μM). Regenerated plantlets were successfully acclimatized and hardened off inside the culture room and then transferred to green house with 100 % survival rate.
The aim of this study was to develop a new micropropagation system for Cassia angustifolia Vahl., an important medicinal legume using root explant as starting material. Root explants taken from 30-day-old aseptic seedlings were cultured on Murashige and Skoog (MS) medium supplemented with different plant growth regulators: 6-benzyladenine (BA), kinetin (Kn), and thidiazuron (TDZ). Organogenic nodular calli obtained on MS ? TDZ (1.0 lM) were transferred to shoot regeneration medium supplemented with different cytokinins (BA, Kn or TDZ) either alone or in combination with auxin:indole-3-acetic acid or a-naphthalene acetic acid. Maximum shoot regeneration frequency (90%) was obtained on MS ? BA (2.5 lM) ? NAA (0.6 lM) wherein a maximum of 42.76 ± 1.47 shoot buds per explant were induced with a maximum conversion rate of 35.63 ± 0.75 shoots per explant and average shoot length of 5.43 ± 0.20 cm. Elongated microshoots were successfully rooted under ex vitro conditions by pulse treatment in 200 lM of indole-3-butyric acid for half an hour. Microshoots were rooted, acclimatized and hardened off simultaneously in sterilized soilrite inside the growth room and then established in pots containing sterilized soil and manure (1:1) and grown under greenhouse condition with 90% survival rate. The histological sections at different developmental stages of shoot buds revealed the organization of nodular meristematic zone leading to the orientation and differentiation of shoot buds in large number and thereafter conversion into healthy shoots.
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