Anupam Kumari*, Sunil Kumar, Alwina Anam, Sami Ahmad and Md. Naseem
Tissue Culture Lab., Univ. Department of Botany, B.R.A. Bihar University, Muzaffarpur-842001
Email: anupamkumarimahto@gmail.com
Received-29.04.2015, Revised-13.05.2015
Abstract: An efficient protocol for in vitro cloning of Rauwolfia serpentina L. was developed using leaf segment (LS), nodal segment (NS) and internodal segments (INS) as explants. The technique involves in vitro shoot regeneration, rooting of microshoots and transplantation of regenerated plantlets under in vivo condition. Sterilized explants were cultured on MS media supplemented with different auxin (IAA, IBA, NAA, & 2,4-D) and cytokinins (Kn & BAP) within a concentration range of 0.5-3.0mg/L used singly or in combination. The best shoot multiplication was obtained from nodal explants on MS medium supplemented with BAP+NAA (1.5+0.5) mg/L along with CW (5% v/v). Excellent rooting of microshoots (4-6cm) was noticed on the medium (1/2 MS salt) fortified with combination of auxins [NAA+IBA, (1+0.5)mg/L]. Compact callus which was hydrated, green and crystalline in appearance was obtained from LS and INS on medium having 1.5mg/L 2,4-D. Nodal explants were superior to internodal as well as leaf explants in response to shoot proliferation. Regenerated plantlets were transferred to pots having mixture of sand:soil:vermicuilite(1:1:1) and little fungicides (Eco fungicide). The survival rate of plantlets was much promising (around 85%) and regenerated plantlets were healthy, green and morphologically identical to mother plants.
Keywords:Rauwolfia serpentina, Callus, Phytohormones, Multiple shoot, Conservation
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