In Vitro Propagation of Withania coagulance through seedling segment (epicotyledonary node)

Bhanwar Lal Jat¹, Girraj Prasad Meena², CR Choudhary³, Raaz K Maheshwari⁴, Garima Jeshwani^5 
1Department of Botany, SBRM Govt PG College, Nagaur, Rajasthan
²Department of Botany, Mewar University, Gangrar, Chittorgarh, Rajasthan
³Pro-President of Mewar University, Gangrar, Chittorgarh, Rajasthan
⁴Department of Chemistry, SBRM Govt PG College, Nagaur, Rajasthan
⁵Department of Physiology, SN Medical College, Jodhpur, Rajasthan, India
Received: 10 May 2014, Accepted: 18 June 2014, Published Online: 27 July 2014

Abstract
The seeds of Withania smnifwa Dun. were germinated which elongated after 4-5 weeks. The seedlings were used as explants. The epicotyledonary nodes of seedling were used to regenerate multiple shoots. The explants were inoculated aseptically on MS medium containing various combination and concentration of plant growth regulators, viz BAP (2.21µM/lit to 22.19µM/lit), kinetin (2.32µM/lit to 23.23µM/lit), IAA (0.57µM/lit to 11.41µM/lit) and NAA (0.53µM/lit to 6.71µM/lit). In a combination of two Cytokinins (BAP and kinetin) with one auxin (IAA) a mean of 50% epicotiledonary explants showed shoot regeneration on BAP (2.21µM/lit), kinetin (2.32µM/lit) and IAA (0.57µM/lit) on this medium a mean of 3 shoots were produced while attained a length of 14mm in 4-5 weeks. The shoot buds achieved from epicotiledonary node explants cultured on MS medium containing BAP (13.31µM/lit), kinetin (13.93µM/lit) and IAA (7.13µM/lit) were further subcultured on the MS medium with same concentration BAP, kinetin and reduced IAA (5.70µM/lit). The excised shoots produced on MS medium containing BAP (13.31µM/lit), kinetin (13.93µM/lit) and IAA (2.85µM/lit) alone were transferred on rooting medium. The medium on which the multiple shoots were produced showed marked influence on subsequent rooting behavior of such shoots originally grown on MS + cytokinins and auxins. The present investigation describes the development of efficient in vitro propagation protocol by seedling segment (epicotyledonary node).
Keywords: Withania coagulance, media, in vitro, field evaluation, epicotyledonary node
List of Abbreviations:
Benzylamino-purine (BAP) Kinetin (KIN) Indole 3-acetic acid (IAA) ά Naphthalene acetic acid (NAA) and 2, 4-Diclorophenoxy acetic acid (2, 4-D)