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Plant Regeneration from Axillary Shoot Segments Derived Callus in wifhania Somnifera (l.) Dunal (ashwagandha)

N. A. Siddique
Institute of Biological Sciences, University of Rajshahi, Rajshahi -6205, Bangladesh.

M. A. Bari
Institute of Biological Sciences, University of Rajshahi, Rajshahi -6205, Bangladesh.

S. Huda
Institute of Biological Sciences, University of Rajshahi, Rajshahi -6205, Bangladesh.

M. E. Haque
Institute of Biological Sciences, University of Rajshahi, Rajshahi -6205, Bangladesh.

Abstract/ Executive Summary:

Callus cultures of Withallia sotnnifera (L.) were initiated from axillary shoots on MS (Murashige and Skoog,1962) medium supplemented with 2,4-D, BAP and Kn. The highest frequency (86%) of organogenic callus induction was observed in MS medium containing 1.0 rng/1 BAP and 2.0 mg/1 Kn. Development of adventitious shoots occurred when the calli were subcultured in MS medium supplemented in the BAP and Kn. Shoots differentiated best (85%) from axillary shoot derived callus on MS medium containing 1.0 mg/l BAP and 2.5 mg/1 Kn. Regenerated shoots rooted best (90%) on MS medium containing IBA and Kn (l mg /1).

Keywords: Plant regeneration, Shoot segments, Callus, Withania Somnifera

Location: University of Rajshahi, Rajshahi

Start Date: 01-01-2002

End Date: 31-12-2002

Program Area: Variety and Species

Non-commodity: Performance

Objectives:

1. To examine the potential of different explants,

2. To respond under in vitro conditions with the possibility of developing a protocol for the in vitro multiplication of W. somnifera.

Methodology:

Explants were collected from in vivo grown of Withania somnifera (L.) Dunal, (Ashwagandha) and axillary shoots arising from the nodes of nodal segments were used for establishing callus. Nodal segments (1-2 crn) were taken from lateral branches and washed thoroughly under running tap water, then treated with a few drops of Tween-80 and 1% Savlon for 10 minutes with constant shaking. This followed by successive three washing with distilled water to make the material free from savlon. Surface sterilization was carried out with 0.1 % HgCI₂ for seven minutes followed by gentle shaking. After surface sterilization, the segmented parts were thoroughly washed for several times with sterile distilled water. Then explants were transferred in 25x150 mm culture tubes with 15 ml basal medium (MS ) supplemented with different concentrations of 2,4-D, Kn and BAP for callus induction. Cultures were incubated with 16 h photo period in the growth chamber at 25 + 2•C under the fluorescent light intensity varied from 2000-3000 lux illumination. The pH of the medium was adjusted to 5.7 prior to autoclaving. Callus from these primary cultures was transferred to MS medium containing different concentrations and combinations of BAP, Kn, NAA and IAA for shoot differentiation and incubated in light. Data on shoot proliferation efficiency were recorded after 8 weeks of culture. Proliferated shoots were transferred to MS medium with different concentrations of Kn, IBA, NAA and IAA for adventitious root formation.

Source of Information: Bangladesh Journal of Agriculture, Vol. 27 & 28, pp. 65-70, 2002 & 2003, ISSN- 0253-5408, Bangladesh Agricultural Research Council.

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The present study was revealed that an efficient and easy to handle protocol has been developed for micropropagation of the endangered medicinal plant (Wifhania somnifera L.). This protocol provides a successful and rapid technique that can be used for ex-situ conservation. As a part of domestication strategy, these plants can be grown and further cultivated in fields. The application of this protocol can help to minimize the pressure on wild populations and contribute to the conservation of the valuable flora of Bangladesh.

Knowledge Management: Journal