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Micropropagation of Centella Asiatica L. an Important Medicinal Herb

R. Das
Department of Genetic Engineering and Biotechnology, University of Rajshahi, Rajshahi-6205, Bangladesh

M. F. Hasan
Department of Genetic Engineering and Biotechnology, University of Rajshahi, Rajshahi-6205, Bangladesh

M. S. Hossain
Department of Genetic Engineering and Biotechnology, University of Rajshahi, Rajshahi-6205, Bangladesh

M. Rahman
Department of Genetic Engineering and Biotechnology, University of Rajshahi, Rajshahi-6205, Bangladesh

Abstract/ Executive Summary:

High frequencies of multiple shoot regeneration were achieved from shoot tip explants cultured on MS media fortified with 4.0 mgl^-1 BAP + 0.1 mgl^-1 NAA. On an average 10.2±0.38 shoots per explant were obtained. The regenerated shoots were rooted in MS medium supplemented with 1.0 mgl^-1 IBA. The in vitro grown plantlets were acclimatized and successfully transferred to natural condition with 80% survival. A reproducible protocol was established for in vitro propagation through multiple shoot induction of Centella asiatica L. an important medicinal herb having high medicinal value.

Keywords: Micropropagation, Shoot tip, Centella asiatica

Location: The Campus of the University of Rajshahi, Bangladesh.

Start Date:

End Date:

Program Area: Development of Host and Medicinal Plants

Commodity: Medicinal Plants

Objectives:

To identify the micropropagation of centella asiatica L. an important medicinal herb

Methodology:

For this investigation, shoot tips of Centella asiatica were collected from the Campus of the University of Rajshahi, Bangladesh. They were washed first under running tap water for 30 minutes and treated with 1% tween 80 for 10 minutes followed by repeated rinsing with sterile distilled water. Further sterilization was done under aseptic conditions in a Laminar Airflow Hood. Explants were surface sterilized with 0.1% (W/V) HgCl₂ for 10 minutes. Finally, the explants were washed thoroughly with autoclaved distilled water for several times to remove the traces of sterilant. The shoot tips were cut into appropriate size and cultured on MS basal medium. Throughout the experiments full strength MS medium with 3% (W/V) sucrose and gelled with 0.8% (W/V) agar was used. The pH of all media was adjusted to 5.8 prior to autoclaving .The cultures were incubated in a culture room at 25±2⁰C with a photoperiod of 16 hour at 3000 lux light intensity provided by cool white fluorescent tubes. The basal medium was supplemented with BAP (1.0-7.0 mgl^-1) and NAA (0.1mgl^-1) of a single concentration. Rooting of shoots was achieved on full strength of MS medium supplemented with IBA at different concentrations. Well developed plantlets were removed from the culture vessels, washed gently under running tap water and planted in plastic pots containing a potting mixture of sand, soil and farmyard manure in the ratio of 1:1:1. The potted plantlets were covered by polythene sheet to maintain suitable humidity. After sufficient acclimatization, the plantlets were transplanted in the field condition, where 80% plants were survived.

Source of Information: Progress. Agric. 19(2) : 51-56, 2008 ISSN 1017-8139

Article Link (Online Journal): http://www.banglajol.info/index.php/PA/article/view/16928/11884

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The highest percentage of multiple shoot induction was 76.67% on the medium augmented with 4.0 mgl-1 BAP+0.1 mgl^-1 NAA followed by 70.00% on the medium consisting of 3.0 mgl^-1 BAP and 0.1 mgl^-1 NAA. On the other hand, the lowest percentage of multiple shoots induction was found to be only 10.00% on the medium supplemented with 7.0 mgl-1 BAP and 0.1 mgl-1 NAA. The highest number of shoots was 10.2 ± 0.38 per explant obtained on the medium having 4.0 mgl^-1 BAP and 0.1 mgl-1 NAA followed by 8.0±0.38 shoots per explant in the medium fortified with. 3.0 mgl^-1 BAP + 0.1 mgl^-1 NAA. On the contrary, the minimum number of shoots was 1.1±0.20 per explant on the medium supplemented with 1.0 mgl-1 BAP and 0.1 mgl-1 NAA. The induced shoots were elongated in the same medium. In the present investigation, MS medium augmented with 4.0 mgl^-1 BAP+0.1 mgl^-1 NAA found to be the best treatment for the highest multiple shoot induction as well as maximum number of shoots per explant. In the present investigation, MS medium augmented with 4.0 mgl^-1 BAP+0.1 mgl^-1 NAA found to be the best treatment for the highest multiple shoot induction as well as maximum number of shoots per explant. Similar results were also reported in several medicinal plants, such as, Smilax zeylanica L.; Celastrus paniculatus; Heracleum candicans); Spilanthes mauritiana; and Coleus blumei.

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