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Mass Propagation of Rauwolfia Serpentina L. Benth

U. Salma
Department of RDDR, Modern Herbal group, Dhaka - 1217, Bangladesh

M.S.M. Rahman
Biotechnology and Genetic Engineering Discipline, Khulna University, Khulna-9208, Bangladesh

S. Islam
Plant Tissue Culture Laboratory, BCSIR, Dhaka - 1205, Bangladesh

N. Haque
Department of RDDR, Modern Herbal group, Dhaka - 1217, Bangladesh

M. Khatun
Biotechnology and Genetic Engineering Discipline, Khulna University, Khulna-9208, Bangladesh

T.A. Jubair
Biotechnology and Genetic Engineering Discipline, Khulna University, Khulna-9208, Bangladesh

B.C. Paul
Biotechnology and Genetic Engineering Discipline, Khulna University, Khulna-9208, Bangladesh

Abstract/ Executive Summary:

A protocol for mass propagation through axillary bud proliferation was established for Rauwolfia serpentina L. Benth. (Apocynaceae). MS medium supplemented with 1.5 mg L^-1 BA and 0.2 mg L^-1 NAA elicited the maximum number of shoots (4 multiple shoots) from nodal explants. These adventitious shoots were best rooted on half strength MS medium supplemented with 1.0 mg L^-1 each of IBA and IAA. The in vitro raised plants were acclimatized in glass house and successfully transplanted to field condition with almost 95% survival.

Keywords: Mass propagation, Adventitious, Heterogeneous, Indole alkaloids, Microcutting

Location: Plant Tissue Culture Laboratory, Biological Research Division, Bangladesh Council of Scientific and Industrial Research (BCSIR), Dhanmondi, Dhaka

Start Date: 00-00-2007

End Date: 00-00-2007

Program Area: Conservation and Biodiversity

Commodity: Plant(s)

Objectives:

To identify most suitable media supporting competence necessary for large-scale propagation scheme to replenish the exhausting supply and to conserve the threatened species.

Methodology:

The experiment was carried out in the Plant Tissue Culture Laboratory, Biological Research Division, Bangladesh Council of Scientific and Industrial Research (BCSIR), Dhanmondi, Dhaka, Bangladesh in the year 2007. Nodal segments containing buds of 3-4 years old plants were collected from the medicinal plant garden of BCSIR. For surface sterilization, explants were cut into small pieces and cleaned thoroughly under a continuous stream of running tap water for 30 min. After that the segments were washed with detergent and kept under running tap water for 15 min. Then treated with 1% Savlon and washed in running tap water for 30 min. The explants were then taken under laminar airflow cabinet and surface sterilized with a 0.1% HgCl₂ for 5 min followed by their washing three times with autoclaved double distill water. Explants of approximately 1 to 2 cm in length were cut and inoculated aseptically onto MS (Murashige and Skoog, 1962) media supplemented with different concentrations and combinations of auxin and cytokinine. The pH of the medium were adjusted to 5.8±0.05 before adding agar and the media were autoclaved at 1.1 kg cm^-1 for 20 min at 121°C. Cultures were incubated at 25±1°C with a photoperiod of 16 h at 3000 lux light intensity of cool white fluorescent light.

All cultures were initiated in 150x25 mm²glass tube containing 15-20 mL of medium. The cultures were regularly subcultured on fresh medium at 4 weeks intervals in glass tubes or 100 mL flasks. Observations were recorded every 5 days following inoculation and subculturing. For inducing adventitious shoots and their development, nodal segments containing buds were cultured on MS medium supplement with 0.5-2.0 mg L^-1 BAP with NAA (0.2, 0.5) mg L^-1. Auxiliary shoots produced within four weeks of culture were isolated and regularly subcultured on fresh medium at four-week intervals. In rooting experiments proliferated shoot of approximately 2-4 cm length were rescued aseptically from culture vessels and transferred to freshly prepared half strength MS macro and micro-nutrients contained in glass tube. IBA and IAA (0.1-1.5) mg L^-1 were used as supplementary with the media for root induction. Culture tubes containing rooted plantlets were kept in a room of normal temperature (30±2°C) and normal daylight for 7 days. Plantlets were then taken out from the culture tubes and washed carefully under running tap water for complete removal of media. Then the plantlets were transplanted to small plastic pots containing garden soil and compost in a ratio of 2:1. The pots were immediately covered with polythene bag to prevent desiccation. After the hardening period the plantlets were transferred to field condition.

Source of Information: Pakistan Journal of Biological Sciences, 11: 1273-1277; ISSN 1028-8880

Article Link (Online Journal): DOI: 10.3923/pjbs.2008.1273.1277; URL: http://scialert.net/abstract/?doi=pjbs.2008.1273.1277

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The ultimate success of in vitro propagation lies in successful establishment of plants in the soil. 80-85% survival of plantlets were recorded in the experiments of previous workers. The high survival rate in the present study indicates that this procedure could be easily adapted for large-scale propagation. As R. serpentina holds the tremendous potentialities for massive propagation towards the commercialization of many invaluable indole alkaloids this present study will be fruitful for this novel application and will help in the conservation study of such threatened species.

Knowledge Management: Journal