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In Vitro Micropropagation of Orchid (vanda Tessellata L.) from Shoot Tip Explant

M S Rahman
Department of Genetic Engineering & Biotechnology, University of Rajshahi, Rajshahi-6205, Bangladesh

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

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

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

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

Abstract/ Executive Summary:

Context: Orchid produces a huge number of minute seeds but the seeds can not germinate easily in nature due to the lack of endosperm in the seeds is an incompatibility barrier that limits its propagation in nature. Objectives: To develop in vitro culture techniques for quick propagation of Vanda tessellate, a commercially important orchid species. Materials and Methods: Shoot tips were used as experimental materials. The explants were surface sterilized and the shoot tips were excised. The isolated shoot tips were cultured in MS medium supplemented with different concentration and combinations of auxin and cytokinin. Results: The combination of 1.5 mgl-1 NAA and 1.0 mgl-1 BAP was proved to be the best medium formulation for multiple shoot formation as well as maximum shoot elongation. The single shoots were isolated from the multiple shoots and subcultured in MS medium having NAA and IBA individually and in combinations for root induction. Maximum root induction was obtained in MS agarified medium having 0.5 mgl-1NAA and 1.0 mgl-1IBA. The well rooted plantlets were hardened successfully in the potting mixture containing coconut husk, perlite, charcoal, brick pieces in the ratio of 2:1:1:1 and eventually established under natural condition. Conclusion: An efficient regeneration protocol for micropropagation in V. tessellata through shoot tip culture has been established.

Keywords: Shoot tip, Micropropagation, Orchid

Location: Rajshahi University Campus, Rajshahi, Bangladesh

Start Date:

End Date:

Program Area: Crop-Soil-Water Management

Non-commodity: Performance

Objectives:

To develop in vitro culture techniques for quick propagation of Vanda tessellate, a commercially important orchid species.

Methodology:

For this experiment, shoot tips collected from in vivo grown V. tessellata plants from Rajshahi University Campus, Rajshahi, Bangladesh, were used as explants. The explants were surface sterilized with 0.1% mercuric chloride solution. The excised shoot tips were cultured in MS medium (Murashige and Skoog 1962) supplemented with different concentration and combinations of auxin and cytokinin. The cluster of multiple shoots was rescued aseptically from the culture vessels and the single shoots with basal segments were separated. Shoot tips were cultured on MS medium supplemented with different concentrations of BAP, NAA, IAA and KIN individually and in combinations with NAA, IAA, BAP and KIN to find out optimum medium formulation for induction of multiple shoot from the isolated shoot tips. The separated single shoots were sub cultured on fresh MS medium supplemented with different types of growth regulators either singly or in combinations for further proliferation and root induction. In all the cases, pH was adjusted to 5.8 and the media were autoclaved at 121°C for 20 minutes under 15 lb inch-2 pressure. The culture was incubated at 25 ± 2°C with a photoperiod of 16 h at 2000-3000 lux light intensity of cool white fluorescent light. Twenty explants were used for each treatment and each experiments replication for thrice.

Source of Information: J. bio-sci. 17: 139-144, 2009 ISSN 1023-8654

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

An efficient regeneration protocol for micropropagation in V. tessellata through shoot tip culture has been established.

Knowledge Management: Journal