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Efficient in Vitro Cultural Techniques for Seeds Germination of Vanda Roxburghii

Md. Rafiqul Islam
Department of Biotechnology and Genetic Engineering, Islamic University, Kushtia 7003, Bangladesh.

Khandakar Md. Rayhanul Kabir
Department of Biotechnology and Genetic Engineering, Islamic University, Kushtia 7003, Bangladesh.

Md. Sabir Hossain
Department of Biochemistry and Molecular Biology, Jahangirnagar University, Savar, Dhaka, Bangladesh.

Md. Fuad Hossain
Department of Agriculture Biology, University of Ruhuna, Sri Lanka.

Md. Ibrahim Khalil
Human Genome Center, School of Medical Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia

Abstract/ Executive Summary:

Since orchid seeds lack endosperm, their germination in nature is very limited. Moreover, ruthless collections, destruction of habitats by reclamation and deforestation, unauthorized trade have led to reduction in natural population of many orchid species. In the meantime some of the orchid species have been extinct and a large number of species have become rare and endangered. It is therefore important to take step for saving this group of plants from extinction and also to support their quick propagation to meet up our demand for their ornamental, medicinal and environmental need. This study was carried out to develop a suitable protocol for in vitro seed germination of an indigenous orchid, Vanda roxburghii in different nutrient media. Murashige and Skoog (MS) media supplemented with 0.1 mg/NAA (á-naphthalene acetic acid) and 15% coconut water (CW) was found the best formation for seed germination and subsequent plantlet development. This media composition also shortens the germination period from 35 days to 22 days only. In conclusion, this protocol could be useful for biodiversity conservation and commercial application for Vanda seed germination.

Keywords: Orchid, Vanda roxburghii, Germination

Location: Rabindranat Kuti Bari Shalidah, Kumarkali, Kushtia, Bangladesh.

Start Date:

End Date:

Program Area: Crop-Soil-Water Management

Commodity: Orchid

Objectives:

To develop a suitable in vitro protocol for seed germination of indigenous orchid, Vanda sp. both for their commercial uses and conservation purposes.

Methodology:

In the present study, one indigenous orchid species viz. Vanda sp. was selected. The plant specimens were collected form Rabindranat Kuti Bari Shalidah, Kumarkali, Kushtia, Bangladesh. Seeds were used as explants of in vitro germination. In order to determine the optimum condition of a specific factor; experiments were conduction with various modification of one factor, keeping the others constant. Most of experiments were conducted following standard tissue culture method. The experiments had 8-15 or 20-30 replicated culture per treatment and they were grow for 1-4 consecutive passages of 2-6 weeks each and data on different parameters were recorded at the end of each passage.

The capsule of Vanda sp were treated with 0.1% (w/v) HgCl₂ for 10 minutes for surface sterilization and thereafter washed 2-3 times with autoclaved distilled water in a laminar airflow cabinet. Sterilization of non- living Articles. The one liter medium routine sterilization procedure of non-living articles such as nutrient medium glass goods, distilled water, instruments (wrapped with brown paper) were sterilized by autoclaving. Capsules opening and Culturing of Seeds were done aseptically under laminar airflow cabinet. The culture vessels with inoculated seeds were incubated in 16:8 h Light:Dark photoperiod at 25±1°C. Sub-cultures were maintained in the same conditions as for in germination. Thus regular sub-culturing was done at every 28 days interval when the masses of seedlings were further thinned.

The stock solution of various constituents of MS media were prepared for ready use. Different concentrations, separate stock solution of macronutrients, micronutrients, organic compound (vitamin and amino acids) for Murashige and Skoog (MS) and growth regulators were prepared. The constituents of stock solution of MS media are given in Appendix. Data were collected on the following parameters for the evaluation of the result of different sets of experiments. The lag period seed germination was counted in days starting from the day of culture inoculation to the first sign of shoot emergence.

Source of Information: World Journal of Agricultural Sciences 10 (4): 163-168, 2014, ISSN 1817-3047

Article Link (Online Journal): DOI: 10.5829/idosi.wjas.2014.10.4.1819

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The best formation for seed germination and subsequent plantlet development. This media composition also shortens the germination period from 35 days to 22 days only. In conclusion, this protocol could be useful for biodiversity conservation and commercial application for Vanda seed germination.

Knowledge Management: Journal