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Regeneration of an Indigenous Orchid, Vanda Teres (roxb.) Lindl. Through in Vitro Culture

Pinaki Sinha
Department of Botany, Jahangirnagar University, Savar, Dhaka-1342, Bangladesh

Shyamal K. Roy
Department of Botany, Jahangirnagar University, Savar, Dhaka-1342, Bangladesh

Abstract/ Executive Summary:

A protocol for regeneration of Vanda teres through long term culture of protocorms obtained from germinated seeds has been achieved in Vacin and Went medium. Seeds germinated within 40 - 45 days in Vacin and Went medium with 1.0 mg/l BAP + 0.5 mg/l NAA + 2% sucrose + 2 g/l peptone. Best shoot regeneration from the protocorm was found on the same medium containing 10% coconut water. For the production of strong elongated shoots and their multiplication 2 g/l banana powder + 100 mg/l casein hydrolysate were added to the medium. Identical plantlets with stout shoot and roots were obtained by subculturing shoots of same size in half strength MS medium without plant growth regulator. Enough quantity of regenerated plantlets were acclimatized in natural environment which flowered at the age of two - three years.

Keywords: Vanda teres, Indigenous orchid, Regeneration

Location: Sylhet district, Bangladesh

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Orchid

Objectives:

To developing and establish an efficient protocol for mass propagation and consequently commercial exploitation and conservation of this important indigenous orchid.

Methodology:

Capsules of Vanda teres (Roxb.) Lindl. were collected from mature plants naturally grown on the trees in the forests of Sylhet district, brought to the laboratory and made ready for culture. They were scrubbed with a soft brush and washed with running tap water. The capsules were then dipped in 0.2% HgCl₂ soln. for 15 min, then submerged in 95% ethanol and flamed rapidly for a few seconds holding with forceps. They were then washed by autoclaved distilled water. The surface sterilized capsules were then splitted longitudinally with a sterile scalpel and the seeds were taken by forceps and transferred to the agar gelled nutrient medium for seed germination. For seed germination and multiplication different nutrient media, namely, MS, KC and VW were used as basal medium. Various concentrations and combinations of BAP, Kn, IAA and NAA were used as growth regulators. Unless otherwise mentioned 2% sucrose, 2 g/l peptone and 0.85% agar (BDH) were used in each medium preparation. The pH of the medium was adjusted to 5.8 before adding agar. The media were autoclaved at 1.1 kg/cm2 for 20 min at 120⁰C. Cultures were incubated at 5,000 lux with cool white fluorescent light at 16/8 h light/dark regime maintained at 24 ± 1⁰C. The cultures were subcultured at four week intervals. Each culture medium repeated thrice with 20 replicates in each.

Source of Information: Plant Tissue Cult. 14(1) : 55-61, 2004 (June)

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The plantlets were sprayed with balanced fertilizers (1 g/l 20N-20P-20k) twice weekly. After six months the plants were transferred to 13 cm orchid pots. The plantlets thus obtained grew vigorously and flowered after two years of transfer in the pot. The original protocorm cultures are still maintained, for the fourth year, through subcultures in the same medium growing PLBs and shoots as described above. Likewise large number of plantlets and flowered plants of Vanda teres could be raised from a single capsule. The protocol described in this paper is quite feasible and repeatable for a long-term high frequency regeneration as well as conservation of this important indigenous tropical orchid. Best shoot regeneration from the protocorm was found on the same medium containing 10% coconut water. For the production of strong elongated shoots and their multiplication 2 g/l banana powder + 100 mg/l casein hydrolysate were added to the medium. Identical plantlets with stout shoot and roots were obtained by subculturing shoots of same size in half strength MS medium without plant growth regulator. Enough quantity of regenerated plantlets were acclimatized in natural environment which flowered at the age of two - three years.

Knowledge Management: Journal