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In Vitro Seed Germination and Micropropagation of Dendrobium Lindleyi Steud. an Indigenous Orchid of Bangladesh

S. K. Bhadra
Department of Botany, University of Chittagong, Chittagong - 4331, Bangladesh.

T. K. Bhowmik
Department of Botany, University of Chittagong, Chittagong - 4331, Bangladesh.

Abstract/ Executive Summary:

The seeds of Dendrobium lindleyi were germinated on 0.8% (w/v) agar solidified Murashige and Skoog (MS) as well as Phytamax (PM) basal media. The germination rate was found to be better on PM medium. The germinated seedlings underwent rapid elongation on Plant Growth Regulators (PGRs) supplemented media and the highest growth was recorded on MS with 3% (w/v) sucrose + 2.0 mg/I 6-Benzyl Amino Purine (BAP) + 0.1 mg/I Indole-3-Butyric Acid (IBA). The seed derived seedlings developed strong and stout roots on half strength MS medium with 1.5% sucrose. For micro propagation the pseudo bulb segments were used as explant and the lower part of pseudo bulb segment produced highest number of multiple shoot buds (9-10/segment) on agar solidified MS medium supplemented with 3% (w/v) sucrose + 2.5 mg/I BAP + 0.1 % (w/v) Activated Charcoal. The highest rate of elongation and rooting was found from multiple shoot bud derived seedlings as well as same seed derived seedlings. Seed and multiple shoot bud derived rooted seedlings were finally transferred to outside natural environment by successive phases of acclimatization.

Keywords: In vitro germination, Micropropagation, Dendrobium lindleyi, Orchid

Location: Botanical Garden of Chittagong University.

Start Date:

End Date:

Program Area: Crop-Soil-Water Management

Commodity: Orchid

Objectives:

To develop an efficient and reproducible protocol for in vitro germination and micro propagation of Dendrobium lindleyi, a commercially important orchid of Bangladesh.

Methodology:

The fruits of Dendrobium lindleyi Steud. were collected from the Botanical Garden of Chittagong University. The fruits were washed with running tap water, rubbed with cotton and further washed with distilled water. These were then surface sterilized by treating with 0.2% (w/v) HgCI₂ for 10 minutes and thereafter washed 2 - 3 times with double distilled sterile water in a laminar airflow cabinet. The fruits were dissected longitudinally with a sterile surgical blade; the seeds were separated from capsule and inoculated on to the surface of sterile germination medium taken in culture vessels. All these operations were done in a laminar airflow cabinet. Before inoculation of seeds, the culture vessels (2.5 cm x 15 cm test tubes or 100/150 ml conical flasks) containing medium were autoclaved at 1.9 kg/cm² of pressure at a temperature of 121°C for 20 minutes to ensure sterilization. The pH of the media was adjusted to 5.8 for MS and 5.4 for PM based media. The temperature of the culture room was maintained at 25 ± 2°C, where a regular cycle of 14h continuous light and 10 h dark w~s maintained. MS (Murashige and Skoog 1962) and PM (Arditti 1977) basal media were used for germination. After germination the seedlings continued its growth on germination medium but the growth rate was very slow. In order to induce rapid elongation, seedlings were transferred on to a wide spectrum of plant growth regulators (PGRs) supplemented media. In elongation media the seedlings produced weak root system. In order to induce strong and stout root system, the seedlings were grown on two different kinds of 0.8% (w/v) agar solidified rooting media namely, (i) half strength MS + 1.5% (w/v) sucrose and (ii) MS + 3% (w/v) sucrose + 0.5 mgl^-1 IAA. Finally the rooted seedlings were taken out of the culture vessels and transferred to outside environment following successive phases of acclimatization. For this purpose, the culture vessels were kept open for one day in the culture room and then kept outside of the culture room for 6 h in the next day. Later on, those were kept outside of the culture room for 12 h. Finally the seedlings were taken out of the culture vessels and rinsed with running tap water for removal of agar attached to the roots. Then the seedlings were transplanted to pots containing moistened coir, saw dust and coal. Transplanted seedlings were watered regularly for about 1 - 2 months where the seedlings established and grew well.

Source of Information: J. Asiat. Soc. Bangladesh, Sci. 41(2): 163-171, December 2015

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

MS + 3 % (w/v) sucrose + 2.0 mg/I BAP + 0.1 mg/I IBA was found most effective for enhancing elongation. Whereas rooting was best on half strength MS + 1.5 % (w/v) sucrose.

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