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In Vitro Mass Scale Propagation and Seedlings Development of Medicinal Orchid Dendrobium Transparens Wall Ex Lindl

Tapash Kumar Bhowmik
Assistant Professor
Department of Botany, University of Chittagong, Chittagong, Bangladesh

Md. Mahbubur Rahman
Professor
Department of Botany, University of Chittagong, Chittagong, Bangladesh

Abstract/ Executive Summary:

In the present investigation, in vitro seed germination and seedlings development of Dendrobium transparens Wall ex Lindl. Was assessed on agar solidified KC; MS; PM and MVW media with three different sources of carbohydrates viz. sucrose, glucose and lactose. PM medium proved to be best for early germination and PLBs formation followed by MS medium. Sucrose supplemented media was found to be best whereas lactose-containing medium gave at least performance. Germinated PLBs were cultured on agar solidified or liquid MS and PM media supplemented with different concentrations and combinations of auxins, cytokinins. Highest shoot growth was found on liquid PM medium supplemented with 1.0 mg/l IAA and 1.0 mg/l BAP (3.58 ± 0.12 cm) followed by agar solidified PM +1.0 mg/l NAA + 1.0 mg/l BAP (3.35 ± 0.13 cm). PM was found better than MS medium and liquid culture was superior to agar solidified condition for elongation of in vitro grown germinated seedlings. BAP, IAA was better in elongation media than Kn, NAA, Picloram. Agar solidified full strength MS with 1.0 mg/l IAA produced highest increased length and number of roots (3.27 ± 0.20 cm/shoot bud and 1.74 ± 0.13 no/shoot bud) followed by half-strength hormone-free MS medium (2.18 ± 0.15 cm/shoot bud and 1.56 ± 0.12 no/shoot bud). The well-developed seedlings were acclimatized subsequently transferred to pots in the greenhouse and watered regularly.

Keywords: Dendrobium transparens, In vitro germination, Medicinal orchid, PGRs, PLBs

Location: Plant Tissue Culture and Biotechnology Laboratory, Department of Botany, University of Chittagong.

Start Date:

End Date:

Program Area: Development of Host and Medicinal Plants

Commodity: Orchid

Objectives:

In the present study in vitro seed germination and seedlings, the development method was investigated for the rapid propagation of medicinally and horticulturally important orchid Dendrobium transparens.

Methodology:

Studies on the seed germination of indigenous orchid species, Dendrobium transparens were carried out in the Plant Tissue Culture and Biotechnology Laboratory, Department of Botany, University of Chittagong. The materials used for the present investigation were the immature green capsules of D. transparens which were collected from Bandarban, Bangladesh. The dust in the pods collected from the naturally grown orchid was removed. The young green capsules were washed with tap water containing few drops of teepol solution for few minutes and washed under running tap water for five minutes. The capsules were surface sterilized by immersing it in the solution of 0.1% HgCl₂ for 10 minutes followed by 70% ethanol for 30 seconds and finally rinsing three times with sterile distilled water. Four basal media namely, KC ^[9]; MS ^[10]; PM ^[11] and MVW ^[12] with three different sources of carbohydrates viz. sucrose, glucose and lactose were used for germination. The pH of all media was adjusted to 5.8 using 0.1N NaOH or HCl before mixing agar. 0.8% (w/v) agar was added as a gelling agent and dissolved by boiling. 100 ml of the media were dispensed into 250 ml culture bottles and autoclaved at 121⁰C for 20 minutes at 15 lbs pressure. The experiment was conducted under aseptic condition and the cultures, incubated at 25 ± 2 ^oC were subjected to 14 hr photoperiod at 4000-5000 lux intensity and 60% humidity level were maintained regularly ^[13]. For subsequent development of the seedlings, they were subcultured on respective media at different intervals. Hormones viz. BAP, Kn, Pic, NAA, IAA and IBA were freshly prepared. For the inoculation of seeds, an immature green capsule was put on a sterile petridish containing sterile filter paper and cut longitudinally using a sharp sterile blade under a laminar airflow cabinet. The very tiny seeds were scooped out with the help of sterile forceps and spread over the surface of the germination media. Eighteen types of full-strength MS and PM based solid & liquid elongation media were prepared using different concentrations and combinations of PGRs. 0.8% (w/v) agar was also used in solid media but in liquid media, no agar was added. For in vitro rooting of D. transparens, half strength MS0 with 1.5% (w/v) sucrose and nine different types of 0.8% (w/v) agar solidified MS medium supplemented with 3% (w/v) sucrose with three auxins viz. IAA, IBA and NAA were used for the induction of strong and stout root system. The well-developed seedlings were taken out of the culture vessels and successfully transferred to outside the culture room following successive phases of acclimatization. Transplanted seedlings were watered regularly for about 2-3 months where the seedlings established and grew well in the Orchidarium.

Source of Information: Journal of Medicinal Plants Studies 2020; 8(3): 141-146

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

PM was found better than KC, MS & MVW media for promoting germination of orchid seeds. Sucrose supplemented media was found to be best whereas lactose-containing medium gave at least performance. For comparing the effectiveness in terms of enhancing seedling, liquid culture was better than agar solidified condition. Increased root length and number of roots are higher in IAA supplemented full strength MS media. However, the ex-situ conservation of this species is highly recommended not only for its conservation but also to best utilized its commercial demand.

Knowledge Management: Journal