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In Vitro Propagation of Eria Tomentosa (koen.) Hook. F.: an Important Indigenous Medicinal Orchid of Bangladesh

MA Kashem
Assistant Professor
Department of Botany University of Chittagong, Chattogram 4331, Bangladesh

TK Bhowmik
Assistant Professor
Department of Botany, University of Chittagong, Chattogram 4331, Bangladesh

MM Rahman
Professor
Department of Botany University of Chittagong Chattogram 4331, Bangladesh

Abstract/ Executive Summary:

Stem and leaf segments of aseptically grown seedlings of Eria tomentosa (Koen.) Hook. f. gave a response to both PGR supplemented agar solidified and liquid MS media and PM media. Both the explants underwent direct organogenesis producing shoot primordia like structures (SPSs) on MS medium fortified with 1.5-3.0 mg/l BAP or Kn either alone or in combination with 0.5-1.0 mg/l IAA or NAA. However, liquid MS + 3% (w/v) sucrose + 0.5 mg/l Pic + 1.0 mg/l BAP took lesser time and was effective for induction of shoot primordial than agar solidified PM. Liquid media were found more effective in SPSs induction. It was revealed that MS medium was more effective than PM medium for SPSs induction. The shoot primordia developed from stem and leaf segments underwent elongation when cultured on both liquid and agar solidified MS elongation media supplemented with different concentrations and combinations of PGRs and the maximum rate of elongation was achieved in MS (liquid) + 2.0 mg/l BAP + 1.0 mg/l IAA. The elongated shoots were produced aerial roots in agar solidified MS media supplemented with different PGRs. Thus an efficient protocol was developed for mass scale propagation of the selected orchid species.

Keywords: Eria tomentosa, In vitro culture, Medicinal orchid

Location: Dhopachari of Chattogram district, Bangladesh.

Start Date:

End Date:

Program Area: Development of Host and Medicinal Plants

Commodity: Orchid

Objectives:

To establish an efficient protocol for in vitro rapid propagation of Eria tomentosa an indigenous medicinal orchid species of Bangladesh.

Methodology:

The plant (Eria tomentosa) was collected from Dhopachari of Chattogram district, Bangladesh. The orchid species was identified and authenticated by consulting relevant literature and critical examination of the herbarium. The voucher specimen has been preserved at the Herbarium of Chittagong University (HCU). The capsule of the disease-free and the fresh plant was used for in vitro culture. Collected fresh samples were cultured in aseptic condition for mass scale propagation following tissue culture method in the laboratory of Plant Tissue Culture and Biotechnology, Department of Botany, University of Chittagong. 2.2 Culture of plant material Protocorms with leafy shoot initials raised from in vitro green pods seeds of the selected species cultured on Murashige and Skoog (MS) and Phytamax (PM) basal medium, were used as the explants. As the protocorm explants were obtained through in vitro cultures, no sterilization was required. 2.3 Effect of plant growth regulators and additives on shoot growth Protocorms were inoculated on 1/2 MS medium supplemented with different concentration and combination of cytokinins [6-benzylaminopurine (BAP), kinetin (Kn)], auxins [Indole-3-acetic acid (IAA), Indole- 3-butyric acid (IBA), a-naphthaleneacetic acid (NAA), 2,4- dichlorophenoxyacetic acid (2,4-D) and picloram (Pic)] and of natural additives [banana pulp (BP) and coconut water (CW)] for shoot growth. 2.4 Culture condition The basal 1/2 MS medium was fortified with 2% sucrose, 100 mg/l myo-inositol and will be solidified with 0.7% (w/v) agar. The pH was adjusted to 5.8 ±0.2 either with 0.1 N NaOH or HCl prior to autoclaving at 121 °C and 1.5 kg/cm2 for 30 min. All cultures were aseptically maintained at 25± 2 °C under 16/8 h (light/dark) photoperiod with a light intensity of 40 μmol m-2 s -1 by white fluorescent light. 2.5 Experimental design and analysis of data Protocorm explants were cultured in 100 ml culture bottles with five explants, each treatment was comprised of three bottles and the experiment will be repeated twice. Regular observations of cultures were made once a week. The total number of protocorms responding with healthy shoots was recorded after 5 weeks and the total number and length of shoots, as well as roots, was recorded after 15 weeks of culture. The average number, length of shoots were analyzed statistically using one-way analysis of variance (ANOVA), and the mean±standard error (S.E.) of triplicates was represented and compared using Duncan’s Multiple Range Test at 5% level of significance using SPSS statistic program software version 22.0. 3.

Source of Information: Journal of Medicinal Plants Studies 2020; 8(4): 206-211

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The invention of in vitro propagation technique has saved many naturally growing orchids and their collection from the wild. The increasing popularity of orchids for cut flower and medicinal purpose has added a new dimension to in vitro propagation technique through which a significant number of identical clones can be raised from a single protocorm or shoot tip explants. Thus methods for rapid multiplication of orchids are essential to meet the commercial demand. The present investigation established a protocol for mass propagation of shoot primordial-like structures (SPSs) of the selected orchid species. Further studies should be initiated to acclimatize the orchid species in natural conditions.

Knowledge Management: Journal