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In Vitro Micropropagation of Cymbidium Aloifolium L. Sw. and Spathoglottis Plicata Bi

A. K. Barua
Department of Botany, University of Chittagong, Chittagong-4331, Bangladesh

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

Abstract/ Executive Summary:

The shoot and leaf segments of in vitro germinated seedlings of Cymbidium aloifolium and Spathoglottis plicata were aseptically grown on 0.8% (w/v) agar solidified three different media, namely Murashige and Skoog (MS), Phytamax (PM) and Modified Vacin and Went (MVW) supplemented with 2 - 3% (w/v) sucrose and different combinations of auxins (IAA, IBA, NAA and 2,4-D) and cytokinin (BAP and zeatin) at different concentrations. The shoot segments having nodal zone of both the species produced multiple shoot buds on all the media with different combinations of hormones. Leaf segments, however, did not give any response to any of these media. In case of C. aloifolium the maximum number of multiple shoot buds/shoot segment was developed on MVW supplemented with 2% (w Iv) sucrose + 0.05 mg/l IAA + 2.0 mg/l zeatin. On the other hand, in case of S. plicata MS medium supplemented with 3% (w Iv) sucrose + 0.1 mg/l IBA + 2.0 mg/l BAP was proved best. The multiple shoot buds of both the species underwent rapid elongation on the same media but did not produce better root system. In order to induce stout and strong root system, the shoot buds at a stage of 2 - 3 cm length were individually cultured on two different 0.8% (w Iv) agar solidified media, namely (i) half strength of MS + 1.5% (w Iv) sucrose and (ii) MS + 3% (w Iv) sucrose + 0.5 mg/l IAA. The former medium was better than the latter in both the species. These in vitro grown seedlings were then successfully transferred to outside natural condition through successive phases of acclimatization.

Keywords: Micropropagation, Cymbidium aloifolium, Spathoglottis plicata

Location: Department of Botany, University of Chittagong, Chittagong

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Orchid

Objectives:

To develop a reliable, reproducible and efficient method of rapid micropropagation of C. aloifolium and S. plicata using their in vitro raised seedlings. Once such propagation system is developed that can be used in our commercial horticultural nurseries.

Methodology:

Two-three months old in vitro germinated seedlings (5 - 7 cm) of Cymbidium aloifolium and Spathoglottis plicata were used. Their shoots were cut into two - three segments eah having a node. The leaves were also cut into small pieces (1 - 2 cm). The shoot and leaf segments were then aseptically cultured on three different types of 0.8% (w/v) agar solidified media (MS, PM and MVW) fortified with different combinations of auxins, IAA (0.1 - 1.0 mg/l), IBA (0.05 - 0.1 mg/l), NAA (1.0 mg/l), 2,4-D (1.0 mg/l) and cytokinins BAP (0.5 - 3.0 mg/l) and zeatin (2.0 mg/l). The media were taken in cultural vessels (test tubes, 2 x 15 cm or conical flax 100/150 mg/l). The explants were aseptically grown on the surface of the media taken in cultural vessels which were maintained in the culture room under a period of 14 h continuous light and 10 h of continuous dark at 15 ± 2°C. Subculture was done regularly at an interval of 20 - 25 days. Within 30 - 35 days of subculture new shoot buds came out from the shoot segments and that underwent rapid elongation on the same media. When the shoot buds attained a height of 2 - 3 cm, they were grown individually on two rooting media, namely (i) 0.8% (w /v) agar solidified half-strength of MS + 1.5% (w/v) sucrose and (ii) 0.8% (w/v) agar solidified MS supplemented with 3% (w/v) sucrose and 0.5 mg/l IAA.

Source of Information: Plant Tissue Culture, Vol: 9, (2), pp: 133-140, Dec 1999

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

On the whole, in vitro cultural techniques could be formulated for micropropagation of C. aloifolium and S. plicata using nodal segments and that offers an opportunity of using these techniques in commercial nurseries as well as for ex situ conservation of these species.

Knowledge Management: Journal