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In Vitro Regeneration of Dendrobium Sp. of Orchid Using Leaf Tip as Explant

K. Goswami
USDA Biotechnology laboratory,Bangladesh Agricultural University, Mymensingh

S. Yasmin
USDA Biotechnology laboratory,Bangladesh Agricultural University, Mymensingh

K. M. Nasiruddin
USDA Biotechnology laboratory,Bangladesh Agricultural University, Mymensingh

F. Khatun
USDA Biotechnology laboratory,Bangladesh Agricultural University, Mymensingh

J. Akte
USDA Biotechnology laboratory,Bangladesh Agricultural University, Mymensingh

Abstract/ Executive Summary:

Plant growth regulators (PGRs) namely, 2,4-D, NAA and BAP were added into Murashige and Skoog (MS) medium to observe the effect of PGRs on the growth and development of Dendrobium sp. orchid. Leaf tips of Dendrobium sp. were used as explants and inoculated on MS medium supplemented with 2, 4 D (0, 0.5, 2.5, 5, 10 mgL⁻¹) for development of PLBs. The maximum PLBs formation (90%) and the maximum number of PLBs (16.00) were observed in 10 mgL⁻¹ 2, 4-D into MS medium after 60 days of culture. Subcultured PLBs were inoculated on MS medium supplemented with different combinations of NAA (0, 0.5, 2.5, 5 mgL⁻¹) and BAP (0, 0.5, 2.5, 5 mgL⁻¹) for shoot regeneration. The maximum number of shoot (11.00), the highest fresh weight (0.6233g) and the highest shoot length (3.613 cm) were observed in 0.5 mgL⁻¹ NAA + 0.5 mgL⁻¹ BAP after 60 days of culture. Even, the maximum number of root (4.00), the maximum root length (1.627cm) and the maximum plantlet regeneration percentage (93.33%) were observed with the combined effect of 0.5 mg NAA and 0.5 mg BAP after 60 days of culture. Finally, regenerated plantlets were transferred into half strength MS medium to obtain plants.

Keywords: Growth regulator; Plantlets; PLBs; Subculture

Location:

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Orchid

Objectives:

To establish a suitable protocol for the proliferation of protocorm like bodies.
To find out effective concentrations of plant growth regulators for proliferation of shoots and formation of roots.

Methodology:

Leaf tips of Dendrobium sp. of orchid were used as explants of this experiment. Explants were placed on the MS (Murashige and Skoog, 1962) medium supplemented with plant growth regulatrors. The pH of the medium was adjusted to 5.2 with 1N KOH or HCl prior to autoclaving for 15 min at 121⁰C. Full strength MS medium was used for the development of PLBs (Protocorm Like Bodies) from leaf tips and shoots regeneration from PLBs. Besides, half strength MS medium was also used for the subculture of PLBs and development of roots. Five different concentrations namely 2, 4-D (0, 0.5, 2.5, 5, 10 mgL⁻¹) were added to MS medium for the development of PLBs from leaf tips. For the development of shoot, we used BAP (0, 0.5, 2.5, 5 mgL⁻¹) and NAA (0, 0.5, 2.5, 5 mgL⁻¹) in different combinations. The culture vials containing the media were autoclaved with 1.16 kg/cm² of pressure at 121⁰c for 20 minutes. All the culture vials were placed in a culture room and allowed to grow at 25±1°C under 16 hour photoperiod illuminated with fluorescent tube of 2000-3000 lux. After 60 days of culture, data (number and length of PLBs, root, shoot) were recorded. MSTATC software was used to analyze the data by ANOVA1 function .

Source of Information: Journal of Environmental Science and Natural Resources, Vol 8, No 2 (2015), Pp. 75-78.

Article Link (Online Journal): http://www.banglajol.info/index.php/JESNR/article/view/26869/17943

Funding Source:

1.

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The present study was undertaken with a view to investigate the effect of different plant growth regulators (PGRs) on growth and development of plantlets derived from PLBs using leaf tips. Leaf tips and PLBs were cultured on MS medium supplemented with different concentrations of 2, 4-D, NAA and BAP. It can be concluded from the obtained result that MS medium supplemented with 10 mgL⁻¹ 2, 4-D could be used for the production of PLBs from leaf tips and MS medium supplemented with 0.5 mgL⁻¹ NAA + 0.5 mgL⁻¹ BAP could be recommended for the large scale production of shoots from PLBs as well as for the highest development of root.

Knowledge Management: Journal