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Efficient Plant Regeneration Through Somatic Embryogenesis from Leaf Base-derived Callus of Kaempferia Galanga L.

M.M. Rahman
Department of Botany, University of Rajshahi, Rajshahi-6205, Bangladesh.

M.N. Amin
Department of Botany, University of Rajshahi, Rajshahi-6205, Bangladesh.

T. Ahamed
Department of Botany, University of Rajshahi, Rajshahi-6205, Bangladesh.

M. R. Ali
Biotechnology and Genetic Engineering Discipline, Khulna University, Khulna-9208, Bangladesh.

A. Habib
Biotechnology and Genetic Engineering Discipline, Khulna University, Khulna-9208, Bangladesh.

Abstract/ Executive Summary:

Somatic embryogenesis and subsequent plantlets regeneration were achieved in callus cultures established from the leaf base explants of Kaempferia galanga L. Callus induction and somatic embryogenesis at various frequencies were observed using different concentrations and combinations of growth regulators. The highest percentage of callus induction was observed on MS medium supplemented with 1.5 mg L^-1 2,4-D+1.0 mg L^-1 BA. After transfer on MS medium supplemented with 2.0 mg L^-1 BA+0.1 mg L^-1NAA, this callus produced small globular embryos first that later developed into plantlets by further subculturing on the same medium. Plantlets were acclimatized and subsequently transferred to the field. Survival rate of the plantlets under ex vitro condition was 85%.

Keywords: Ginger, Plant regeneration, Somatic embryogenesis, Biotechnology

Location: Department of Botany, University of Rajshahi, Rajshahi-6205, Bangladesh.

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Ginger

Objectives:

To establish a suitable plantlet regeneration protocol for Kaempferia galanga using in vitro techniques.

Methodology:

Leaf base segments were excised aseptically from in vitro grown cultures, which were originally established from the rhizome buds of field grown mature plants. The sheathing basal parts of excised leaves were chopped into pieces (1-1.5 mm) and segments were placed on the agar-gelled medium. The leaf base explants were ultimately cultured on MS medium supplemented with various concentrations of 2, 4-D alone and in combination with BA for callus induction. Seven weeks old callus was thus sub cultured on MS medium containing different combinations of BA with NAA or IBA for somatic embryogenesis. The embryogenic calluses were sub cultured at monthly intervals on same medium for somatic embryos formation and development of complete plantlets with shoots and roots. The pH of the culture media was adjusted to 5.7±0.1 before addition of agar and sterilized by autoclaving for 20 min at 1.1 kg cm^-2 pressure at 121°C. For solidifying the medium, 7-8 g L^-1 agar or 2.0 g L^-1gelrite was used. The tubes or flasks containing explants were incubated on culture racks in the growth chamber. The cultures were maintained at 25±2°C under the cool white fluorescent lights for 16 h photoperiod with a photon flux density of about 70 μ mol m^-2s^-1.

Source of Information: Asian Journal of Plant Sciences Volume 3, Number 6, 675-678, 2004

Article Link (Online Journal): DOI: 10.3923/ajps.2004.675.678, URL: http://scialert.net/abstract/?doi=ajps.2004.675.678

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Among different combinations of the auxin and cytokinin tested, 2, 4-D + BA was found highly effective for induction of callus. It was mostly soft, faster growing, light creamy-white and friable. The highest frequency of callus induction (85%) was recorded at the end of 7 weeks on medium containing 1.5 mg L^-1 2, 4-D with 1.0 mg L^-1 BA. Similar combination of auxin with cytokinin for callus induction has been reported in the past by Malamug on ginger, Vincent et al. on Kaempferia galanga, Muthukumar et al.on Datura metel. Callus induction from leaf base explant has also been achieved in cardamom using 2, 4-D+ Kn in combination.

Knowledge Management: Journal