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In Vitro Propagation of a Medicinal Plant - Mango Ginger (curcuma Amada Roxb.)

M. M. Ferdous
Plant Tissue Culture Laboratory, Department of Botany, University of Rajshahi, Rajshahi-6205, Bangladesh

M. Shahinozzaman
Plant Tissue Culture Laboratory, Department of Botany, University of Rajshahi, Rajshahi-6205, Bangladesh

M. O. Faruq
Plant Tissue Culture Laboratory, Department of Botany, University of Rajshahi, Rajshahi-6205, Bangladesh

S. P. Paul
Plant Tissue Culture Laboratory, Department of Botany, University of Rajshahi, Rajshahi-6205, Bangladesh

M. A. K. Azad
Plant Tissue Culture Laboratory, Department of Botany, University of Rajshahi, Rajshahi-6205, Bangladesh

M. N. Amin
Plant Tissue Culture Laboratory, Department of Botany, University of Rajshahi, Rajshahi-6205, Bangladesh

Abstract/ Executive Summary:

This paper describes efficient in vitro propagation protocol of Curcuma amada Roxb., an important aromatic zingiberaceous plant in the countries of Indian subcontinent, using rhizome explants. Rhizome tip explants showed better response than rhizome bud explants in terms of shoot proliferation and hence excised rhizome tip explants were inoculated aseptically on Murashige and Skoog’s (MS) medium supplemented with different 6-Benzyl adenine (BA) or 6-furfurylamino purine (Kn) concentrations (0.0, 2.0, 4.0, 6.0, 8.0, 10.0 μM) alone or in combinations with α- Naphthalene acetic acid (NAA) and Indole-3-butyric acid (IBA) (1.0, 1.5, 2.0 μM). Optimum proliferation was obtained on MS medium having 8.0 μM BA + 1.0 μM NAA. Shoot proliferation was maximal (99.8%) with 10.6 ± 0.26 shoots per explant after 6 weeks of culture. The successfully proliferated shoots with an average height of 38 cm were transferred to rooting medium augmented with NAA and IBA at four different concentrations (2.0, 4.0, 6.0, 8.0 μM) and 4.0 μM IBA was found to be best for in vitro adventitious rooting. Rooted plantlets were then transferred to plastic cups containing autoclaved garden soil and compost (1:1), gradually acclimatized and finally transferred to the field condition with 80% survival rate.

Keywords: Mango ginger, Rhizome explants, In vitro propagation, Medicinal plant

Location: Plant Tissue Culture Laboratory, Department of Botany, University of Rajshahi, Rajshahi

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Ginger

Objectives:

To develop the protocol reported here could be used for the conservation, large-scale propagation and for further research on the biochemical composition and medicinal importance of this valuable medicinal herb.

Methodology:

Plant material - Healthy rhizome with active buds were collected from the rhizome of C. amada maintained in the nursery bed of experimental garden of Plant Tissue Culture Laboratory, department of Botany, Rajshahi University. They were cut into 1.5 to 2 cm length with active buds intact and categorized into two separate groups- rhizome tip and rhizome bud. These rhizome explants were initially washed with Savlon (Chlorhexidine gluconate 0.3% w/v + cetrimide 3% w/v solution) for 10 minutes and rinsed several times under running tap water. These excised explants were transferred to laminar air flow cabinet and surface sterilized with 0.1% HgCl₂ for 10 minutes and then washed with sterile distilled water thoroughly 3 to 4 times with sterile distilled water and soaked with sterile blotting paper and used as explants for in vitro cultures before the inoculation in to sterilized nutrient agar medium (MS). Culture medium and growth conditions - The sterilized blotted explants were implanted on to the Murashige and Skoog’s agar-gelled medium fortified with various concentrations/combinations of growth regulators. For shoot induction, the medium was supplemented with 0.0, 2.0, 4.0 6.0 8.0 and 1.0 μM BA and Kn alone or in combination with 1.0, 1.5 and 2.0 μM NAA and IBA. For root induction, in vitro raised shoots grown in multiplication medium were cultured on MS medium supplemented with either NAA or IBA in concentration of 2.0, 4.0, 6.0 and 8.0 μM. The pH of the medium was adjusted to 5.8 before autoclaving at 1.2 kg/cm² pressure and 121°C temperature for 15 - 20 minute. Molten medium of 20 ml was dispensed into the culture tube and plugged with nonabsorbent cotton wrapped in one layer of cheesecloth. All the cultures were incubated in the culture room at 25 ± 2⁰C under a 16h light and 8h dark cycle with the light intensity of 2000 – 3000 lux provided by cool-white fluorescent tubes (36W). Each treatment had 15 culture tubes and the experiment was repeated thrice. The cultures were maintained by regular subcultures at 2 weeks intervals on fresh medium with the same compositions. Hardening and acclimatization - Plantlets having a well-developed root system were removed from the culture tubes and the roots were washed under running tape water to remove agar. Then the plantlets were transferred to sterile small plastic cups containing autoclaved garden soil and compost (1:1) and maintained inside growth chamber set at temperature 28⁰C and 70-80% relative humidity. After four weeks they were kept under laboratory conditions for additional two weeks before transferring them to the Botanical Evaluation Garden and maintained there for acclimatization. Culture observation and presentation of results - Fifteen cultures were used per treatment and each experiment was repeated three times. The data pertaining to percentage of cultures showing proliferation, average number of shoots/culture, percentage of rooting, mean number of roots and average length of roots were statistically analyzed by the Duncan’s multiple Range Test at the P < 0.05 level of significance using a statistical software- SPSS.

Source of Information: International Journal of Biosciences (IJB); ISSN: 2220-6655 (Print) 2222-5234 (Online); Vol. 2, No. 11, p. 166-172, 2012

Article Link (Online Journal): http://www.innspub.net

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

In the present investigation, in-vitro micro-plantlet multiplication system of C. amada has been optimized through rhizome tip culture. The described protocol is the first report for this species which can be used to produce a higher amount of propagules around the year. Production of in-vitro plantlets would be a suitable methodology for direct regeneration of shoot lets as a source of disease free quality planting material that could be stored and transported easily and a step forward towards commercial scale of propagule production in mango ginger.

Knowledge Management: Journal