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An Efficient Method for in Vitro Clonal Propagation of a Newly Introduced Sweetener Plant (stevia Rebaudiana Bertoni.) in Bangladesh

M.B. Ahmed
Plant Breeding and Gene Engineering Laboratory, Department of Botany, Rajshahi University, Rajshahi-6205, Bangladesh

M. Salahin
Plant Breeding and Gene Engineering Laboratory, Department of Botany, Rajshahi University, Rajshahi-6205, Bangladesh

R. Karim
Plant Breeding and Gene Engineering Laboratory, Department of Botany, Rajshahi University, Rajshahi-6205, Bangladesh

M.A. Razvy
Plant Breeding and Gene Engineering Laboratory, Department of Botany, Rajshahi University, Rajshahi-6205, Bangladesh

M.M. Hannan
Plant Breeding and Gene Engineering Laboratory, Department of Botany, Rajshahi University, Rajshahi-6205, Bangladesh

R. Sultana
Plant Breeding and Gene Engineering Laboratory, Department of Botany, Rajshahi University, Rajshahi-6205, Bangladesh

M. Hossain
Plant Breeding and Gene Engineering Laboratory, Department of Botany, Rajshahi University, Rajshahi-6205, Bangladesh

R. Islam
Plant Breeding and Gene Engineering Laboratory, Department of Botany, Rajshahi University, Rajshahi-6205, Bangladesh

Abstract/ Executive Summary:

Shootlets were regenerated from nodal explants of Stevia rebaudiana Bertoni through axillary shoot proliferation. The induction of multiple shoots from nodal segments was the highest in MS medium supplemented with 1.5 mg l BA + 0.5 mg l Kn. For rooting different concentrations of IBA, NAA and IAA 1 1 were used and highest rooting percentage (97.66%) was recorded on MS medium with 0.1 mg l IAA. The 1 rooted plantlets were hardened and successfully established in soil.

Keywords: Stevia rebaudiana Bertoni, Node, In vitro, Sweetner

Location: Plant Breeding and Gene Engineering Laboratory, Department of Botany, Rajshahi University, Rajshahi-6205, Bangladesh

Start Date:

End Date:

Program Area: Development of Host and Medicinal Plants

Commodity: Medicinal Plants

Objectives:

The present study was carried out to standardize a suitable protocol for in vitro propagation of S. rebaudiana Bertoni.

Methodology:

The twigs (about 5-6 cm) of shoots of pot grown S. rebaudiana plants were collected from the Sugarcane Research Institute, Ishwardi. The twigs with node explants were washed in running tap water and then washed again thoroughly by adding a few drops of Tween-20 to remove the superficial dust particles as well as fungal and bacterial spores. They were then surface sterilized with 0.1% mercuric chloride for 5 min followed by rinsing them five times with double distilled water inside the Laminar Air flow chamber. Nodal segments (with a single axillary bud) about 0.5-0.8 cm were prepared aseptically and were implanted vertically on MS medium fortified with specific concentrations of growth regulators (BA, KIN and NAA) singly or in combination adding 30 g l sugar (market sugar) and 0.7% Difco Bacto-agar. 1 The pH of the medium was adjusted to 5.7 with 0.1 NaOH before autoclaving at 1.06 kg cm and 121°C for 20 min. 2 The cultures were incubated at a constant temperature of 26±1°C with 16 h photoperiod (2000 lux). Subcultures were done every 21 days interval. Nodal segments from the proliferated shoots were subcultured again for further multiple shoot induction. Regenerated multiple shoots were cut and individual shoots were placed in MS medium containing different concentrations of IBA, NAA and IAA for root induction.

Source of Information: American-Eurasian Journal of Scientific Research 2 (2): 121-125, 2007 ISSN 1818-6785

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

In vitro rooted shoots were kept under normal growth room conditions for 4 weeks until the induced roots became partially brown. The shoots were then taken out from the growth room, kept under room temperature for 15 days andthen taken out from the culture tubes carefully and the medium attached to the roots was gently washed out with running tap water. The plantlets were transplanted to small polythene bags containing garden soil: compost (2:1) treated with 0.1% Agrason (fungicide). The transplanted plantlets were kept under shade for 15 days and then were transferred to normal environmental conditions. Through this process of acclimatization almost 70% survival was achieved. By using the method described above, hundreds of clonal plants can be produced from one nodal explant by continuous subculturing of shoot propagules. The multiplication rate that was achieved was not significantly large to be commercially significant, but our results provide a basis for further research in micropropagation of other genotypes of S. rebaudiana.

Knowledge Management: Journal