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Micropropagation of Stevia Plant from Nodal Segments

M. A. Karim
Department of Crop Botany, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

R. Jannat
Department of Crop Botany, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

M. S. Rahman
Department of Crop Botany, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

M. S. Haque
Department of Biotechnology, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Abstract/ Executive Summary:

The experiment was conducted to develop and establish a reproducible protocol for plantlet regeneration in Stevia. Nodal cutting explants were cultured in Murashige and Skoog (MS) medium supplemented with different concentrations and combinations of α-Napthaleneacetic acid (NAA) and 6-Benzylamino purine (BAP). The combination of NAA at 1.0 mgl^-1 and BAP at 1.0 mgl^-1 resulted in the highest percentage (100%) of callus initiation. The maximum shoot regeneration and development of shoot was observed at the same combination. The developed shoots from nodal cuttings, upon transfer to the MS medium containing indole butyric acid (IBA) at 0.1mgl-1 resulted in best rooting within 8 days.

Keywords: Micropropagation, plantlet regeneration, Stevia, Nodal segment

Location: Department of Biotechnology, Bangladesh Agricultural University, Mymensingh, Bangladesh

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Stevia

Objectives:

a. To develop and establish a reproducible protocol for plantlet regeneration, and to determine the suitable concentration.

Methodology:

An experiment on in vitro callus induction and subsequent plantlet regeneration of Stevia was conducted at the Laboratory of the Department of Biotechnology, Bangladesh Agricultural University, Mymensingh in order to fulfill the objectives set in this research programme. Murashige and Skoog (MS) medium was used. Proper sterilization of MS medium, test tubes, pipette, petridishes, beakers, scalpels, forceps, needles and other glass wares and growth chamber were made. Incubation chamber was properly maintained. The nodal explants were cut into small pieces (about 0.5-0.6 cm long) and then treated with 2% savlon with constant shaking for 5-6 minutes and washed thoroughly with distilled water. Then the explants were taken under Laminer Air Flow Cabinet. The surface sterilization of the explants was done with 0.5% mercuric chloride solution for 5 minutes under aseptic condition followed by washing 5-6 times with sterilized distilled water. Explants were cultured on MS medium supplemented with different concentrations and combinations of BAP at (0, 1, 2.5 and 5 mgl^-1) and NAA at (0, 1, 2.5 and 5 mgl^-1) for callus initiation and proliferation. Calli from nodes were subcultured on the same medium. The calli were again cultured on MS medium supplemented with different concentrations and combinations of BAP at (0, 1, 2.5, 5 and 10 mgl^-1) and NAA at (0 & 1 mgl^-1) for shoot proliferation. Media supplemented with different concentrations of IBA (0, 0.1, 0.5 and 1.0 mgl^-1) and NAA (0, 0.1, 0.5 and 1.0 mgl^-1) were used for root induction. Culture vessels were incubated at 25 ± 1⁰ C with 16 h photoperiod with light intensity of approximately 2000 lux.

Source of Information: Progress. Agric. 19(2) : 21-26, 2008 ISSN 1017-8139

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

It may be concluded that maximum calli induction, shoot regeneration were achieved with the combination of NAA at 1.0 mgl^-1 and BAP at 1.0 mgl^-1. Shoots from node explants produced maximum number of roots containing 0.1 mgl^-1 IBA in the medium. A protocol for the micropropagation Stevia has been developed in this study. The maximum shoot regeneration and development of shoot was observed at the same combination. The developed shoots from nodal cuttings, upon transfer to the MS medium containing indole butyric acid (IBA) at 0.1mgl^-1 resulted in best rooting within 8 days.

Knowledge Management: Journal