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In Vitro Rapid Clonal Propagation of Rauvolfia Serpentina (linn.) Benth

R. Baksha
Biotechnology Division, BRRI, Gazipur -1701

Miskat Ara Akhter Jahan
Biological Research Division, BCSIR Laboratories, Dhaka-1205, Bangladesh

Rahima Khatun
Biological Research Division, BCSIR Laboratories, Dhaka-1205, Bangladesh

John Liton Munshi
Biological Research Division, BCSIR Laboratories, Dhaka-1205, Bangladesh

Abstract/ Executive Summary:

A protocol was developed for in vitro clonal propagation of Rauvolfia serpentina through direct regeneration from shoot tip explants. Multiple shoots (eight shoots per explant) induction were obtained on MS medium supplemented with 4.0 mg/l BAP and 0.5 mg/l NAA within 10-15 days. The elongated shoots rooted well in half strength MS medium with 0.5 mg/l NAA. The in vitro raised plantlets were acclimatized in glass house and successfully transplanted to field condition with 80% survival. The results indicated that large scale commercial micropropagation of Rauvolfia serpentina is technically feasible.

Keywords: In vitro, Rapid clonal propagation, Rauvolfia serpentina

Location: Biological Research Division in Bangladesh Council of Scientific and Industrial Research (BCSIR), Dhaka, Bangladesh

Start Date: 00-00-2004

End Date:

Program Area: Development of Host and Medicinal Plants

Non-commodity: Performance

Objectives:

The present study was undertaken to develop a more efficient protocol for rapid in vitro multiplication of Rauvolfia serpentina using shoot tip explant as an initial plant material.

Methodology:

The experiment was conducted at Biological Research Division in Bangladesh Council of Scientific and Industrial Research (BCSIR), Dhaka, Bangladesh during the period of 2004. The apical shoots with leaf primordia of two nodes (2-4 mm long) were collected from the field grown three months old of Rauvolfia serpentina and washed thoroughly under running tap water. For surface sterilization the shoot tips were then treated with 1 % savlon for 10 minutes with constant shaking. Then these were washed four times with distilled water. Finally the shoot tips were treated with 0.1 % HgCl₂ solution for 10 minutes under laminar air flow cabinet and washed three times with double distilled water to remove any trace of HgCl₂. After surface sterilization the shoot tips of 2-4 mm long were excised and placed on MS (Murashige and Skoog, 1962) medium supplemented with different combinations of auxin and cytokinin to identify appropriate combinations for multiple shoot regeneration of Rauvolfia serpentina. Media consisted of 3 % sucrose, 0.6 % agar, pH was adjusted to 5.7 before addition of agar and autoclaved at 120^OC for 15 minutes. All cultures were inoculated in 250 ml flasks containing 150 ml medium. Explants were kept at 25 ± 2^OC under 16 hours photoperiod regime at 2000 lux light intensity of cool white fluorescent light. Elongated healthy micro shoots were then excised and implanted to the rooting medium consisting of half strength MS supplemented with IBA and NAA in different concentrations. After initiation of root the plantlets were carefully separated from the medium to avoid damaging and washed with tap water to remove the agar adhering to them. The plantlets were then transplanted to polythene bags containing a mixture of soil and sand (2:1). The plantlets were kept in a glass house for one week with normal light and temperature. New leaves emerged from the plantlets after 10-15 days in the glass house; they were then placed in the normal environment for 1h and assessed for signs of wilting. The exposure time was increased daily until the plants were fully acclimatized. When the plantlets were established fully under normal environmental conditions, they were transferred into the field for their normal growth. The cultures were regularly subcultured on fresh medium at four weeks intervals and observations were recorded. Each experiment consisted of 25 replicates and repeated thrice. The means and standard errors of the results were also calculated.

Source of Information: Bangladesh J. Sci. Ind. Res. 42(1), 37-44, 2007

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

In the present investigation, a high frequency and rapid shoot induction was observed in Rauvolfia serpentina through shoot tip explants with BAP (4.0 mg/l) and combination of NAA (0.5 mg/l). For root induction NAA(0.5 mg/l) was found to be ideal growth regulator. For commercial exploitation this method for rapid micropropagation of Rauvolfia serpentina (a valuable medicinal plant) is reliable and promising.

Knowledge Management: Journal