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In Vitro Regeneration Protocol of Rauvolﬁa Serpentina L.

S. Khan*
Plant Tissue Culture Section, Biological Research Division, Bangladesh Council of Scientific and Industrial Research, Dhanmondi, Dhaka-1205

T. A. Banu
Plant Tissue Culture Section, Biological Research Division, Bangladesh Council of Scientific and Industrial Research, Dhanmondi, Dhaka-1205

S. Akter
Plant Tissue Culture Section, Biological Research Division, Bangladesh Council of Scientific and Industrial Research, Dhanmondi, Dhaka-1205

B. Goswami
Plant Tissue Culture Section, Biological Research Division, Bangladesh Council of Scientific and Industrial Research, Dhanmondi, Dhaka-1205

M. Islam
Plant Tissue Culture Section, Biological Research Division, Bangladesh Council of Scientific and Industrial Research, Dhanmondi, Dhaka-1205

U. Hani
Plant Tissue Culture Section, Biological Research Division, Bangladesh Council of Scientific and Industrial Research, Dhanmondi, Dhaka-1205

A. Habib
Plant Tissue Culture Section, Biological Research Division, Bangladesh Council of Scientific and Industrial Research, Dhanmondi, Dhaka-1205

Abstract/ Executive Summary:

An efficient in vitro regeneration system was developed for Rauvolfia serpentina L. through direct and indirect organogenesis from nodal and leaf explants. Among the different growth regulators, MS medium supplemented with 2.0 mg/l BAP, 0.5mg/l IAA and 0.02mg/l NAA found best for the multiple shoot formation from nodal segments. In this combination 98% explants produced multiple shoots and the average number of shoots per explants is 13.4. The frequency of callus induction and multiple shoot induction from leaves was highest 88% in MS medium supplemented with 2.0 mg/l BAP, where mean number of shoots/explants was 12.5. The highest frequency of root induction (80%) and mean number of roots/plantlets (10) were obtained on half strength of MS medium containing 0.2 mg/l IBA. The rooted plantlets were transferred for hardening following acclimatization and finally were successfully established in the field.

Keywords: Rauvolﬁa serpentina L.; Nodal segment; Leaf explants; Multiple shoot; Callus induction

Location: Plant tissue culture section, Biological Research Division, BCSIR Laboratories Dhaka

Start Date:

End Date:

Program Area: Development of Host and Medicinal Plants

Commodity: Medicinal Plants

Objectives:

The present study was undertaken to develop an efficient protocol for rapid in vitro regeneration of R. serpentina using node and leaf explant as an initial plant material by optimizing the growth regulators such as auxins and cytokinins.

Methodology:

Two types of explants namely, nodal segment and young leaf were used in the present experiment. The explants were collected from R. serpnetina grown in the research fields of BCSIR, Dhaka. The experiment was conducted at Plant tissue culture section, Biological Research Division, BCSIR Laboratories Dhaka. After collection, the explants were primarily kept under running tap water for 5 minutes. The primary surface sterilization was carried out by treating the samples with mild detergent (Tween-20) and washed with tap water for 30 minutes. After that, explants were surface sterilized with 70% (v/v) ethanol for 1 min, followed by 0.1% HgCl₂ through gentle shaking for 5 min and rinsed three times with autoclaved distilled water. Isolated explants were inoculated and cultured on MS (Murashige and Skoog, 1962) media containing BAP, NAA, IAA and Kn singly or in combinations for direct and indirect (through callus) in vitro regeneration of shoots. In vitro regenerated shoots were sub-cultured regularly to fresh medium at an interval of 21-28 days for further multiplication. Elongated shoots were separated and cultured on rooting medium for root formation. About 2-3 cm long shoots were separated and cultured on rooting medium containing full and half strengths of MS without hormonal supplement or with different concentrations of IBA. All in vitro grown cultures were maintained under illumination on a 16 h photoperiod at 25 ± 2^oC. The well rooted plantlets were then kept in room temperature for 2-3 days and transferred to plastic pot containing garden soil and compost in a ratio of 2:1 and was moistened them adequately for proper hardening.

Source of Information: Bangladesh J. Sci. Ind. Res. 53(2), 133-138, 2018

Article Link (Online Journal): DOI: http://dx.doi.org/10.3329/bjsir.v53i2.36674

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

In conclusion, it can be mentioned that, the present study describes a reproducible and efficient protocol for mass propagation of this important medicinal plant species. It would also ensure a continuous supply of R. serpentina plants in limited time and space for large scale commercial production thereby ruling out the enslavement on natural source to execute the growing demands for the pharmaceutical industry.

Knowledge Management: Journal