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Protocol Establishment for Multiplication and Regeneration of Ocimum Sanctum Linn. an Important Medicinal Plant with High Religious Value in Bangladesh

L. A. Banu
Institute of Biological Sciences, University of Rajshahi, Rajshshi 6205, Bangladesh.

M. A. Bari
Biotechnology Lab, Institute of Biological Sciences, University of Rajshahi, Rajshshi 6205, Bangladesh.

Abstract/ Executive Summary:

An experiment was conducted at the Plant Biotechnology Laboratory of Institute of Biological Sciences, University of Rajshahi, Bangladesh. Shoot tip and leaf explants of Ocimum sanctum Linn. were cultured on different concentrations and combinations of growth regulators (BAP, Kin, 2, 4-D, IAA and IBA) in MS medium to observe shoot multiplication, callus induction, callus regeneration and root induction. Among the different concentrations and combinations of growth regulators, the highest percentage of shoot formation and highest average number of shoots were observed 90 and 5.88%, respectively in 0.2 mg L^-1 BAP from shoot tip explants. Callus induction was obtained within 12-15 days of culture from leaf explants. The highest frequency (90.00%) of organic callus induction was observed in MS medium containing 1.0 mg L^-1 NAA. Shoot regeneration occurred when the calli were sub cultured in Ms medium supplemented in BAP formulation. The highest percentage of shoot regeneration was obtained 90.00 in 0.2 mg L^-1 BAP. In vitro grown shoots rooted best on MS medium containing 0.1 mg L^-1 NAA. The in vitro grown plantlets were transferred to pots containing sand and soil mixture, acclimatized in a culture room and finally rooted plants were transferred to soil.

Keywords: In vitro, Tulsi, Regeneration, Multiplication, Shoot tip, Callus, Ocimum sanctum

Location: Plant Biotechnology Laboratory of Institute of Biological Sciences, Rajshahi, Bangladesh.

Start Date:

End Date:

Program Area: Development of Host and Medicinal Plants

Commodity: Medicinal Plants

Objectives:

To establish a in vitro regeneration protocol for the plant Ocimum sanctum (Tulsi) using all possible explants for direct regeneration

Methodology:

Shoot tip and leaf explants (1-2 cm) of Ocimum sanctum Linn. (Tulsi) were collected from matured plants and used for establishing shoot formation and callus induction. The explants were washed thoroughly under tap water. Then treated with a few drops of Tween-80 and 1% savlon for 20 min. with constant shaking washed thoroughly with distilled water to remove gummy substant and savlon. Explants were surface sterilized with 0.1% HgCl₂ for 6 min followed by gentle shaking in a laminar air flow cabinet. After surface sterilization the explants were thoroughly washed for several times with sterile distilled water. Shoot tip explants then cut into pieces of 0.7-1.0 cm and transferred to 25 x 150 mm culture tubes with 15 mL agar-gelled MS (Murashige and Skoog, 1962) basal medium supplemented with different concentrations of BAP, Kin and combination with NAA for shoot initiation. The leaf explants cut into pieces of 1 cm and cultured on agar-gelled MS medium supplemented with different concentrations of 2, 4-D and NAA for inducing callus. In vitro grown leaves were also cultured on MS medium containing same hormone. All media were supplemented with 30% sugar, adjusted to pH 5.8, gelled with 0.6% agar (BDH) and autoclaved at 120°C for 20 min under 1.1 kg cm^-1 pressure. Cultures were incubated at 25±2°C under the warm fluorescent light with intensity varied from 2000-3000 lux. The calli was then transferred on MS medium containing different concentrations of BAP for differentiating shoots and incubated in light. Proliferated shoots were transferred to MS with different concentrations of NAA, IAA and IBA for root formation. Data were recorded after 28 days of culture.

Source of Information: Journal of Plant Sciences, 2(5): 530-537, 2007, ISSN 1816-4951.

Article Link (Online Journal): DOI: 10.3923/jps.2007.530.537; URL: http://scialert.net/abstract/?doi=jps.2007.530.537

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Among the all concentrations of IAA, 50% shoots produced root in 0.1 mg L^-1 IAA. Maximum average number of root per culture was 3.20 and average length of root was 3.10 cm in 0.1 mg L^-1 IAA. On the other hand, among the all concentrations of IBA, 0.1 mg L^-1 IBA was effective for root induction than other concentration used. Maximum number of root per culture was 3.75 and maximum length of root was 4.06 cm in 0.1 mg L^-1 IBA. The in vitro plantlets were successfully established on the soil and 75% of them survived.

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