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In Vitro Rapid Clonal Propagation of Ocimum Basilicum L.

F. Begum
Department of Botany, University of Rajshahi, Rajshahi-6205, Bangladesh

M. N. Amin
Department of Botany, University of Rajshahi, Rajshahi-6205, Bangladesh

M. A. K. Azad
Department of Botany, University of Rajshahi, Rajshahi-6205, Bangladesh

Abstract/ Executive Summary:

In vitro regenerated shoots of Ocimum basilicum L. were excised, sectioned into nodal pieces and subcultured individually in the nutrient medium which produced in an average of eight multiple shoots per transfer. For rooting, in vitro grown shoots were excised from the culture flask and implanted individually on root induction medium containing half strength of MS salts and 0.1 - 1.0 mg/l NAA, IBA or IAA. The highest frequency and healthy rooting was observed on MS medium containing 1.0 mg/l NAA. In vitro regenerated plantlets were transferred on to specially made plastic tray containing coco-peet as potting mix and thereafter successfully established under ex vitro condition. The survival percentage of transplanted plantlets was 75.

Keywords: Ocimum basilicum, Regeneration, Nodal segments, Shoot tip

Location: Department of Botany, University of Rajshahi, Rajshahi, Bangladesh

Start Date:

End Date:

Program Area: Development of Host and Medicinal Plants

Commodity: Medicinal Plants

Objectives:

To establish a suitable regeneration protocol for O. basilicum growing in Bangladesh using tissue culture technique.

Methodology:

Nodal and shoot tip segments from mature plants of Ocimum basilicum L. were used in the present study. The explants were collected, washed thoroughly under running tap water for 15 min, treated in 1% Savlon for five min and were surface sterilized with 01.% HgCl₂ for different lengths of time to ensure contamination-free culture. Thereafter the explants were washed three - four times with autoclaved distilled water to remove traces of HgCl₂ inside a laminar air flow cabinet. The surface sterilized shoots were cut into 1.0 - 1.5 cm long segments, each containing a single node. The cut segments were then cultured individually on half strength of MS medium containing different levels (0.1 - 2.0 mg/l) of BAP or Kn. For regeneration of complete plantlets, isolated shoots were rooted on half strength of MS medium with NAA, IBA or IAA either singly or in combination. Both proliferation and rooting media contained 3% sucrose and gelled with 0.8% agar (Hi-Media, India). The pH was adjusted to 5.7 ± 0.1. All media were steam sterilized under 1.1 kg/cm² pressure at 121⁰C. Cultures were grown at 26 ± 1⁰C under 16 h photoperiod with a light intensity of 2000 - 3000 lux.

Source of Information: Plant Tissue Cult. 12(1) : 27-35, 2002 (June)

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

Among the three types of auxin used, NAA was found to be the best for root induction. The findings are in agreement with those observed in other plant species such as Adhatoda vasica, Caphaelis ipecacuanha, Ocimum sanctum, Plantago ovata, Rehum emodi, Ruscus hypophyllum. About 70 - 80% of the regenerated plantlets could tolerate and survive under ex vitro environment or field conditions. A number of plantlets were lost due to damping off and necrosis during acclimatization in ex vitro condition. Loss of regenerates due to such symptoms was also observed in Eucalyptus tereticornis, Solanum nigrum, Rauvolfia serpentina and Rosa damascena. Due to the delicate nature of in vitro regenerated plantlets, special arrangement such as controlled green house conditions, use of soil free potting mix like perlite, vermiculite, peat plugs and application of fungicides are needed for easy and successful acclimatization of plantlets.

Knowledge Management: Journal