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In Vitro Direct and Indirect Plantlets Regeneration from Nodal Segment of Boerhaavia Repens L.

A. ROWNAQ
Department of Genetic Engineering and Biotechnology, University of Rajshahi, Rajshahi-6205, Bangladesh

M. S. HOSSAIN
Department of Genetic Engineering and Biotechnology, University of Rajshahi, Rajshahi-6205, Bangladesh

Abstract/ Executive Summary:

An efficient protocol for in vitro propagation of Boerhaavia repens L. was established using nodal segment. The highest direct shoot regeneration from nodal segment was noted (94%) in the MS medium having 2.0 mg/l BAP+0.2 mg/l NAA. The mean number of shoots per culture (12.51) and mean length of shoot per culture (5.10 cm) were also highest in the same medium. Callus induction and plantlet regeneration were observed using different concentrations and combinations of growth regulators. The highest frequency (100%) of organogenic callus was observed in the MS medium containing 1.0 mg/l NAA+1.0 mg/l BAP. Again, the highest (91%) shoot was produced in the MS medium in the combination with 2.0 mg/l BAP+1.5 mg/l Kn+0.5 mg/l NAA was proved to be most suitable in shoot formation from callus. The elongated shoots developed maximum (100%) roots on a half strength MS medium containing 1mg/l IBA. The well rooted plantlets were successfully established in natural condition, where the survival rate of plantlets was 80%

Keywords: In vitro regeneration, Nodal segment, Boerhaavia repens L.

Location: Professor Ali Md. Eunus laboratory, Department of Genetic Engineering and Biotechnology, Rajshahi University, Bangladesh

Start Date:

End Date:

Program Area: Crop-Soil-Water Management

Commodity: Tree

Objectives:

To examine the in vitro regeneration technique that can be used as a potential tool for large scale production of this valuable plant.

Methodology:

The experiment was carried out at the Professor Ali Md. Eunus laboratory, Department of Genetic Engineering and Biotechnology, Rajshahi University, Bangladesh. Nodal segments of Punornova (Boerhavia repens L.) were collected from field grown six months old plants from Rajshahi University campus and washed thoroughly under running tap water and treated with 1% Tween-80 for eight minutes. This was followed by three successive washing with distilled water to make the material free from Tween-80. Further sterilization was done under aseptic condition in a Laminar Airflow Hood. Surface sterilization was carried out with 0.1% HgCl₂ for 3.5 minutes followed by gentle shaking. The material then washed sterilized doubled distilled water for 4-5 times to remove HgCl₂. Explants with single nodal segment of 1 cm in length was cut from the sterilized shoot. Then explants were transferred to 25×150 mm culture tube with 15 ml MS medium supplemented with different hormonal concentrations for direct shoot regeneration. The pH was adjusted to 5.7 prior to autoclaving. The cultures were maintained at 25 ± 2°C under the cool white fluorescent tubes at 3000 lux with a photoperiod of 16 hours. Direct shoot regeneration from nodal segment was cultured in medium with different combinations of BAP (Benzylamino purine), Kn (Kinetin), NAA (1- Naphthalene acetic acid) and IAA (Indole-3-acetic acid). Callus induction from nodal segment was suitable in combinations of BAP and NAA. Shoot formation from callus was cultured in medium with combinations of BAP, Kn and NAA. Well developed shoots were excised and sub cultured on MS medium with different concentrations of NAA, IBA and IAA for root induction. After development of sufficient root system the regenerated plantlets were considered ready to transfer to soil. The roots of the plantlets are gently washed under running tap water to remove agar attached to the root zone. Immediately after that they were transferred to small pots containing a mixture of sand, soil and humus in the ratio of 1:2:2. After sufficient acclimatization, the plantlets were transferred into the larger pot containing soil, where 80% plantlets were survived.

Source of Information: Int. J. Sustain. Crop Prod. 5(4):5-10 (2010)

Article Link (Online Journal):

Funding Source:

1. Government

Budget:

Total Budget (Tk.) :

Achievement/Findings:

In the investigation it was observed that the callus induction and regeneration ability of the nodal explants has the potential for future biotechnological studies for the improvement of Boerhaavia repens L.

Knowledge Management: Journal