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Callus Induction and Plant Regeneration from Shoot Tips of Chakunda (cassia Obtusifolia L.)

M. F. Hasan
Department of Genetic Engineering and Biotechnology, University of Rajshahi, Rajshahi-6205, Bangladesh

R. Das
Department of Genetic Engineering and Biotechnology, University of Rajshahi, Rajshahi-6205, Bangladesh

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

M. H. Rashid
Department of Botany, University of Rajshahi, Rajshahi-6205, Bangladesh

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

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

Abstract/ Executive Summary:

The present investigation was conducted at Professor Ali Md. Eunus Laboratory, Depertment of Genetic Engineering and Biotechnology, University of Rajshahi, Rajshahi, Bangladesh, during the period of April to December 2007. The experiment was carried out to develop an efficient protocol for regeneration of Chakunda (Cassia obtusifolia L.) via callus culture from shoot tips, rooting of callus derived shoots and establishment onto the natural condition. Different concentrations and combinations of auxin and cytokinin were used in full strength of medium to observe the callus induction, shoot regeneration and root proliferation. The highest percentage of callus induction was 96.6% observed in the medium supplemented with 2.0 mgL^-1 2,4-D. The highest number of shoots was 5.0 found in the medium having 2.0 mgL^-1 2.4-D+0.2 mgL^-1 Kn. The highest percentage of root induction was 80.0% and the highest number of roots per shoot was 5.0 observed in MS medium consisting of 2.0mgL^-1 NAA. Well rooted plantlets were acclimatized and successfully established onto the natural condition with 70% survival. Regenerated plantlets were morphologically uniform having normal leaf shape and growth.

Keywords: Callus induction, Plant regeneration, Shoot tips, Chakunda

Location: Professor Ali Md. Eunus Laboratory, Depertment of Genetic Engineering and Biotechnology, University of Rajshahi, Rajshahi

Start Date: 31-03-2007

End Date: 30-11-2007

Program Area: Development of Host and Medicinal Plants

Commodity: Medicinal Plants

Objectives:

Objective of this study was to examine the in vitro propagation technique that can be used as a potential tool for large-scale production of this valuable medicinal plant.

Methodology:

The shoot tips of chakunda were collected from the Campus of Rajshahi University, Rajshahi, Bangladesh. They were washed first under running tap water for 30 minutes and treated with 1% Tween-80 for 10 minutes followed by repeated rinsing with autoclave-distilled water. Further sterilization was done under aseptic conditions in a Laminar Airflow Hood. Explants were surface sterilized with 0.1% (W/V) HgCl₂ for 10 minutes. Finally, the explants were washed thoroughly with autoclave-distilled water for several times to remove traces of HgCl₂. The shoot tips were cut into convenient size (1 cm) and cultured on MS medium consisting of different concentrations and combinations of auxin and cytokinin. Throughout the experiments full strength MS medium with 3% (W/V) sucrose and gelled with 0.8% (W/V) agar was used. The pH of all media (supplemented with respective growth regulators) was adjusted to 5.8 with 1 N NaOH or 1N HCl prior to autoclaving (21 min). The cultures were incubated in a culture room at 25±2^oC with a photoperiod of 16 hour at 3000-lux light intensity provided by cool white fluorescent tubes. In this investigation, the basal medium was supplemented with different concentrations and combinations of auxin and cytokinin for callus induction. Once the callus developed, they were further cultured for regeneration and elongation in the medium having different concentrations and combinations of auxin and cytokinin. Root induction from the callus-derived shoots was achieved on MS medium supplemented with NAA/IBA at different concentrations. Well developed plantlets were removed from the culture vessels, washed gently under running tap water and planted in plastic pots containing a potting mixture of sand, soil and farmyard manure in the ratio of 1:1:1. The potted plantlets were covered by polythene bag to maintain suitable humidity. After 30-35 days of bagging, the plantlets were transplanted in the natural condition, where 70% plants were survived.

Source of Information: Int. J. Sustain. Crop Prod. 3(6):6-10, 2008

Article Link (Online Journal):

Funding Source:

1. Government

Budget:

Total Budget (Tk.) :

Achievement/Findings:

In this investigation, a reproducible protocol for plant regeneration was established through callus induction from nodal explants in C. obtusifolia L. It is expected that a standard protocol to induce callus and rapid proliferation of shoots through in vitro culture would provide a more homogeneous source of medicine.

Knowledge Management: Journal