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Comparison of the Effects of Growth Regulators on in Vitro Regeneration of Ridge Gourd and Sponge Gourd Through Shoot

E Nahar
Department of Genetic Engineering and Biotechnology, University of Rajshahi, Rajshahi 6205, Bangladesh

ME Haque
Department of Genetic Engineering and Biotechnology, University of Rajshahi, Rajshahi 6205, Bangladesh

B Sikdar
Department of Genetic Engineering and Biotechnology, University of Rajshahi, Rajshahi 6205, Bangladesh

Abstract/ Executive Summary:

Context: Luffa cylindrica and Luffa acutangula are highly cross pollinated crops and propagated mainly by seeds. Genetic stability cannot be maintained easily by seed propagation. It can be maintain by developing special vegetative technique through tissue culture. Objectives: To compare the effects of growth regulators between two species of Luffa using shoot tips for develop the rapid, simple and efficient in vitro regeneration protocol. Materials and Methods: Shoot tips used were collected from in vivo grown plants. They were excised from plants and surface sterilized by HgCl₂ treatment. The isolated tips were cultured on semisolid MS medium supplemented with different concentrations and combinations of different growth regulators. Results: The highest result of direct shoot multiplication of ridge gourd was observed using 2 mg/l BAP + 0.2 mg/l GA and in case of sponge gourd it was 1.5 mg/l BAP. For callus induction significant result was found using 4 mg/l BAP + 0.2 mg/l NAA in ridge gourd and 3 mg/l BAP + 0.2 mg/l NAA in sponge gourd. Indirect regeneration was performed by subculturing organogenic callus of sponge gourd on MS with 1.5 mg/l BAP + 0.2 mg/l GA₃ and the callus of ridged gourd on MS + 1.5 mg/l BAP + 0.2 mg/l NAA + 0.2 mg/l nicotinic acid. Regenerated shoots of both species were rooted well on MS containing NAA at low concentration. Conclusion: Hormonal differences and simple rapid in vitro regeneration protocol of L. cylindrica and L. acutangula have been established.

Keywords: Growth regulators, In vitro regeneration, Ridge gourd, Sponge gourd.

Location:

Start Date:

End Date:

Program Area: Crop-Soil-Water Management

Commodity: Ridge gourd, Sponge gourd

Objectives:

To compare the effects of growth regulators between two species of Luffa using shoot tips for develop the rapid, simple and efficient in vitro regeneration protocol.

Methodology:

Shoot tips were collected from in vivo grown plants. The explants were cut into pieces carefully, kept in a conical flask and thoroughly washed under running tap water for 20 min. to remove loose contaminates. They were then treated with 1% savlon for 5 min. and washed five times with sterile distilled water under aseptic condition. For finally surface sterilization, explants were taken in laminar airflow bench and treated with 0.05% aqueous HgCl₂ solution for 2.5 min. The explants were further trimmed to remove excess tissues. Then they were cultured on MS fortified with different concentrations and combinations of BAP (0.5 - 4 mg/l), KIN (0.1 - 0.3 mg/l), NAA (0.1-1.5 mg/l), IBA (0.1- 0.8 mg/l), 2,4-D (0.1-1.5 mg/l), GA₃ (0.1-0.3 mg/l), nicotinic acid (0.2 mg/l) along with 3% sucrose and 0.7% agar. The pH of the medium was adjusted to 5.7 using 0.1 N NaOH or 0.1 N HCl before autoclaving and adding agar. About 10 ml of the medium were taken in each test tube and covered with non-absorbent cotton plug prior to autoclaving at 121°C for 21 min. at 1.5 kg/cm² pressure. Cultures were incubated at 25 ± 2°C under the warm fluorescent light intensity varied from 2000-3000 lux.

Source of Information: J. bio-sci. 18: 88-93, 2010, ISSN 1023-8654

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Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

Hormonal differences and simple rapid in vitro regeneration protocol of L. cylindrica and L. acutangula have been established which will help in conservation and propagation of these two important species.

Knowledge Management: Journal