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Efficient Direct Organogenesis from Shoot Tips and Nodal Segments of Ash Gourd (benincasa Hispida L.)

M. Kausar
Professor Joarder DNA and Chromosome Research Lab. Department of Genetic Engineering and Biotechnology University of Rajshahi, Rajshahi 6205, Bangladesh

S. Parvin
Professor Joarder DNA and Chromosome Research Lab. Department of Genetic Engineering and Biotechnology University of Rajshahi, Rajshahi 6205, Bangladesh

M.E. Haque
Professor Joarder DNA and Chromosome Research Lab. Department of Genetic Engineering and Biotechnology University of Rajshahi, Rajshahi 6205, Bangladesh

M. Khalekuzzaman
Professor Joarder DNA and Chromosome Research Lab. Department of Genetic Engineering and Biotechnology University of Rajshahi, Rajshahi 6205, Bangladesh

B. Sikdar
Professor Joarder DNA and Chromosome Research Lab. Department of Genetic Engineering and Biotechnology University of Rajshahi, Rajshahi 6205, Bangladesh

M.A. Islam*
Professor Joarder DNA and Chromosome Research Lab. Department of Genetic Engineering and Biotechnology University of Rajshahi, Rajshahi 6205, Bangladesh

Abstract/ Executive Summary:

The effect of external application of phytohormone on explants viz., shoot tips and nodal segments of ash gourd (Benincasa hispida L.) was tested on the frequency of shoot and root induction. Shoot tips and nodal segments were cultured on MS medium supplemented with different concentrations and combinations of cytokinins (BAP, Kinetin), auxin (IBA, NAA) and gibberellic acid (GA3) for multiple shoot formation and root induction. The highest number (up to 90%) of multiple shoot formation was obtained from the shoot tips in MS medium fortified with 1.5 mg/l BAP + 0.2 mg/l GA3, where average number of shoots per culture was 5.55. In case of nodal segment, better response (up to 78%) for shoot multiplication was found in MS with 2.0 mg/l BAP + 0.2 mg/l GA3. The concentration of 1.0 mg/l IBA was found to be most effective for root initiation in microshoot developed from both types of explants. Rooted plantlets were acclimatatized and established in sandy soil with good survival rate

Keywords: Organogenesis; Ash gourd; MS medium; Shoot tips; Nodal segments

Location: Department of Genetic Engineering and Biotechnology, University of Rajshahi, Bangladesh

Start Date:

End Date:

Program Area: Seed Technology

Commodity: Ash gourd

Objectives:

The present work was designed to develop an efficient protocol for in vitro regeneration of ash gourd which could be used further in genetic transformation system for its development.

Methodology:

The work was carried out at Professor Joarder DNA and Chromosome Research Laboratory, Department of Genetic Engineering and Biotechnology, University of Rajshahi, Bangladesh. Shoot tips and nodal segments were collected from mature healthy in vivo grown plants of Benincasa hispida and the explants were washed thoroughly in running tap water for 15 min. Then these were treated with 1% Tween-80 for eight minutes and again washing with distilled water to make the materials free from Tween-80. Further sterilization was done under aseptic condition in a laminar airflow cabinet. Surface sterilization was carried out with 0.05% HgCl2 for 2 minutes by gentle shaking and rinsed 4-5 times in sterilized doubled distilled water to remove HgCl2. Explants were cut into 1 cm in length and transferred in culture tube with 15 ml agar semisolified with 1.0% Difco Bacto agar MS (Murashige and Skoog, 1962) medium. The medium was supplemented with 3% sucrose and different hormonal concentrations BAP (0.5-3 mg/l), GA3 (0.1-0.2mg/l), Kinetin (0.1-0.2mg/l), IBA (0.5-2.0 mg/l), NAA ((0.1- 0.2mg/l and 0.5-2.0 mg/l) for direct organogenesis. pH of the medium was adjusted to 5.7 and was autoclaved by stream sterilization at 121°C for 22 min at 1.0 kg/cm3 pressure. The cultures were maintained at 25 ± 2°C under the cool white fluorescent tubes at 3000 lux with a photoperiod of 16 hours. The shoot formation was checked by 3 times of culture in same medium. Then in vitro developed shoots were subcultured in root induction medium with different concentrations of IBA and NAA. After development of sufficient roots, the micro plantlets were considered ready to transfer in soil. The roots of the plantlets are gently washed by distilled water to remove agar attached to the root zone. Immediately these were transferred to small pots containing a mixture of sand, soil and humus in the ratio of 1:2:2 for acclimatization.Thenewly transplanted plantlets were covered by polythene paper and kept in growth chamber for 7 days. Polythene covering was removed and the plantlets were transferred into the larger pot containing soil. Each treatment had 10 replicates of culture and experiments were repeated twice

Source of Information: J. Life Earth Sci., Vol. 8: 17-20, 2013

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

Micropropagation of ash gourd has been achieved through in vitro culture of nodal segments and shoot tips as explants. The technique described appears to be readily adaptable for large scale micro propagation of ash guard.

Knowledge Management: Journal