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In Vitro Propagation of Pumpkin and Ash Gourd Through Nodal Segments

M E Haque
Department of Genetic Engineering & Biotechnology, University of Rajshahi, Rajshahi 6205, Bangladesh

M A R Sarkar
Department of Genetic Engineering & Biotechnology, University of Rajshahi, Rajshahi 6205, Bangladesh

M A Mahmud
Department of Genetic Engineering & Biotechnology, University of Rajshahi, Rajshahi 6205, Bangladesh

D Rezwana
Department of Genetic Engineering & Biotechnology, University of Rajshahi, Rajshahi 6205, Bangladesh

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

Abstract/ Executive Summary:

The present study was undertaken with a view to develop an efficient protocol for in vitro multiple shoot formation and subsequent root induction considering various cultural aspects using nodal segments of Cucurbita maxima and Benincasa hispida. The best hormone for shoot multiplication of pumpkin was BAP (2 mg/l), incase of ash-gourd BAP was 1.5 mg/l. For callus induction BAP+2,4-D was best combination for pumpkin and it was 2.0+0.1 mg/l when in ash gourd BAP+NAA was the best combination. IBA + ½MS medium were used for induction of shoot bud root. In both of pumpkin and ash gourd 1.5 mg/l IBA was found best for induction of roots.

Keywords: In vitro, MS medium, Nodal segments, Multiplication, Rooting, Pumpkin and ash gourd.

Location: Department of Genetic Engineering & Biotechnology, University of Rajshahi

Start Date:

End Date:

Program Area: Crop-Soil-Water Management

Commodity: Ash gourd, Pumpkin

Objectives:

The present paper describes rapid, simple and comparatively efficient shoot regeneration and root induction from nodal segments of pumpkin and ash-gourd and it describes the hormonal differences between two species in different aspects of tissue culture.

Methodology:

In both case of pumpkin and ash gourd nodal segments were used as plant material. The segments were cut into pieces carefully with the help of a forcep and dissecting blade. These explants were taken into a conical flask and thoroughly washed under running tap water for 20 minutes to remove loose contaminates attached to explants. Then the explants were washed with distilled water containing 1% savlon (v/v) and two drops of tween-80 for 5 minutes to remove gummy substances. This was followed by successively 3 washing with distilled water to make the materials free from savlon. Subsequently the materials were transferred to laminar airflow cabinet. Washed explants were treated by different concentration of HgCl₂ at different periods of time. After 7 days of incubation, 80% of the explants were free from contamination and tissue killing when pumpkin explants with 0.03% HgCl₂ (w/v) at 3.0 minutes and ash gourd explants with 0.03% HgCl₂ (w/v) at 3.5 minutes. Agar solidifying MS media (1962) containing sucrose 3% and different concentrations and combinations of BAP (0 - 4.5 mg/l), NAA (0.1- 4.5 mg/l), 2,4-D (1.0 -2.5 mg/l) were used for shoot multiplication and callus induction. The pH of the medium was adjusted to 5.7 for pumpkin and 5.6 for ash gourd prior to autoclaving. Initially, the medium (10 ml) was dispensed into test tubes covered with non-absorbent cotton plugs and stream sterilized by autoclaving the medium at 121ºC for 21 minutes at 1.0 Kg/cm² pressure. Cultures were incubated at 25 ± 2ºC under the warm fluorescent light intensity varied from 2000-3000 lux. Rooting of in vitro developed shoots of pumpkin and ash gourd was done by growing on half strength MS medium supplemented with IBA (0.5-4.5 mg/l). The newly transferred cultures were kept in dim light for 3 days, and then they were kept in full light as in shooting.

Source of Information: J. bio-sci. 16: 67-71, 2008 ISSN 1023-8654

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

For induction of roots in pumpkin and ash gourd, half MS + IBA was used. In both cases 1.5 mg/l IBA was the best hormonal dose but the result was not same. In pumpkin mean number of root per explant after 21 days were 6.5 and mean length of the longest root per explant after 21 days 5.5 cm. but in ash gourd mean number of root was 6.5 and mean length of the longest root 5.8 cm. after 21 days.

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