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In Vitro Regeneration of Bitter Gourd (momordica Charantia L.) Through Callus Culture.

J. Halder
Professor
Plant Breeding Division, Bangladesh Institute of Nuclear Agriculture, Mymensingh-2202

S. C. Das
Professor
Department of Biotechnology, Bangladesh Agricultural University, Mymensingh-2202

K. R. Das
Professor
Plant Breeding Division, Bangladesh Institute of Nuclear Agriculture, Mymensingh-2202

M. A. Karim
Professor
Department of Botany, Bangladesh Agricultural University, Mymensingh-2202

M. S. Haque
Department of Biotechnology, Bangladesh Agricultural University, Mymensingh-2202

Abstract/ Executive Summary:

Explant from leaf, node and internode of Momordica charantia were cultured on MS medium supplement with different concentrations of auxin and cytokininin in combination. The best callus induction was observed from all three explants (node, leaf and internodes) on the medium supplemented with 1.5 mg l^-l 2, 4-D with 0.5 mg l^-l BAP and 0.9 mg l^-l NAA with 2.5 mg l^-l BAP. A combination of 1.5 mg/l 2,4-D and 0.5 mg/l BAP produced the highest callus frequency in node (95.00%), leaf (82.50%) and internode (77.50%) segment. In this combination, node segments required minimum number of days (8) while internode segments required the maximum about14 days for callus induction. A combination of 0.9 mg l^-l NAA with 2.5 mg l-l BAP produced the highest callus frequency in node (90.00%), leaf (67.50%) and internode (87.50%) segments. In this combination node segments required minimum number of days (8) while internodes required 12 days for callus induction, which was maximum. Among the explants, the highest average shoot length (3.21 cm) was recorded 0.4 mg l^-l IAA with 2.0 mg l^-l BAP from node segments.

Keywords: In vitro regeneration, Bitter gourd, Nodal segments, Leaf segments, Internode segments

Location: Bangladesh Institute of Nuclear Agriculture, Mymensingh, Bangladesh

Start Date:

End Date:

Program Area: Crop-Soil-Water Management

Commodity: Bitter gourd

Objectives:

To develop and establish an efficient protocol for in vitro plant regeneration, and
To find out most suitable growth regulator combination for regeneration of bitter gourd.

Methodology:

Three explants (Leaf, Node and Internode segments) were collected from field grown plants and were washed with 70% ethanol, 0.1% HgCl2 (w/v) for 3 minutes and rinsed with sterile distilled water for 3 times. Explants were cultured on MS medium fortified with various concentrations and combinations of 2,4-D (0.0, 1.0, 1.5, 2.0 and 2.5) and BAP (0.0, 0.5, 1.0, 1.5 and 2.0) for callus induction. For multiple shoot regeneration, calli derived from all three types of explants were cultured on MS medium strengthened with various concentrations and combinations of 2,4-D, BAP, NAA (0.0, 0.3, 0.6, 0.9 and 1.2), IAA (0.2, 0.4 and 0.6). The culture room was initially cleaned by gentle washing with a detergent followed by wiping with 70% ethanol; the process of sterilization was repeated at regular intervals. The pH of the medium was adjusted to 5.8 ± 0.1 before addition of 0.8% (w/v) agar for solidification. All inoculations and aseptic manipulations were carried out in a Laminar Air Flow Cabinet. Following inoculation, all the cultures were maintained in a 16 h photoperiod, having light intensity of 2000-3000 lux from white fluorescent light at temperature 25±1°C. Sub culturing was carried out at regular intervals. The experiment was laid out in a Completely Randomized Design (CRD) as it was conducted in laboratory condition. The data were analyzed and the mean values were compared with Duncan's New Multiple Range Test (DMRT) (Gomez and Gomez 1984) considering its relative advantages over other tests.

Source of Information: Bangladesh J. Seed Sci. & Tech. 14 (1 &2): 249-255 (2010), ISSN 1029 - 8800

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

From the result, it could be concluded that growth regulators had significant effects on plant regeneration through callus formation from any of the three explants of bitter gourd. However, nodal segments were found most suitable as explants for plant regeneration. The callus formation and regeneration ability of the explants have the potential for future biotechnological studies for general improvement of bitter gourd.

Knowledge Management: Journal