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In Vitro Propagation of Bitter Gourd (momordica Charantia L.) from Nodal and Root Segments

M. A. Z. Al Munsur
Department of Biotechnology, Bangladesh Agricultural University, Mymensingh

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

K. M. Nasiruddin
Department of Biotechnology, Bangladesh Agricultural University, Mymensingh

M. S. Hossain
Department of Biotechnology, Bangladesh Agricultural University, Mymensingh

Abstract/ Executive Summary:

Explants of nodal and root segments of bitter gourd were cultured on MS supplemented with various concentrations of BAP in combination with either 2,4‐D or NAA. Nodal segments produced the highest percentage (93.75) of callus in MS supplemented with 1.0 mg/l 2,4‐D and 1.0 mg/l BAP whereas, root segments produced the highest (85.00%) callus in 0.6 mg/l NAA and 2.5 mg/l BAP combination. A combination of 1.0 mg/l 2,4‐D and 1.0 mg/l BAP exhibited 75.00% shoot regeneration from nodal segments. The highest shoot length (5.15 cm) was recorded with 2.5 mg/l BAP and 0.2 mg/l IAA from nodal segments. No sign of regeneration of shoot was found from root segments in any of the combinations. However, some combinations produced only roots.

Keywords: In vitro propagation, Bitter gourd, Nodal segments, Root segments

Location: Mymensingh

Start Date: 01-02-2006

End Date: 30-11-2006

Program Area: Variety and Species

Commodity: Bitter gourd

Objectives:

The present study was carried out to select the most suitable explants and growth regulators for in vitro regeneration and to develop an efficient protocol for in vitro plant regeneration

Methodology:

The experiment was carried out during February to November, 2006 in the laboratory of the Department of Biotechnology, Bangladesh Agricultural University, Mymensingh. Seeds of a hybrid variety of bitter gourd (Momordica charantia L.) marketed by the East West Seed (Bangladesh) Ltd. were purchased from the local market. For better germination, collected seeds were treated with hot water (55 ‐ 57°C) for 15 min by using “Vegetable Seed Treating device” developed by IPM Lab., Department of Plant Pathology, Bangladesh Agricultural University. Then they were dried overnight under shade. Surface sterilization of seeds was done with 0.1% (w/v) HgCl₂ solution for five minutes. The seeds were then rinsed for three ‐ four times by sterile distilled water to remove traces of the HgCl₂_. Then the seeds were placed on hormone free half strength MS medium. Seeds started germination within four days of inoculation and subsequently developed into rooted plantlets within ten to 12 days. Nodal and root segments were dissected from the plantlets and were used as explants. Explants were cultured on MS supplemented with various concentrations and combinations of 2,4‐D (0.0, 0.25, 0.50, 1.0 and 5.0 mg/l), BAP (0.2, 0.5, 1.0, 2.0, 2.5 and 3.0 mg/l) and NAA (0.1, 0.3 and 0.6 mg/l) for callus induction. For shoot regeneration, calli derived from both types of explants were cultured on MS fortified with various concentrations and combinations of 2,4‐D, BAP, NAA, IAA (0.1, 0.2, 0.6 mg/l), IBA and GA3 ( 0.1 and 0.2 mg/l). The pH of the medium was adjusted to 5.8 ± 0.1 before addition of 0.8% (w/v) agar for solidification. All the cultures were maintained in 16 hr photoperiod, with light intensity of 2000 ‐ 3000 lux at 25 ± 1°C. Subcultures were carried out at regular intervals. After sufficient development of shoot and root systems, the small plantlets were taken out from the culture vessels and washed to remove excess of agar around the roots. Then it was placed in normal environment for acclimatization. Then they were transplanted to small pots. The experiments were arranged in CRD with four replications. Data recorded for different parameters under study were statistically analyzed. The significance of difference between the pair of means was evaluated at 5% level of significance by DMRT (Gomez and Gomez 1984).

Source of Information: Plant Tissue Cult. & Biotech. 19(1): 45-52, 2009 (June)

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The highest number of roots per callus (7.55) was found in 3.0 mg/l BAP and 0.1 mg/l NAA and lowest 4.25 was found in 3.0 mg/l BAP and 0.30 mg/l NAA. The highest root length (2.75 cm) was found in 3.0 mg/l BAP and 0.10 mg/l NAA and lowest was found in 2.0 mg/l BAP and 0.3 mg/l NAA. It was observed that the highest number (8.25) of root per callus and the highest length of root (2.75 cm) were obtained from 0.5 mg/l NAA.After sufficient development of shoot and root, the small plantlets were transplanted in small pots containing soil. When the plantlets grew to a height of 5 ‐ 6 cm, they were transferred to plastic pots. The growth of plantlets was satisfactory. The rate of success of plantlet survival was recorded to be above 75 per cent. The established plants in pots were kept in natural condition to grow them with proper care.

Knowledge Management: Journal