Amdadul Huq
Plant Tissue Culture Section, Biological Research Division, Bangladesh Council of Scientific and Industrial Research, Dhanmondi, Dhaka-1205, Bangladesh.
Shahina Akter
Plant Tissue Culture Section, Biological Research Division, Bangladesh Council of Scientific and Industrial Research, Dhanmondi, Dhaka-1205, Bangladesh.
Shahina Islam
Plant Tissue Culture Section, Biological Research Division, Bangladesh Council of Scientific and Industrial Research, Dhanmondi, Dhaka-1205, Bangladesh.
Salim Khan
Plant Tissue Culture Section, Biological Research Division, Bangladesh Council of Scientific and Industrial Research, Dhanmondi, Dhaka-1205, Bangladesh.
In vitro multiplication, Shoot regeneration, Pointed gourd, Transplantation, Acclimatization
Variety and Species
The experiment was conducted at Plant Tissue Culture Section, Bangladesh Council of Scientific and Industrial Research (BCSIR), Dhanmondi, Dhaka. Healthy and profusely growing vines of pointed gourd were collected from various places of Bangladesh such as Magura, Kushtia and Manikgang and used as sources of explants for this experiment. Shoot tips and stem nodes with a single axillary bud were used for this experiment. The explants were surface sterilized with soft detergent for three times followed by washing with a few drops of Tween 20 and thoroughly washed in running tap water for 20-25 minutes. Then the explants were transferred in autoclaved plastic pot and treated with 0.1% mercuric chloride (HgCl2) for 5 minutes for surface sterilization. The sterilized explants were then rinsed 4-5 times with sterile distilled water inside the clean bench to remove all traces of HgCl2. The sterilized explants were excised in the laminar airflow cabinet aseptically using a fine sterile forceps and scalpel. The excised explants were then inoculated in MS (Murashige and Skoog, 1962) medium supplemented with alone or combination of BAP (0.5, 1.0, 1.5, 2.0, 2.5 & 3.0 mg/l), Kn (0.5, 1.0, 1.5, 2.0, 2.5, 3.0 mg/l), NAA (0.1, 0.2, 0.3, 0.4, 0.5 mg/l), IAA (0.1, 0.3, 0.5, 1.0 mg/l) and IBA (0.1, 0.3, 0.5, 1.0 mg/l) for shoot regeneration and multiplication. The pH of the medium was adjusted to 5.7 ± 0.1 using 0.1N sodium hydroxide (NaOH) or 0.1N HCI. In order to solidify the media, laboratory grade agar of 5.5g (0.55%) was added to the solution. The culture tubeswere plugged with aluminum foil and marked with glass marker pen to indicate specific hormonal supplement. The culture tubes were sterilized at 1.09 kg/cm2 pressure at 121oC for 15 minutes in an autoclave. After autoclaving, the culture media were taken out and allowed to cool and solidify. The cultures were maintained in the temperature set on 26 ± 1oC with a light intensity of 2000-3000 lux from fluorescent tubular lamps. The maintained photo period was 16 hours light and 8 hours dark (16 L/8 D) and relative humidity of 60-70%. Successful shoot formations become evident when small green fresh leaves began to emerge. Subcultures carried out regularly at an interval of 4-5 weeks. The percentage of explant induced shoot, days to shoot initiation and number of shoots per explant have been recorded after four weeks of culture. In vitro shoots of pointed gourd were cultured in ½ MS medium supplemented with different concentrations of NAA (0.1, 0.2, 0.3, 0.4, 0.5 and 1.0 mg/1), IAA (0.1, 0.3, 0.5, 1.0 mg/l) and IBA (0.1, 0.3, 0.5, 1.0 mg/l) for root initiation. The well rooted plant lets were then kept in room temperature for 2-3 days and transferred to plastic pot containing garden soil and compost in ratio of 2:1 and moist them adequately for proper hardening.
Bangladesh J. Sci. Ind. Res. 47(2), 217-222, 2012
Journal