Apical meristems and vegetative buds of different sizes were used as explants. Explants were obtained from pot grown plants from greenhouse of Department of Botany, Lahore College for Women University, Lahore, Pakistan. These were washed thoroughly with tap water and then with house hold detergent to remove all the traces of dust particles. The explants were then immersed in 20% aqueous solution of sodium hypochlorite for 20 min and were thoroughly washed. Sodium hypochlorite solution was decanted and explants were rinsed three times with autoclaved distilled water to remove all the traces of sterilant. While dealing with Gerbera, it was recognized that sterilization was very tedious and the above mentioned sterilization method was not satisfactory. So to get the explants free of contamination, the following method was opted: The explants were surface sterilized with the method described above. Then, these were shifted into a mixture of MS medium (100 ml) without sugar. From this MS (100 ml) mixture, 5% mixture was removed and replaced with 5% plant preservative mixture [PPM (5-chloro-2-methyl-3(2H)-isothiazolone + 2-methyl-3(2H)-isothiazolone + inert ingredients)]. Then, the flask (containing MS mixture + explants) was placed on a shaker for 3 h at 122 rpm at 37°C for
removal of all types of contamination. Inoculation was carried out in laminar air flow cabinet. It was cleaned by scrubbing with 70% ethanol solution and was irradiated with UV radiations for 25 min before use.
Apical meristems and vegetative buds of different sizes were used as explants. Explants were obtained from pot grown plants from greenhouse of Department of Botany, Lahore College for Women University, Lahore, Pakistan. These were washed thoroughly with tap water and then with house hold detergent to remove all the traces of dust particles. The explants were then immersed in 20% aqueous solution of sodium hypochlorite for 20 min and were thoroughly washed. Sodium hypochlorite solution was decanted and explants were rinsed three times with autoclaved distilled water to remove all the traces of sterilant. While dealing with Gerbera, it was recognized that sterilization was very tedious and the above mentioned sterilization method was not satisfactory. So to get the explants free of contamination, the following method was opted: The explants were surface sterilized with the method described above. Then, these were shifted into a mixture of MS medium (100 ml) without sugar. From this MS (100 ml) mixture, 5% mixture was removed and replaced with 5% plant preservative mixture [PPM (5-chloro-2-methyl-3(2H)-isothiazolone + 2-methyl-3(2H)-isothiazolone + inert ingredients)]. Then, the flask (containing MS mixture + explants) was placed on a shaker for 3 h at 122 rpm at 37°C forremoval of all types of contamination. Inoculation was carried out in laminar air flow cabinet. It was cleaned by scrubbing with 70% ethanol solution and was irradiated with UV radiations for 25 min before use.
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