Abstract: The alternative technique for sterilization of culture
medium to replace autoclaving was carried out. For sterilization of
culture medium without autoclaving, some commercial pure essential
oils, bergamot oil, betel oil, cinnamon oil, lavender oil and turmeric
oil, were tested alone or in combinations with some disinfectants,
10% povidone-iodine and 2% iodine + 2.4% potassium iodide. Each
essential oil or combination was added to 25-mL Murashige and
Skoog (MS) medium before medium was solidified in a 120-mL
container, kept for 2 weeks before evaluating sterile conditions.
Treated media, supplemented with essential oils, were compared to
control medium, autoclaved at 121 degree Celsius for 15 min. In
vitro sterile conditions were found 20 – 100% from these treated
media compared to 100% sterile condition from autoclaved medium.
Treated media obtained 100% sterile conditions were chosen for
culturing chrysanthemum shoots. It was found that 10% povidoneiodine
in combination with cinnamon oil (3:1) and 2% iodine + 2.4%
potassium iodide in combination with lavender oil (1:3) at the
concentration of 36 3L/25 mL medium provided the promising
growth of shoot explants.
Abstract: Plant tissue culture is an important in vitro technology applied for agricultural and industrial production. A sterile condition of culture medium is one of the main aspects. The alternative technique for medium sterilization to replace autoclaving was carried out. For sterilization of plant tissue culture medium without autoclaving, ten commercial pure essential oils and 5 disinfectants were tested. Each essential oil or disinfectant was added to a 20-mL Murashige and Skoog (MS) medium before medium was solidified in a 120-mL container, kept for 2 weeks before evaluating sterile conditions. Treated media, supplemented with essential oils or disinfectants, were compared to control medium, autoclaved at 121 degree Celsius for 15 min. Sterile conditions of MS medium were found 100% from betel oil or clove oil (18 mL/20 mL medium), cinnamon oil (36 mL/20 mL medium), lavender oil or holy basil oil (108 mL/20 mL medium), and lemon oil or tea tree oil or turmeric oil (252 mL/20 mL medium), compared to 100% sterile condition from autoclaved medium. For disinfectants, 2% iodine + 2.4% potassium iodide, 2% merbromine solution, 10% povidone-iodine, 6% sodium hypochlorite or 0.1% thimerosal at 36 mL/20 mL medium provided 100% sterile conditions. Furthermore, growth of new shoots from chrysanthemum node explants on treated media (fresh weight, shoot length, root length and number of node) were also reported and discussed in the comparison of those on autoclaved medium.