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Sterilisation of in vitro Culture Medium of Chrysanthemum by Plant Essential Oils without Autoclaving

Authors: Chockpisit Thepsithar, Aree Thongpukdee, Apichya Daorat

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.

Keywords: Sterilizing agents, essential oils, disinfectants, MS medium, in vitro culture, chrysanthemum, sterilization of medium without autoclaving

Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1086765

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