In vitro Culture Medium Sterilization by Chemicals and Essential Oils without Autoclaving and Growth of Chrysanthemum Nodes
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.
Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1079020Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 5595
 A. D. Russell, "Similarities and differences in the responses of microorganisms to biocides.” Journal of Antimicrobial Chemotherapy, vol. 52, no. 5, pp. 750–763, 2003.
 United State Patent 5750402. "Compositions and methods to prevent microbial contamination of plant tissue culture media.”
[Online]. Available; http://www. freepatentonline.com/ 5750402.html.
 U. Habiba, S. Reza, M. L. Saha, M. R. Khan, and S. Hadiuzzaman, "Endogenous bacterial contamination during in vitro culture of table banana: Identification and prevention.” Plant Tissue Culture, vol. 12, no. 2, pp. 117–124, 2002.
 S. L. Teixeira, "The development of a new protocol that uses sodium hypochlorite to replace the autoclaving procedure for establishing axenic in vitro banana (Musa sp.) culture.” Agricell Report, vol. 47, no. 3, pp. 17–18, 2006.
 T. Yanagawa, M. Nagai, T. Ogino, and M. Maeguchi, "Application of disinfection to orchid seeds, plantlet and media as a means to prevent in vitro contamination.” Lindleyana, vol. 10, pp. 33–36, 2006.
 M. Chansean, and I. Syoichi, "Conservation of Wild Orchids in Cambodia by Simple Aseptic Culture Method.” Proceedings of NIOC, pp. 13–19, April, 2007.
 M. M. Hoque, S. Rattila, M. A. Shishir, M. L. Bari, Y. Inatsu, and S. Kawamoto, "Antibacterial activity of ethanol extract of betel leaf (Piper betle L.) against some food borne pathogens.” Bangladesh Journal of Microbiology, vol. 28, no. 1, 58–63, 2011.
 M. Seema, S. S. Sreenivas, N. D. Rekha, and N. S. Devaki, "In vitro studies of some plant extracts against Rhizoctonia solani Kuhn infecting FCV tobacco in Karnataka Light Soil, Karnataka, India.” Journal of Agricultural technology, vol. 7, no. 5, pp. 1321–1329, 2011.
 D. Srichana, A. Phumruang, A. and Chongkid, B. "Inhibition effect of betel leaf extract on the growth of Aspergillus flavus and Fusarium verticillioides.” Thammasat International Journal of Science and Technology, vol. 14, no. 3, pp. 74–77, 2009.
 T. S. A. T. Kamazeri, O. A. Samah, M. Taher, D. Susanti, and H. Qaralleh, "Antimicrobial activity and essential oils of Curcuma aerugonosa, Curcuma mangga and Zingiber cassumunar from Malaysia.” Asian Pacific Journal of Tropical Medicine, vol. 5, no. 3, 202–209, March, 2012.
 B. Joshi, G. P. Sah, B. B. Basnet, M. R. Bhatt, D. Sharma, K. Subedi, J. Pandey, and R. Malla, "Phytochemical extraction and antimicrobial properties of different medicinal plants: Ocimum sanctum (tulsi), Eugenia caryophyllata (clove), Achyranthes bidentata (datiwan) and Azadirachta indica (neem).” Journal of Microbiology and Antimicrobials, vol. 3, no. 1, pp. 1–7, January, 2011.
 I. N. Fit, G. Rapuntean, S. Rapuntean, F. Chirila, and G. C. Nadas, "Antibacterial effect of essential essential vegetal extracts on Staphylococcus aureus compared to antibiotics.” Notulae Botanicae Horti Agrobotanici Cluj-Napoca, vol. 37, 117–223, 2009.
 L. Hui, L. He, L. Huan, L. Xiaolan, and Z. Aiguo, "Chemical composition of lavender essential oil and its antioxidant activity and inhibition against rhinitis related bacteria.” African Journal of Microbiology Research, vol. 4, pp. 309–313, 2010.
 F. G. Kirbaslar, A. Tavman, B. Dulger, and G. Turker, "Antimicrobial activity of Turkish Citrus peel oils.” Pakistan Journal of Botany, vol. 41, pp. 3207–3212, 2009.
 S. S. Allawi, J. M. Auda, H. Q. Hameed, and T. I. Ali, "The effect of Curcuma longa (turmeric) rhizomes extracts on pathogenic bacteria in comparison with standard antibiotics.” Journal of Biotechnology Research Center 3, pp. 15–20, 2009.
 T. Murashige, and F, Skoog, "A revised medium for rapid growth and bioassays with tobacco tissue culture.” Physiologia Plantarum, vol. 15, no. 3, pp. 473–497, 1962.
 D. B. Duncan, "Multiple range and multiple F test.” Biometrics, vol. 11, pp. 1–42, 1995.