IN VITRO CULTURE OF Sequoia sempervirens L. ON NUTRITIVE MEDIA STERILIZED WITH SODIUM HYPOCHLORITE
DOI:
https://doi.org/10.5902/198050982749Keywords:
autoclaving, chemical sterilization, contamination.Abstract
The autoclaving used for sterilization of glassware, culture media and surgical materials in laboratory is a costly operation, due to the high cost of the equipment and the equally high consumption of energy. For these reasons, the substitution of this sterilization technique for another less costly one, such as chemical sterilization, would be highly desirable. The present study aimed to compare the techniques of sterilization of plant tissue culture media with sodium hypochlorite and that of autoclaving, in Sequoia sempervirens culture, in order to develop a less costly technique in the sterilization of glassware and nutrient media for plant tissue culture. In the trial with Sequoia sempervirens, the concentrations of sodium hypochlorite added to the culture media were (w/v): 0% (autoclaved); B) 0.002%; C) 0.003%; D) 0.004% and E) 0% (without autoclaving). It was observed that the concentrations equal to or higher than 0.003% of total chlorine added to the nutrient media resulted in complete sterilization, as well as in plants with larger numbers and shoots lengths.
Downloads
References
BIOACTIVOS QUÍMICOS CBQ CENTRO. Diaz Martinez Grisell; Conzalez Bedia Martha Mayra; Salazar Yera Eloisa; Castanedo Cancio Nilo Ramon; Cueto Sanchez Madaisy de la Ca; Gonzalez Lorenzo Carlos Manuel; Machado Rodriguez Rosa Margari; Martin Triana Esther Lilia; Ramirez Dieguez Alain. Microcide composition. AO1N43/00. CUEP 09208 04. 23 jun. 1997. Disponível em < http://v3.espacenet.com/texto?AB>. Acesso em: 23 abr. 2005.
EMMANUEL, E. et al. Toxicological effects of disinfestations using sodium hypochlorite on aquatic organisms and its contribution to AOX formation in hospital wastewater. Environment International, Oxford, v. 30, p. 891-900, 2004.
GONZÁLES, E. A. J. Generalidades del cultivo in vitro . In: PONCE, J. N. P. (Ed.). Propagación y mejora genética de plantas por biotecnología. Villa Clara: Instituto de Bioteconologia de las Plantas, 1998. p. 13-24.
MACEDO, J. A. B. de. Uso de derivados clorados orgânicos no processo de desinfecção de água para abastecimento público. In: CONGRESSO BRASILEIRO DE QUÍMICA, 44., 2004, Fortaleza. Anais... Sociedade Brasileira de Química, 2004. Disponível em: http:// www.jorgemacedo.pro.br/CBQ%202004%20O%20USO%20DE%20DERIVADOS%20 CLORADOS%20ORGANICOS.pdf>. Acesso em: 15 jun. 2009.
MEYER, S. T. O uso do cloro na desinfecção de águas, a formação de trihalometanos e os riscos potenciais à saúde pública. Caderno de Saúde Pública, Rio de Janeiro, v. 10, n. 1, p. 99-110, 1994.
MURASHIGE, T.; SKOOG, F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum, Copenhagen, v. 15, p. 473-497, 1962.
PÉREZ, P. A. O. Introduccion a la propagación masiva. GONZÁLES, E. A. J. Generalidades del cultivo in vitro . In: PONCE, J. N. P. (Ed.). Propagación y mejora genética de plantas por biotecnología. Villa Clara: Instituto de Bioteconologia de las Plantas, 1998. p. 125-133.
GURI, A. Z.; PATEL, K. N. Compositions and methods to prevent microbial contamination of plant tissue culture media. C12N005/00; C12N005/02. 2 jun. 1995, 5,750,402 12 maio 1998. Disponível em: http://www. kitchenculturekit.com/ppmPatent5,750,402.pdf> Acesso em: 23 abr. 2005.
PONCE, J. N. P.; CASTELLÁ, M. S.; PÉREZ, P. O. Possibilidades y potencial de la propagación masiva de plantas en Cuba. Biotecnología Vegetal, .Cuba, v. 1, p. 3-12, 2000.
SHERBONDY, A. L. et al.Variability in catheter microwave sterilization techniques in a single clinic population. Journal of Urology, Linthicum, v. 168, p. 562-564, 2002.
TAIZ, L.; ZEIGER, E. Fisiologia vegetal. 3. ed. Porto Alegre: Artmed. 2004. 719 p.
TEIXEIRA, S. L.; SOUSA, R. T. S.; TEIXEIRA, M. T. Esterilização de meios nutritivos para cultura de tecidos vegetais em forno de microondas. Revista Ceres, Viçosa, v. 52, n. 302, p. 499-507, 2004.
TEIXEIRA, S.,L. et al. Sterilization of nutrient medium for plant tissue culture, by combining chemical sterilants with microwave oven. Revista Ceres, Viçosa, v. 52, n. 301, p. 343-349, 2005.
TEIXEIRA, S. L.; RIBEIRO, J. M.; TEIXEIRA, M. T. Influence of NaClO on nutrient medium sterilization and on pineapple (/Ananas comosus/ cv Smooth cayenne) behavior. Plant Cell, Tissue and Organ Culture, Holanda, v. 86, n. 3, p. 375-378, 2006.
TEIXEIRA, S. L.; RIBEIRO, J. M.; TEIXEIRA, M. T. Utilização de hipoclorito de sódio na esterilização de meio de cultura para multiplicação in vitro de Eucalyptus pellita. Ciência Florestal, Santa Maria, v. 8, p. 185-191, 2008.
YANAGAWA, T. et al. Application of desinfectants to orchid seeds, plantelets and media as a means to prevent /in vitro/ contamination. Lindleyana, Palm Beach, v. 10, n. 1, p. 33-36, 1995.
WHITE, P. R. Nutrient deficiency studies and an improved inorganic nutrient medium for cultivation of excised tomato roots. Growth, Lakeland, v. 7, p. 53-65, 1943.