Protocol Optimization for Surface Sterilization of Sugarcane Variety (B-52/298)

Abstract

The aim of this study was to optimize conditions for surface sterilization of ex-plants of B-52/298 sugarcane cultivar variety. Experiment was carried out to determine the optimum concentrations of surface sterilizing agents and at what time they optimally sterilize. This experiment was conducted at MIT Plant Tissue Culture and Micro-propagation Laboratory Center, situated in Mekelle City. Three chemisterilants (Sodium Hypochlorite, Mercuric Chloride and Ethanol) were taken for the investigation. Three of the sterilizing agents were used in different concentrations at various time intervals and each fungal and bacterial contamination was evaluated in intervals of three days from the initiation time for two weeks. The numbers of each contamination, clean cultures and dead ones were counted. Most of the contaminations were found to be fungal, while there was considerable numbers of cultures that were of bacterial contamination. Only a few of the cultures were observed as dead. Among these sterilizing chemicals, 60% Ethanol was found to be a better sterilizing agent when acted on the ex-plants for 6.5 minutes, resulting in 33.3% clean cultures.