Abstract

Tilahun Mekonnen, Mulugeta Diro and Manju Sharma

Regardless of its serious health effect, mercury chloride is frequently utilized for surface sterilization to mitigate microbial contamination in sugarcane tissue culture. The current study aimed at finding an alternative safer and cost effective sterilization method to substitute mercury chloride. In the study, sugarcane shoot tip blocks were treated with three concentrations (1, 3, and 5% active ingredient of chlorine) of local bleach (Berekina) for varying exposure time (10, 15, 20, 25 and 30 min). Surface sterilization with 0.1% mercury chloride for 10 min was used as standard check. Combinations of the surface sterilization treatments were applied to explants of two sugarcane genotypes in completely randomized design. Data were collected on contamination and survival percentage of explants after 15 days of in vitro culturing on Murashige and Skoog (MS) medium supplemented with 2 mg l-1 6- benzylamino purine + 0.5 mg l-1 indole-3- butyric acid. Data were subjected to three way analysis of variance. The study verified that surface sterilization with Berekina 5% ingredients of chlorine for 25 min exposure time is optimal for sugarcane shoot tip decontamination and this treatment combination can replace sterilization with 0.1% mercury chloride for 10 min.