Explants Sterilization Protocol for In-vitro Propagation of Elite Enset (Ensete ventricosum(Welw.) Chessman) Cultivars

Previous Enset in vitro micropropagation attempts were hindered by explants microbial contamination and for successful in vitro propagation explants specific standard sterilization protocol have to be developed. Therefore, this study was designed to standardized sterilization protocol for shoot tip and leaf sheath explants of three elite Enset (Ensete ventricosum (Welw.) Chessman) cultivar. Sodium hypochlorite under different concentrations (1, 2 and 3% in double sterilization for shoot tip explants and 0.5, 1 and 2% for 10 min immersion for leaf sheath explants) was used for surface sterilization of explants. Explants sterilized with 70% ethanol without NaOCl was used as control. Sterilized explants were cultured on 500 mg/L cefotaxim containing MS medium to evaluate response of explants sterilization to different NaOCl concentration. Results of shoot tip sterilization experiment showed that 2% NaOCl was found to be significantly superior compared to control, 1 and 3% concentration at p<0.05 for obtaining contamination free and growing explants. Among leaf sheath explants sterilization treatments, 1% NaOCl was 100% effective for cultivar Yanbule, while 0.5% NaOCl was found to be best with regard to achieving highest percentage of contamination free healthy leaf sheath culture of Mesena and Endale cultivars.

Author(s): Dejene Zinabu*, Endale Gebre and Jiregna Daksa

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