Surface Sterilization Protocol for Nodal Cultures of Potato (Solanum tuberosum L.)

Abstract

The process of surface sterilizing plant materials is a difficult step in constructing a plant tissue culture technique. Nodal explants from potted plants of Solanum tuberosum L. grown in the screenhouse were given various surface sterilization procedures before being introduced into sterile Murashige and Skoog (MS) culture. A total of 16 treatments were performed with varied periods of immersion in two sterilant successions: 70% (v/v) ethanol (20 s, 1 min, 3 min, and 5 min) and 10% (w/v) calcium hypochlorite (Ca(ClO)2) granules (5, 10, 15, and 20 min) at varying times, followed by three sterile distilled water rinses. After 4 weeks in culture, 70 % ethanol (20 s)+10 % Ca(ClO)2 (15 min) and 70 % ethanol (3 min)+10 % Ca(ClO)2 (20 min) treatments yielded 100 % clean culture. 70 % ethanol (20 seconds) Plus 10% calcium (ClO) 2 (15 min) is recommended as the optimum surface sterilizing protocol since it produced 90% growth compared to 60% growth in 70% ethanol (3 min) + 10% Ca(ClO)2 (20 min). This is a preliminary study, and more research is needed to develop an efficient, successful, and repeatable methodology for surface sterilization of various explants utilized for tissue culture of S. biafrae acquired from the open field.

Keywords: calcium hypochlorite (Ca(ClO)2, Murashige & Skoog, nodal explant, Solanum tuberosum L., surface sterilization.

DOI: 10.7176/JBAH/11-13-03

Publication date:July 31^st 2021