Surface sterilization is a vital step in preparation of healthy and viable explants in tissue culture because plants in the field are highly exposed to microbial contamination. Most surface contaminants, such as bacteria and fungi, can be eliminated by surface sterilization with a suitable sterilizing agent. In this study three surface sterilizing agents were used: plant preservative mixture (PPM), mercuric chloride (HgCl₂) and sodium hypochlorite (NaOCl). Different concentrations and time exposure were tested for decontamination and shoot development of the nodal segments of Cestrum nocturnumL. in vitro. The results showed that among the three sterilizing agents used in this study, PPM treatment at concentrations (4,6%) for 4 hours of constant shaking were effective in terms of decontamination percentage (100), survival and viability of the nodal segments and shoot development percentage (70%). The PPM treatment at all concentrations (1,2,4,6%) for 12 hours of constant shaking gave all dead nodal segments, thus viability of the nodal segments was affected. Sterilization with HgCl2 at all concentrations (0.05,0.1,0.2%) for 7 minutes of continuous rinsing showed good results in terms of decontamination (90-100%) and survival and shoot development percentages (50-60%), while (NaOCl) at higher concentration of (50%) for 5 minutes of continuous rinsing showed satisfactory result of viability and shoot development (50%). The numbers and percentages of contaminated and survived nodal segments were recorded after 30 days. All Viable nodal segments developed to shoots. It had been concluded that the best surface sterilization agent was PPM.