Published Paper Details:

Abstract During In-vitro studies, there was an investigation on the effect of the use of different surface sterilizing agents on nodal explants of Dendrocalamus hamiltonii. Surface sterilization is the most vital step during the preparation of healthy and viable explants for the micropropagation technique. The collection of explants was done from CPTs, that were highly exposed to microbial contaminants. Most contaminants such as bacteria and fungi are discarded by surface sterilization with the help of appropriate sterilizing agents. There was the use of HgCl2 and NaOCl, and Ca (OCl)2 a sterilizing agent during in-vitro culture. The leaf sheath covering the nodal segments containing axillary buds (1.5-2.00cm) was carefully removed and cleaned with 70% ethanol using sterilized cotton. These segments were pre-treated with 0.5% Bavistin as fungicides for 15 min. After pre-treatment segments were rinsed with Savlon and fresh running tap water at least three times. Further surface sterilization of explants was done under a laminar airflow cabinet with different treatments of HgCl2 and Ca (OCl)2 at different concentrations. Two methods of surface sterilization were used under a laminar airflow cabinet, (a) the explants were treated with 0.1% HgCl2 solution for 10,15, and 20 minutes. It was thoroughly washed with sterile distilled water. It was then 30sec.dip within 70% ethanol and again rinsed in sterile distilled water. (b) the explants were treated for 10, 15, and 2o min with Ca (OCl)2 with a few drops of tween-20. After this treatment, explants were washed with sterile distilled water, and they were then given a 30 sec. dip in 70% ethanol and again rinsed in sterile distilled water. After these sterilizing agents were used for the satisfactory result of the aseptic culture, viability, and bud break; sterilization with 0.1% HgCl2 for 20 min and 30min continuous rinsing showed 68% Aseptic buds with 52% bud break whereas Ca (OCl)2 (10min.) and similarly rinsing for 30min. showed 17% Aseptic buds with 12% Bud-break.