Sterilization Method for Reducing Microbial Contamination and Phenolic Compounds present in Coconut (Cocos Nucifera L.) Leaf Culture

Abstract

The aim of the study was to investigate efficient sterilization methods for reducing microbial contamination and phenolic compound of coconut (Cocos nucifera L.) leaf culture. The non-chlorophyllous immature coconut leaves explant used were taken from unopened spear leaves tissue of the coconut seedling, from the apical growing regions close to the meristem of the palm sucker of about 15 months old. Murashige and Skoog (MS medium) supplemented with 2,4-Dichlorophenoxyacetic acid (2,4-D) at concentration of 30 mg/L and 6-Benzyl amino purine (BAP) at concentration of 1.5 mg/L were used for morphologic responses. Mercuric chloride, ethanol, calcium hypochlorite and sodium hypochlorite were usedto sterilize the explants at concentrations of 0.1 %, 0.2 %, 0.3 % and 0.4 % and 70-95 % of ethanol for 5 minutes. This was followed by rinsing the explants with distilled water four successive times. The sterilized explants were inoculated on MS media and were incubated at 25±2^oC in the dark. Results showed that contamination was less in the cultures, particularly in explants sterilized with 70 % ethanol. Although, all the steriliants did well, but ethanol is more preferable than the rest steriliants, in solving both problems.