Sterilization of non-autoclavable vessels and culture media by sodium hypochlorite for in vitro culture.

摘要

As an alternative for sterilization by autoclaving, disinfection protocols for vessels and media using sodium hypochlorite (NaOCl) were developed for banana (Musa spp.) shoot and Agrobacterium tumefaciens cultures. The NaOCl concentration of 0.002% was sufficient to prevent microbial growth when the disinfested vessels were incubated with a modified MS medium for banana shoot cultures. However, instead of the MS medium, when they were incubated with a MYA medium for Agrobacterium cell cultures, bacterial contamination appeared, and a higher concentration of NaOCl was necessary to have an efficient disinfection. Culture media disinfected by NaOCl were successfully achieved for banana but not for Agrobacterium cell cultures. For plant tissue cultures, non-autoclavable vessels are suggested to be disinfected by the solution of 0.01% NaOCl at pH 5.4, and culture media by supplementing with 0.002% NaOCl. For Agrobacterium cell cultures, vessel disinfections by 0.01% NaOCl are suggested, but media could not be disinfected by the NaOCl supplementation.