Methods of asepsis for in vitro establishment and germination of Eucalyptus grandis

Technical and scientific studies have been conducted for the cultivation of Eucalyptus, with special attention tolarge-scale clonal propagation. However, in some studies are necessary to obtain aseptic material from seeds, andas far as we are aware efficient protocols of asepsis for this purpose are still not available. Therefore, this studyaimed to define a protocol for sterilization of seeds of Eucalyptus grandis for in vitro establishment andgermination. Four methods of sterilization were tested using different application sequences of the disinfectants, asbenomyl fungicide, hydroalcoholic solution (70% v/v) and sodium hypochlorite (NaOCl). The experiment wasconducted in completely randomized design with five repetitions. The percentage of establishment andcontamination were evaluated at the end of the experiment. Washing with sterile water and 70% alcohol for 30 s,2.5% sodium hypochlorite for 20 min, 1% benomyl for 20 min and three washes with sterile water was the bestmethod found, accounting to 87.5% of establishment.