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

Abstract

Technical and scientific studies have been conducted for the cultivation of Eucalyptus, with special attention to large-scale clonal propagation. However, in some studies are necessary to obtain aseptic material from seeds, and as far as we are aware efficient protocols of asepsis for this purpose are still not available. Therefore, this study aimed to define a protocol for sterilization of seeds of Eucalyptus grandis for in vitro establishment and germination. Four methods of sterilization were tested using different application sequences of the disinfectants, as benomyl fungicide, hydroalcoholic solution (70% v/v) and sodium hypochlorite (NaOCl). The experiment was conducted in completely randomized design with five repetitions. The percentage of establishment and contamination 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 best method found, accounting to 87.5% of establishment.