Evaluation of Antibiotics and Antimicrobial Agents for the Suppression of Contamination in in vitro Cannabis (Cannabis sativa L.) Seedlings

Interest in Cannabis (Cannabis sativa L.) plants for secondary metabolite production has increased in recent years. Although Cannabis plant tissue culture methods are crucial for molecular breeding and micropropagation, the lack of an established protocol offering acceptable quality tissue culture methods for aseptic in vitro Cannabis seed germination is a major obstacle. Here, we established a combination of hydrogen peroxide (H₂O₂) and sodium hypochlorite (NaClO) solutions and examined the effects of several antibiotics, specifically carbenicillin, Augmentin, and Timentin, on the successful suppression of contamination in Cannabis plant tissue cultures. Cannabis seeds were treated with a 1% H₂O₂ solution for one, two, three, and four days and with 0.04%, 0.2%, 0.4%, and 0.8% NaClO for 15 min. A culture medium containing 300 mg/L carbenicillin, 300 mg/L Augmentin, and 300 mg/L Timentin was also used to suppress contamination efficiently. The greatest amount of suppression was obtained upon a treatment with the 1% H₂O₂ solution for two days and the 0.2% NaClO solution for 15 min; subsequent incubation in a culture medium containing 300 mg/L Timentin was an effective antibiotic for minimizing contaminants without negative effects on Cannabis seedling growth.