A Robust Protocol for Managing Microbial Contamination of In-vitro Banana Plants

In-vitro regeneration of banana (Musa spp) is a crucial technique in banana improvement via modern biotechnology like virus indexing, genetic transformation, and genome editing. However, in-vitro banana plants are prone to microbial contamination from the environment, leading to the loss of important lines and germplasm. Protocols for disinfecting banana plants before their in-vitro culture have been reported; however, there is limited information on strategies for disinfecting in-vitro contaminated banana plants, which are more sensitive to most disinfectants. Thus, this study aimed to establish an efficient disinfection protocol that is effective against contaminants and safe on in-vitro plants. Contaminated in-vitro banana plants (cv. Grand naine) were subjected to commercial bleach (Jik®; 3.85% w/v NaClO) at different exposure times (2, 4, 6, and 8 min) with or without rinsing then reinitiated in-vitro. Jik® exposure time of 2 min with or without rinsing preserved the plant viability by 100% but led to >75% fungal contamination. At 4 min of Jik® exposure, the viability of the plants remained at 100%, but >33% fungal contamination was observed. No contamination was observed at 8 min of Jik® exposure, but the plants' viability was reduced to below 85%. Notably, 6 min of Jik® exposure preserved the viability of the plants by 100% while destroying all the contaminants and is, therefore, recommended as the most efficient treatment. This protocol can save time and other resources and should be applied in banana genetic transformation laboratories.