| The current investigation assessed the impact of five natural essential oils on the in vitro sterilization effectiveness of different explants. The different explants were subjected to three concentrations (1%, 2%, 3% v/v) and three exposure times (1, 2, and 3 min). In addition, the sterilization rates were assessed after 14 days. Results explained that sterilization effectiveness was clearly affected by both oil type and treatment factors. At 1% concentration, eucalyptus and neem attained the highest early sterilization in Bougainvillea (83.3% and 76.0%, respectively), though peppermint and clove confirmed lower activity (56.0% and 66.7%, respectively). The results indicated that increasing the exposure time to 3 min usually increased sterilization rates, with clove, eucalyptus, peppermint, thyme, and neem oils succeeding in 89 to 100% sterilization through species but caused loss of viability. At 2% concentration and 2 min exposure, most oils efficiently sterilized explants while maintaining tissue viability; for example, A. julibrissin explants reached 83.3 to 100% sterilization without death, whereas 3% concentration often caused explant death, mainly in Bougainvillea and R. chinensis minima. Thyme and neem oils reliably confirmed the highest and most reliable activity, attaining 100% sterilization through concentrations and exposure times in R. chinensis minima and A. julibrissin, emphasizing their broad-spectrum antimicrobial potential. Clove required greater concentrations to reach similar efficacy due to its eugenol content, whereas eucalyptus and peppermint performed moderately well at lower concentrations. Based on these results, a recommended sterilization protocol for in vitro culture means using 2% essential oil concentration with 2-minute exposure time. |
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