Evaluation of Natural Essential Oils as Sustainable Disinfectants for the Control of Microbial Contaminants in Vitro Plant Tissue Culture

Document Type : Research Paper

Authors

1 Department of Horticulture and Landscape Design, College of Agriculture, Kirkuk University, Kirkuk, Iraq

2 Ministry of Health, Sulaymaniyah General Directorate of Health, Blood Bank Directorate, Sulaymaniyah, Iraq

3 Agricultural Botany Department, Faculty of Agriculture, (Saba Basha), Alexandria University, Alexandria, Egypt

10.22034/jmpb.2026.371791.2121

Abstract

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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Journal of Medicinal plants and By-products
Volume 15, Issue 1
January and February 2026
Pages 146-152

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