Optimization and characterization of essential oil-loaded nanoemulsions for enhanced antiviral activity against tobacco mosaic virus (TMV) in tobacco plants

Tobacco mosaic virus (TMV) is a significant viral pathogen that infects various economically important crops, including tobacco, tomatoes, and cucumbers. The current study aimed to develop and optimize essential oilloaded nanoemulsions using Response Surface Methodology (RSM) to enhance antiviral activity against TMV in Nicotiana tabacum plants. Essential oils from tea tree (Melaleuca alternifolia) and lime peel (Citrus latifolia) were chemically analyzed using gas chromatography-mass spectrometry (GC-MS). Terpinene-4-ol (70.92 %) and transisolimonene (33.82 %) were identified as the major components in tea tree and lime peel oils, respectively. RSM was used to optimize the nanoemulsion formulations based on oil concentration, hydrophilic-lipophilic balance (HLB), and stirring speed. Optimal tea tree oil (TTEO) and lime peel oil (LPEO) nanoemulsions were developed and characterized using Fourier-transform infrared (FTIR) spectroscopy, scanning electron microscopy (SEM), and particle size analysis. The antiviral efficacy of the optimized formulations was tested against TMV using RTqPCR analysis and physiological measurements. TTEO nanoemulsions significantly inhibited TMV expression by 89.78%, while LPEO nanoemulsions achieved a 42.22% reduction after six days. Additionally, these essential oils (EOs) enhanced plant defense mechanisms by modulating the activities of phenylalanine ammonia-lyase (PAL) and peroxidase (POD), along with maintaining chlorophyll levels. This study demonstrates that TTEO and LPEO nanoemulsions offer promising eco-friendly antiviral agents for TMV management in tobacco plants. The research underscores the potential of essential oil-based formulations in mitigating viral infections in crops, providing a sustainable alternative to synthetic chemicals.