# High-Energy Emulsified Clove Essential Oil Nanoemulsion as a Natural Herbicidal Product: Germination Suppression and Seed Structure Alteration in Echinochloa crus-galli

## TL;DR

Clove essential oil nanoemulsions can act as eco-friendly herbicides by suppressing weed seed germination and damaging seed structures.

## Contribution

Development of high-energy emulsified clove essential oil nanoemulsions with herbicidal effects on Echinochloa crus-galli.

## Key findings

- Clove EO nanoemulsions inhibited E. crus-galli germination and seedling growth.
- Nanoemulsions damaged seed structures, particularly the endosperm, as observed via SEM-EDS.
- Optimized nanoemulsions had stable droplet sizes and remained stable for at least four weeks.

## Abstract

Clove (Syzygium aromaticum (L.) Merr. & L.M. Perry) essential oil (EO)-based nanoemulsions may have a promising future in eco-friendly herbicide development. Clove EO was found to have a high eugenol content of 87.27%. Organic-solvent-free nanoemulsions using clove EO as a bioactive ingredient were fabricated using ultrasonication and microfluidization emulsification methods. Fourier-transform infrared spectroscopy confirmed that both emulsification methods did not affect the EO components. The droplet size of optimized nanoemulsions was determined using dynamic light scattering. The smallest size of 66.9 nm was obtained by microfluidization at 20,000 psi and eight passes. Additionally, the smallest droplet size for a sonicated nanoemulsion was 103.9 nm, obtained by ultrasonication at 20% for 6 min. Transmission electron microscopy confirmed the droplet sizes of both optimized nanoemulsions. In a storage test, both optimized nanoemulsions were stored at 4 °C for at least four weeks. Finally, both nanoemulsions were evaluated on pre-emergence herbicidal activities against Echinochloa crus-galli. The results showed that both nanoemulsions inhibited E. crus-galli germination and seedling growth, and additionally, inhibited seed imbibition and α-amylase activity. Micro-morphological and ultrastructural analysis was observed using a scanning electron microscope and an energy dispersive X-ray spectrometer (SEM-EDS). SEM-EDS micrographs of the treated seeds showed that the seed structure was damaged, especially the endosperm, leading to the inhibition of seed germination.