The Exosome-like nanoparticles (ELNs) are essential for both intra- and inter-kingdom communication. The presence and function of plant ELNs discharged into the environment remain unknown, despite their significant biological significance. In the present study, polymer precipitation method was used to isolate the ELNs from Actinidia deliciosa followed by characterization and antifungal activity evaluation against F. oxysporum, F. solani, and B. cinerea in the term of germination and density of fungal spores' formation. Root-released EVs (extracellular vesicles) were found to have a size ranged from 30 to 150 nm, according to biophysical assessments using dynamic light scattering, and transmission electron microscope. Thirty-three unique proteins were identified using LC-MS/MS of ELNs. Several of these proteins are known to be associated with interactions between plants and microbes. The presence of multiple plant defense-related proteins in these EVs raises the possibility that they are important elements of plant immunity.