The food industry is interested in replacing artificial sweeteners with natural sugars that possess zero calories and carbohydrates and do not cause spikes in blood sugar levels. The steviosides leaves, synthesized at Stevia rebaudiana Bertoni, are 300 times sweeter than common table sugar. Stevia propagation is limited due to the poor viability of the seeds, the long time and low germination rate, and the poor rooting ability of vegetative cuttings. Because of this, an alternative biotechnological method for its reproduction is being studied, such as multiple shoot production through direct organogenesis using nanofibers, formed from a derivative of amino acid valine as a carrier of the biologically active agent silver atoms/particles (NF-1%Ag and NF-2%Ag). The stevia explants were cultured on a medium containing NF-1%Ag and NF-2%Ag at concentrations of 1, 10, 50, and 100 mg L⁻¹. The NF-1%Ag and NF-2%Ag treatment caused hormetic effects on stevia plantlets. At low concentrations of from 1 to 50 mg L⁻¹ of nanofibers, the stimulation of plant growth was observed, with the maximum effect being observed at 50 mg L⁻¹ nanofibers. However, at the higher dose of 100 mg L⁻¹, inhibition of the values of parameters characterizing plant growth was recorded. The presence of nanofibers in the medium stimulates stevia root formatting.