Abstract. The application of nanotechnology in agriculture is gaining increasing significance. The use of graphene oxide (GO)-based nanoensembles as stabilizers of functional groups or plant growth inducers, such as phytohormones, represents a promising direction for further research. This study aimed to conduct a comparative analysis and examine the effects of a GO-based nanoensemble with the phytohormone kinetin on the morphophysiological and biochemical parameters of Gossypium hirsutum L. Agdash-3 cultivar seedlings. Morphophysiological analysis revealed that treatment with kinetin + GO stimulated seed germination and increased stem length compared to the control and free kinetin treatment. Biochemical studies demonstrated an increase in polyphenol oxidase activity, the concentration of photosynthetic pigments (chlorophyll a and carotenoids), as well as proline content in leaf tissues upon nanoensemble treatment. In contrast, root length, catalase and guaiacol peroxidase activity, along with chlorophyll b and proline concentrations in roots, remained at levels comparable to those observed with kinetin alone, suggesting no additional effect of GO on these parameters. The obtained results indicate a synergistic effect of GO in combination with kinetin on specific morphophysiological and biochemical characteristics of cotton seedlings, highlighting potential avenues for further research.