In the present study, cellulose nanofibers composite films were manufactured based on thermoplastic starch. Nanofibers were extracted from rice straw employing a developed chemo-mechanical method. In the chemical step, almost all of non-cellulosic components were removed and a white pulp of cellulose microfibers was obtained. Then, a diluted suspension of fibers was ultrasonicated to destruct intermolecular hydrogen bonds achieving nanofibers networks. Afterward, bio-nanocomposites were prepared by film casting. In order to study the effect of nanofibers content on the composite properties, the mechanical and dynamic mechanical properties, morphology, humidity absorption, and transparency of films were investigated. The yield strength and Young modulus of nanocomposites were satisfactorily enhanced compared to the pure thermoplastic starch film. The glass transition temperature of films was shifted to higher temperatures by increasing nanofibers contents. The uniform dispersion of the nanofibers was investigated using SEM images. The humidity absorption resistance of films was significantly enhanced by using 10 wt% cellulose nanofibers. The transparency of the nanocomposites was reduced compared to the pure starch films.