Inspired by nature, cellulose extracted from plant wastes has been explored, due to its great potential as an alternative for synthetic fiber and filler that contributes to structural performance. The drive of this study was to extract, treat, and evaluate the characteristics of rice straw (RS) (Oryza sativa L.) cellulose as a biodegradable reinforcement to be utilized in polymer base materials. Two routes of extraction and treatment were performed via the pulping (Route 1) and chemo-mechanical methods (Route 2), in order to discover comparative characteristics of the synthesized cellulose fiber. Comprehensive characterization of RS cellulose was carried out to determine crystallinity, surface morphology, and chemical bonding properties, using X-ray diffraction (XRD), field emission scanning electron microscopy (FESEM), and Fourier transform infra-red (FTIR), respectively. The XRD test results showed that the crystallinity index (CI) of cellulose powder (CP) decreased after the surface modification treatment, Route 2, from 64.50 to 50.10% CI for modified cellulose powder (MCP), due to the surface alteration of cellulose structure. From Route 1, the crystallinity of the fibers decreased up to 33.5% (dissolve cellulose, DC) after the pulp went through the surface modification and dissolution processes, resulting from the transformation of cellulose phase into para-crystalline structure. FESEM micrographs displayed a significant reduction of raw RS diameter from 7.78 µm to 3.34 µm (treated by Route 1) and 1.06 µm (treated by Route 2). The extracted and treated cellulose via both routes, which was considerably dominated by cellulose II because of the high percentage of alkaline used, include the dissolve cellulose (DC). The dissolution process, using NMMO solvent, was performed on the pulp fiber produced by Route 1. The fiber change from cellulose I to cellulose II after undergoes the process. Thus, the dissolution process maintains cellulose II but turned the pulp to the cellulose solution. The acquired characteristics of cellulose from RS waste, extracted by the employed methods, have a considerably greater potential for further application in numerous industries. It was concluded that the great achievement of extracted RS is obtained the nanosized fibers after surface modification treatment, which is very useful for filler in structural composite applications.