Rice straw (RS) is a waste by-product from rice paddy fields that is often produced at a large-scale and typically found in many Southeast Asian countries. Chemical pre-treatment methods emerge as the most effective means for extracting pure cellulose derived from lignocellulosic biomass, as they aid in the removal of hemicellulose and lignin. This research focuses on exploring an environmentally friendly technique for extracting cellulose from RS using low-concentration and sustainable chemicals. The cellulose yield from RS reached 21.35% through alkaline delignification and formic acid hydrolysis. Characterization techniques including FESEM, FTIR, and XRD were used to examine the structural properties of the raw RS, de-waxed RS, delignified RS, acid-treated RS, and pure RS cellulose. In this research, the cellulose content in this biomass is used as a base for producing a novel biopolymer-based hydrogel. Crosslinking with 1% glutaraldehyde method is employed to fabricate hydrogel from the extracted RS cellulose, incorporating polyvinyl alcohol (PVA) at varying cellulose concentrations (1, 2, and 3% w/w). The swelling ratio achieved ~ 85% at crosslinked-PVA hydrogel with 3% cellulose after 17 h. This research supports the development of an eco-friendly extraction process for cellulose from RS, promotes the advancement of sustainable processing technologies, and enables the effective utilization of lignocellulosic agricultural waste in various applications.