Objective: Casuarina junghuhniana Miq. is a multipurpose tree, belongs to the family Casuarinaceae. It has a symbiotic association with actinomycete Frankia which makes the tree to grow vigorously on varied environmental conditions. Not much work has been documented on secondary metabolites and therapeutic approach of this species in India. Therefore, the objective of the present study was to explore the potential phytochemicals in the root extracts and its therapeutic values.Methods: Casuarina junghuhniana root samples were collected from the State Forest Research Institute, Kolappakam, Chennai. The qualitative and quantitative phytochemicals screening were carried out using standard procedures. Antibacterial assay was tested against different pathogens using well diffusion method. Antioxidant activity was carried out using reducing power, hydrogen peroxide scavenging and 1, 1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging methods. The anti-inflammatory assay was tested using protein denaturation method. Further, the phytochemicals in the root extract were evaluated using GC-MS studies. The functional role of the phytoconstituent was confirmed by docking against the target using mcule software.Results: The qualitative screening revealed the presence of various phytoconstituents. The quantitative analysis revealed the presence of maximum phenols (68.95±0.46 mg/g), flavonoids (34.15±0.33 mg/g) and terpenoids (3.9%) in methanolic root extract with respective standards. Antibacterial activity showed a maximum zone of inhibition against Bacillus subtilis (19±0.2 mm) followed by Staphylococcus sp (14±0.3 mm). The antioxidant and anti-inflammatory activity was compared with standard ascorbic acid and the extract showed activities in a dose-dependent manner. Several peaks were obtained in the GC-MS analysis which indicates the presence of different secondary metabolites. The specific phytocompound showed prominent binding affinity against the target enzyme in docking studies.Conclusion: The presence of potent phytochemicals in the Casuarina junghuhniana root with antioxidant and anti-inflammatory activities could be used as a promising source for developing novel plant-based therapeutic agents.Â