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Melaleuca polandii, commonly known as "Gold-tipped Bottlebrush", is widely grown in Indian gardens and parks as an ornamental plant. In the present study, the chemical composition, antioxidant, antimicrobial and cytotoxic activities of M. polandii leaf essential oil (MPLEO) have been investigated. GC-MS analysis of MPLEO isolated from plants grown along the coastal plains of Eastern India revealed the presence of 50 constituents with α-pinene (46.17%), 3Z-hexenyl acetate (10.10 %), α-terpineol (5.43%), limonene (3.63%) and ρ-cymene (2.44 %) as the major constituents. MPLEO exhibited moderate antioxidant capacity in DPPH, ABTS and Reducing ability assays with IC50 values of 7.94 ± 0.34 μg/mL, 5.71 ± 0.21 μg/mL and EC50 7.09 ± 0.35 μg/mL, respectively. Of the 5 bacterial strains tested, the EO exhibited highest sensitivity against Bacillus subtilis with the ZOI of 12.66 ± 0.57 mm and strong antimicrobial activity against Staphylococcus saprophyticus with MIC value of 6.25 μg/mL. MPLEO exhibited significant cytotoxic effects against Human alveolar lung adenocarcinoma (A549), Human hepato-cellular carcinoma (HepG2), Human cervix adenocarcinoma (HeLa) and normal Human Embryonic Kidney cell (HEK-293) as evaluated through MTT assay with IC50 values of 9.5 ± 0.38 μg/mL, 22.73 ± 0.95 μg/mL, 20.41 ± 0.82 μg/mL and 237.68 ± 9.26 μg/mL, respectively. The findings of the present study demonstrated that MPLEO could be considered as a potential bioresource having applications in pharmaceutical industries as an antibacterial and antiproliferative agent. [ABSTRACT FROM AUTHOR]