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The current research study focuses to formulate the biosynthesized silver nanoparticles for the first time from silver acetate using methanolic root extracts of Diospyros sylvatica,a member of family Ebenaceae.TEM analysis revealed the average diameter of Ag NPs around 8 nm which is in good agreement with the average crystallite size (10 nm) calculated from X-ray Diffraction (XRD) analysis.Further the study has been extended to the antimicrobial activity against test pathogenic Gram (+) ve,Gram (-) ve bacterial and fungal strains.The bioinspired Ag-NP showed promising activity against all the tested bacterial strains and the activity was enhanced with increased dosage levels.