In this study, reduced graphene oxide nanosheets (rGO) were biosynthesized using the methanolic extract of Iraqi Rhus coriaria (L.) fruits with substantial bioreduction capabilities. The GC-MS analysis of the methanolic extract of Iraqi Rhus coriaria (L.) fruits was used to determine the synthesis mechanism of rGO nanosheets. The synthesized graphene oxide (GO) and the biosynthesized methanolic extract of Iraqi Rhus coriaria (L.) fruits-rGO nanosheets (MERCF-rGO) were characterized using UV-Vis at 226 nm, and 238 nm, respectively. FTIR explained the functional groups of GO and MERCF-rGO nanosheets, XRD spectra of the GO and MERCF-rGO show the sizes at 10.42 nm, and 4.07 nm, TEM observed the size of GO and MERCF-rGO at 15.54 nm and 9.6 nm. As well as FESEM of GO and MERCF-rGO was carried out to explain sheet shape about them. Raman spectroscopy of GO and MERCF-rGO displayed the two peaks D at (1353 cm-1, 1336 cm-1) and G at (1597 cm-1, 1594 cm-1) respectively. MERCF-rGO nanosheets showed activity against antibacterial gram-positive (Staphylococcus aureus) and gram-negative (Escherichia coli), cytotoxicity by MTT assay against breast cancer MCF-7 cell line IC50 at 251.99 μg/mL, genotoxicity fragmented DNA of MCF-7 by comet assay. Comprehensively, the green synthesis of rGO is safe, the lowest cost and developable biogenic nano-formulation of Iraqi Rhus coriaria (L.)–rGO owns antibacterial and anticancer therapeutic applications.