In this report, copper nanoparticles (Cu NPs) are successfully prepared using biosynthesis from stem extract of Holoparasitic plant (Orobanche aegyptiaca). The structural and morphological properties of NPs were determined by using X-ray Diffraction (XRD), Fourier-Transform Infrared Spectroscopy, Scanning Electron Microscopy, energy dispersive x-ray spectroscopy (EDX) and Transmission Electron Microscopy (TEM). The optical study has been analyzed using Ultraviolet–Visible Spectroscopy and optical band gap was calculated 4.65 eV. The XRD and TEM patterns confirmed the formation of single phase Cu NPs having an average particle size less than 50 nm. Notable antimicrobial activity of Cu NPs was observed against E. coli & S. aureus and nematicidal properties was confirmed against Meloidogyne incognita in-vitro. This is the first report on biosynthesis of Cu NPs for its enhanced nematicidal activity.