Role of defect states of thermally evaporated molybdenum trioxide (MoO3−x) on electrical conductivity was investigated via low temperature current–voltage and capacitance–voltage measurements. To clarify the charge transport phenomena through MoO3−x, a 15 nm thin layer of MoO3−x film was used as an interface layer between gold and n-type Silicon (n-Si). The formation of an interface dipole between n-Si and MoO3−x exhibits a rectifying behaviour of Au/MoO3−x/n-Si Schottky barrier diode (SBDs). The rectifying nature of the SBDs shown up to 175 K due to proper electron extraction from valence band to conduction band via the defect states; however at ≤165 K the rectifying nature was not observed due to insulating behaviour of MoO3−x layer. Oxygen deficiency as a formation of defects was determined by x-ray photoelectron spectroscopy (XPS). Consequences of these defects as a function of current conduction across the MoO3−x was also confirmed by low temperature photoluminescence (PL)measurement.