Background and Aims: This study aims to investigate the cytotoxicity and DNA protective effects of semi-arid, arid, or desert truffles such as the Terfezia and Picoa species. Methods: The DNA protective effects of the Terfezia and Picoa species was evaluated using plasmid pBR322 DNA treated with UV and H 2 O 2 . The cytotoxic effects of water and methanol extracts of the Terfezia and Picoa species on H1299 and HUVEC cells were evaluated using the MTT assay. Results: Higher concentrations of T. olbiensis and P. lefebvrei extracts have demonstrated a DNA protective effect. The water extract of P. juniperi only at a 40 mg/mL concentration demonstrated a DNA-protective effect, whereas T. boudieri extract did not show any DNA-protective effect at all concentrations tested. H1299 cells showed more sensitivity to the water extract of T. olbiensis (4.66%), P. juniperi (12.04%), P. lefebvrei (21.93%), and the methanol extract of T. boudieri (20.93%). In general, the water and methanol extracts of the Terfezia and Picoa species demonstrated the least cytotoxic effects on HUVEC cells, except for the water extract of P. juniperi (38.46%). Conclusions: In conclusion, results obtained from this study show that the Terfezia and Picoa truffle species have potential DNA protective and cancer prevention properties.