Background:Desert truffles (Terfezia species) are known by their vital nutritional benefits as they are considered as rich sources of vitamins, fatty acid, minerals and proteins.Methods:The chemical constituents of the different solvent extracts of Terfezia species were isolated and identified by column chromatography, spectroscopic and GC-MS analyses. Also, the ethyl acetate and acetone extracts of different fungal isolates, associated Terfezia, after grown on rice medium were screened for their antimicrobial, anticancer and antioxidant activities via disc agar plate, micro culture tetrazolium (MTT) and 2,2-azino-di-[3-ethylbenzo-thiazolin-sulphonate] (ABTS) assays, respectively. The promising fugal strains were molecularly identified by 18SrRNA tool.Results:Bio-guided separation of methylene chloride, ethyl acetate and n-butanol fractions of Terfezia species led to identification of nine compounds namely; (R)-4,8-dihydroxy-7-hydroxymethyl-6- methoxy isochroman-1-one (1), 4-deoxy-4α-phorbal-12-(2,3-dimethyl)butyrate-13-isobutyrate (2), oxyphylline B (3), terfezien A (4), latilagascene D (5), amaiouine (6), senbusine acetate (7), terfezien B (8) and marinoquinoline D (9). Moreover, sixteen compounds were identified in the n-hexane extract via GC-MS analysis, accounting for 93.69% of the total detected components in the extract. While, twenty five components were detected in the methylene chloride extract, representing 43.86% from total detected components in the extract. Eight fungal strains were isolated from Terfezia sp., powder by serial dilution methods and these fungi were cultivated on solid rice medium. Also, their ethyl acetate and acetone extracts were subjected to biological studies including antimicrobial, antioxidant and anticancer activities. The three potent fungal strains (1M, 4M and 8M) were identified by the molecular technique 18SrRNA as Aspergillus niger 1M-EGY-IQ, Penicillium crustosum 4M-EGY-IQ, and Fusarium proliferatum 8M-EGY-IQ for 1M, 4M and 8M, respectively.Conclusion:Terfezia sp., comprise a rich source of bioactive compounds and could be considered as an interesting candidate for the treatment of infectious diseases.