Ascomata of two truffle species, Tuber borchii and T. asa-foetida, both naturally growing in woodlands of the Basilicata region (southern Italy), were identified on the basis of ascospore morphology and compared under a volatile organic compound profile to determine the particular volatile organic compounds that characterize each taxon. Solid-phase microextraction-gas chromatography-mass spectrometry analysis of the samples showed the presence of 1-methyl-1,3-butadiene as a primary component in both truffles. T. asa-foetida showed a compound, toluene, not present in T. borchii, which creates the penetrating "solvent" smell of the truffle.