Studies have revealed that some Xylaria species were closely associated with fungus-growing termite nests. However this relationship rarely had been investigated and the host specificity of termite-associated Xylaria was not yet clearly established. Eighteen Xylaria rDNA-ITS sequences were obtained from fungus combs belonging to 11 Macrotermitinae species from eight regions. Low diversity was found between isolates, and nine sequences were retrieved. Termite-associated Xylaria were shown to be monophyletic, with three main clades, all including strains from various termite hosts and geographical localities. This new molecular study shows no species specificity with respect to fungus-growing termites, which suggests that there might be substrate specialization.