Fungus-growing termites (subfamily Macrotermitinae) cultivate the symbiotic basidiomycete fungus Termitomyces in their fungus comb to digest cellulosic materials and to supply nitrogen-rich fungal diet. In Japan, the fungus-growing termite Odontotermes formosanus is found on the Yaeyama Islands and Okinawa Island, Okinawa Prefecture. Odontotermes formosanus is thought to have been recently and artificially introduced to Okinawa Island as its distribution is discontinuous and restricted to small areas. Previous DNA analyses revealed that two types of Termitomyces, namely Termitomyces sp. Type A and Termitomyces sp. Type B, whose fruiting bodies correspond to Termitomyces microcarpus-like pseudorhiza-lacking small mushroom and Termitomyces intermedius, respectively, are cultivated by O. formosanus on the Yaeyama Islands. However, information about the Termitomyces types cultivated by O. formosanus on Okinawa Island is limited. To define the fungal types cultivated by O. formosanus on Okinawa Island, I developed a diagnostic polymerase chain reaction method using primer sets specific to the nuclear ribosomal DNA sequences consisting of the internal transcribed spacers (ITS1 and ITS2) and 5.8S rDNA of Termitomyces not using fungal mycelium, but using the termite gut metagenome including fungal DNA as a template. The results indicated that the same two Termitomyces types from Iriomote Island are cultivated by O. formosanus on Okinawa Island. The distribution pattern of Termitomyces types on Okinawa Island showed that Termitomyces sp. Type A is limited to the mountainous side of Sueyoshi Park, despite Termitomyces sp. Type B being widely distributed in the area in which O. formosanus is found. This finding implies that O. formosanus on Okinawa Island was recently introduced from Iriomote Island to Sueyoshi Park.
