Bioaerosol particles including bacteria, fungi, and virus are originated from marine and terrestrial environments. The airborne microorganisms are transported for long distance through the free troposphere and are thought to influence the downwind ecosystems and human life. However, microbial communities in the free troposphere have not been understood in detail because the direct sampling of microbial cells at high altitude requires sophisticated sampling techniques. In this study, for the investigation of microbial species compositions in the free troposphere, air sampling using an aircraft was performed over the Noto Peninsula in Japan, where the tropospheric winds carry aerosol particles from continental areas. Two air samples were collected at 3,000 m on March 27, 2010, when air mass was carried from the Gobi Desert to Japan area. Microorganisms from one air sample grew in culture media containing up to 15 % NaCl, suggesting that halotolerant bacteria maintain their viabilities in the free troposphere. DGGE analysis revealed that the amended cultures were dominated by Bacillus subtilis, and the isolates obtained from the amended cultures were identical to B. subtilis. Furthermore, the 16S rDNA clone library (culture-independent survey) of the other air sample grew was composed of three phylotypes belonging to Firmicutes, Bacteroidetes, and Proteobacteria with the sequences of Firmicutes phylotype corresponding to that of the cultured B. subtilis sequence. Microscopic observation using FISH method indicated that B. subtilis particles occupied 80 % of total eubacterial particles on the mineral particles. The halotolerant bacteria identical to B. subtilis would dominate at high altitudes over Noto Peninsula where the prevailing westerly wind was blowing.