Fungi produce a wide variety of volatile organic compounds (VOCs), chemical cues with central roles in mediating interspecific interactions. Here we present a platform consisting of 14 cuvettes and 2 types of mass spectrometers, that allows for the first time automated, real-time analysis of various microbial VOCs. We used this platform to comprehensively analyze the VOCs of 43 fungal species. The further computational analyses reveal fundamental links between VOCs and ecological functions of fungi.