The Automobile HVAC system is a habitat for odor-associated fungal communities. We investigated the odorassociated fungal community in an automobile HVAC system using a high-throughput DNA sequencing method. The fungal community structure was evaluated via metagenome analysis. At the phylum level, Ascomycota and Basidiomycota were detected, accounting for 43.41% and 56.49% of the fungal community in the HVAC system, respectively. Columnosphaeria (8.31%), Didymella (5.60%), Davidiella (5.50%), Microxyphium (4.24%), unclassified Pleosporales (2.90%), and Cladosporium (2.79%) were abundant at phylum of Ascomycota and Christiansenia (36.72%), Rhodotorula (10.48%), and Sporidiobolus (2.34%) were abundant at phylum of Basidiomycota. A total of 22 genera of fungi were isolated and identified from the evaporators of the HVAC systems which support fungal growth and biofilm formation. Among them, Cladosporium, Penicillium, Aspergillus and Alternaria are the most representative odor-associated fungi in HVAC systems. They were reported to form biofilm on the surface of HVAC systems with other bacteria by hypha. In addition, they produce various mVOCs such as 3-methyl-1-butanol, acetic acid, butanoic acid, and methyl isobutyl ketone. Our findings may be useful for extending the understanding of odor-associated fungal communities in automobile HVAC systems.