Abstract
Fungus-farming termites cultivate a Termitomyces fungus monoculture in enclosed gardens (combs) free of other fungi, except during colony declines, where Pseudoxylaria spp. stowaway fungi appear and take over combs. Here, we determined Volatile Organic Compounds (VOCs) of healthy Macrotermes bellicosus nests in nature and VOC changes associated with comb decay during Pseudoxylaria takeover. We identified 443 VOCs and unique volatilomes across samples and nest volatilomes that were mainly composed of fungus comb VOCs with termite contributions. Few comb VOCs were linked to chemical changes during decay, but longipinocarvone and longiverbenone were only emitted during comb decay. These terpenes may be involved in Termitomyces defence against antagonistic fungi or in fungus-termite signalling of comb state. Both comb and Pseudoxylaria biomass volatilomes contained many VOCs with antimicrobial activity that may serve in maintaining healthy Termitomyces monocultures or aid in the antagonistic takeover by Pseudoxylaria during colony decline. We further observed a series of oxylipins with known functions in the regulation of fungus germination, growth, and secondary metabolite production. Our volatilome map of the fungus-farming termite symbiosis provides new insights into the chemistry regulating complex interactions and serves as a valuable guide for future work on the roles of VOCs in symbioses.