Amplicon-Based Profiling of Fungal Communities Associated with Scots Pine Bark Beetles: Selective Antagonism and Monoterpene Tolerance

Bark beetle–fungus associations are essential for nutrition, detoxification, and host colonisation, but their composition and function vary across developmental stages and environmental contexts. Hence, we characterised the fungal communities associated with two pine-feeding bark beetles, Ips sexdentatus (ISX) and Ips acuminatus (IAC), across developmental stages and compared wild-collected and laboratory-bred populations using ITS2 amplicon sequencing. Both beetle species maintained a stable core mycobiome dominated by Kuraishia, Ogataea, Ophiostoma, Graphilbum, and Cyberlindnera. These taxa have been earlier reported to be associated with nutrient provisioning, detoxification of host secondary metabolites, and chemical signalling. Adult beetles showed species-specific community differences, whereas wild-collected beetles, particularly IAC, harboured higher fungal diversity than laboratory populations, indicating a strong environmental effect. Beetles shared more fungal taxa with control wood than with gallery wood, suggesting possible fungal acquisition during feeding and concurrent restructuring of the wood mycobiome during infestation. Monoterpene bioassays with selected yeast symbionts showed differential growth responses to α-pinene, 3-carene, and terpinolene, and their mixture, with the mixture producing stronger inhibition than individual compounds. These yeast symbionts further displayed selective antagonistic activity in vitro against selected filamentous fungi, including entomopathogenic taxa, along with detectable lytic and digestive enzyme activities. Together, our findings highlight a link between community structure, predicted functions, and observed interaction phenotypes, providing a strong basis for future mechanistic studies of beetle–fungus–conifer interactions.