摘要:SummaryInsights into the genomic consequences of symbiosis for basidiomycete fungi associated with social insects remain sparse. Capitalizing on viability of spores from centuries-old herbarium specimens of free-living, facultative, and specialist termite-associatedPodaxisfungi, we obtained genomes of 10 specimens, including two type species described by Linnaeus >240 years ago. We document that the transition to termite association was accompanied by significant reductions in genome size and gene content, accelerated evolution in protein-coding genes, and reduced functional capacities for oxidative stress responses and lignin degradation. Functional testing confirmed that termite specialists perform worse under oxidative stress, while all lineages retained some capacity to cleave lignin. Mitochondrial genomes of termite associates were significantly larger; possibly driven by smaller population sizes or reduced competition, supported by apparent loss of certain biosynthetic gene clusters. Our findings point to relaxed selection that mirrors genome traits observed among obligate endosymbiotic bacteria of many insects.Graphical abstractDisplay OmittedHighlights•Germination of spores from 250-year-old herbarium specimens described by Linnaeus•Comparative genomics of 10Podaxisspecimens—free-living and termite specialists•In vitrotests of hypotheses based on genomic analysis•Identification of traits linked to termite-specialist lifestyleBiological sciences; Genetics; Genomics