Forty-four herbarium specimens of the hepatic Barbilophozia hatcheri, collected from across the entire range of the species in Antarctica (54–68 °S), were examined for fungal colonisation. Mycothalli, consisting of hyaline hyphae in rhizoids or hyphal coils in rhizoid bases and stem medullary cells, were present in 42 specimens. The frequencies of mycothalli declined at higher latitudes and were associated positively with annual air temperatures. In contrast, the frequency of dark septate hyphae on stems increased at higher latitudes. Sequencing of ITS regions of fungal rRNA genes amplified from B. hatcheri from 54 °S, 61 °S and 68 °S indicated that Cladophialophora species colonised plants at higher latitudes, but that most fungi had >99 % homology with members of Sebacinales clade B, with best matches to mycobionts of B. lycopodioides, Lophoziopsis excisa and Protolophozia crispata. We propose the use of herbarium specimens of mycothalli as proxies for climate change in Antarctica.