Advancements in molecular genetics have revealed that hybridization may be common among plants, animals, and fungi, playing a role in evolutionary dynamics and speciation. While hybridization has been well-documented in pathogenic fungi, the effects of these processes on speciation in fungal lineages with different life histories and ecological niches are largely unexplored. Here we investigated the potential influence of hybridization on the emergence of morphologically and reproductively distinct asexual lichens. We focused on vagrant forms (growing obligately unattached to substrates) within a clade of rock-dwelling, sexually reproducing species in the Rhizoplaca melanophthalma (Lecanoraceae, Ascomycota) species complex. We used phylogenomic data from both mitochondrial and nuclear genomes to infer evolutionary relationships and potential patterns of introgression. We observed multiple instances of discordance between the mitochondrial and nuclear trees, including the clade comprising the asexual vagrant species R. arbuscula, R. haydenii, R. idahoensis, and a closely related rock-dwelling lineage. Despite well-supported phylogenies, we recovered strong evidence of a reticulated evolutionary history using a network approach that incorporates both incomplete lineage sorting and hybridization. these data suggest that the rock-dwelling western north American subalpine endemic R. shushanii is potentially the result of a hybrid speciation event, and introgression may have also played a role in other taxa, including vagrant species R. arbuscula, R. haydenii and R. idahoensis. We discuss the potential roles of hybridization in terms of generating asexuality and novel morphological traits in lichens. furthermore, our results highlight the need for additional study of reticulate phylogenies when investigating species boundaries and evolutionary history, even in cases with wellsupported topologies inferred from genome-scale data. open Scientific RepoRtS | (2020) 10:1497 | https://doi.org/10.1038/s41598-020-58279-x www.nature.com/scientificreports www.nature.com/scientificreports/ nuclear markers, which in turn can influence the genome in both adaptive and maladaptive ways 5,[19][20][21][22] . Adaptive introgression of mitochondrial DNA may play an important role in speciation and phylogeography 23,24 .While hybridization has been well-documented among pathogenic fungi 21,[25][26][27][28][29][30] , the role of hybridization on the process of speciation of fungal lineages with different life histories and ecological functions is not well understood, including among lichen-forming fungi. The role of gene flow and hybridization in lichen-forming fungal evolution has been a long-standing question [31][32][33][34][35] . Species boundaries in fungi, including symbiotic fungi such as lichen formers, have received substantial attention and become more robust with molecular sequence data 36 . However, processes involved in speciation in lichen-forming fungi have received far less attention. With a poor fossil record 37,38 and pote...