Leaf-associated fungi, the fungi that depend on leaves to sporulate, have a rich Cenozoic record, however their earlier diversity is poorly characterized. Here we describe Harristroma eboracense gen. et sp. nov., a Middle Jurassic leaf-associated fungus colonizing the leaf cuticle of Nilssonia tenuicaulis (cycadophyte). To place our newly described species into a picture of the diversification of Mesozoic fungi, we reassess fossils with leaf-associated stromata in the context of fungal molecular phylogeny. Being melanized, with radiate stromata, and on leaves, H. eboracense and other fossils from the Jurassic and earlier periods are probably related to filamentous Ascomycota in the superclass Leotiomyceta. Characters needed for further resolution of leaf-associated fungal biology and classification, such as the presence of an ostiole for spore discharge and appressoria for entry into leaf tissue first appear in the Mesozoic fossil record. Among Early Cretaceous fossils, Spataporthe taylorii represents the oldest unambiguous evidence of perithecial Sordariomycetes while Protographum luttrellii and Bleximothyrium ostiolatum are the oldest Dothideomycetes thyriothecia. Environmental observations show that broad leaved gymnosperms (especially cycadophytes) growing in warm temperate wet forests might have been the first environment for the radiation of Leotiomyceta.