Abstract We provide an expanded and updated, 2-locus phylogeny (mtSSU, nuLSU) of the lichenized fungal family Trypetheliaceae , with a total of 196 ingroup OTUs, in order to further refine generic delimitations and species concepts in this family. As a result, the following 15 clades are recognized as separate genera, including five newly established genera: Aptrootia , Architrypethelium , Astrothelium (including the bulk of corticate species with astrothelioid ascospores; synonyms: Campylothelium , Cryptothelium , Laurera ), Bathelium s. str. (excluding B. degenerans and relatives which fall into Astrothelium ), the reinstated Bogoriella (for tropical, lichenized species previously placed in Mycomicrothelia ), Constrictolumina gen. nov. (for tropical, lichenized species of Arthopyrenia ), Dictyomeridium gen. nov. (for a subgroup of species with muriform ascospores previously placed in Polymeridium ), Julella (provisionally, as the type species remains unsequenced), Marcelaria ( Laurera purpurina complex), Nigrovothelium gen. nov. (for the Trypethelium tropicum group), Novomicrothelia gen. nov. (for an additional species previously placed in Mycomicrothelia ), Polymeridium s. str., Pseudopyrenula , Trypethelium s. str. ( T. eluteriae group), and Viridothelium gen. nov. (for the Trypethelium virens group). All recognized genera are phenotypically characterized and a discussion on the evolution of phenotypic features in the family is given. Based on the obtained phylogeny, species delimitations are revised and the importance of characters such as thallus morphology, hymenial inspersion, and secondary chemistry for taxonomic purposes is discussed, resulting in a refined species concept.