Few molecular-based studies have tested the monophyly of the Lithosiini subtribes proposed by Bendib and Minet (2000), and those have been limited by meager taxon sampling. Several studies have suggested some subtribes are not monophyletic as they are currently defined. We conducted a molecular phylogenetic study of representatives of the North American lithosiine fauna which are currently classified within the subtribes Acsalina, Lithosiina, Cisthenina and Eudesmiina, the latter having never been included in a molecular-based analysis before. Based on analyses of cytochrome oxidase subunit I (COI), ribosomal protein S5 (RPS5) and the large subunit 28S ribosomal DNA (28S), we assign some of the North American genera to subtribe for the first time and re-assign others contrary to previous placements. Then, we discuss the morphological characters that Bendib and Minet (2000) proposed to define subtribes and re-consider them in the context of our inferred phylogeny. We report high support for a monophyletic Lithosiina+ Agylla + Inopsis + Gnamptonychia, three genera being unplaced or of uncertain placement (Agylla) by Bendib and Minet (2000). We remove Gardinia from the subtribe Lithosiina and place it in the Cisthenina, along with Eudesmia, formerly placed in its own subtribe, the Eudesmiina. Two other genera, Bruceia and Ptychoglene, not previously assigned to a subtribe are found to be members of the Cisthenina. We remove Clemensia from the Cisthenina and report it, along with the neotropical Pronola, as part of undefined clade. After these changes, our phylogeny shows strong support for the monophyly of Cisthenina + Gardinia + Eudesmia + Bruceia + Ptychoglene. We find Acsala anomala occurs on a long branch by itself, confirming the uniqueness of this species and its placement in a monotypic subtribe. Finally, we confirm that Afrida exegens, sometimes considered a member of the Lithosiini, is not even an erebid, but rather it is a member of the Nolidae, as proposed by Holloway (1998) and Kitching and Rawlins (1998).

Keywords: Arctiinae, Lithosiini, Lichen Tiger Moths, Cisthenina, Lithosiina, Eudesmiina, Acsalina, Nolidae, Afridinae