A thorough chemical, molecular and morphological study has been carried out on a problematic group in the genus Lepraria characterized by lobed, granular thalli with a raised rim and a preference for mosses, rocks and mineral soils in exposed, xeric habitats. The material examined was collected in seven countries of the Mediterranean-Macaronesian region, from chalk, carbonate rocks and derived soils (A-thalli), and from siliceous rocks and derived soils (B-thalli). The results of a phylogenetic analysis based on 93 ITS sequences (29 of which newly obtained) support the identification and the segregation of two taxa at species level, provisionally identified as Lepraria isidiata s. lat., (most of the A-thalli), and L. santosii s. lat. (B-thalli plus the remaining A-thalli), as the observed genetic variability is quite high. Three chemotypes were detected in L. isidiata s. lat., and seven in L. santosii s. lat. The two taxa are morphologically well characterized: in L. isidiata s. lat. the thallus is thicker and the propagules larger than in L. santosii s. lat. The first stages of thallus development from single propagules are described in both species. Some critical remarks are made about the increasing use of the species rank for taxa of Lepraria, which are morphologically and genetically scarcely characterized.