Two new species, Hesperis anatolica A. Duran and H. hamzaoglui A. Duran (Brassicaceae), are described and illustrated from Anatolia, Turkey. Hesperis anatolica grows in serpentine pebbly slopes and open pine forest and scrub in Pozantı and Karsantı (Adana province). It is closely related to H. persica Boissier, an endemic confined to South Anatolia. Hesperis hamzaoglui grows in granitic rocky sites in a forest-steppe ecotone from Zorkun Yayla (Osmaniye province). It is closely related to H. podocarpa Boissier, an endemic confined to the Amanos Mountains. Diagnostic morphological characters are discussed, as well as the ecology, biogeography, and conservation status of both new species. In addition, the pollen characteristics and seed coat surface of H. anatolica, H. persica, H. hamzaoglui, and H. podocarpa are examined by SEM. Hesperis anatolica is diploid with a new report of the chromosome number of 2n = 12.