The Santa Rosa range presently contains one of the few moderately preserved vegetation remnants in the state of Guanajuato. Unfortunately, this state has been characterized by the degradation of its vegetation cover since colonial times. In this floristic study we recorded 21 infraspecific taxa, 496 species, 273 genera and 93 families of vascular plants. Among these, Asteraceae comprises the largest number of species, followed by Poaceae and Lamiaceae, respectively. Three species cited in the Mexican Norm of Endangered Species (Selaginella porphyrospora, Zigadenus virescens and Gentiana spathacea) were recorded. Despite the reduced area of this mountain range, which represents around 0.45% of the area covered by the Flora of the Bajío and of Adjacent Regions project (about 50,000 km2), its floristic richness represents 8.7% of the total flora expected for that region.