A new newt species, Tylototriton daloushanensis Zhou, Xiao & Luo, sp. nov., is described from Guizhou, China, based on phylogenetic and morphological analyses. Phylogenetic analyses based on mitochondrial 16S and ND2 genes show that the new species is an independent lineage closely related to T. maolanensis, T. broadoridgus, T. dabienicus, T. anhuiensis, and T. wenxianensis. Morphologically, the new species is distinguished from its congeners by a combination of the following morphological characters: (1) large body size (SVL 64.7–83.6 mm in male and 70.5–100.3 mm in female); (2) head longer than wide, prominent bony ridges present; (3) snout rounded in dorsal view; (4) tail length less than snout-vent length in both males and females; (5) dark black body coloration, except for bright orange cranial region, palms and soles, vent region, and ventral ridge of tail; (6) relative length of toes III > IV > II > I > V; (7) fingertips reaching to between the eyes and nostrils when the forelimbs are stretched forward; (8) distal tip of the limbs greatly overlapping when the fore and hind limbs are pressed along the trunk; (9) presence of gular fold; (10) vertebral ridge slightly segmented, slightly flattened rib nodules, indistinct interspaces between rib nodules; and (11) lacking orange color markings on posterior parotoids and rib nodules. The found of the new species suggests that the species diversity of the genus Tylototriton may be underestimated.