Two previously unknown species of Rhinogobius have been discovered in the streams of the Upper Youshui River, within the Yuan River Basin, Xiushan County, Chongqing, China. These new species are named as Rhinogobius sudoccidentalis and Rhinogobius lithopolychroma. Phylogenetic analysis based on mitochondrial genomes revealed that R. sudoccidentalis is genetically closest to R. reticulatus, while R. lithopolychroma shares the greatest genetic similarity with R. leavelli. Morphological distinctions allow for the clear differentiation of these species. Rhinogobius sudoccidentalissp. nov. is characterized by having VI–VII rays in the first dorsal fin and I, 8–9 rays in the second dorsal fin. The longitudinal scale series typically consists of 22–24 scales, while the transverse scale series comprises 7–8 scales. Notably, the predorsal scale series is absent and the total vertebrae count is 12+17=29. Rhinogobius lithopolychromasp. nov. can be distinguished from other species by the presence of 13–15 rays on the pectoral fin. Its longitudinal scale series ranges from 30 to 33 scales, with no scales in the predorsal area. The total vertebral count is 30, with 12 precaudal and 18 caudal vertebrae. The head and body of this species are light gray with irregular orange markings on the cheeks and opercle. Through morphological and molecular analyses, it has been confirmed that R. lithopolychroma and R. sudoccidentalis represent novel species within the Rhinogobius genus.