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A new species of the genus Leptobrachella, L. guinanensis sp. nov., is described in this study based on morphological, molecular, and bioacoustic data. The species was discovered in the Shiwandashan National Nature Reserve in Shangsi County, Guangxi, China. Phylogenetically, L. guinanensis sp. nov. is closely related to L. ventripunctata. However, there are distinct morphological differences between L. guinanensis sp. nov. and L. ventripunctata, as well as three other sympatric species (L. shangsiensis, L. shiwandashanensis, and L. sungi). These differences include body size (SVL 30.5–32.5 mm in males; 38.7–41.8 mm in females in the new species vs 25.5–28.0 mm in males, 31.5–35.0 mm in females in L. ventripunctata), the absence of brown spots on the ventral surface (vs chest and belly creamy white with many scattered brown spots in L. ventripunctata), 1/3 toe webbing and wide toe lateral fringes (vs no toe webbing and no lateral fringes in L. ventripunctata), and distinct dermal ridges under toes (vs absent in L. ventripunctata). Furthermore, the dominant vocal frequencies of the new species range from 7.3 to 8.3 kHz, which is unique compared to other Leptobrachella species and represents the highest dominant frequencies ever recorded. The Shiwandashan National Nature Reserve is now home to four known sympatric species of Leptobrachella.