Abstract: Leaf phenotypic variation among 10 populations of three Polyspora species was analyzed using multiple comparisons, nested variance analysis, correlation analysis, principal component analysis (PCA), and cluster analysis. Results revealed: (1) Significant differences in leaf phenotypic traits both within and among Polyspora populations (P<0.01). The average coefficient of variation was 17.03%, with trait-specific variation ranging from 0.05% to 31.37%. Mean variance components for leaf traits were higher within populations (180.79) than among populations (82.63), indicating greater differentiation within populations. (2) Dominant contributors to phenotypic variation included leaf width, petiole length, leaf area, leaf base angle, leaf roundness, and petiole index. Significant correlations among leaf traits were identified, highlighting the strong influence of environmental factors on leaf phenotypic variation. Cluster analysis grouped the 10 populations into three distinct groups based on leaf phenotypic traits. The results provide valuable insights into the adaptive evolution and exploitation of Polyspora species.