Abstract: To explore the relationship between leaf morphological characteristics and epiphyllous bud development of tropical water lilies, the viviparous and non-viviparous leaves of Nymphaea ‘Ruby’ and N.‘Blue Bird’ were used as materials, and the non-viviparous leaf of N. ‘Colorata’ was as control. The leaf morphological parameters of three cultivars were measured and compared, and the leaf anatomical structure during the leaf-rolling stage and leaf-expansion stage was observed using paraffin section. The results showed that all three cultivars had typical characteristics of tropical water lilies, the viviparous leaves of N. ‘Ruby’ and N. ‘Blue Bird’, except for the epiphyllous bud, their leaf length, leaf width, and leaf area were slightly lower than non-viviparous leaves, but there were no significant differences（P＞0.05）, while the anatomical structural parameters of the leaves changed with the leaf development. Although the upper and lower epidermis, sponge tissue of the leaves of viviparous N. ‘Ruby’ and N. ‘Blue Bird’ were thinner at the leaf-expansion stage, both were higher than that of non-viviparous leaves. However, the palisade tissue and the ratio of palisade to sponge increased at the leaf-expansion stage, and were lower than that of non-viviparous leaves, indicating that the development of the viviparous buds resulted in a decrease in the structural compactness of the palisade tissue, while the looseness of sponge tissue structure increased, but it was not related to the leaf thickness. The ratio of palisade to sponge and cell structure compactness of the non-viviparous variety N. ‘Colorata’ were significantly higher than those of the viviparous variety N. ‘Ruby’. In addition, the plasticity variation of each indicator was relatively high, and there was a certain correlation and an obvious co-evolution. Principal component analysis showed that the thickness of the upper and lower epidermis, spongy tissue, palisade tissue, the ratio of palisade to spongy, and the compactness and looseness of the leaf tissue structure might be regarded as the main indicators to reflect the anatomical structure characteristics of the viviparous leaves of water lilies. Viviparous leaves responded to epiphyllous bud development by changing the structure of mesophyll tissue.