Bamboo is one of the essential ornamental plants that is widely used as a decorative landscape element in gardens. Phyllostachys violascens cv. Viridisulcata has a unique internode color phenotype with yellow culm and green sulcus, but their structural and development differences remain unknown. In the current study, we analyzed the histological analysis of internode cross-sections through SEM and microscopy. These results revealed that the vascular bundles distributed in the culm were organized in oblique rows and multiple lines. In contrast, the vascular bundles’ distribution in the sulcus was much more random. The distribution density, maximum length, and maximum width of vascular bundles were also differentiated between the sulcus and the culm. Further, the cell wall thickness of fiber cells in the culm was more than 30% thicker than the sulcus. The FT-IR analysis identified that the culm and sulcus had similar structural properties. The total lignin content measurement revealed that lignin accumulated more in the sulcus than in the culm. Additionally, we identified the lignin biosynthesis pathway genes, Pv4CL and PvC4H, which were differentially expressed between the culm and sulcus through transcriptomic data and qPCR analyses. In conclusion, our results identified that the vascular bundles’ structure differed between the culm and sulcus, and Pv4CL and PvC4H genes might play an essential role in their development.