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Calli derived from young leaves of Aesculus turbinata contained tracheary elements with large pores that resembled perforations of vessel elements. The differentiation of tracheary elements in vitro provides a useful system for detailed analysis of xylem cell differentiation. To examine the mechanism of formation of cell wall structures, new differentiation systems are required that allows us to induce highly organized structures, such as perforations. In this study, we developed such a system in which we were able to induce formation o