Pathways of extra- and intercellular diffusion of colored substances in the blackened xylem of Diospyros kaki

Some species of Diospyros form the black wood that is known as ebony. Infrequently, D. kaki forms wood with black patterning that is called “kurogaki” in Japan and is highly valued. To understand the mechanism of formation of the blackened xylem, we attempted to clarify the pathways of diffusion of colored substances from the site of their biosynthesis to their deposition in secondary xylem. We observed colored substances in pits and intercellular spaces and we recorded the deposition of colored substances in the cell walls of wood fibers. In gray and black regions, we found colored substances in the pits between xylem cells, in blind pits and in intercellular spaces. At the boundary between whitish and black regions, areas of coloration in the cell walls of wood fibers gradually increased in size from the whitish to the black regions. Heavy deposition of colored substances was observed in the inner region of the secondary walls of wood fibers. Furthermore, coloration of the outer region of the secondary walls and compound middle lamella (CML) of wood fibers gradually became stronger toward the black regions. Our observations suggest that pit-pairs and intercellular spaces might provide the pathways for inter- and extracellular diffusion of colored substances, respectively. In addition, colored substances might penetrate the cell walls of wood fibers, moving from the lumen into the cell wall.