Developmental Dynamics of Intercalary Meristem and Pith Cavity in Rice Stems.

In rice, internode elongation is a critical aspect of plant development and agricultural productivity. Previous morphological and histochemical studies using [³H]thymidine have visualized the cell division zone (intercalary meristem) in internodes. However, it has remained unclear how the intercalary meristem forms during stem development. In addition, while a pith cavity forms in the central part of the rice stem, the spatiotemporal relationship between pith cavity formation and intercalary meristem development is not well understood. Therefore, we performed histological analysis of intercalary meristem and pith cavity development using C9285, a deepwater rice variety that shows internode elongation from the vegetative growth stage. We classified the developmental stages of the stem into four stages based on the analysis of pith cavity formation using Trypan blue, Calcein-AM, and MitoRed staining, and visualized dividing cells using the Click-iT EdU imaging assay. In Stage 1, no pith cavity was formed. Vertical cell rows were observed between above the axillary bud attachment and the upper node, suggesting anticlinal divisions that lead to internode formation in the early stage of stem development. In Stage 2, the first pith cavity formed in the pith of the foot, which is the region of axillary bud attachment. Compared to cell division in the internode, that in the foot was significantly activated resulting in slight elongation from Stage 1 to Stage 2. In Stage 3, cell division in the foot ceased, while active cell division at the base of the internode led to significant vertical elongation. The second pith cavity formed due to cell death in the pith of the internode. In Stage 4, the two pith cavities connected to form a single large pith cavity. Although the intercalary meristem maintained cell division activity, the number of cell divisions decreased. Based on these results, we propose a model for stem development that involves two phases of elongation regulation: primary elongation involving slight elongation in the foot, and secondary elongation involving significant internode elongation due to the activation of cell division and cell elongation in the intercalary meristem. This is the first study to anatomically elucidate the spatiotemporal relationship between intercalary meristem development and pith cavity formation in rice stem development. It provides new insights for future research on rice stem development and studies of other grass species.