Morphogenesis of flattened unifacial leaves in Juncus prismatocarpus (Juncaceae)

To reveal the mode of morphogenesis of flattened unifacial leaves, we analysed the cell division direction and distribution on the leaf blade of Juncus prismatocarpus. Using the pulse-chase 5-ethynyl-2'-deoxyuridine method, we quantified and mapped the cell division direction on the leaf blade of J. prismatocarpus and compared the distribution of thickening cell divisions with the expression pattern of DROOPING LEAF (DL), a key gene involved in leaf blade thickening. Thickening cell divisions were the most abundant (> 45%) among all cell division directions on the leaf blade of J. prismatocarpus from the early plastochron 2 stage through the plastochron 3 stage. Mapping of cell divisions indicated that cell divisions in a particular direction were not restricted to a particular domain but were distributed diffusely throughout the entire cross-sectional area of the leaf blade. Gradient analysis indicated that the distribution of thickening cell divisions of the adaxial domain was denser than that of the abaxial domain. Contrary to the prolonged and diffuse distribution of thickening cell divisions, DL expression was transient and restricted in a narrow band. Our results suggest that a diffuse 'thickening meristem' plays the key role in the development of flattened unifacial leaves.