National Key Facility for Crop Gene Resources and Genetic Improvement, Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing, China.
Methods Mol Biol. 2022;2382:131-140. doi: 10.1007/978-1-0716-1744-1_8.
Rice leaves have a consistent spatial and temporal organization of cell division and expansion, which leads to typical cell length profiles along the longitudinal axis. The growth of rice leaves is usually studied during a steady-state period when leaf elongation rate is constant and the spatial distribution of cell length is temporally invariable. In this chapter, we define the steady-state period by analyzing the leaf elongation rate of leaf three in rice. During steady growth of leaf three, we determine the meristem size by identifying the epidermal cell files next to the stomatal files which are the distal position of meristem zone with confocal laser scanning microscopy. Meanwhile, we plot the cell length profiles along the longitudinal axis from which we directly determine the length of growing zone and mature cell size. Other cell division and expansion parameters such as cell division rate, cell cycle duration, and stain rate are calculated through indirect kinematic analysis.
水稻叶片具有一致的细胞分裂和扩张时空组织,这导致了沿着纵轴的典型细胞长度分布。水稻叶片的生长通常在稳定状态期间进行研究,此时叶片伸长率保持恒定,细胞长度的空间分布在时间上是不变的。在这一章中,我们通过分析水稻三叶的叶片伸长率来定义稳定状态期。在三叶的稳定生长期间,我们通过共聚焦激光扫描显微镜识别靠近气孔列的表皮细胞列来确定分生组织的大小,这些表皮细胞列是分生组织区的远轴位置。同时,我们绘制了沿纵轴的细胞长度分布,从中我们可以直接确定生长区的长度和成熟细胞的大小。其他细胞分裂和扩张参数,如细胞分裂率、细胞周期持续时间和染色率,都是通过间接运动学分析来计算的。