Department of Plant Biotechnology and Bioinformatics, Ghent University, Ghent, Belgium.
VIB Center for Plant Systems Biology, Ghent, Belgium.
Methods Mol Biol. 2022;2382:155-179. doi: 10.1007/978-1-0716-1744-1_10.
Oriented cell divisions are crucial throughout plant development to define the final size and shape of organs and tissues. As most of the tissues in mature roots and stems are derived from vascular tissues, studying cell proliferation in the vascular cell lineage is of great importance. Although perturbations of vascular development are often visible already at the whole plant macroscopic phenotype level, a more detailed characterization of the vascular anatomy, cellular organization, and differentiation status of specific vascular cell types can provide insights into which pathway or developmental program is affected. In particular, defects in the frequency or orientation of cell divisions can be reliably identified from the number of vascular cell files. Here, we provide a detailed description of the different clearing, staining, and imaging techniques that allow precise phenotypic analysis of vascular tissues in different organs of the model plant Arabidopsis thaliana throughout development, including the quantification of cell file numbers, differentiation status of vascular cell types, and expression of reporter genes.
定向细胞分裂在植物发育过程中至关重要，可决定器官和组织的最终大小和形状。由于成熟根和茎中的大多数组织都来源于维管束组织，因此研究维管束细胞谱系中的细胞增殖具有重要意义。尽管维管发育的扰动通常在整个植株宏观表型水平上已经可见，但对特定维管束细胞类型的维管解剖、细胞组织和分化状态进行更详细的特征描述，可以深入了解受到影响的途径或发育程序。特别是，从维管束细胞层数中可以可靠地识别出细胞分裂频率或方向的缺陷。在这里，我们提供了详细的描述，介绍了不同的透明化、染色和成像技术，这些技术可用于在拟南芥模型植物的不同器官中对维管组织进行精确的表型分析，包括细胞层数的定量、维管束细胞类型的分化状态和报告基因的表达。
