Pasternak Taras, Pérez-Pérez José Manuel
Centre for BioSystems Analysis, BIOSS Centre for Biological Signalling Studies University of Freiburg, Institute of Biology II/Molecular Plant Physiology, 79104 Freiburg, Germany.
Instituto de Bioingeniería, Universidad Miguel Hernández, 03202 Elche, Spain.
Plants (Basel). 2021 Nov 6;10(11):2399. doi: 10.3390/plants10112399.
When dealing with plant roots, a multiscale description of the functional root structure is needed. Since the beginning of 21st century, new devices such as laser confocal microscopes have been accessible for coarse root structure measurements, including three-dimensional (3D) reconstruction. Most researchers are familiar with using simple 2D geometry visualization that does not allow quantitative determination of key morphological features from an organ-like perspective. We provide here a detailed description of the quantitative methods available for 3D analysis of root features at single-cell resolution, including root asymmetry, lateral root analysis, cell size and nuclear organization, cell-cycle kinetics, and chromatin structure analysis. Quantitative maps of the root apical meristem (RAM) are shown for different species, including (L.), Heynh, L., L., and (L.) P. Beauv. The 3D analysis of the RAM in these species showed divergence in chromatin organization and cell volume distribution that might be used to study root zonation for each root tissue. Detailed protocols and possible pitfalls in the usage of the marker lines are discussed. Therefore, researchers who need to improve their quantitative root biology portfolio can use them as a reference.
在研究植物根系时，需要对根系功能结构进行多尺度描述。自21世纪初以来，诸如激光共聚焦显微镜等新设备已可用于粗根结构测量，包括三维（3D）重建。大多数研究人员熟悉使用简单的二维几何可视化方法，这种方法无法从类似器官的角度对关键形态特征进行定量测定。我们在此详细描述了可用于单细胞分辨率下根系特征三维分析的定量方法，包括根的不对称性、侧根分析、细胞大小和细胞核组织、细胞周期动力学以及染色质结构分析。展示了不同物种根尖分生组织（RAM）的定量图谱，包括（L.）、Heynh、L.、L.和（L.）P. Beauv。对这些物种RAM的三维分析表明，染色质组织和细胞体积分布存在差异，这可能用于研究每个根组织的根分区。讨论了标记系使用中的详细方案和可能存在的问题。因此，需要完善其定量根系生物学研究组合的研究人员可以将其作为参考。
