Fiorani Fabio, Beemster Gerrit T S
Department of Plant Systems Biology, Flanders Interuniversity Institute for Biotechnology (VIB)/University of Ghent, Technologiepark 927, Ghent, Belgium.
Plant Mol Biol. 2006 Apr;60(6):963-79. doi: 10.1007/s11103-005-4065-2.
At the molecular level regulatory interactions between cell cycle genes are being uncovered rapidly, but less progress is made in unravelling how these molecular events regulate growth processes at the level of cells and of the whole organism. The main obstacle is the absence of a set of analytical tools that are powerful enough to determine pertinent parameters and, at the same time, relatively easy to use by non-specialized laboratories. Appropriate methodology to obtain this type of data has been pioneered in the first half of the last century and is now commonly defined as 'kinematic analysis'. Unfortunately, the laborious nature of these analyses and the relatively complex numerical methods used, have limited their use to only a handful of specialized research groups. In this article we attempt to present an accessible entry to this methodology, particularly in terms of the mathematical framework. We start describing the simplest possible system, i.e., a virtually homogenous cell suspension culture. Then, we outline the analysis of dicotyledonous leaves, root tips, monocotyledonous leaves, and finally shoot apical meristems. For each of these systems we discuss the details of the calculation of cell division parameters such as cell cycle duration, size of the meristem and number of cells contained in it, which enables answering fundamental questions about the relative contribution of differences in cell production and cell size to variation in growth. In addition, we discuss the assumptions and limitations of these and alternative methodologies with the aim to facilitate the choice of appropriate analyses depending on the specific research question.
在分子水平上，细胞周期基因之间的调控相互作用正迅速被揭示，但在阐明这些分子事件如何在细胞水平和整个生物体水平上调节生长过程方面进展较小。主要障碍是缺乏一套分析工具，这些工具既要强大到足以确定相关参数，又要相对易于非专业实验室使用。获取这类数据的适当方法在上世纪上半叶就已开创，现在通常被定义为“运动学分析”。不幸的是，这些分析的繁琐性质以及所使用的相对复杂的数值方法，使得它们仅被少数专业研究小组使用。在本文中，我们试图提供一种易于理解的该方法入门介绍，特别是在数学框架方面。我们首先描述最简单的可能系统，即几乎均匀的细胞悬浮培养物。然后，我们概述双子叶植物叶片、根尖、单子叶植物叶片，最后是茎尖分生组织的分析。对于这些系统中的每一个，我们都讨论细胞分裂参数计算的细节，如细胞周期持续时间、分生组织的大小及其所含细胞的数量，这有助于回答关于细胞产生差异和细胞大小对生长变化的相对贡献的基本问题。此外，我们讨论这些方法和替代方法的假设和局限性，目的是根据具体研究问题促进选择合适的分析方法。
