| A Pulse–chase EdU Method for Detection of Cell Division Orientation in Arabidopsis and Juncus prismatocarpus Leaf Primordia
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Author:
Date:
2021-01-05
[Abstract] In plants, the morphological diversity of leaves is largely determined by cell division, especially cell division orientation. Whereas cell division itself is easily monitored, the detection and quantification of cell division orientation are difficult. The few existing methods for detection and quantification of cell division orientation are either inefficient or laborious. Here, we describe a pulse-chase strategy using a 5-ethynyl-2’-deoxyuridine (EdU) labeling assay. Plant tissues are first incubated with EdU for a short period (pulse), followed by a long incubation without EdU (chase). Using this method, the positions of daughter cells are easily detected and can be used to quantify cell division orientation. Our protocol is rapid and very efficient for quantitative analysis of ...
[摘要] [摘要]在植物中，叶片的形态多样性在很大程度上取决于细胞分裂，尤其是细胞分裂方向。尽管细胞分裂本身很容易监测，但是细胞分裂方向的检测和定量却很困难。现有的几种检测和定量细胞分裂方向的方法要么效率低下要么费力。在这里，我们描述了使用5-乙炔基-2'-脱氧尿苷（EdU ）标记测定的脉冲追踪策略。首先将植物组织与EdU一起短时间（脉冲）孵育，然后在没有EdU的情况下长时间孵育（追逐）。使用这种方法，子细胞的位置易于检测，可用于量化细胞分裂方向。我们的协议可以快速有效地定量分析细胞分裂方向，并且可以同时应用于模型植物和非模型植物。
图形摘要：
通过脉冲追逐EdU方法清晰可见的植物细胞分裂对
[背景]植物细胞通过细胞壁彼此附接，并且不能迁移。因此，在叶片发育的早期，组织化，定向的细胞分裂在很大程度上决定了成熟叶片的形状。迄今为止，还没有报道用于有效和快速检测和定量细胞分裂取向的方法。现有方法包括使用ap CYCB1; 1 :: GUS （β-葡萄糖醛酸糖苷酶）报告基因线（末期）可视化子核（末期）（Horiguchi et al。，2011）或使用4'，6-diamidino可视化纺锤状赤道（中期） -2-苯基吲哚（DAPI）染色（Fukushima et ...
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| DNA Fragmentation Analysis
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Author:
Date:
2014-08-05
[Abstract] DNA fragmentation with length corresponding to multiple integer of approximately 180 base pairs is a distinct feature of apoptosis in animals and programmed cell death in plants. This feature can simply be detected by DNA gel electrophoresis followed by ethidium bromide staining, although in some cases it is difficult to distinguish the DNA laddering. We herein describe a protocol to detect a programmed cell death-associated DNA laddering of plant tissues. After agarose-gel electrophoresis of genomic DNA, Southern hybridization using DIG-labeled genomic DNA probe is performed, that improves detection of DNA laddering.
[摘要] 具有对应于大约180个碱基对的多个整数的长度的DNA片段是动物中凋亡和植物中程序性细胞死亡的独特特征。 这个特征可以简单地通过DNA凝胶电泳随后溴化乙锭染色检测，尽管在一些情况下难以区分DNA梯状。 我们在本文中描述了用于检测植物组织的程序性细胞死亡相关DNA梯状的方案。 在基因组DNA的琼脂糖凝胶电泳后，进行使用DIG标记的基因组DNA探针的Southern杂交，其改善DNA梯状的检测。
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