| A Method to Analyze Local and Systemic Effects of Environmental Stimuli on Root Development in Plants
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Author:
Date:
2016-09-05
[Abstract] Root development in vascular plants is innately tied to the environment. However, relatively little attention has been paid toward understanding the spatial scales at which the root perceives and responds to external stimuli. While some environmental signals elicit global responses that affect root system architecture, others may have more localized effects. We have observed that various developmental processes can be induced or suppressed along the circumference of the main root depending on local contact with available water in a process termed hydropatterning (Bao et al., 2014). Our studies of hydropatterning indicate that the root can detect and respond to certain external stimuli at the resolution of the diameter of a single organ. In order to characterize developmental ...
[摘要] 维管植物中的根发育与环境固有关系。然而，相对较少的关注已经被用于理解根部感知和响应外部刺激的空间尺度。虽然一些环境信号引起影响根系统结构的全局响应，但其他环境信号可能具有更局部化的效应。我们已经观察到，根据在称为水图案化的过程中与可用水的局部接触，可以沿着主根的圆周诱导或抑制各种发育过程（Bao等人，2014）。我们对水印的研究表明根部可以检测和响应某些外部刺激在单个器官的直径的分辨率。为了表征在该空间尺度的发育图案化，我们开发了使用琼脂培养基在体外改变穿过根的周向轴的环境输入的程序。根在"三明治"中的两块琼脂培养基之间生长。局部环境条件可以根据任一侧的介质的组成而变化。局部作用的刺激可以与基于根的发育反应系统作用的刺激相区分。在这里我们描述了整体的方法，并提供了一个例子，它如何可以用于分析外部水势梯度响应中玉米Zea mays（玉米）的侧根图案。我们还讨论了该方法如何可以更广泛地用于其他植物物种和环境处理。
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| In vitro CLE Peptide Bioactivity Assay on Plant Roots
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Author:
Date:
2015-12-20
[Abstract] Plant CLAVATA3/ESR (CLE)-related proteins play diverse roles in plant growth and development including regulating the development of root meristem. The mature functional forms of CLE peptides are typically 12-13 amino acids (aa) in length that are derived from the conserved C-termini of their precursor proteins. Genes encoding small secreted peptides sharing similarity to plant CLE proteins have recently been cloned from plant-parasitic nematodes, pests that infect many important crops. It is demonstrated that exogenous application of synthetic 12-14 aa CLE peptides corresponding to the CLE domain of their precursor proteins can suppress plant root growth. This protocol is to evaluate the bioactivity of CLE peptides originated from plant-parasitic nematodes by measuring the growth of ...
[摘要] 植物CLAVATA3/ESR（CLE）相关蛋白质在植物生长和发育中发挥多种作用，包括调节根分生组织的发育。 CLE肽的成熟功能形式通常为12-13个氨基酸（aa）长度，其衍生自其前体蛋白的保守C-末端。 最近从植物寄生线虫，感染许多重要作物的害虫克隆了编码与植物CLE蛋白相似的小分泌肽的基因。 证明外源应用对应于其前体蛋白的CLE结构域的合成12-14aa CLE肽可以抑制植物根生长。 该协议是通过测量CLE肽处理后植物根的生长或根尖分生组织（RAM）的大小来评价源自植物寄生线虫的CLE肽的生物活性。 用于研究的植物包括拟南芥和马铃薯。
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| Analysis of in vivo Cellulose Biosynthesis in Arabidopsis Cells by Spinning Disk Confocal Microscopy
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Author:
Date:
2015-10-05
[Abstract] Cellulose is a main component of plant cell walls. Tools to analyze cellulose mainly rely on analytical chemistry, which yields information about cellulose amounts and structure, but cannot be applied to intact tissues. Moreover, these methods measure total cellulose and cannot be used to assay cellulose synthesis per se. Live cell imaging of the catalytic subunits of the cellulose synthesis complex (CSC) conjugated to fluorescent proteins is an important tool to understand the dynamics of the cellulose biosynthesis process (Paredez et al., 2006). This method can be used in various genetic backgrounds (Sorek et al., 2014) or with different chemical inhibitors (Brabham and Debolt, 2012). Here we describe in detail the procedure to visualize the movement of CSCs ...
[摘要] 纤维素是植物细胞壁的主要成分。用于分析纤维素的工具主要依赖于分析化学，其产生关于纤维素量和结构的信息，但不能应用于完整组织。此外，这些方法测量总纤维素，并且不能用于测定纤维素合成本身。与荧光蛋白结合的纤维素合成复合物（CSC）的催化亚基的活细胞成像是理解纤维素生物合成过程的动力学的重要工具（Paredez等人，2006）。该方法可用于各种遗传背景（Sorek等人，2014）或使用不同的化学抑制剂（Brabham和Debolt，2012）。在这里我们详细描述的过程可视化CSCs在质膜的运动。由于CSC的运动可能是由葡聚糖合成和挤出进入细胞壁引起的，因此CSC速度的活细胞分析提供了一种直接测量体内纤维素合成的方法。
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