Histochemical Staining of Arabidopsis thaliana Secondary Cell Wall Elements
Histochemical Staining of Arabidopsis thaliana Secondary Cell Wall Elements
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DOI：
10.3791/51381
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发表时间：
2014-05-01
影响因子：
1.2
通讯作者：
Loque, Dominique
中科院分区：
文献类型：
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作者：
Mitra, Prajakta Pradhan;Loque, Dominique
拟南芥（Arabidopsis thaliana）是了解和操纵植物各种细胞过程的模式生物，被广泛应用于次生细胞壁形成的研究。次生细胞壁沉积发生在初代细胞壁形成之后，这一过程仅由形成血管和纤维组织的特化细胞进行。大多数次生细胞壁由纤维素（40-50%）、半纤维素（25-30%）和木质素（20-30%）组成。一些影响次生细胞壁生物合成的突变已经被分离出来，相应的突变可能会或可能不会表现出明显的生化成分变化或视觉表型，因为这些突变可能被代偿反应所掩盖。染色程序历来用于显示细胞基础上的差异。这些方法完全是可视的分析手段；然而，它们在快速和批判性分析中的作用是非常重要的。刚果红和钙氟白是用于检测多糖的染色剂，而Maule和间苯三酚通常用于测定木质素的差异，甲苯胺蓝O用于区分染色多糖和木质素。切片、染色和成像这些看似简单的技术对初学者来说可能是一个挑战。本研究从拟南芥模型的样品制备开始，详细介绍了各种染色方法的方案，这些方法可以很容易地用于观察植物次生细胞壁的细胞和组织组织。
Arabidopsis thaliana is a model organism commonly used to understand and manipulate various cellular processes in plants, and it has been used extensively in the study of secondary cell wall formation. Secondary cell wall deposition occurs after the primary cell wall is laid down, a process carried out exclusively by specialized cells such as those forming vessel and fiber tissues. Most secondary cell walls are composed of cellulose (40-50%), hemicellulose (25-30%), and lignin (20-30%). Several mutations affecting secondary cell wall biosynthesis have been isolated, and the corresponding mutants may or may not exhibit obvious biochemical composition changes or visual phenotypes since these mutations could be masked by compensatory responses. Staining procedures have historically been used to show differences on a cellular basis. These methods are exclusively visual means of analysis; nevertheless their role in rapid and critical analysis is of great importance. Congo red and calcofluor white are stains used to detect polysaccharides, whereas Maule and phloroglucinol are commonly used to determine differences in lignin, and toluidine blue O is used to differentially stain polysaccharides and lignin. The seemingly simple techniques of sectioning, staining, and imaging can be a challenge for beginners. Starting with sample preparation using the A. thaliana model, this study details the protocols of a variety of staining methodologies that can be easily implemented for observation of cell and tissue organization in secondary cell walls of plants.