Max-Planck-Institut für Molekulare Pflanzenphysiologie, Potsdam, Germany.
Methods Mol Biol. 2020;2156:255-268. doi: 10.1007/978-1-0716-0660-5_17.
The cell wall has a crucial influence on the mechanical properties of plant cells. It therefore has a strong impact on the freezing behavior and very likely also the freezing tolerance of plants. However, not many studies have addressed the question how cell wall composition and structure impact plant freezing tolerance and cold acclimation. In this chapter, we describe a comprehensive workflow to extract total cell wall material from leaves of Arabidopsis thaliana and to separate this material into fractions enriched in crystalline cellulose, pectins, and hemicelluloses by sequential fractionation. We further describe methods for the analysis of chemical structure, monosaccharide composition, and cellulose and uronic acid contents in the total cell wall material and the fractions in response to cold acclimation. Structural properties of cell wall material are analyzed by attenuated total reflectance-Fourier-transform infrared spectrometry (ATR-FTIR) and monosaccharide composition by gas chromatography-mass spectrometry (GC-MS) after isolation of alditol acetate derivatives of the sugars.
细胞壁对植物细胞的机械性能有至关重要的影响。因此，它对植物的冻结行为，很可能还有抗冻性有强烈的影响。然而，目前还没有多少研究涉及细胞壁组成和结构如何影响植物的抗冻性和耐寒性。在本章中，我们描述了一种从拟南芥叶片中提取总细胞壁物质并通过连续分级分离将该物质分离为富含结晶纤维素、果胶和半纤维素的级分的综合工作流程。我们还描述了用于分析总细胞壁物质和级分中化学结构、单糖组成以及纤维素和糖醛酸含量的方法，以响应低温驯化。细胞壁物质的结构特性通过衰减全反射傅里叶变换红外光谱（ATR-FTIR）进行分析，单糖组成通过气相色谱-质谱（GC-MS）进行分析，方法是在分离糖的糖醇乙酸酯衍生物后进行。
