| Rapid Determination of Cellulose, Neutral Sugars, and Uronic Acids from Plant Cell Walls by One-step Two-step Hydrolysis and HPAEC-PAD
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Author:
Date:
2016-10-20
[Abstract] The plant cell wall is primarily composed of the polysaccharides cellulose, hemicellulose and pectin. The structural and compositional complexity of these components are important for determining cell wall function during plant growth. Moreover, cell wall structure defines a number of functional properties of plant-derived biomass, such as rheological properties of foods and feedstock suitability for the production of cellulosic biofuels. A typical characterization of cell wall chemistry in the molecular biology lab consists of a mild acid hydrolysis for the quantification of hemicellulose and pectin-derived monomers and a separate analysis of cellulose by the Updegraff method. We have adopted a streamlined ‘one-step two-step’ hydrolysis protocol that allows for the simultaneous ...
[摘要] 植物细胞壁主要由多糖纤维素，半纤维素和果胶组成。这些组分的结构和组成复杂性对于确定植物生长期间的细胞壁功能是重要的。此外，细胞壁结构限定了植物来源的生物质的多种功能性质，例如食品的流变性质和用于生产纤维素生物燃料的原料适用性。分子生物学实验室中细胞壁化学的典型表征包括用于定量半纤维素和果胶衍生单体的温和酸水解和通过Updegraff方法对纤维素的单独分析。我们采用了一个简化的"一步两步"水解方案，允许通过配对的脉冲安培检测（HPAEC-PAD）的高效阴离子交换层析同时测定纤维素含量，中性糖和糖醛酸样品。在我们的工作中，该方案已经在很大程度上替代了Updegraff纤维素定量和用2μMTFA水解以在微量级上测定基质多糖组成。
[背景] 是基于在121℃下在4％（w/v）硫酸中水解的样品的配对分析。一组样品首先用72％（w/w）硫酸预处理以使纤维素膨胀并使其易于稀释酸水解（图1中的Saeman水解; ...
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| Analysis of Monosaccharides in Total Mucilage Extractable from Arabidopsis Seeds
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Author:
Date:
2016-05-05
[Abstract] The Arabidopsis thaliana seed coat epidermis produces copious amounts of mucilage polysaccharides (Haughn and Western, 2012). Characterization of mucilage mutants has identified novel genes required for cell wall biosynthesis and modification (North et al., 2014). The biochemical analysis of seed mucilage is essential to evaluate how different mutations affect cell wall structure (Voiniciuc et al., 2015c). Here we describe a robust method to screen the monosaccharide composition of Arabidopsis seed mucilage using ion chromatography (IC). Mucilage from up to 48 samples can be extracted and prepared for IC analysis within 24 h (only 4 h hands-on). Furthermore, this protocol enables fast separation (31 min per sample), automatic detection and ...
[摘要] 拟南芥种皮表皮产生大量的粘液多糖（Haughn和Western，2012）。 粘液突变体的表征已经鉴定了细胞壁生物合成和修饰所需的新基因（North等人，2014）。 种子粘液的生物化学分析对于评估不同突变如何影响细胞壁结构是必要的（Voiniciuc等人，2015c）。 在这里我们描述了使用离子色谱（IC）筛选拟南芥种子粘液的单糖组成的有力方法。 在24小时内（仅4小时手动）可以提取并准备来自多达48个样品的粘蛋白并进行IC分析。 此外，该协议使快速分离（每个样品31分钟），自动检测和定量的中性和酸性糖。
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| Quantification of the Mucilage Detachment from Arabidopsis Seeds
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Author:
Date:
2016-05-05
[Abstract] The Arabidopsis thaliana seed coat produces large amounts of cell wall polysaccharides, which swell out of the epidermal cells upon hydration of the mature dry seeds. While most mucilage polymers immediately diffuse in the surrounding solution, the remaining fraction tightly adheres to the seed, forming a dense gel-like capsule (Macquet et al., 2007). Recent evidence suggests that the adherence of mucilage is mediated by complex interactions between several cell wall components (Griffiths et al., 2014; Voiniciuc et al., 2015a). Therefore, it is important to evaluate how different cell wall mutants impact this mucilage property. This protocol facilitates the analysis of monosaccharides in sequentially extracted mucilage fractions, and quantifies the ...
[摘要] 拟南芥种子产生大量的细胞壁多糖，其在成熟干种子水合时从表皮细胞中膨胀。 虽然大多数粘液聚合物立即在周围溶液中扩散，但剩余部分紧密粘附于种子，形成致密的凝胶状胶囊（Macquet等人，2007）。 最近的证据表明粘液的粘附由几个细胞壁组分之间的复杂相互作用介导（Griffiths等人，2014; Voiniciuc等人，2015a）。 因此，重要的是评估不同的细胞壁突变体如何影响这种粘液性质。 该协议有助于分析顺序提取的粘液部分中的单糖，并且量化每种成分从种子中的脱离。
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