Profiling Cell Wall Monosaccharides and Nucleotide-Sugars from Plants

Q1 Agricultural and Biological Sciences Current protocols in plant biology Pub Date : 2019-06-12 DOI:10.1002/cppb.20092

Carsten Rautengarten, Joshua L. Heazlewood, Berit Ebert

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引用次数: 6

Abstract

The cell wall is an intricate mesh largely composed of polysaccharides that vary in structure and abundance. Apart from cellulose biosynthesis, the assembly of matrix polysaccharides such as pectin and hemicellulose occur in the Golgi apparatus before being transported via vesicles to the cell wall. Matrix polysaccharides are biosynthesized from activated precursors or nucleotide sugars. The composition and assembly of the cell wall is an important aspect in plant development and plant biomass utilization. The application of anion-exchange chromatography to determine the monosaccharide composition of the insoluble matrix polysaccharides enables a complete profile of all major sugars in the cell wall from a single run. While porous carbon graphite chromatography and tandem mass spectrometry delivers a sensitive and robust nucleotide sugar profile from plant extracts. Here we describe detailed methodology to quantify nucleotide sugars within the cell and profile the non-cellulosic monosaccharide composition of the cell wall. © 2019 by John Wiley & Sons, Inc.

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植物细胞壁单糖和核苷酸糖分析

细胞壁是一个复杂的网状结构，主要由不同结构和丰度的多糖组成。除了纤维素的生物合成外，基质多糖如果胶和半纤维素的组装在通过囊泡运输到细胞壁之前发生在高尔基体中。基质多糖是由活化的前体或核苷酸糖生物合成的。细胞壁的组成和组装是植物发育和生物量利用的一个重要方面。应用阴离子交换色谱法来确定不溶性基质多糖的单糖组成，可以从一次运行中获得细胞壁中所有主要糖的完整轮廓。而多孔碳石墨色谱和串联质谱提供了一个敏感和强大的核苷酸糖谱从植物提取物。在这里，我们描述了详细的方法来量化细胞内的核苷酸糖和描述细胞壁的非纤维素单糖组成。©2019 by John Wiley &儿子,Inc。

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Current protocols in plant biology Agricultural and Biological Sciences-Plant Science

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期刊介绍： Sound and reproducible laboratory methods are the foundation of scientific discovery. Yet nuances that are critical for an experiment''s success are not captured in the primary literature but exist only as part of a lab''s oral tradition. Current Protocols in Plant Biology provides reproducible step-by-step instructions for protocols relevant to plant research. Furthermore, Current Protocols content is thoughtfully organized by topic for optimal usage and to maximize contextual knowledge. Quarterly issues allow Current Protocols in Plant Biology to constantly evolve to keep pace with the newest discoveries and developments. Current Protocols in Plant Biology is the comprehensive source for protocols in the multidisciplinary field of plant biology, providing an extensive range of protocols from basic to cutting edge. Coverage includes: Extraction and analysis of DNA, RNA, proteins Chromosome analysis Transcriptional analysis Protein expression Metabolites Plant enzymology Epigenetics Plant genetic transformation Mutagenesis Arabidopsis, Maize, Poplar, Rice, and Soybean, and more.