Department of Cell Biology, John Innes Centre for Plant Science Research, Norwich Research Park, Colney Lane, Norwich, NR4 7UH, United Kingdom.
Plant Physiol. 1992 Dec;100(4):1940-7. doi: 10.1104/pp.100.4.1940.
Highly reproducible Fourier transform infrared (FTIR) spectra from both single onion (Allium cepa) cell walls and their constituent polymers were obtained under a variety of sampling conditions. The specificity of the chemical extraction sequence used in the preparation of the material was confirmed: pectins only are extracted by cyclohexanediaminetetraacetic acid and sodium carbonate, whereas xyloglucans are extracted by increasing concentrations of potassium hydroxide. There was very little contamination of the first potassium hydroxide extract with residual pectin. The low abundance of both phenolics and protein was also confirmed. The first sodium carbonate extraction almost completely removes esters remaining in the cell wall. We have demonstrated that FTIR spectroscopy can detect large conformational changes in pectic polymers on removal from the cell wall and on drying. FTIR spectroscopy provides a powerful and rapid assay for wall components and putative cross-links by identifying polymers and functional groups nondestructively in muro. The availability of micro-sampling and data acquisition techniques that permit subtraction of the blanket absorption of water make FTIR spectroscopy particularly suitable for studies of cell wall architecture. The use of polarizers with the microscope accessory permits determination of the orientation of particular functional groups with respect to the direction of cell elongation in carrot suspension cells.
从各种采样条件下获得了单个洋葱(Allium cepa)细胞壁及其组成聚合物的高度可重现的傅里叶变换红外(FTIR)光谱。所使用的化学提取序列的特异性在材料制备中得到了确认:仅用环己二胺四乙酸和碳酸钠提取果胶,而用氢氧化钾的浓度增加提取木葡聚糖。第一氢氧化钾提取物中残留果胶的污染非常少。还证实了酚类和蛋白质的含量非常低。第一次碳酸钠提取几乎完全去除细胞壁中残留的酯。我们已经证明,FTIR 光谱可以检测到果胶聚合物从细胞壁上除去和干燥时的大构象变化。FTIR 光谱通过在细胞壁内非破坏性地识别聚合物和官能团,提供了一种用于细胞壁成分和假定交联的强大而快速的分析方法。微采样和数据采集技术的可用性允许减去水的 blanket 吸收,这使得 FTIR 光谱特别适合于细胞壁结构的研究。使用显微镜附件中的偏振器可以确定胡萝卜悬浮细胞中特定官能团相对于细胞伸长方向的取向。