Ratković S
Department of Technology and Chemical Research, Maize Research Institute.
Glas Srp Akad Nauka Med. 1992(42):157-70.
We review here the recent achievements in high resolution NMR spectroscopy of four biologically most important nuclei (1H, 13C, 15N, 31P) applied on plant seeds in vivo or on extracts from the same tissue. Most often used nuclei in NMR analysis of seeds were protons (1H). The application of low resolution NMR for nondestructive analysis of moisture and oil in seeds had a long history--this was actually the only true nondestructive chemical analysis of genetic material. The combination of high resolution 1H NMR with magic angle sample spinning (MAS) enabled some authors to obtain a method for in vivo study of fatty acids composition of seed lipids. A promising technique for in vivo studies of biochemical composition and physiological processes in seeds is 13C NMR, particularly 1H-13C cross-polarization NMR combined with MAS technique. This enabled analysis of lipids in intact seeds, but also obtaining of high resolution spectra from the solid components of the seed matrix (starch, sugars, proteins). An interesting application of 13C NMR was monitoring the biochemical events following seed germination in vivo. Low natural ability of the NMR accessible 15N nucleus restricts the application of 15N NMR for in vivo seed analysis. Nevertheless, some sophisticated experiments combining double cross-polarization 15N/13C with the MAS technique were reviewed. 31P NMR is a promising technique in biological studies. Some recent achievements included detection of energetically important molecules of cells (ADP; ATP, oligosaccharides) as well as compartmentation of the inorganic phosphate (cytoplasmic, vacuolar). There were some indications of the studies made on intact seeds, but most of the 31P NMR work has been done on the extracts from seed material.
在此,我们回顾了高分辨率核磁共振光谱技术在四种对生物学最为重要的原子核(氢-1、碳-13、氮-15、磷-31)上的最新研究成果,这些研究是针对植物种子活体或同一组织的提取物进行的。在种子的核磁共振分析中,最常使用的原子核是质子(氢-1)。低分辨率核磁共振技术用于种子水分和油脂的无损分析已有很长历史——这实际上是对遗传物质唯一真正的无损化学分析。高分辨率氢-1核磁共振与魔角样品旋转(MAS)相结合,使一些研究人员得以开发出一种用于体内研究种子脂质脂肪酸组成的方法。碳-13核磁共振是一种用于体内研究种子生化组成和生理过程的有前景的技术,特别是氢-1-碳-13交叉极化核磁共振与MAS技术相结合。这不仅能够分析完整种子中的脂质,还能从种子基质的固体成分(淀粉、糖类、蛋白质)中获得高分辨率光谱。碳-13核磁共振的一个有趣应用是在体内监测种子萌发后的生化事件。氮-15原子核的低自然丰度限制了氮-15核磁共振在种子体内分析中的应用。尽管如此,本文还是回顾了一些将双交叉极化氮-15/碳-13与MAS技术相结合的复杂实验。磷-31核磁共振是生物学研究中的一项有前景的技术。近期的一些成果包括检测细胞中具有重要能量意义的分子(二磷酸腺苷;三磷酸腺苷、寡糖)以及无机磷酸盐的区室化(细胞质、液泡)。虽然有一些关于完整种子的研究迹象,但大多数磷-31核磁共振工作是在种子材料的提取物上完成的。