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对包埋于LR白色树脂中的成熟亚麻纤维的显微镜研究:细胞壁基质多糖的免疫金定位

Microscopic studies on mature flax fibers embedded in LR white: immunogold localization of cell wall matrix polysaccharides.

作者信息

His I, Andème-Onzighi C, Morvan C, Driouich A

机构信息

UMR CNRS 6037, IFRMP23, Centre Commun de Microscopie Electronique, Université de Rouen, Mont Saint Aignan, France.

出版信息

J Histochem Cytochem. 2001 Dec;49(12):1525-36. doi: 10.1177/002215540104901206.

Abstract

Flax fibers have been the subject of many biochemical studies, which revealed that cellulose and pectins are the major constituents of their walls. In contrast, little is known about the location of these polymers within the walls of mature fibers by microscopic methods. This has been technically hampered by the very thick secondary wall of fibers, resulting in inadequate tissue preservation unsuitable for immunogold microscopy. In this study, we adapted the basic chemical fixation, dehydration and infiltration methods to achieve a good preservation of the cell structures of mature fibers and reduced damage to antigens. We were able to apply postembedding immunocytochemical techniques to map the location of various pectic epitopes within the walls of mature fibers. Our immunolabeling data show that homogalacturonans were exclusively found in the middle lamellae and the cell junctions but were not detectable in the secondary wall. In contrast, rhamnogalacturonan I (RG I)-associated epitopes, as well as galactan and arabinan epitopes, were abundantly distributed over the secondary wall of mature fibers.

摘要

亚麻纤维一直是众多生物化学研究的对象,这些研究表明纤维素和果胶是其细胞壁的主要成分。相比之下,通过显微镜方法对这些聚合物在成熟纤维细胞壁内的位置了解甚少。这在技术上受到纤维非常厚的次生壁的阻碍,导致组织保存不充分,不适用于免疫金显微镜观察。在本研究中,我们调整了基本的化学固定、脱水和渗透方法,以实现对成熟纤维细胞结构的良好保存,并减少对抗原的损伤。我们能够应用包埋后免疫细胞化学技术来绘制各种果胶表位在成熟纤维细胞壁内的位置。我们的免疫标记数据表明,同型半乳糖醛酸仅存在于中层和细胞连接处,而在次生壁中未检测到。相比之下,鼠李半乳糖醛酸聚糖I(RG I)相关表位以及半乳聚糖和阿拉伯聚糖表位在成熟纤维的次生壁上大量分布。

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