Eslick Enid M, Beilby Mary J, Moon Anthony R
Department of Physics and Advanced Materials, University of Technology, Sydney, Broadway, NSW 2007, Australia.
Microscopy (Oxf). 2014 Apr;63(2):131-40. doi: 10.1093/jmicro/dft083. Epub 2014 Jan 23.
A substantial proportion of the architecture of the plant cell wall remains unknown with a few cell wall models being proposed. Moreover, even less is known about the green algal cell wall. Techniques that allow direct visualization of the cell wall in as near to its native state are of importance in unravelling the spatial arrangement of cell wall structures and hence in the development of cell wall models. Atomic force microscopy (AFM) was used to image the native cell wall of living cells of Ventricaria ventricosa (V. ventricosa) at high resolution under physiological conditions. The cell wall polymers were identified mainly qualitatively via their structural appearance. The cellulose microfibrils (CMFs) were easily recognizable and the imaging results indicate that the V. ventricosa cell wall has a cross-fibrillar structure throughout. We found the native wall to be abundant in matrix polysaccharides existing in different curing states. The soft phase matrix polysaccharides susceptible by the AFM scanning tip existed as a glutinous fibrillar meshwork, possibly incorporating both the pectic- and hemicellulosic-type substances. The hard phase matrix producing clearer images, revealed coiled fibrillar structures associated with CMFs, sometimes being resolved as globular structures by the AFM tip. The coiling fibrillar structures were also seen in the images of isolated cell wall fragments. The mucilaginous component of the wall was discernible from the gelatinous cell wall matrix as it formed microstructural domains over the surface. AFM has been successful in imaging the native cell wall and revealing novel findings such as the 'coiling fibrillar structures' and cell wall components which have previously not been seen, that is, the gelatinous matrix phase.
尽管已经提出了一些细胞壁模型,但植物细胞壁的大部分结构仍然未知。此外,关于绿藻细胞壁的了解更少。能够直接观察接近天然状态的细胞壁的技术,对于阐明细胞壁结构的空间排列以及细胞壁模型的建立至关重要。原子力显微镜(AFM)被用于在生理条件下对活的伞藻(Ventricaria ventricosa,V. ventricosa)细胞的天然细胞壁进行高分辨率成像。细胞壁聚合物主要通过其结构外观进行定性鉴定。纤维素微纤丝(CMFs)很容易识别,成像结果表明伞藻细胞壁整体具有交叉纤维结构。我们发现天然细胞壁富含处于不同固化状态的基质多糖。易受AFM扫描尖端影响的软相基质多糖以粘性纤维状网络的形式存在,可能包含果胶和半纤维素类型的物质。产生更清晰图像的硬相基质显示出与CMFs相关的卷曲纤维结构,有时被AFM尖端解析为球状结构。在分离的细胞壁片段图像中也可见到卷曲纤维结构。壁的粘液成分从凝胶状细胞壁基质中可分辨出来,因为它在表面形成了微观结构域。AFM成功地对天然细胞壁进行了成像,并揭示了诸如“卷曲纤维结构”和以前未见的细胞壁成分(即凝胶状基质相)等新发现。