Raimundo Sandra Cristina, Pattathil Sivakumar, Eberhard Stefan, Hahn Michael G, Popper Zoë A
Botany and Plant Science and Ryan Institute for Environmental, Marine and Energy Research, School of Natural Sciences, National University of Ireland Galway, Galway, Ireland.
Complex Carbohydrate Research Center, University of Georgia, Athens, GA, 30602, USA.
Protoplasma. 2017 Mar;254(2):997-1016. doi: 10.1007/s00709-016-1007-6. Epub 2016 Aug 25.
LAMP is a cell wall-directed monoclonal antibody (mAb) that recognizes a β-(1,3)-glucan epitope. It has primarily been used in the immunolocalization of callose in vascular plant cell wall research. It was generated against a brown seaweed storage polysaccharide, laminarin, although it has not often been applied in algal research. We conducted in vitro (glycome profiling of cell wall extracts) and in situ (immunolabeling of sections) studies on the brown seaweeds Fucus vesiculosus (Fucales) and Laminaria digitata (Laminariales). Although glycome profiling did not give a positive signal with the LAMP mAb, this antibody clearly detected the presence of the β-(1,3)-glucan in situ, showing that this epitope is a constituent of these brown algal cell walls. In F. vesiculosus, the β-(1,3)-glucan epitope was present throughout the cell walls in all thallus parts; in L. digitata, the epitope was restricted to the sieve plates of the conductive elements. The sieve plate walls also stained with aniline blue, a fluorochrome used as a probe for callose. Enzymatic digestion with an endo-β-(1,3)-glucanase removed the ability of the LAMP mAb to label the cell walls. Thus, β-(1,3)-glucans are structural polysaccharides of F. vesiculosus cell walls and are integral components of the sieve plates in these brown seaweeds, reminiscent of plant callose.
LAMP是一种细胞壁导向的单克隆抗体(mAb),可识别β-(1,3)-葡聚糖表位。它主要用于维管植物细胞壁研究中胼胝质的免疫定位。它是针对一种褐藻贮藏多糖海带多糖产生的,尽管它在藻类研究中并不常用。我们对墨角藻(墨角藻目)和掌状海带(海带目)这两种褐藻进行了体外研究(细胞壁提取物的糖组分析)和原位研究(切片的免疫标记)。尽管糖组分析未用LAMP单克隆抗体给出阳性信号,但该抗体在原位清楚地检测到了β-(1,3)-葡聚糖的存在,表明该表位是这些褐藻细胞壁的一个组成部分。在墨角藻中,β-(1,3)-葡聚糖表位存在于所有叶状体部分的整个细胞壁中;在掌状海带中,该表位仅限于传导元件的筛板。筛板壁也用苯胺蓝染色,苯胺蓝是一种用作胼胝质探针的荧光染料。用内切β-(1,3)-葡聚糖酶进行酶消化消除了LAMP单克隆抗体标记细胞壁的能力。因此,β-(1,3)-葡聚糖是墨角藻细胞壁的结构多糖,是这些褐藻筛板的组成部分,类似于植物胼胝质。