Occurrence and characterization of arabinogalactan-like proteins and hemicelluloses in Micrasterias (Streptophyta)

The cell wall of the green alga Micrasterias denticulata Breb. ex Ralfs (Desmidiaceae, Zygnematophyceae, Streptophyta) was investigated to obtain information on the composition of component polysaccharides and proteoglycans to allow comparison with higher plants and to understand cell wall functions during development. Various epitopes currently assigned to arabinogalactan-proteins (AGPs) of higher plants could be detected in Micrasterias by immuno TEM and immunofluorescence methods, but the walls did not bind the beta-D-glycosyl-Yariv (beta-GlcY) reagent. Secretory vesicles and the primary wall were labeled by antibodies against AGPs (JIM8, JIM13, JIM14). Dot and Western blot experiments indicated a proteoglycan nature of the epitopes recognized, which consisted of galactose and xylose as major sugars by high performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD). Epitopes of alkali-soluble polysaccharides assigned to noncellulosic polysaccharides in higher plants could be detected and located in the wall during its formation. The polyclonal anti-xyloglucan (anti-XG) antibody labeled primary and secondary wall of Micrasterias, whereas the monoclonal antibody CCRC-M1, directed against the fucose/galactose side chain of xyloglucan (XyG), did not recognize any structures. Labeling by anti-XG antibody at the trans-sites of the dictyosomes and at wall material containing vesicles indicated that secretion of the epitopes occurred similar to higher plants. The presence of (1 -> 3, 1 -> 4)-beta-glucan (mixed linked glucan) in the secondary cell wall but not in the primary cell wall of Micrasterias could be demonstrated by an antibody recognizing this glucan type, whereas (1 -> 3)-beta-glucan (callose) could not be detected. The analytical results revealed that alkali-soluble polysaccharides in the secondary wall of Micrasterias consist mostly of (1 -> 3, 1 -> 4)-beta-D-glucan.