The Polygonal Cell Shape and Surface Protein Layer of Anaerobic Methane-Oxidizing Methylomirabilislanthanidiphila Bacteria

Methylomirabilis bacteria perform anaerobic methane oxidation coupled to nitrite reduction via an intra-aerobic pathway, producing carbon dioxide and dinitrogen gas. These diderm bacteria possess an unusual polygonal cell shape with sharp ridges that run along the cell body. Previously, a putative surface protein layer (S-layer) was observed as the outermost cell layer of these bacteria. We hypothesized that this S-layer is the determining factor for their polygonal cell shape. Therefore, we enriched the S-layer from M. lanthanidiphila cells and through LC-MS/MS identified a 31 kDa candidate S-layer protein, mela_00855, which had no homology to any other known protein. Antibodies were generated against a synthesized peptide derived from the mela_00855 protein sequence and used in immunogold localization to verify its identity and location. Both on thin sections of M. lanthanidiphila cells and in negative-stained enriched S-layer patches, the immunogold localization identified mela_00855 as the S-layer protein. Using electron cryo-tomography and sub-tomogram averaging of S-layer patches, we observed that the S-layer has a hexagonal symmetry. Cryo-tomography of whole cells showed that the S-layer and the outer membrane, but not the peptidoglycan layer and the cytoplasmic membrane, exhibited the polygonal shape. Moreover, the S-layer consisted of multiple rigid sheets that partially overlapped, most likely giving rise to the unique polygonal cell shape. These characteristics make the S-layer of M. lanthanidiphila a distinctive and intriguing case to study.

Localization of xyloglucan epitopes in the gelatinous layer of developing and mature gelatinous fibers of European aspen (Populus tremula L.) tension wood

There is controversy concerning the presence of xyloglucans in gelatinous (G) layers of Populus spp. tension wood, particularly in mature G-fibers. Transmission electron microscopy (TEM) immunogold localization combined with LM15 antibody (recognizes XXXG-motif of xyloglucans, heptasaccharide) was used to investigate the distribution of xyloglucan epitopes in both transverse and radial sections of P. tremula tension wood. Results provided clear evidence for the presence of xyloglucans in both mature and developing G-layers. Developmental decrease of LM15 epitope localization in G-layers was also detected during G-fiber maturation. High magnification TEM observations showed specific localization of LM15 epitopes on newly synthesized cellulose macrofibrils present in the innermost layer of developing G-layers adjacent to the cell lumen, suggesting linkage between xyloglucans and cellulose macrofibrils. Possible mechanisms were discussed for developmental changes of xyloglucan with respect to the different results reported in the literature.

Chemical and ultrastructural changes of ash wood thermally modified (TMW) using the thermo-vacuum process: II. Immunocytochemical study of the distribution of noncellulosic polysaccharides

Following structural and cytochemical studies (Part I) on thermally modified ash wood (TMW) by the thermo-vacuum (Termovuoto) process, changes in the distribution of noncellulosic polysaccharides have been investigated in TMW treated for 3 h at 220°C (TMW3 h, 220°C) by means of immunogold localization methods. Pectins (homogalacturonan, rhamnogalacturonan-I) and xyloglucan were significantly degraded in compound middle lamella (CML), including the middle lamella cell corner regions (CMLcc), of all xylem cells after thermal modification. Xylan and mannan degradation were also visible in fiber cell walls. In particular, degradation of mannan was very significant and showed variation between cell wall regions even within the same cell wall. The degradation of pectins was more significant than that of hemicelluloses. In summary, results suggest that each noncellulosic polysaccharide may have a different degradation process in ash TMWs.