Microscopic studies on Thermosipho globiformans implicate a role of the large periplasm of Thermotogales
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Thermosipho globiformans is a member of Thermotogales, which contains rod-shaped, Gram-negative, anaerobic (hyper)thermophiles. These bacteria are characterized by an outer sheath-like envelope, the toga, which includes the outer membrane and an amorphous layer, and forms large periplasm at the poles of each rod. The cytoplasmic membrane and its contents are called “cell”, and the toga and its contents “rod”, to distinguish between them. Optical cells were constructed to observe binary fission of T. globiformans. High-temperature microscopy of rods adhering to optical cells' coverslips showed that the large periplasm forms between newly divided cells in a rod, followed by rod fission at the middle of the periplasm, which was accompanied by a sideward motion of the newly generated rod pole(s). Electron microscopic observations revealed that sessile rods grown on a glass plate have nanotubes adhered to the glass, and these may be involved in the sideward motion. Epifluorescence microscopy with a membrane-staining dye suggested that formation of the septal outer membrane is distinct from cytokinesis. Transmission electron microscopy indicated that the amorphous layer forms in the periplasm between already-divided cells. These findings suggest that the large periplasm is the structure in which the septal toga forms, an event separate from cytokinesis.
KeywordsAnaerobic bacteria Cell division Deep-sea thermophiles Nanotubes Toga
Anaerobic thermophile observation chamber
Field-emission scanning electron microscopy
Transmission electron microscopy
We thank Drs. Shigeru Deguchi and Sada-atsu Mukai of the Japan Agency for Marine-Earth Science and Technology for microscopic observations using their HTM system during the initial stages of this study, and Dr. Haruyo Yamaguchi for helping us with electron microscopy techniques. We are also grateful to Mr. Akitomo Kawasaki for some TEM work.
Supplementary material 2: Movie S2 (MPG 1647 kb)
Supplementary material 3: Movie S1 (MPG 4586 kb)
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