Journal: Front Microbiol / Year: 2020 Title: Cryo-Electron Tomography Reveals the Complex Ultrastructural Organization of Multicellular Filamentous () Bacteria. Authors: Vasil A Gaisin / Romain Kooger / Denis S Grouzdev / Vladimir M Gorlenko / Martin Pilhofer / Abstract: The cell biology of is poorly studied. We applied cryo-focused ion beam milling and cryo-electron tomography to study the ultrastructural organization of thermophilic and , and mesophilic ". ...The cell biology of is poorly studied. We applied cryo-focused ion beam milling and cryo-electron tomography to study the ultrastructural organization of thermophilic and , and mesophilic ". Viridilinea mediisalina." These species represent the three main lineages within a group of multicellular filamentous anoxygenic phototrophic bacteria belonging to the order. We found surprising structural complexity in the . As with filamentous cyanobacteria, cells of and ". Viridilinea mediisalina" share the outer membrane-like layers of their intricate multilayer cell envelope. Additionally, cells of and ". Viridilinea mediisalina" are connected by septal channels that resemble cyanobacterial septal junctions. All three strains possess long pili anchored close to cell-to-cell junctions, a morphological feature comparable to that observed in cyanobacteria. The cytoplasm of the bacteria is crowded with intracellular organelles such as different types of storage granules, membrane vesicles, chlorosomes, gas vesicles, chemoreceptor-like arrays, and cytoplasmic filaments. We observed a higher level of complexity in the mesophilic strain compared to the thermophilic strains with regards to the composition of intracellular bodies and the organization of the cell envelope. The ultrastructural details that we describe in these bacteria will motivate further cell biological studies, given that the function and evolution of the many discovered morphological traits remain enigmatic in this diverse and widespread bacterial group.
Focused ion beam - Instrument: OTHER / Focused ion beam - Ion: OTHER / Focused ion beam - Voltage: 5 kV / Focused ion beam - Current: 0.035 nA / Focused ion beam - Duration: 300 sec. / Focused ion beam - Temperature: 125 K / Focused ion beam - Initial thickness: 1000 nm / Focused ion beam - Final thickness: 300 nm Focused ion beam - Details: The value given for _emd_sectioning_focused_ion_beam.instrument is Helios. This is not in a list of allowed values set(['DB235', 'OTHER']) so OTHER is written into the XML file.
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Electron microscopy
Microscope
FEI TITAN KRIOS
Image recording
Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Average electron dose: 120.0 e/Å2
Electron beam
Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron optics
Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company
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Image processing
Final reconstruction
Algorithm: BACK PROJECTION / Software - Name: IMOD / Number images used: 61
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