Ultrastructure of the Exosporium Enveloping Spores Ofbacillus Cereus

Gerhardt, Philipp (The University of Michigan, Ann Arbor), and Edgar Ribi. Ultrastructure of the exosporium enveloping spores of Bacillus cereus. J. Bacteriol. 88:1774-1789. 1964.-Structural details in the outer envelope of spores, such as those of Bacillus cereus and B. anthracis, were studied by electron microscopy and by X-ray diffraction analysis. Procedures were developed for isolating homogeneous fragments of the membrane with minimal damage to or germination of the spore proper. Exosporium of B. cereus appeared to embody two main layers. An outer layer was made up of a nap of hairlike projections, irregularly distributed and about 250 A deep; these arose from an intermediate covering, about 60 A in depth and similarly lead-stainable. An inner basal layer had a hexagonally perforate surface pattern of holes, averaging 76 A from center to center, and was made up of four lamellae, which could fragment into crystal-like elements. The intact basal membrane was about 190 A thick and the thinnest elements, 45 A. Microscopic observations of a crystal-like nature of the exosporium basal membrane were confirmed by X-ray diffraction analysis; the pattern of reflection lines in powder diagrams of exosporium fragments or paracrystals, or intact spores, corresponded to a hexagonal, close-packed crystal structure. The unit cell was calculated to have dimensions of 7.6 A along the a axis and 11.9 A along the c axis of the space lattice.