The Klebsiellu aerogenes ureE gene product was previously shown to facilitate assembly of the urease metallocenter (Lee, M.H., et al., 1992, J. Bacteriol. 174,4324-4330). UreE protein has now been purified and characterized. Although it behaves as a soluble protein, UreE is predicted to possess an amphipathic &strand and exhibits unusually tight binding to phenyl-Sepharose resin. Immunogold electron microscopic studies confirm that UreE is a cytoplasmic protein. Each dimeric UreE molecule (M, = 35,000) binds 6.05 + 0.25 nickel ions (Kd of 9.6 * 1.3 p M ) with high specificity according to equilibrium dialysis measurements. The nickel site in UreE was probed by X-ray absorption and variable-temperature magnetic circular dichroism spectroscopies. The data are most consistent with the presence of Ni(I1) in pseudo-octahedral geometry with 3-5 histidyl imidazole ligands. The remaining ligands are nitrogen or oxygen donors. UreE apoprotein has been crystallized and analyzed by X-ray diffraction methods. Addition of nickel ion to apoprotein crystals leads to the development of fractures, consistent with a conformational change upon binding nickel ion. We hypothesize that UreE binds intracellular nickel ion and functions as a nickel donor during metallocenter assembly into the urease apoprotein.

Pankratz, H. S., and C. C. Bowen. (Iowa State U., Ames.) Cytology of blue‐green algae. I. The cells of Symploca muscorum. Amer. Jour. Bot. 50(4): 387–399. Illus. 1963.—The cellular morphology of Symploca muscorum is described, based upon electron micrographs utilizing improved techniques of specimen preparation. Except for a limiting plasma membrane, ribosomes, and Feulgen‐positive chromatin, the cells have little resemblance to those of higher organisms. The longitudinal components of the cellular envelope consist of a 200–300 mμ fibrous sheath and a complex inner investment about 35 mμ thick which includes at least 3 distinctly layered wall elements in addition to the typical 7‐mμ unit membrane forming the plasma membrane. A row of very small elongate “pores” pierce the inner investment on each side of, and immediately adjacent to, the junction of the longitudinal walls and the crosswalls. Crosswalls vary in thickness from 3 to 20 mμ, depending upon their age, and arise as elaborations of the inner layers of the longitudinal inner investment. The photosynthetic lamellar component of the cytoplasm consists of flattened sacs formed from unit membranes. The lamellae are concentrated in the peripheral region of the cell and usually are parallel to the longitudinal wall. These often extend from one crosswall to the next but, except for a few cases, are not continuous with the plasma membrane at either end. The Feulgen‐positive nucleoplasm appears as an anastomosing system of lightstaining regions containing fibrils 2–5 mμ in diameter. The morphology and interrelationship of a number of other cellular elements are described: (1) structured granules range up to 0.5μ in diameter and occur near crosswalls; (2) polyhedral bodies, 0.2–0.5μ in diameter, are closely associated with the nucleoplasm; (3) “cylindrical bodies” characteristically consist of 2 concentric cylinders, are about 13 mμ in diameter and up to lμ in length; (4) “α granules” are spherical or somewhat elongate elements about 30 mμ in diameter and characteristically associated with the photosynthetic lamellae and structured granules; (5) “β granules” are spherical, highly osmiophilic granules which range from 30 to 90 mμ in diameter; (6) ribosomes, 10–15 mμ, in diameter, are most numerous near the nucleoplasm; (7) plasmodesms penetrate the crosswalls between adjacent cells. The cells of this organism can best be described as being in a “steady state” of division, and there is no evidence of any kind of organized distribution of the nucleoplasm to daughter cells during the constant progress of cytokinesis.

Abstract.We examined the development of the aquatic N2-fixing symbiosis between Rhizobium sp. (Neptunia) and roots of Neptunia natans L. f. (Druce) (previously N. oleracea Lour.) under natural and laboratory conditions. When grown in its native marsh habitat, this unusual aquatic legume does not develop root hairs, the primary sites of rhizobial infection for most temperate legumes. Under natural conditions, the aquatic plant floats and develops nitrogen-fixing nodules at emergence of lateral roots on the primary root and on adventitious roots at stem nodes, but not from the stem itself. Cytological studies using various microscopies revealed that the mode of root infection involved an intercellular route of entry followed by an intracellular route of dissemination within nodule cells. After colonizing the root surface, the bacteria entered the primary root cortex through natural wounds caused by splitting of the epidermis and emergence of young lateral roots, and then stimulated early development of nodules at the base of such roots. The bacteria entered the nodule through pockets between separated host cells, then spread deeper in the nodule through a narrower intercellular route, and eventually evoked the formation of infection threads that penetrated host cells and spread throughout the nodule tissue. Bacteria were released from infection droplets at unwalled ends of infection threads, became enveloped by peribacteroid membranes, and transformed into enlarged bacteroids within symbiosomes. In older nodules, the bacteria within symbiosomes were embedded in an unusual, extensive fibrillar matrix. Cross-inoculation tests of 18 isolates of rhizobia from nodules of N. natans revealed a host specificity enabling effective nodulation of this aquatic legume, with lesser affinity for Medicago sativa and Ornithopus sp., and an inability to nodulate several other crop legume species. Acetylene reduction (N2 fixation) activity was detected in nodules of N. natans growing in aquatic habitats under natural conditions in Southern India. These studies indicate that a specific

Five types of dormant Bacillus spores, between and within species, were selected representing a 600-fold range in moist-heat resistance determined as a

The interaction between Rhizobium lipopolysaccharide (LPS) and white clover roots was examined. The Limulus lysate assay indicated that Rhizobium leguminosarum bv. trifolii (hereafter called R. trifolii) released LPS into the external root environment of slide cultures. Immunofluorescence and immunoelectron microscopy showed that purified LPS from R. tnifolii 0403 bound rapidly to root hair tips and infiltrated across the root hair wall. Infection thread formation in root hairs was promoted by preinoculation treatment of roots with R. trifolii LPS at a low dose (up to 5 ,ug per plant) but inhibited at a higher dose. This biological activity of LPS was restricted to the region of the root present at the time of exposure to LPS, higher with LPS from cells in the early stationary phase than in the mid-exponential phase, incubation time dependent, incapable of reversing inhibition of infection by N03 or NH4', and conserved among serologically distinct LPSs from several wild-type R. trifolii strains (0403, 2S-2, and ANU843). In contrast, infections were not increased by preinoculation treatment of roots with LPSs from R. leguminosarum bv. viciae strain 300, R. meliloti 102F28, or members of the family Enterobacteriaceae. Most infection threads developed successfully in root hairs pretreated with R. trifolii LPS, whereas many infections aborted near their origins and accumulated brown deposits if pretreated with LPS from R. meliloti 102F28. LPS from R. leguminosarum 300 also caused most infection threads to abort. Other specific responses of root hairs to infection-stimulating LPS from R. trifolii included acceleration of cytoplasmic streaming and production of novel proteins. Combined gas chromatography-mass spectroscopy and proton nuclear magnetic resonance analyses indicated that biologically active LPS from R. trifolii 0403 in the early stationary phase had less fucose but more 2-0-methylfucose, quinovosamine, 3,6-dideoxy-3-(methylamino)galactose, and noncarbohydrate substituents (0-methyl, N-methyl, and acetyl groups) on glycosyl components than did inactive LPS in the mid-exponential phase. We conclude that LPS-root hair interactions trigger metabolic events that have a significant impact on successful development of infection threads in this Rhizobium-legume symbiosis.Establishment of an effective Rhizobium-legume symbiosis can be viewed as a process of cellular recognition and compatibility between bacterial and plant cells. The infection process involves bacterial attachment, root hair deformation, bacterial penetration of the root hair wall, formation and sustained development of the infection thread, bacterial release from infection threads within emerging root nodule cells, and bacterial differentiation into nitrogen-fixing bacteroids.The lipopolysaccharides (LPS) of rhizobia are likely to be involved in the infection process. They are major glycoconjugates on the surface of Rhizobium leguminosarum biovars * Corresponding author. t Present address: Laboratoire des Relationes Plantes-Microorganismes,