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E L Schmidt

Publications and source records attributed to E L Schmidt.

At least 37 records · Page 2Linked to original sources

Serological Diversity of Nitrobacter spp. from Soil and Aquatic Habitats.

Serotypic diversity among Nitrobacter spp. isolates is greater than previously reported. Typing with fluorescent antibodies prepared against 11 Nitrobacter spp. cultures isolated from soil and water placed the isolates into six serogroups. When these fluorescent antibodies were applied to a group of 16 additional isolates, 8 were identifiable by cross-reaction to 3 of the 11 fluorescent antibodies. Some nitrite-oxidizing enrichment cultures from different habitats contained cross-reacting strains of Nitrobacter spp.

Journal Article↗

Plasmid transfer within and between serologically distinct strains of Rhizobium japonicum, using antibiotic resistance mutants and auxotrophs.

Methionine-requiring and pantothenic acid-requiring auxotrophs of Rhizobium japanicum USDA 31, as well as highly antibiotic-resistant mutants of R. japonicum strains USDA 31, USDA 110, USDA 138, and Webster 48, were isolated. These mutants were used to transfer the P-1 group plasmids R68.45 and RP4 within and between strains USDA 31, USDA 110, and Webster 48. Attempts to demonstrate transfer of either plasmid to strain USDA 138 were unsuccessful.

Ampicillin↗

Localization and partial characterization of soybean lectin-binding polysaccharide of Rhizobium japonicum.

Immunoelectron microscopy was combined with partial characterization of isolated exopolysaccharide to study binding of soybean lectin by Rhizobium japonicum strain USDA 138. Lectin-binding activity resided in two forms of exopolysaccharide produced during growth: an apparently very high-molecular-weight capsular form and a lower-molecular-weight diffusible form. At low-speed centrifugation, the capsular form cosedimented with cells to form a viscous, white, cell-gel complex which was not diffusible in 1% agar, and the diffusible form remained in the cell-free supernatant. Electron microscopic observation of the cell-gel complex after labeling with soybean lectin-ferritin conjugate revealed that capsular polysaccharides, frequently attached to one end of the cells, were receptors for lectin. The outer membrane of the cell bound no lectin. Various preparations of exopolysaccharide isolated from the culture supernatant were tested for lectin binding, interaction with homologous somatic antigen, and the presence of 2-keto-3-deoxyoctonate and were chromatographed in Sepharose 4B and 6B gel beds. Lectin binding was restricted to a polysaccharide component designated as lectin-binding polysaccharide. This polysaccharide, as present in the cell-free culture supernatant, was a diffusible acidic polysaccharide devoid of 2-keto-3-deoxyoctonate, with a molecular weight of 2 X 10(6) to 5 X 10(6). It was concluded that the soybean lectin-binding component of R. japonicum is an extracellular polysaccharide and not a lipopolysaccharide and that the diffusible lectin-binding polysaccharide probably differs from the very high-molecular-weight lectin-binding polysaccharide of the loose capsule (slime) only in the degree of polymerization.

Lectins↗

Accumulation of Soybean Lectin-Binding Polysaccharide During Growth of Rhizobium japonicum as Determined by Hemagglutination Inhibition Assay.

A hemagglutination inhibition assay was used to estimate the presence of soybean lectin-binding polysaccharide in whole culture, culture supernatant, and isolated exopolysaccharide of Rhizobium japonicum USDA 138. The occurrence of 0.1 to 0.2 mug of lectin-binding polysaccharide could be detected within 2 h with a 0.5-ml total sample. Lectin-binding polysaccharide was detected in all preparations during both exponential and stationary growth phases. The formation of lectin-binding polysaccharide was not, whereas that of total exopolysaccharide was, markedly affected by culture conditions.

Journal Article↗

Production of nitrous oxide by ammonia-oxidizing chemoautotrophic microorganisms in soil.

Gas chromatographic studies showed that nitrous oxide was produced in each instance when sterilized (autoclaved) soil was incubated after treatment with ammonium sulfate and inoculation with pure cultures of ammonia-oxidizing chemoautotrophic microorganisms (strains of Nitrosomonas, Nitrosospira, and Nitrosolobus). Production of N(2)O in ammonium-treated sterilized soil inoculated with Nitrosomonas europaea increased with the concentration of ammonium and the moisture content of the soil and was completely inhibited by both nitrapyrin and acetylene. Similar effects of nitrapyrin, acetylene, ammonium concentration, and soil moisture content were observed in studies of factors affecting N(2)O production in nonsterile soil treated with ammonium sulfate. These observations support the conclusion that, at least under some conditions, most of the N(2)O evolved from soils treated with ammonium or ammonium-producing fertilizers is generated by chemoautotrophic nitrifying microorganisms during oxidation of ammonium to nitrite.

Journal Article↗

Further examination of presumptive Rhizobium trifolii mutants that nodulate Glycine max.

Two recent reports described the isolation of derivatives of a Rhizobium trifolii strain that had gained the ability to nodulate Glycine max and Vigna radiata and that had demonstrated altered patterns of carbon source utilization, free-living nitrogen fixation, and hydrogen uptake. More extensive characterization of these strains now supports the conclusion that these strains are R. japonicum and are not derived from the putative parent R. trifolii.

Journal Article↗

Population Densities of Rhizobium japonicum Strain 123 Estimated Directly in Soil and Rhizospheres.

Rhizobium japonicum serotype 123 was enumerated in soil and rhizospheres by fluorescent antibody techniques. Counting efficiency was estimated to be about 30%. Indigenous populations of strain 123 ranged from a few hundred to a few thousand per gram of field soil before planting. Rhizosphere effects from field-grown soybean plants were modest, reaching a maximum of about 2 x 10 cells of strain 123 per g of inner rhizosphere soil in young (16-day-old) plants. Comparably slight rhizosphere stimulation was observed with field corn. High populations of strain 123 (2 x 10 to 3 x 10 cells per g) were found only in the disintegrating taproot rhizospheres of mature soybeans at harvest, and these populations declined rapidly after harvest. Pot experiments with the same soil provided data similar to those derived from the field experiments. Populations of strain 123 reached a maximum of about 10 cells per g of soybean rhizosphere soil, but most values were lower and were only slightly higher than values in wheat rhizosphere soil. Nitrogen treatments had little effect on strain 123 densities in legume and nonlegume rhizospheres or on the nodulation success of strain 123. No evidence was obtained for the widely accepted theory of specific stimulation, which has been proposed to account for the initiation of the Rhizobium-legume symbiosis.

Journal Article↗

Diversity in the ammonia-oxidizing nitrifier population of a soil.

Multiple genera of ammonia-oxidizing chemoautotrophic nitrifiers in a soil were detected, isolated, and studied by means of modified most-probable-number (MPN) techniques. The soil examined was a Waukegon silt loam treated with ammonium nitrate or sewage effluent. The genera Nitrosomonas and Nitrosospira were found to occur more commonly than the genus Nitrosolobus. Three different MPN media gave approximately the same overall ammonia oxidizer counts within statistical error after prolonged incubation but differed markedly in ratios of Nitrosomonas to Nitrosospira. Selectivity and counting efficiency of MPN media were studied by observing the growth response of representative pure cultures isolated from the soil. Selectivity was evident in each medium with respect to all strains tested, and the media differed greatly in incubation times required to obtain maximum counts.

Journal Article↗

Serological diversity within a terrestrial ammonia-oxidizing population.

Fluorescent antibodies (FAs) prepared against 16 ammonia-oxidizing nitrifying bacteria were examined as to cross-reactivity in heterologous FA staining tests. Virtually all cross-reactions to the seven Nitrosomonas FAs were confined to the Nitrosomonas isolates. The five Nitrosospira, three Nitrosolobus, and one Nitrosovibrio FAs stained isolates only within their respective genera. Within each genus shared antigens were more common among Nitrosomonas and Nitrosolobus than among Nitrosospira isolates. Isolates obtained from a single soil were stained to examine the effectiveness of the suite of FAs for study of a given ammonia-oxidizing population. Of the seven Nitrosomonas isolates, five were stained effectively through the use of four FAs; six of twelve Nitrosospira isolates were identifiable through use of four different FAs. The one Nitrosolobus isolate was stained well only by one (its homologous) FA.

Journal Article↗

Nitrification in the intertidal zone: influence of effluent type and effect of tannin on nitrifiers.

Nitrification by intertidal sediments was measured by using a tide simulator that approximated the cycle of seawater on tidal flats. Sediments were chosen from sites affected by industrial and municipal effluents and pastoral seepage and runoff. The ability of sediments from different sites to nitrify endogenous nitrogen varied markedly. All sites exhibited an initial lag before activity commenced. The duration of this lag and the rate of nitrate production were different at each site. The sediments were also capable of oxidizing NH3-N supplied to them in seawater. This "nitrification potential" was highest at sites receiving nitrogenous effluents (slaughterhouse and sewage), but was also substantial in sediments affected by bark extract effluent and pasture runoff. The lowest potential and the longest lag were exhibited by sediments in an apple cannery effluent area. Enrichment cultures of nitrifying microorganisms were obtained from all sites using NH4+ as a source of energy, but enrichments for nitrite oxidizers were unsuccessful. Concentrated pine bark tannins, similar in origin to those in effluents at the well-nitrifying chipmill site, were tested for toxicity to pure cultures of nitrifying bacteria. Two Nitrobacter strains and one Nitrosomonas strain were unaffected by tannins even at 5 mg/ml. A Nitrosolobus and a Nitrosospira strain were inhibited partially at 5 mg/ml and only slightly or not at all at 1 mg/ml.

Ammonia↗

Viability of Rhizobium bacteroids.

Bacteroids prepared from nodules of soybean and bean were tested for viability. Contrary to the prevailing view that bacteroids are nonviable, it was found that bacteroids averaged 90% viability, irrespective of Rhizobium strain, nodule age, or nodule environment.

Cell Division↗

Application of the fluorescent antibody technique to the study of an isolate of Beijerinckia in soil.

Fluorescent antibody was prepared against a temperate-soil isolate of Beijerinckia obtained from a rhizosphere of rice growing in Camargue (France). The antibody did not cross-react with any of 6 species of Azotobacter, 4 species of Beijerinckia, or 44 unidentified soil bacteria isolated from a spectrum of rhizospheres, but strongly stained the homologous Beijerinckia isolate. The isolate grew well in autoclave Camargue soil, but increased in numbers only slightly in nonsterile soil during 9 days. Preliminary examination of rice plants grown in the laboratory in soil from which the Beijerinckia was originally isolated did not show detectable Beijerinckia in the rhizosphere. The fluorescent antibody was sufficiently sensitive and specific to permit more extensive study of Beijerinckia in relation to nitrogen fixation in the rhizospher of rice.

Azotobacter↗

Immunofluorescence studies of Nitrobacter populations in soils.

Certain steps of a protocol to enumerate a bacterium directly in soil by immunofluorescence were studied with respect to the enumeration of Nitrobacter in soils of diverse properties. Maximal counts of Nitrobacter were obtained by varying factors involved in the release of bacteria from the soil. Differences with respect to these factors were related to soil-colloidal properties. Enumeration protocols modified with regard to soil properties were used with strain-specific fluorescent antibodies (FA) to enumerate Nitrobacter populations (a) in soil during storage, (b) in comparison with most probable number (MPN) enumeration, and (c) occurring in a spectrum of soil samples. FA counting was rapid and precise and gave counts generally 10- to 100-fold higher than obtained by MPN. Nitrobacter cells of the two serotypes studied, designated as agilis and Winogradsky, occurred in each soil at levels of 10(4)-10(5) per gram.

Bacteriological Techniques↗

Polarity in the exponential-phase Rhizobium japonicum cell.

Highly distinctive aspects of the exponentail-phase Rhizobium japonicum cell were disclosed by means of thin sections, freeze etching, fluorescent antibodies, and ruthenium red staining. Polarity was expressed in the form of reserve polymer distribution near one end of the cell and as cytoplasmic localization near the opposite end. In addition, exocellular polysaccharide (EPS) accumulated preferentially around the cytoplasmic end, and the feature described previously as an "immunofluorescent polar tip" was seen clearly as an extracellular polar body (EPB) on the tip of the cell at the reserve polymer end. Compartmentalization of cytoplasm and reserves were consistent features of nearly all exponential cells of the two strains studied; strain 31, however, formed little EPS and had a high incidence of a large, tightly bound EPB, while strain 138 formed EPS extensively and had a low incidence of EPB. Extracellular polysaccharides of strain 138 reacted with soybean lectin in gel diffusion tests, so that the EPS seen in electron micrographs is tentatively considered to include the lectin-binding material. Extracellular polar bodies were accumulations of granular and fibrillar material with properties consistent with the presence of polysaccharide and lipopolysaccharide. The role of EPB in cell to cell attachment was confirmed by electron microscopy.

Antigens, Bacterial↗

Lipids of Nitrobacter and effects of cultural conditions on fatty acid composition.

The nitrite-oxidizing autotroph, Nitrobacter was studied with respect to fatty acid composition and lipids. One fatty acid, shown to be cis-11-actadecenoic acid (cis-vaccenic) accounted for almost 96% of the total fatty acids of the extractable lipids of Nitrobacter agilis, Nitrobacter winogradskyi and each of several isolates from Minnesota and Moroccan soils studied. The cis-vaccenic acid was high in all organisms, ranging from 85 to 95% when grown at 27degreesC in the log growth phase, the other major acid was plamitic (16 : 1). All converted ces-vaccenic acid to a 19-carbon cyclopropanecarboxylic acid upon entering the stationary growth phase. The 11-carbon cyclopropanecarboxylic acid was not degraded when stationary phase cells were reinoculated into fresh medium. In N. agilis the levels of cis-vaccenic acid ranged from 86.8% when grown at 33degreesC to 95.6% when grown at 19degreesC. Addition of acetate or casein hydrolyzate to the inorganic medium had virtually no effect on the fatty acid composition of N. agilis, while propionate effected both qualitative and quantitative changes. In all organisms phosphatidylcholine made up a large portion of the extractable lipids. The distribution was phosphatidylcholine, 54%; phosphatidylethanolamine, 23%; phosphatidylglycerol, 10%; and neutral lipids, 11% for N. agilis.

Carboxylic Acids↗