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Biomedical subjects

B B Bohlool

Publications and source records attributed to B B Bohlool.

At least 37 records · Page 2Linked to original sources
PubMed1983

Competition Among Rhizobium leguminosarum Strains for Nodulation of Lentils (Lens esculenta).

Thirty-one cultures of Rhizobium leguminosarum were screened for effectiveness (C(2)H(2) reduction) on lentils (Lens esculenta). Fluorescent antibodies prepared against three of the most effective strains (Hawaii 5-0, Nitragin 92A3, and Nitragin 128A12) exhibited a high degree of strain specificity; the antibodies reacted strongly with their homologous rhizobia in culture and with bacteroids in nodules. They did not cross-react with one another, and only weakly with 5 of the 47 other R. leguminosarum cultures tested. In competition studies in the growth chamber, whenever strain Nitragin 92A3 was included in the inoculum mixture, it consistently (but not always significantly, P = 0.05) occupied the majority of nodules on all four cultivars used. However, some degree of strain X cultivar interaction was apparent: Hawaii 5-0 was of equal competitiveness (P = 0.05) with Nitragin 92A3 on three of the varieties (Commercial, Tekoa, and Benewah), but inferior (P = 0.01) on the Chilean variety; Nitragin 92A3 completely dominated (P = 0.01) Nitragin 128A12 on all cultivars; and Hawaii 5-0 was of equal competitiveness (P = 0.05) to Nitragin 128A12 on the Chilean variety and more competitive (P = 0.01) on the commercial variety and less so on the other two varieties. In field experiments, Hawaii 5-0 proved of equal competitiveness (P = 0.01) with Nitragin 92A3 in one soil (an Inceptisol) and superior (P </= 0.05) to it in another (an Oxisol). Incidence of double-strain occupancy of nodules varied from 0 to 36% in vermiculite, depending on the strains in the mixture and the host variety, and from 0 to 38% in the field, depending on the strains in the mixture and the soil type. The results suggest a close relationship between the competitiveness of a strain and its occurrence in doubly infected nodules.

Journal Article↗
PubMed1983

Competition of Rhizobium japonicum Strains in Early Stages of Soybean Nodulation.

The effects of preexposure of soybean (Glycine max L. Merrill) roots to Rhizobium japonicum strains and subsequent establishment of other strains in the nodules were investigated by using combinations of effective strains (USDA 110 and USDA 138) and effective-ineffective strains (USDA 110 and SM-5). Strain USDA 110 was a better competitor than either USDA 138 or SM-5 on cultivars Lee and Peking. However, when either of the two less-competitive strains was inoculated into 2-day-old seedlings before USDA 110 was, their nodule occupancy increased significantly on both cultivars. With USDA 138 as the primary inoculum and USDA 110 delayed for 6, 48, and 168 h, the incidence of USDA 138 nodules increased on cultivar Peking from 6% (at zero time) to 28, 70, and 82% and on cultivar Lee from 17% (at zero time) to 32, 88, and 95% for the three time delays, respectively. Preexposure of 2-week-old roots of cultivar Lee to USDA 138 had essentially the same effect: the incidence of USDA 138 nodules increased from 23% at zero time to 89 and 97% when USDA 110 was delayed for 24 and 72 h, respectively. When the ineffective strain SM-5 was used as the primary inoculum, followed by USDA 110 72 h later, the percentage of nodules containing SM-5 increased from 7 to 76%. These results indicate that the early events in the nodulation process of soybeans are perhaps the most critical for competition among R. japonicum strains.

Journal Article↗
PubMed1983

Possible involvement of a megaplasmid in nodulation of soybeans by fast-growing rhizobia from china.

Several isolates from a newly described group of fast-growing acid-producing soybean rhizobia, Rhizobium japonicum, were analyzed for plasmid content. All contained from one to four plasmids with molecular weights of 100 x 10 or larger. Although most of the isolates shared plasmids of similar size, the restriction endonuclease (BamHI, EcoRI, and HindIII) patterns of the plasmids from three of the isolates were vastly different. Growth in the presence of acridine orange was effective in producing mutants cured of the largest plasmid in one of the strains. These mutants also lost the ability to form nodules on soybeans. High-temperature curing of a smaller plasmid in another strain did not lead to loss of nodulating ability or alteration of symbiotic effectiveness on soybean cultivars. The identities of all of the isolates and mutants were ascertained by immunofluoresence and immunodiffusion. The new fast-growing strains of R. japonicum may provide a better genetic system for the study of the soybean symbiosis than the slow-growing R. japonicum, not all of which can be shown to contain plasmids.

Journal Article↗
PubMed1982

Effect of salinity on Rhizobium growth and survival.

This study examines the effect of salinity on the growth and survival of Rhizobium spp. in culture media and soil. Eleven isolates from saline and nonsaline environments were compared. The growth (mean doubling time) of all strains and species tested decreased when the electrical conductivity of the culture medium (yeast extract-mannitol) was raised from 1.2 mS cm to 6.7 mS cm (15% seawater equivalent) or to 13.1 mS cm (28% seawater equivalent). Three of eleven strains failed to grow at 13.1 mS cm. Although growth was affected by salinity, four strains selected from the growth rate study could survive in extremely high concentrations of salt. Two strains with growth rates sensitive to salt and two strains with growth rates relatively unaffected by salt were inoculated into solutions with electrical conductivities of up to 43.0 mS cm (92% seawater equivalent). Not only did all four strains survive the initial osmotic shock (at 5 h after inoculation), but it was not until 27 days after inoculation that the sensitive strains exhibited a significant reduction in viable numbers. The salt-tolerant strains survived for more than 65 days with no reduction in viable counts. The interaction between soil moisture tension and soil salinity in relation to Rhizobium survival in gamma-irradiated soil was also examined. Six treatment combinations were used, ranging from -0.1 bars and 0.2 mS cm to -15 bars and 12 mS cm. Sensitive strains declined from 10 to 10 organisms per g of soil after 84 days of incubation at -15 bars and 12 mS cm. Tolerant strains survived for the same period with no loss in viable numbers. The results of these experiments indicate that many strains of Rhizobium can grow and survive at salt concentrations which are inhibitory to most agricultural legumes. The emphasis of research concerning the effects of salinity on symbiotic nitrogen fixation should, therefore, be directed to aspects of the symbiosis other than the survival of the Rhizobium spp.

Journal Article↗
PubMed1981

Release of Rhizobium spp. from Tropical Soils and Recovery for Immunofluorescence Enumeration.

Limitations associated with immunofluorescence enumeration of bacteria in soil derive largely from the efficiency with which cells can be separated from soil particles and collected on membrane filters for staining. Many tropical soils fix added bacteria tightly, resulting in low recoveries. Eight soils, representative of three of the major soil orders found in the tropics (oxisols, vertisols, and inceptisols), were tested for recovery of added Rhizobium strains. All except one Hawaiian andept (Typic Eutrandept) yielded recoveries ranging from <1 to 13%. Recovery from the andept was 100%. In soil-sand mixtures, addition of only a small amount of soil caused a dramatic decrease in recovery of added rhizobia. Increasing the soil content of the mixture from 0% (10 g of sand) to 50% (5 g of soil-5 g of sand) reduced recoveries from >90 to <1%. Varying the ionic strength and pH of the extracting solution did not cause marked increases in recovery. Protein solutions, ethylenediaminetetraacetate, and NaHCO(3), on the other hand, improved release of bacteria. We report a modification to the usual membrane filter immunofluorescence procedure which yielded consistently high and reproducible recovery (coefficient of variation, 30%) of rhizobia from several tropical soils. In the modified procedure, partially hydrolyzed gelatin, diluted in ammonium phosphate, was used to suspend the soil. This caused dispersion of the soil and release of the bacteria from soil flocs. The efficiency of recovery of Rhizobium spp. from several tropical and two temperate soils remained high as the content of these soils in soil-sand mixtures was increased from 0 to 100%. The modified membrane filter immunofluorescence procedure was used to follow the growth of a strain of chickpea (Cicer arietinum) Rhizobium in a sterilized oxisol. The results showed a close agreement with viable counts at different stages during the growth cycle. Diluent for the hydrolyzed gelatin also had a marked effect on recovery. The efficiency of release of Rhizobium spp. from an oxisol was in the following order for the diluents used: 0.1 M (NH(4))(2)HPO(4) > 0.1 M Na(2)HPO(4) = 0.1 M sodium-phosphate-buffered saline (pH 7.2) > 0.2 M NH(4)Cl > 0.2 KCl > NaCl = LiCl > water.

Journal Article↗
PubMed1979

Colorimetric enzyme-linked immunosorbent assay for the identification of strains of Rhizobium in culture and in the nodules of lentils.

An indirect enzyme-linked immunosorbent assay has been developed to identify strains of Rhizobium in culture and in lentil nodules. The test can be used on cells from both fresh and frozen nodules obtained from plants grown either in a growth chamber or in the field. Test results were confirmed by immunofluorescence. The enzyme-linked immunosorbent assay technique can be used for field studies and requires less antisera than other serological techniques.

Journal Article↗
PubMed1978

Nitrogen-fixing communities in an intertidal ecosystem.

Nitrogen gixation (acetylene reduction) associated with various communities in the intertidal sand and mud flat was measured in situ. Areas which were colonized by algae, plants, and animals generally had significantly higher activities than areas which were visibly uncolonized. The highest activities were measured at sites colonized by a mixed bloom of Oscillatoria-Euglena-photosynthetic bacteria. These areas occupied only about 1% of the surface of the sediments, yet contributed nearly 50% of the biologically fixed nitrogen. Enteromorpha communities also exhibited relatively high activities. Sites were grouped according to the statistical significance of the mean of their activities: group I: uncolonized sand, mud and gravel flats, Eelgrass (Zostera) communities, and snail (amphibola) beds, with an estimated contribution of 1 g N ha-1 day-1; group II: Cockel (Chione) beds, 3 g N ha-1 day-1; group III: Salicornia, Ulva, Juncus, and Enteromorpha, 10 g N ha-1 day-1; and group IV: Oscillatoria-Euglena-photosynthetic bacteria association, 200 g N ha-1 day-1. Enteromorpha and Oscillatoria-Euglena-photosynthetic bacteria communities showed significantly lower rates of nitrogen fixation in the dark than in the light.

Acetylene↗
PubMed1977

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↗
PubMed1976

Immunofluorescent polar tips of Rhizobium japonicum: possible site of attachment or lectin binding.

Rhizobium japonicum USDA 31 demonstrated marked polarity by binding homologous fluorescent antibody (FA) heavily on one end of the cell. FA prepared against R. japonicum strains 110 and 138, and against R. trifolii TA1 cross-reacted with strain 31 only in the polar tip region. No polar immunofluorescing tips could be seen with FA against two other strains of R. japonicum or with those against several unrelated microorganisms. Common antigens localized only in a polar region were seen in many rhizobia stained with R. japonicum 31 FA: 22 of 23 strains of R. japonicum, 10 of 17 strains of R. trifolii, 3 of 7 strains of R. melitolii, 3 of 6 strains of R. phaseoli, and 3 of 9 strains of R. leguminosarum had some cells with detectable polar tips. The proportion of R. japonicum 31 cells with polar tips was high throughout the growth cycle. Polar tip staining was not affected by drastic cell treatments. A function was proposed for the polar tip region as a site for attachment. R. japonicum 31 cells attached to each other in a tip-to-tip fashion and endwise to fungal hyphae with the polar tip in contact with the hyphal wall. Binding of fluorescein isothiocyanate-labeled soybean lectin to certain strains of R. japonicum gave additional evidence of polarity. Polar binding of both antibody and lectin may provide insights into relationships between rhizobia and roots of host legumes.

Antibodies, Bacterial↗
PubMed1974

Autecological study of the chemoautotroph Nitrobacter by immunofluorescence.

Fluorescent antibodies (FA) prepared for Nitrobacter agilis and N. winogradskyi were highly reactive in homologous staining. Low-level cross-reactions between the two species were removed by adsorption. All 15 pure-culture isolates of Nitrobacter tested reacted strongly with either N. agilis FA or N. winogradskyi FA. All pure-culture isolates from soils were determined to be N. winogradskyi; those from Mammoth Cave sediments and a cattle waste oxidation ditch were N. agilis. No cross-reaction was found in extensive tests that included five isolates of Nitrosomonas europaea and 668 heterotrophic aerobic and anaerobic bacteria isolated from soil, sewage, and cave sites. The FA preparations were used to detect Nitrobacter species in Mammoth Cave sediments, in a cattle waste oxidation ditch, and in surface waters and sediments of a river and to observe that N. winogradskyi can outgrow N. agilis in enrichment culture.

Aerobiosis↗
PubMed1974

Immunofluorescence approach to the study of the ecology of Thermoplasma acidophilum in coal refuse material.

Specific immunofluorescence staining was applied to the study of the localization, distribution, and growth of Thermoplasma acidophilum in its natural habitat, the coal refuse pile. Different antigenic groups of T. acidophilum could be isolated from the same refuse pile, and the same antigenic groups were isolated from piles from different geographical areas. No correlation could be established between the antigenic groups and the pH or temperature of the habitats. Brightly fluorescing cells of T. acidophilum were detected on microscope slides buried in contact with the coal refuse material or immersed in the water in the stream draining a refuse pile. T. acidophilum grew when inoculated into either coal refuse material and/or an aqueous extract of coal refuse when incubated at its optimal temperature of 55 C, but not when incubated at room temperature or 37 C. The coal refuse pile appears to be a primary habitat for T. acidophilum.

Antigens, Bacterial↗
PubMed1974

Growth rates of Sulfolobus acidocaldarius in nature.

Turnover times for water passing through several Sulfolobus acidocaldarius-containing springs were determined by measuring the dilution rates of small amounts of sodium chloride that were added to the springs. Chloride was diluted out exponentially, while concentrations of the bacteria remained constant. Additionally, temperature, pH, and chemical composition of the springs also remained constant during the time that the chloride was being diluted. The springs are thus steady-state systems, and since the rates of bacterial growth must be at least equal to the chloride dilution rates, minimal doubling times for the bacterial populations can be calculated. Half-times for chloride dilution, equivalent to bacterial doubling times, were on the order of 10 to 20 h for springs ranging in volume from about 20 to 2,000 liters, but approximately 30 days for two larger springs of about 1 million liters. Formaldehyde-fixed cells of a serologically distinguishable strain of S. acidocaldarius were also added as markers to four of the smaller springs, and the dilution rates of these bacteria were compared with the chloride dilution rates. The rates agreed reasonably well, thus verifying the growth rates obtained from the chloride dilution rates. In three springs, exponential growth was studied by draining the springs and allowing them to refill with bacteria-free water. Exponential doubling times were on the order of a few hours, much more rapid than steady-state doubling times. The methods used in this work may have wider utility in aquatic environments.

Antigens, Bacterial↗