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1.
Surface wood samples obtained from a Douglas fir log (Pseudotsuga menziesii) in a Pacific Northwest stream were incubated in vitro with [C]lignocellulose in a defined mineral salts medium supplemented with 10 mg of N liter of N-labeled NO(3) (50 atom% N). Evolution of CO(2), distribution and isotopic dilution of N, filtrate N concentrations, and the rates of denitrification, N(2) fixation, and respiration were measured at 6, 12, and 18 days of incubation. The organic N content of the lignocellulose-wood sample mixture had increased from 132 mug of N to a maximum of 231 mug of N per treatment after 6 days of incubation. Rates of [C]lignocellulose decomposition were greatest during the first 6 days and then began to decline over the remaining 12 days. Total CO(2) evolution was also highest at day 6 and declined steadily over the remaining duration of the incubation. Filtrate NH(4)-N increased from background levels to a final value of 57 mug of N per treatment. Filtrate NO(3) N completely disappeared by day 6, and organic N showed a slight decline between days 12 and 18. The majority of the N that could be recovered appeared in the particulate organic fraction by day 6 (41 mug of N), and the filtrate NH(4) N fraction contained 11 mug of N by day 18. The N enrichment values of the filtrate NH(4) and the inorganic N associated with the particulate fraction had increased to approximately 20 atom% N by 18 days of incubation, whereas the particulate organic fraction reached its highest enrichment by day 6. Measurements of N(2) fixation and denitrification indicated an insignificant gain or loss of N from the experimental system by these processes. The data show that woody debris in stream ecosystems might function as a rapid and efficient sink for exogenous N, resulting in stimulation of wood decomposition and subsequent activation of other N cycling processes. 相似文献
2.
Contributions of Autotrophic and Heterotrophic Nitrifiers to Soil NO and N2O Emissions 总被引:4,自引:2,他引:2
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Soil emission of gaseous N oxides during nitrification of ammonium represents loss of an available plant nutrient and has an important impact on the chemistry of the atmosphere. We used selective inhibitors and a glucose amendment in a factorial design to determine the relative contributions of autotrophic ammonium oxidizers, autotrophic nitrite oxidizers, and heterotrophic nitrifiers to nitric oxide (NO) and nitrous oxide (N2O) emissions from aerobically incubated soil following the addition of 160 mg of N as ammonium sulfate kg−1. Without added C, peak NO emissions of 4 μg of N kg−1 h−1 were increased to 15 μg of N kg−1 h−1 by the addition of sodium chlorate, a nitrite oxidation inhibitor, but were reduced to 0.01 μg of N kg−1 h−1 in the presence of nitrapyrin [2-chloro-6-(trichloromethyl)-pyridine], an inhibitor of autotrophic ammonium oxidation. Carbon-amended soils had somewhat higher NO emission rates from these three treatments (6, 18, and 0.1 μg of N kg−1 h−1 after treatment with glucose, sodium chlorate, or nitrapyrin, respectively) until the glucose was exhausted but lower rates during the remainder of the incubation. Nitrous oxide emission levels exhibited trends similar to those observed for NO but were about 20 times lower. Periodic soil chemical analyses showed no increase in the nitrate concentration of soil treated with sodium chlorate until after the period of peak NO and N2O emissions; the nitrate concentration of soil treated with nitrapyrin remained unchanged throughout the incubation. These results suggest that chemoautotrophic ammonium-oxidizing bacteria are the predominant source of NO and N2O produced during nitrification in soil. 相似文献
3.
Use of Aliphatic n-Alkynes To Discriminate Soil Nitrification Activities of Ammonia-Oxidizing Thaumarchaea and Bacteria 总被引:1,自引:0,他引:1
Anne E. Taylor Neeraja Vajrala Andrew T. Giguere Alix I. Gitelman Daniel J. Arp David D. Myrold Luis Sayavedra-Soto Peter J. Bottomley 《Applied and environmental microbiology》2013,79(21):6544-6551
Ammonia (NH3)-oxidizing bacteria (AOB) and thaumarchaea (AOA) co-occupy most soils, yet no short-term growth-independent method exists to determine their relative contributions to nitrification in situ. Microbial monooxygenases differ in their vulnerability to inactivation by aliphatic n-alkynes, and we found that NH3 oxidation by the marine thaumarchaeon Nitrosopumilus maritimus was unaffected during a 24-h exposure to ≤20 μM concentrations of 1-alkynes C8 and C9. In contrast, NH3 oxidation by two AOB (Nitrosomonas europaea and Nitrosospira multiformis) was quickly and irreversibly inactivated by 1 μM C8 (octyne). Evidence that nitrification carried out by soilborne AOA was also insensitive to octyne was obtained. In incubations (21 or 28 days) of two different whole soils, both acetylene and octyne effectively prevented NH4+-stimulated increases in AOB population densities, but octyne did not prevent increases in AOA population densities that were prevented by acetylene. Furthermore, octyne-resistant, NH4+-stimulated net nitrification rates of 2 and 7 μg N/g soil/day persisted throughout the incubation of the two soils. Other evidence that octyne-resistant nitrification was due to AOA included (i) a positive correlation of octyne-resistant nitrification in soil slurries of cropped and noncropped soils with allylthiourea-resistant activity (100 μM) and (ii) the finding that the fraction of octyne-resistant nitrification in soil slurries correlated with the fraction of nitrification that recovered from irreversible acetylene inactivation in the presence of bacterial protein synthesis inhibitors and with the octyne-resistant fraction of NH4+-saturated net nitrification measured in whole soils. Octyne can be useful in short-term assays to discriminate AOA and AOB contributions to soil nitrification. 相似文献
4.
Inhibition of Chemoautotrophic Nitrification by Sodium Chlorate and Sodium Chlorite: a Reexamination 总被引:3,自引:5,他引:3
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The oxidation of NH4+ by Nitrosomonas europaea was insensitive to 10 mM NaClO3 (sodium chlorate) but was strongly inhibited by NaClO2 (sodium chlorite; Ki, 2 μM). The oxidation of NO2− by Nitrobacter winogradskyi was inhibited by both ClO3− and ClO2− (Ki for ClO2−, 100 μM). N. winogradskyi reduced ClO3− to ClO2− under both aerobic and anaerobic conditions, and as much as 0.25 mM ClO2− was detected in the culture filtrate. In mixed N. europaea-N. winogradskyi cell suspensions, the oxidation of both NH4+ and NO2− was inhibited in the presence of 10 mM ClO3− after a 2-h lag period, despite the fact that, under these conditions, ClO2− was not detected in the filtrate. The data are consistent with the hypothesis that, in mixed culture, NH4+ oxidation is inhibited by ClO2− produced by reduction of ClO3− by the NO2− oxidizer. The use of ClO3− inhibition of NO2− oxidation in assays of nitrification by mixed populations necessitates cautious interpretation unless it can be shown that the oxidation of NH4+ is not affected. 相似文献
5.
Sub-Parts-Per-Billion Nitrate Method: Use of an N2O-Producing Denitrifier to Convert NO3− or 15NO3− to N2O
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A more sensitive analytical method for NO3− was developed based on the conversion of NO3− to N2O by a denitrifier that could not reduce N2O further. The improved detectability resulted from the high sensitivity of the 63Ni electron capture gas chromatographic detector for N2O and the purification of the nitrogen afforded by the transformation of the N to a gaseous product with a low atmospheric background. The selected denitrifier quantitatively converted NO3− to N2O within 10 min. The optimum measurement range was from 0.5 to 50 ppb (50 μg/liter) of NO3− N, and the detection limit was 0.2 ppb of N. The values measured by the denitrifier method compared well with those measured by the high-pressure liquid chromatographic UV method above 2 ppb of N, which is the detection limit of the latter method. It should be possible to analyze all types of samples for nitrate, except those with inhibiting substances, by this method. To illustrate the use of the denitrifier method, NO3− concentrations of <2 ppb of NO3− N were measured in distilled and deionized purified water samples and in anaerobic lake water samples, but were not detected at the surface of the sediment. The denitrifier method was also used to measure the atom% of 15N in NO3−. This method avoids the incomplete reduction and contamination of the NO3− -N by the NH4+ and N2 pools which can occur by the conventional method of 15NO3− analysis. N2O-producing denitrifier strains were also used to measure the apparent Km values for NO3− use by these organisms. Analysis of N2O production by use of a progress curve yielded Km values of 1.7 and 1.8 μM NO3− for the two denitrifier strains studied. 相似文献
6.
Microcystin Production by Microcystis aeruginosa in a Phosphorus-Limited Chemostat 总被引:4,自引:0,他引:4
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The production of microcystins (MC) from Microcystis aeruginosa UTEX 2388 was investigated in a P-limited continuous culture. MC (MC-LR, MC-RR, and MC-YR) from lyophilized M. aeruginosa were extracted with 5% acetic acid, purified by a Sep-Pak C18 cartridge, and then analyzed by high-performance liquid chromatography with a UV detector and Nucleosil C18 reverse-phase column. The specific growth rate (μ) of M. aeruginosa was within the range of 0.1 to 0.8/day and was a function of the cellular P content under a P limitation. The N/P atomic ratio of steady-state cells in a P-limited medium varied from 24 to 15 with an increasing μ. The MC-LR and MC-RR contents on a dry weight basis were highest at μ of 0.1/day at 339 and 774 μg g−1, respectively, while MC-YR was not detected. The MC content of M. aeruginosa was higher at a lower μ, whereas the MC-producing rate was linearly proportional to μ. The C fixation rate at an ambient irradiance (160 microeinsteins m−2 s−1) increased with μ. The ratios of the MC-producing rate to the C fixation rate were higher at a lower μ. Accordingly, the growth of M. aeruginosa was reduced under a P limitation due to a low C fixation rate, whereas the MC content was higher. Consequently, increases in the MC content per dry weight along with the production of the more toxic form, MC-LR, were observed under more P-limited conditions. 相似文献
7.
Influence of Starvation on Potential Ammonia-Oxidizing Activity and amoA mRNA Levels of Nitrosospira briensis 总被引:1,自引:0,他引:1
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Annette Bollmann Ingo Schmidt Aaron M. Saunders Mette H. Nicolaisen 《Applied microbiology》2005,71(3):1276-1282
The effect of short-term ammonia starvation on Nitrosospira briensis was investigated. The ammonia-oxidizing activity was determined in a concentrated cell suspension with a NOx biosensor. The apparent half-saturation constant [Km(app)] value of the NH3 oxidation of N. briensis was 3 μM NH3 for cultures grown both in continuous and batch cultures as determined by a NOx biosensor. Cells grown on the wall of the vessel had a lower Km(app) value of 1.8 μM NH3. Nonstarving cultures of N. briensis showed potential ammonia-oxidizing activities of between 200 to 250 μM N h−1, and this activity decreased only slowly during starvation up to 10 days. Within 10 min after the addition of fresh NH4+, 100% activity was regained. Parallel with activity measurements, amoA mRNA and 16S rRNA were investigated. No changes were observed in the 16S rRNA, but a relative decrease of amoA mRNA was observed during the starvation period. During resuscitation, an increase in amoA mRNA expression was detected simultaneously. The patterns of the soluble protein fraction of a 2-week-starved culture of N. briensis showed only small differences in comparison to a nonstarved control. From these results we conclude that N. briensis cells remain in a state allowing fast recovery of ammonia-oxidizing activity after addition of NH4+ to a starved culture. Maintaining cells in this kind of active state could be the survival strategy of ammonia-oxidizing bacteria in nature under fluctuating NH4+ availability. 相似文献
8.
Steady-State Growth and Chemical Composition of the Marine Chlorophyte Dunaliella tertiolecta in Nitrogen-Limited Continuous Cultures 总被引:2,自引:2,他引:0
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The marine chlorophyte Dunaliella tertiolecta was grown in continuous cultures under NH4+-N, NO2−-N, NO3−-N, and urea-N limitations. The effect of the nitrogen cell quota (Qn) on the steady-state growth rate (μ) was the same regardless of the N source. The relationship between μ and Qn was well described by the Droop equation, but only up to the true maximum growth rate ^μ (= cell washout rate). The ratio between the minimum cell quota (kQ) and the maximum cell quota (Qm) was 0.19. Hence, there is no substitute for determining ^μ experimentally. That there was no difference in growth response to different N sources suggests that no internal pooling of inorganic nitrogen occurred. Both the carbon (Qc) and phosphorus (Qp) cell quotas under N limitation increased with increasing μ in a threshold fashion: virtually no change in either cell quota up to ~0.8 ^μ, followed by a rapid and large increase up to ^μ. In addition, in the region of low μ, there was an increase in Qp with a decreasing medium N/P ratio of between 15 and 5 (by atoms). The results generally indicate the physiological limits in cellular constituency under N limitation. The usefulness of this information, however, in describing the response of natural populations of marine phytoplankton to transient nutrient exposures on the temporal and spatial microscales that most likely exist is of limited value. 相似文献
9.
Kinetic parameters for nitrate reduction in intact sediment cores were investigated by using the acetylene blockage method at five sites along the Swale-Ouse river system in northeastern England, including a highly polluted tributary, R. Wiske. The denitrification rate in sediment containing added nitrate exhibited a Michaelis-Menten-type curve. The concentration of nitrate for half-maximal activity (Kmap) by denitrifying bacteria increased on passing downstream from 13.1 to 90.4 μM in the main river, but it was highest (640 μM) in the Wiske. The apparent maximal rate (Vmaxap) ranged between 35.8 and 324 μmol of N m−2 h−1 in the Swale-Ouse (increasing upstream to downstream), but it was highest in the Wiske (1,194 μmol N m−2 h−1). A study of nitrous oxide (N2O) production at the same time showed that rates ranged from below the detection limit (0.05 μmol of N2O-N m−2 h−1) at the headwater site to 27 μmol of N2O-N m−2 h−1 at the downstream site. In the Wiske the rate was up to 570 μmol of N2O-N m−2 h−1, accounting for up to 80% of total N gas production. 相似文献
10.
Simultaneous determinations of nitrogen gas production, ammonia, and particulate organic nitrogen formation in the coastal sediments of Mangoku-Ura, Simoda Bay, and Tokyo Bay were made by using the 15N-label tracer method. The rate of nitrogen gas production in the sediment surface layer was about 10−2 μg atom of N per g per h, irrespective of the location of the sediments examined. [15N]ammonia and -particulate organic nitrogen accounted for 20 to 70% of the three products, and after several hours of incubation, the major fraction of nondenitrified 15N in Mangoku-Ura and Simoda Bay sediments was recovered as ammonia. In Tokyo Bay sediments, particulate organic nitrogen was produced at a greater rate than was ammonia. The reduction rate data suggest that the pathway of nitrate reduction to ammonia is important in coastal sediments. 相似文献
11.
Studies with 15N indicate that appreciable generation of NH4+ from endogenous sources accompanies the uptake and assimilation of exogenous NH4+ by roots. To identify the source of NH4+ generation, maize (Zea mays L.) seedlings were grown on 14NH4+ and then exposed for 3 d to highly labeled 15NH4+. More of the entering 15NH4+ was incorporated into the protein-N fraction of roots in darkness (approximately 25%) than in the light (approximately 14%). Although the 14NH4+ content of roots declined rapidly to less than 1 μmol per plant, efflux of 14NH4+ continued throughout the 3-d period at an average daily rate of 14 μmol per plant. As a consequence, cumulative 14NH4+ efflux during the 3-d period accounted for 25% of the total 14N initially present in the root. Although soluble organic 14N in roots declined during the 3-d period, insoluble 14N remained relatively constant. In shoots both soluble organic 14N and 14NH4+ declined, but a comparable increase in insoluble 14N was noted. Thus, total 14N in shoots remained constant, reflecting little or no net redistribution of 14N between shoots and roots. Collectively, these observations reveal that catabolism of soluble organic N, not protein N, is the primary source of endogenous NH4+ generation in maize roots. 相似文献
12.
A New Sensitive Bioassay for Determination of Microbially Available Phosphorus in Water 总被引:18,自引:3,他引:15
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Markku J. Lehtola Ilkka T. Miettinen Terttu Vartiainen Pertti J. Martikainen 《Applied microbiology》1999,65(5):2032-2034
The content of assimilable organic carbon has been proposed to control the growth of microbes in drinking water. However, recent results have shown that there are regions where it is predominantly phosphorus which determines the extent of microbial growth in drinking waters. Even a very low concentration of phosphorus (below 1 μg of P liter−1) can promote extensive microbial growth. We present here a new sensitive method to determine microbially available phosphorus concentrations in water down to 0.08 μg of P liter−1. The method is a bioassay in which the analysis of phosphorus in a water sample is based on maximum growth of Pseudomonas fluorescens P17 when the energy supply and inorganic nutrients, with the exception of phosphorus, do not limit bacterial growth. Maximum growth (CFU) in the water sample is related to the concentration of phosphorus with the factor 373,200 ± 9,400 CFU/μg of PO4-P. A linear relationship was found between cell growth and phosphorus concentration between 0.05 to 10 μg of PO4-P liter−1. The content of microbially available phosphorus in Finnish drinking waters varied from 0.1 to 10.2 μg of P liter−1 (median, 0.60 μg of P liter−1). 相似文献
13.
Differences in SOM Decomposition and Temperature Sensitivity among Soil Aggregate Size Classes in a Temperate Grasslands 总被引:4,自引:0,他引:4
The principle of enzyme kinetics suggests that the temperature sensitivity (Q10) of soil organic matter (SOM) decomposition is inversely related to organic carbon (C) quality, i.e., the C quality-temperature (CQT) hypothesis. We tested this hypothesis by performing laboratory incubation experiments with bulk soil, macroaggregates (MA, 250–2000 μm), microaggregates (MI, 53–250 μm), and mineral fractions (MF, <53 μm) collected from an Inner Mongolian temperate grassland. The results showed that temperature and aggregate size significantly affected on SOM decomposition, with notable interactive effects (P<0.0001). For 2 weeks, the decomposition rates of bulk soil and soil aggregates increased with increasing incubation temperature in the following order: MA>MF>bulk soil >MI(P <0.05). The Q10 values were highest for MA, followed (in decreasing order) by bulk soil, MF, and MI. Similarly, the activation energies (Ea) for MA, bulk soil, MF, and MI were 48.47, 33.26, 27.01, and 23.18 KJ mol−1, respectively. The observed significant negative correlations between Q10 and C quality index in bulk soil and soil aggregates (P<0.05) suggested that the CQT hypothesis is applicable to soil aggregates. Cumulative C emission differed significantly among aggregate size classes (P <0.0001), with the largest values occurring in MA (1101 μg g−1), followed by MF (976 μg g−1) and MI (879 μg g−1). These findings suggest that feedback from SOM decomposition in response to changing temperature is closely associated withsoil aggregation and highlights the complex responses of ecosystem C budgets to future warming scenarios. 相似文献
14.
Macroalgae has bloomed in the brackish lake of Shenzhen Bay, China continuously from 2010 to 2014. Gracilaria tenuistipitata was identified as the causative macroalgal species. The aim of this study was to explore the outbreak mechanism of G. tenuistipitata, by studying the effects of salinity and nitrogen sources on growth, and the different nitrogen sources uptake characteristic. Our experimental design was based on environmental conditions observed in the bloom areas, and these main factors were simulated in the laboratory. Results showed that salinity 12 to 20 ‰ was suitable for G. tenuistipitata growth. When the nitrogen sources'' (NH4
+, NO3
−) concentrations reached 40 µM or above, the growth rate of G. tenuistipitata was significantly higher. Algal biomass was higher (approximately 1.4 times) when cultured with NH4
+ than that with NO3
− addition. Coincidentally, macroalgal bloom formed during times of moderate salinity (∼12 ‰) and high nitrogen conditions. The NH4
+ and NO3
− uptake characteristic was studied to understand the potential mechanism of G. tenuistipitata bloom. NH4
+ uptake was best described by a linear, rate-unsaturated response, with the slope decreasing with time intervals. In contrast, NO3
− uptake followed a rate-saturating mechanism best described by the Michaelis-Menten model, with kinetic parameters Vmax = 37.2 µM g−1 DM h−1 and Ks = 61.5 µM. Further, based on the isotope 15N tracer method, we found that 15N from NH4
+ accumulated faster and reached an atom% twice than that of 15N from NO3
−, suggesting when both NH4
+ and NO3
− were available, NH4
+ was assimilated more rapidly. The results of the present study indicate that in the estuarine environment, the combination of moderate salinity with high ammonium may stimulate bloom formation. 相似文献
15.
We compared growth kinetics of Prorocentrum donghaiense cultures on different nitrogen (N) compounds including nitrate (NO3
−), ammonium (NH4
+), urea, glutamic acid (glu), dialanine (diala) and cyanate. P. donghaiense exhibited standard Monod-type growth kinetics over a range of N concentraions (0.5–500 μmol N L−1 for NO3
− and NH4
+, 0.5–50 μmol N L−1 for urea, 0.5–100 μmol N L−1 for glu and cyanate, and 0.5–200 μmol N L−1 for diala) for all of the N compounds tested. Cultures grown on glu and urea had the highest maximum growth rates (μm, 1.51±0.06 d−1 and 1.50±0.05 d−1, respectively). However, cultures grown on cyanate, NO3
−, and NH4
+ had lower half saturation constants (Kμ, 0.28–0.51 μmol N L−1). N uptake kinetics were measured in NO3
−-deplete and -replete batch cultures of P. donghaiense. In NO3
−-deplete batch cultures, P. donghaiense exhibited Michaelis-Menten type uptake kinetics for NO3
−, NH4
+, urea and algal amino acids; uptake was saturated at or below 50 μmol N L−1. In NO3
−-replete batch cultures, NH4
+, urea, and algal amino acid uptake kinetics were similar to those measured in NO3
−-deplete batch cultures. Together, our results demonstrate that P. donghaiense can grow well on a variety of N sources, and exhibits similar uptake kinetics under both nutrient replete and deplete conditions. This may be an important factor facilitating their growth during bloom initiation and development in N-enriched estuaries where many algae compete for bioavailable N and the nutrient environment changes as a result of algal growth. 相似文献
16.
17.
Biodegradation of the Hexahydro-1,3,5-Trinitro-1,3,5-Triazine Ring Cleavage Product 4-Nitro-2,4-Diazabutanal by Phanerochaete chrysosporium
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Diane Fournier Annamaria Halasz Jim Spain Ronald J. Spanggord Jeffrey C. Bottaro Jalal Hawari 《Applied microbiology》2004,70(2):1123-1128
Initial denitration of hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) by Rhodococcus sp. strain DN22 produces CO2 and the dead-end product 4-nitro-2,4-diazabutanal (NDAB), OHCNHCH2NHNO2, in high yield. Here we describe experiments to determine the biodegradability of NDAB in liquid culture and soils containing Phanerochaete chrysosporium. A soil sample taken from an ammunition plant contained RDX (342 μmol kg−1), HMX (octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine; 3,057 μmol kg−1), MNX (hexahydro-1-nitroso-3,5-dinitro-1,3,5-triazine; 155 μmol kg−1), and traces of NDAB (3.8 μmol kg−1). The detection of the last in real soil provided the first experimental evidence for the occurrence of natural attenuation that involved ring cleavage of RDX. When we incubated the soil with strain DN22, both RDX and MNX (but not HMX) degraded and produced NDAB (388 ± 22 μmol kg−1) in 5 days. Subsequent incubation of the soil with the fungus led to the removal of NDAB, with the liberation of nitrous oxide (N2O). In cultures with the fungus alone NDAB degraded to give a stoichiometric amount of N2O. To determine C stoichiometry, we first generated [14C]NDAB in situ by incubating [14C]RDX with strain DN22, followed by incubation with the fungus. The production of 14CO2 increased from 30 (DN22 only) to 76% (fungus). Experiments with pure enzymes revealed that manganese-dependent peroxidase rather than lignin peroxidase was responsible for NDAB degradation. The detection of NDAB in contaminated soil and its effective mineralization by the fungus P. chrysosporium may constitute the basis for the development of bioremediation technologies. 相似文献
18.
Initial organic products of assimilation of [N]ammonium and [N]nitrate by tobacco cells cultured on different sources of nitrogen
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Glutamine is the first major organic product of assimilation of 13NH4+ by tobacco (Nicotiana tabacum L. cv. Xanthi) cells cultured on nitrate, urea, or ammonium succinate as the sole source of nitrogen, and of 13NO3− by tobacco cells cultured on nitrate. The percentage of organic 13N in glutamate, and subsequently, alanine, increases with increasing periods of assimilation. 13NO3−, used for the first time in a study of assimilation of nitrogen, was purified by new preparative techniques. During pulse-chase experiments, there is a decrease in the percentage of 13N in glutamine, and a concomitant increase in the percentage of 13N in glutamate and alanine. Methionine sulfoximine inhibits the incorporation of 13N from 13NH4+ into glutamine more extensively than it inhibits the incorporation of 13N into glutamate, with cells grown on any of the three sources of nitrogen. Azaserine inhibits glutamate synthesis extensively when 13NH4+ is fed to cells cultured on nitrate. These results indicate that the major route for assimilation of 13NH4+ is the glutamine synthetase-glutamate synthase pathway, and that glutamate dehydrogenase also plays a role, but a minor one. Methionine sulfoximine inhibits the incorporation of 13N from 13NO3− into glutamate more strongly than it inhibits the incorporation of 13N into glutamine, suggesting that the assimilation of 13NH4+ derived from 13NO3− may be mediated solely by the glutamine synthetase-glutamate synthase pathway. 相似文献
19.
Estimating Bacterioplankton Production by Measuring [3H]thymidine Incorporation in a Eutrophic Swedish Lake 总被引:3,自引:0,他引:3
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Bacterioplankton abundance, [3H]thymidine incorporation, 14CO2 uptake in the dark, and fractionated primary production were measured on several occasions between June and August 1982 in eutrophic Lake Norrviken, Sweden. Bacterioplankton abundance and carbon biomass ranged from 0.5 × 109 to 2.4 × 109 cells liter−1 and 7 to 47 μg of C liter−1, respectively. The average bacterial cell volume was 0.185 μm3. [3H]thymidine incorporation into cold-trichloroacetic acid-insoluble material ranged from 12 × 10−12 to 200 × 10−12 mol liter−1 h−1. Bacterial carbon production rates were estimated to be 0.2 to 7.1 μg of C liter−1 h−1. Bacterial production estimates from [3H]thymidine incorporation and 14CO2 uptake in the dark agreed when activity was high but diverged when activity was low and when blue-green algae (cyanobacteria) dominated the phytoplankton. Size fractionation indicated negligible uptake of [3H]thymidine in the >3-μm fraction during a chrysophycean bloom in early June. We found that >50% of the 3H activity was in the >3-μm fraction in late August; this phenomenon was most likely due to Microcystis spp., their associated bacteria, or both. Over 60% of the 14CO2 uptake in the dark was attributed to algae on each sampling occasion. Algal exudate was an important carbon source for planktonic bacteria. Bacterial production was roughly 50% of primary production. 相似文献
20.
New Insights into Methyl Bromide Cooxidation by Nitrosomonas europaea Obtained by Experimenting with Moderately Low Density Cell Suspensions
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Khrystyne N. Duddleston Peter J. Bottomley Angela J. Porter Daniel J. Arp 《Applied microbiology》2000,66(7):2726-2731
We examined the rates and sustainability of methyl bromide (MeBr) oxidation in moderately low density cell suspensions (~6 × 107 cells ml−1) of the NH3-oxidizing bacterium Nitrosomonas europaea. In the presence of 10 mM NH4+ and 0.44, 0.22, and 0.11 mM MeBr, the initial rates of MeBr oxidation were sustained for 12, 12, and 24 h, respectively, despite the fact that only 10% of the NH4+, 18% of the NH4+, and 35% of the NH4+, respectively, were consumed. Although the duration of active MeBr oxidation generally decreased as the MeBr concentration increased, similar amounts of MeBr were oxidized with a large number of the NH4+-MeBr combinations examined (10 to 20 μmol mg [dry weight] of cells−1). Approximately 90% of the NH3-dependent O2 uptake activity and the NO2−-producing activity were lost after N. europaea was exposed to 0.44 mM MeBr for 24 h. After MeBr was removed and the cells were resuspended in fresh growth medium, NO2− production increased exponentially, and 48 to 60 h was required to reach the level of activity observed initially in control cells that were not exposed to MeBr. It is not clear what percentage of the cells were capable of cell division after MeBr oxidation because NO2− accumulated more slowly in the exposed cells than in the unexposed cells despite the fact that the latter were diluted 10-fold to create inocula which exhibited equal initial activities. The decreases in NO2−-producing and MeBr-oxidizing activities could not be attributed directly to NH4+ or NH3 limitation, to a decrease in the pH, to the composition of the incubation medium, or to toxic effects caused by accumulation of the end products of oxidation (NO2− and formaldehyde) in the medium. Additional cooxidation-related studies of N. europaea are needed to identify the mechanism(s) responsible for the MeBr-induced loss of cell activity and/or viability, to determine what percentages of cells damaged by cooxidative activities are culturable, and to determine if cooxidative activity interferes with the regulation of NH3-oxidizing activity. 相似文献