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1.
When Azotobacter chroococcum cells grown in batch culture under N2-fixing conditions were transferred to a medium lacking a nitrogen source, the cellular C/N ratio, the amount of alginic acid released into the external medium and the rate of endogenous respiration increased appreciably after 6 h to the exclusion of dinitrogen, whereas nitrogenase activity did not undergo any significant change. Nitrogen deficiency caused a decrease in the ammonium inhibition of nitrogenase activity from 95% inhibition at zero time to 14% after 6 h incubation under dinitrogen starvation, with no difference in the rate of ammonium utilization by N2-fixing and N2-starved cells being observed. This suggests that a balance of nitrogen and carbon assimilation is necessary for the ammonium inhibition of nitrogenase activity in A. chroococcum to take place.  相似文献   

2.
Hydrogenase-negative (Hup-) mutants of Azorhizobium caulinodans ORS571 were isolated by means of Tn5 mutagenesis. The colony test used for screening for Hup- strains was based on the absence of reduction of triphenyltetrazolium chloride with hydrogen. Suspensions from cultures of the mutant strains grown under derepressing conditions did not use hydrogen with methylene blue or oxygen as the hydrogen acceptor. The mutants were shown to carry single Tn5 insertions at different locations in the A. caulinodans genome. Molar growth yields (corrected for poly--hydroxybutyrate formation) in chemostat cultures of the mutants were similar to those of the wild type. Molar growth yields of the mutants were not increased by passing additional hydrogen through chemostat cultures, which is in agreement with the hydrogenase-negative phenotype of the mutants. H2/N2 ratios (mol H2 formed per mol N2 fixed) were calculated from the hydrogen content of the effluent gas and the N-content of the bacterial dry weight. Low H2/N2 ratios (between 1.2 and 1.9) were found in both energy-limited (oxygen or succinate) cultures and in cultures limited by the supply of an anabolic substrate (Mg2+). ATP/2e values (mol ATP used at the transport of 2e to nitrogen or H+) were calculated from the H2/N2 ratios and the molar growth yields of nitrogen-fixing and ammonia-assimilating cultures. ATP/2e values were between 7 and 11. It was concluded that the calculated ATP/2e values comprise not only 4 mol ATP used at the transport of 2e through nitrogenase but also energy equivalents needed for reversed electron flow from NADH to the low-potential hydrogen donor used by nitrogenase.  相似文献   

3.
The nitrate reductase activity (NR) of selected uptake hydrogenase-positive (hup +) and uptake hydrogenase-negative (hup -) strains of Bradyrhizobium japonicum were examined both in free-living cells and in symbioses with Glycine max L. (Marr.) cv. Williams. Bacteria were cultured in a defined medium containing either 10 mM glutamate or nitrate as the sole nitrogen source. Nodules and bacteriods were isolated from plants that were only N2-dependent or grown in the presence of 2 mM KNO3. Rates of activity in nodules were determined by an in vivo assay, and those of cultured cells and bacteriods were assayed after permeabilization of the cells with alkyltrimethyl ammonium bromide. All seven strains examined expressed NR activity as free-living cells and as symbiotic forms, regardless of the hup genotype of the strain used for inoculation. Although the presence of nitrate increased nitrate reduction by cultures cells and nodules, no differences in NR activity were observed between bacteroids isolated from nodules of plants fed with nitrate or grown on N2-fixation exclusively. Cultured cells, nodules and bacteriods of strains with hup - genotype (USDA 138, L-236, 3. 15B3 and PJ17) had higher rates of NR activity than those with hup + genotype (USDA 110, USDA 122 DES and CB1003). These results suggest that NR activity is reduced in the presence of a genetic determinant associated with the hup region of B. japonicum.Abbreviations EDTA ethylene-diamine tetraacetic acid - Hup hydrogen uptake - MOPS 3-(N-morpholino)-propane sulfonic acid - NR nitrate reductase - PVP polyvinyl-polypyrrolidone - Tris Tris(hydroxymethyl)-aminomethane  相似文献   

4.
A methanogenic bacterial consortium was obtained after inoculation of benzoate medium under N2/CO2 atmosphere with intertidal sediment. A hydrogen donating organotroph andMethanococcus mazei were isolated from this enrichment. H2-utilising sulphate reducing bacteria were isolated under H2/CO2 in the absence of organic electron donors. TheMethanococcus was able to produce methane in yeast extract medium under N2/CO2 if the H2 donating organism was present, and sulphate reduction occurred if the hydrogen utilising sulphate reducing bacteria were grown with the H2 donating organism. The ability of the H2 utilising sulphate reducing bacteria to inhibitMethanococcus competitively was shown in cultures containing both of these H2 utilising bacteria.Abbreviations HDO hydrogen donating organism - SRB sulphate reducing bacteria - HSRB hydrogen utilising sulphate reducing bacteria  相似文献   

5.
Plants from four cultivars of Lycopersicon esculentum were grown under different conditions, in controlled environment chambers. Low light intensity, long photoperiod (16 h), 25° C/17°C temperature alternance (day/night) were found to be the most convenient conditions for obtaining viable protoplasts. The use of myo-inositol as an osmoticum in the digestion medium and the adjustment of the pH to 6.5, instead of the usual 5.8, for this medium increased the yield of viable protoplasts and enhanced their stability. Under these conditions neither pretreatment (dark and cold treatments), nor preplasmolysis of leaf tissues, were required before protoplast isolation. The concentrations of ammonium nitrate, calcium chloride, myo-inositol, and sucrose were found to be critical for the success of protoplast culture. A medium containing 5 mM ammonium nitrate, 40 mM calcium chloride, 10 mg l-1 adenine sulfate, 0.5% myo-inositol and 6% sucrose gave sustained protoplast divisions. Under these conditions, plating efficiency ranged from 5% for the cultivar Lukulus to 15% for the cultivar Golden Sunrise.Abbreviations BA benzylaminopurine - CaCl2 calcium chloride, 2,4,-D-2,4-dichlorophenoxyacetic acid - EDTA ethylene diamine tetraacetic acid - KCl potassium chloride - MES-2-N morpholino ethane sulfonic acid - MgCl2 magnesium chloride - NH4NO3 ammonium nitrate - NAA naphthalene acetic acid, p-protoplasts  相似文献   

6.
Summary Seed inoculation with Rhizobium and soil inoculation withGlomus fasciculatum increased nodulation, nitrogen and phosphorus concentration in plants and yield of chickpea (Cicer arietinum) var. BG 212 in pots containing unsterilized soil especially with 50kgP2O5 ha−1 in the form of superphosphate. Inoculation with Rhizobium orG. fasciculatum separately or in combination significantly increased the N2 fixed in straw and grain than uninoculated controls as determined by15N atom percent excess of plants grown in soil amended with labelled ammonium sulphate (15NH4)2SO4) at the rate of 20kg N ha−1. These increases were most pronounced when P was applied at 50kgP2O5 ha−1.  相似文献   

7.
A release of ammonium by non-nitrogen-fixing Anabaena cylindrica (grown on NH4Cl) in the presence of MSX (methionine sulfoximine) and absence of any external nitrogen source was found. In the light the release was maximal at 0.2 mM MSX, a concentration which did not affect net CO2 fixation nor the glycollate excretion, but inhibited the glutamine synthetase activity and the reassimilation of ammonium. It is suggested that the major source of the ammonium released is the photorespiratory conversion of glycine to serine as (1) the release was stimulated by increase in light intensity, (2) high CO2 (3%) lowered the release, if not given as a longer pretreatment (as CO2 or HCO 3 - ) when a stimulation was observed, (3) glyoxylate and glutamate stimulated the release, the latter compound particularly under nitrogen-deficient conditions and (4) isonicotinic acid hydrazide caused a reduced release of ammonium. Furthermore, a substantial part of the ammonium released by N2-fixing A. cylindrica in presence of MSX may thus originate from the glycollate pathway. The data show that in the light the glycine to serine conversion is active in cyanobacteria with a concomitant production of ammonium which is assimilated by glutamine synthetase.Abbreviations MSX L-methionine-Dl-sulfoximine - INH isonicotinic acid hydrazide - RuDP ribulose 1,5-diphosphate - Hepes N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid - GS glutamine synthetase - GOGAT glutamate synthase - DTT Dl-dithiothreitol  相似文献   

8.
Seventeen strains of nitrogen-fixing bacteria, isolated from different habitats on hydrogen and carbon dioxide as well as on other substrates, morphologically resembled each other. All strains, including Mycobacterium flavum 301, grew autotrophically with hydrogen. The isolate strain 6 was sensitive to oxygen when dependent on N2 as nitrogen source, a consequence of the sensitivity of its nitrogenase towards oxygen. At the same time, strain 6 was sensitive to hydrogen when growing autotrophically on N2 as nitrogen source, but hydrogen did not affect acetylene reduction by these cells.Abbreviations MPN Most probable number - BS medium basal salts medium  相似文献   

9.
Several blue-green algae were surveyed for the occurrence of the hydrogenase which was assayed by the oxyhydrogen or Knallgas reaction in the intact organisms. In aerobically grown cultures, the reaction was detectable in Anabaena cylindrica, Nostoc muscorum and in two Anabaena variabilis species, whereas virtually no activity was observed in Anacystis nidulans and Cyanophora paradoxa. In these latter two algae, the reaction was, however, found after growth under molecular hydrogen for several days, which drastically increased the activity levels with all the algae tested. In the nitrogen fixing species, the activity of the Knallgas reaction was enhanced when all combined nitrogen was omitted from the media. H2 and hydrogenase could not significantly support the CO2-fixation in photoreduction experiments with all blue-green algae investigated here. Hydrogenase was assayed by the dithionite and methyl viologen dependent evolution of hydrogen and was found to be present with essentially the same specific activity levels in preparations of both heterocysts and vegetative cells from Anabaena cylindrica. Na2S2O4 as well as H2 supported the C2H2-reduction of the isolated heterocysts. The H2-dependent C2H2-reduction did not require the presence of oxygen but was strictly light-dependent where H2 served as an electron donor to photosystem I of these cells. It is concluded that hydrogen can be utilized by two different pathways in blue-green algae.Abbreviations Chl chlrophyll - CP creatine phosphate - CP kinase creatine phosphokinase - DCMU N-(3,4-dichlorophenyl)N,N-dimethylurea  相似文献   

10.
The effect of various nitrogen sources on the synthesis and activity of nitrogenase was studied in the marine, non-heterocystous cyanobacterium Trichodesmium sp. NIBB1067 grown under defined culture conditions. Cells grown with N2 as the sole inorganic nitrogen source showed light-dependent nitrogenase activity (acetylene reduction). Nitrogenase activity in cells grown on N2 was not suppressed after 7 h incubation with 2 mM NaNO3 or 0.02 mM NH4Cl. However, after 3 h of exposure to 0.5 mM of urea, nitrogenase was inactivated. Cells grown in medium containing 2 mM NaNO3, 0.5 mM urea or 0.02 mM NH4Cl completely lacked the ability to reduce acetylene. Western immunoblots tested with polyclonal antisera against the Fe-protein and the Mo–Fe protein, revealed the following: (1) both the Fe-protein and the Mo–Fe protein were synthesized in cells grown with N2 as well as in cells grown with NaNO3 or low concentration of NH4Cl; (2) two bands (apparent molecular mass of 38 000 and 40 000) which cross-reacted with the antiserum to the Fe-protein, were found in nitrogen-fixing cells; (3) only one protein band, corresponding to the high molecular mass form of the Fe-protein, was found in cells grown with NaNO3 or low concentration of NH4Cl; (4) neither the Fe-protein nor the Mo–Fe protein was found in cells grown with urea; (5) the apparent molecular mass of the Fe-protein of Trichodesmium sp. NIBB1067 was about 5000 dalton higher than that of the heterocystous cyanobacterium, Anabaena cylindrica IAM-M1.  相似文献   

11.
Summary Studies were conducted to evaluate the uptake of mercury by wheat (Triticum aestivum L. runar) and beans (Phaseolus vulgaris L. marshal) growth on an oxisol with different levels of 2-methoxyethylmercury chloride (Aretan) and mercuric chloride. Dry matter and grain yields of wheat were little affected by either Aretan or mercuric chloride, although Aretan at 50 mg Hg/kg soil delayed germination by four to five days. Germination of beans grown with both compounds at the 50 mg Hg/kg soil failed completely, even after repeated sowing. Yields were somewhat, though not significantly, decreased by mercury chloride up to 5 mg Hg/kg soil.The concentration of Hg in wheat straw and grain increased significantly with increased levels of Aretan and HgCl2 application, with more Hg taken up by the plants grown with HgCl2 than with those grown with Aretan. Translocation of Hg to grain was greater in the plants grown with HgCl2.The concentration of Hg in bean straw, but not grain, increased significantly with increasing levels of Aretan and HgCl2 application, and was greater in plants grown with HgCl2. Translocation to grain was low, with little difference between plants grown with Aretan or HgCl2.  相似文献   

12.
The relationship between ammonium assimilation and ammonium export has been studied in free-living, N2-fixing Rhizobium sp. 32H1. After 55 to 67 h of microaerobic growth under a gas phase of 0.2% O2 – 1.0% CO2 – 98.8% Ar high levels of nitrogenase were observed concomitant with a slightly adenylylated glutamine synthetase (GSI) and some glutamine synthetase (GSII) activity. However, after growth of 89 h, or longer, GSI became adenylylated and the level of GSII had decreased. When the gas phase was shifted to 0.2% O2 – 1.0% CO2 – 98.8% N2, a lag was observed before ammonium export could be detected in the 55 to 67 h cultures. No lag in ammonium export was observed in the cultures previously grown for 89 h. The onset of ammonium export in the 55 to 67 h cultures was found to correlate with the adenylylation state of GSI. There appeared to be no correlation between the level of GSII and the export of ammonium. Neither an increase in the adenylylation level of GSI nor ammonium export was observed when the 55 to 67 h cultures were maintained under the Ar gas mixture.Abbreviations GOGAT Glutamate synthase - GS glutamine synthetase - BES [N,N-bis(2-hydroxyethyl)-2-aminoethanesulfonic acid] - CTAB cetyltrimethylammonium bromide - MES [2-(N-morpholino)-ethane sulfonic acid]  相似文献   

13.
The primary steps of N2, ammonia and nitrate metabolism in Klebsiella pneumoniae grown in a continuous culture are regulated by the kind and supply of the nitrogenous compound. Cultures growing on N2 as the only nitrogen source have high activities of nitrogenase, unadenylated glutamine synthetase and glutamate synthase and low levels of glutamate dehydrogenase. If small amounts of ammonium salts are added continuously, initially only part of it is absorbed by the organisms. After 2–3 h complete absorption of ammonia against an ammonium gradient coinciding with an increased growth rate of the bacteria is observed. The change in the extracellular ammonium level is paralleled by the intracellular glutamine concentration which in turn regulates the glutamine synthetase activity. An increase in the degree of adenylation correlates with a repression of nitrogenase synthesis and an induction of glutamate dehydrogenase synthesis. Upon deadenylation these events are reversed.—After addition of nitrate ammonia appears in the medium, probably due to the action of a membrane bound dissimilatory nitrate reductase.—Addition of dinitrophenol causes transient leakage of intracellular ammonium into the medium.  相似文献   

14.
The utilization of some amino acids, added at 1 mM and 10 mM concentrations, as the sole combined nitrogen sources by Frankia sp. strain CpI1, has been investigated. Glutamine, like NH 4 + , provided rapid growth without N2 fixation. Histidine at 1 mM yielded poor N2-fixing activity but better cell growth than N2. Aspartate, glutamate, alanine, proline, each at 1 mM concentration, supported similar levels of N2 fixation and growth. Growth on 10 mM glutamate, proline, or histidine resulted in poor N2-fixing activity and poor cell growth. Cells grown on 10 mM alanine had about half the N2-fixing activity of cells grown on N2 but growth was good. Aspartate at 10 mM concentration, however, stimulated N2-fixing activity dramatically and promoted faster growth. Enzyme analysis suggested that asparate is catabolized by glutamate-oxaloacetate transaminase (GOT), since GOT specific activity was induced, and aspartase activity was not detected, in cells grown on aspartate as the sole combined nitrogen source. Thinlayer chromatography (TLC) of metabolites extracted from N2-grown cells fed with [14C]-aspartate showed that label was rapidly accumulated mainly on aspartate and/or glutamate, depending on the cells' physiological state, without detectable labeling on fumarate or oxaloacetate (OAA). These findings provide evidence that aspartate is catabolized by GOT to OAA which, in turn, is rapidly converted to -ketoglutarate through the TCA cycle and then to glutamate by GOT or by glutamate synthase (GOGAT). The stimulation of N2 fixation and growth by aspartate is probably caused by an increased intracellular glutamate pool.  相似文献   

15.
Nitrate, nitrite and nitrous oxide were denitrified to N2 gas by washed cells ofRhizobium japonicum CC706 as well as by bacteroids prepared from root nodules ofGlycine max (L.) Merr. (CV. Clark 63). Radiolabelled N2 was produced from either K15NO3 or Na15NO2 by washed cells ofRh. japonicum CC705 grown with either nitrate only (5 mM) or nitrate (5 mM) plus glutamate (10 mM). Nitrogen gas was also produced from N2O. Similar results were obtained with bacteroids ofG. max. The stoichiometry for the utilization of15NO 3 - or15NO 2 - and the produciton of15N2 was 2:1 and for N2O utilization and N2 production it was 1:1. Some of the15N2 gas produced by denitrification of15NO 3 - in bacteroids was recycled via nitrogenase into cell nitrogen.  相似文献   

16.
Soaking seeds of cucumber and pumpkin with an extract of Westiellopsis prolifica, an N2-fixing cyanobacterium, promoted germination and their subsequent growth and development. An extract of Lyngbya sp., a non-N2-fixing cyanobacterium, had no significant effect.The authors are with the Algal Physiology Laboratory, Department of Botany, Berhampur University, Berhampur-760007, Orissa, India.  相似文献   

17.
Activation state of ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPCO) is an important parameter determining the rate of net photosynthesis (P N) in situ for which no information is available with reference to altitude. We analyzed activation state along with P N in three plant species and their cultivars grown at low (LA, 1 300 m) and high (HA, 4 200 m) altitudes. No significant change in P N and the initial activity of RuBPCO was obtained with reference to altitude. However, activation state of RuBPCO was reduced significantly in the HA plants as compared to the LA ones. Hence low partial pressure of CO2 prevailing at HA might be responsible for the lower activation state of RuBPCO.  相似文献   

18.
When a new strain of Pseudomonas aeruginosa was grown aerobically and then transferred to anaerobic conditions, cells reduced NO 3 quantitatively to NO 2 in NO 3 -respiration. In the absence of nitrate, NO 2 was immediately reduced to NO or N2O but not to N2 indicating that NO 2 -reductase but not N2O-reductase was active. The formation of the products NO or N2O depended on the pH in the medium and the concentration of NO 2 present. When P. aeruginosa was grown anaerobically for at least three davs N2O-reductase was also active. Such cells reduced NO to N2 via N2O. The new strain generated a H+-gradient and grew by reducing N2O to N2 but not by converting NO to N2O. For comparison, Azospirillum brasilense Sp7 showed the same pattern of NO-reduction. In contrast, Paracoccus denitrificans formed 3.5 H+/NO during the reduction of NO to N2O in oxidant pulse experiments but could not grow in the presence of NO. Thus the NO-reduction pattern in P. denitrificans on one side and P. aeruginosa and A. brasilense on the other was very different. The mechanistic implications of such differences are discussed.  相似文献   

19.
Syntrophomonas wolfei and Syntrophus buswellii were grown with butyrate or benzoate in a defined binary coculture with Methanospirillum hungatei. Both strains also grew independent of the partner bacteria with crotonate as substrate. Localization of enzymes involved in butyrate oxidation by S. wolfei revealed that ATP synthase, hydrogenase, and butyryl-CoA dehydrogenase were at least partially membrane-associated whereas 3-hydroxybutyryl-CoA dehydrogenase and crotonase were entirely cytoplasmic. Inhibition experiments with copper chloride indicated that hydrogenase faced the outer surface of the cytoplasmic membrane. Suspensions of butyrate-or benzoate-grown cells of either strain accumulated hydrogen during oxidation of butyrate or benzoate to a low concentration that was thermodynamically in equilibrium with calculated reaction energetics. The protonophore carbonylcyanide m-chlorophenyl-hydrazone (CCCP) and the proton-translocating ATPase inhibitor N,Ndicyclohexylcarbodiimide (DCCD) both specifically inhibited hydrogen formation from butyrate or benzoate at low concentrations, whereas hydrogen formation from crotonate was not affected. A menaquinone was extracted from cells of S. wolfei and S. buswellii grown syntrophically in a binary methanogenic culture. The results indicate that a proton-potential-driven process is involved in hydrogen release from butyrate or benzoate oxidation.Abbreviations BES Bromoethanesulfonate - CCCP Carbonyl cyanide-m-chlorophenyl-hydrazone - DCCD N,Ndicyclohexylcarbodiimide - DCPIP Dichlorophenol indophenol - PMS Phenazine methosulfate  相似文献   

20.
It was shown that tRNA fromAzotobacter vinelandii grown in the presence of ammonium chloride lacks ribothymidine while that grown in the absence of the ammonium salt contains this modified nucleoside. [32P]-Labelled tRNA from this organism grown in a medium containing the ammonium salt was digested with RNase T1 and the pseudouridinecontaining tetranucleotide, common to all tRNAs was isolated and analysed for the nucleoside replacing the ribothymidine. It was found to be uridine. Cells previously labelled with [32P]-phosphate in the ammonium salt medium were washed and incubated in the ammonium saltfree medium to test whether ribothymidine would be formed upon removal of the ammonium ions. Methylation of the uridine did not take place.  相似文献   

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