The role of formate metabolism in nitrogen fixation in Rhizobium Spp.

Formate metabolism supported nitrogen-fixation activity in free-living cultures of Rhizobium japonicum. However, formate0dependent nitrogense activity was observed only in the presence of carbon sources such as glutamate, ribose or aspartate which by themselves were unable to support nitrogenase activity. Formate-dependent nitrogenase activity was not detected in the presence of carbon sources such as malate, gluconate or glycerol which by themselves supported nitrogenase activity. A mutant strain of R. japonicum was isolated that was unable to utilise formate and was shown to lack formate dehydrogenase activity. This mutant strain exhibited no formate-dependent nitrogenase activity. Both the wild-type and mutant strains nodulated soybean plants effectively and there were no significant differences in the plant dry weight or total nitrogen content of the respective plants. Furthermore pea bacteroids lacked formate dehydrogenase activity and exogenously added formate had no stimulatory effect on the endogenous oxygen uptake rate. The role of formate metabolism in symbiotic nitrogen fixation is discussed.Abbreviation FDH formate dehydrogenase     

Fed-Batch Cultivation of Arthrospira platensis Using Carbon Dioxide from Alcoholic Fermentation and Urea as Carbon and Nitrogen Sources

To reduce CO₂ emissions from alcoholic fermentation, Arthrospira platensis was cultivated in tubular photobioreactor using either urea or nitrate as nitrogen sources at different light intensities (60 μmol m^?2 s^?1?≤?I?≤?240 μmol m^?2 s^?1). The type of carbon source (pure CO₂ or CO₂ from fermentation) did not show any appreciable influence on the main cultivation parameters, whereas substitution of nitrate for urea increased the nitrogen-to-cell conversion factor (Y _X/N ), and the maximum cell concentration (X _m ) and productivity (P _X ) increased with I. As a result, the best performance using gaseous emissions from alcoholic fermentation (X _m ?=?2,960?±?35 g m^?3, P _X ?=?425?±?5.9 g m^?3 day^?1 and Y _X/N ?=?15?±?0.2 g g^?1) was obtained at I?=?120 μmol m^?2 s^?1 using urea as nitrogen source. The results obtained in this work demonstrate that the combined use of effluents rich in urea and carbon dioxide could be exploited in large-scale cyanobacteria cultivations to reduce not only the production costs of these photosynthetic microorganisms but also the environmental impact associated to the release of greenhouse emissions.     

The Influence of Urea and Other Nitrogen Sources on Growth Rate of Scots Pine Seedlings

The growth rate and water content of urea-fed seedlings of Pinus silvestris L. were compared with those of nitrate-and ammonium-fed seedlings grown in continuously renewed nutrient solutions, in which the hydrolysis of urea to ammonia and carbon dioxide was minimized. The growth rate of seedlings grown in an ammonium nutrient solution, in an urea nutrient solution and in a nitrate nutrient solution was about 90 per cent, 75 per cent and 60 per cent, respectively, of that of seedlings grown in a mixture of ammonium and nitrate. Seedlings with urea as the sole nitrogen source developed very severe chlorosis of the needles, the old roots were dark-coloured, the whole root system was very fragile, and the lateral roots of the third order were missing. Urea-grown seedlings had the highest nitrogen contents, closely followed by the ammonium and the ammonium + nitrate seedlings. The lowest nitrogen level was in nitrate seedlings. The low growth rate and the chlorosis of urea-fed seedlings were suggested to be the result of a hydrolysis of urea inside the root, causing an increase in pH and an accumulation of ammonia in the root.     

Dipeptides as Nitrogen Sources for Drosera rotundifolia in Aseptic Culture

The growth of Drosera rotundifolia was studied in aseptic cultures with 17 dipeptides as the only nitrogen source. About half of the dipeptides were well or partly utilized. Compounds containing glycine, alanine, glutamic or aspartic acid are clearly more favourable than dipeptides containing proline. Arginyl-aspartic acid (1.25 mM) promoted growth more than inorganic nitrogen (1.25 mM of NH₄NO₃). Glycyl-alanine gave about the same growth response as NH₄NO₃. The inocules died rapidly in medium containing leucyl-tyrosine and dipeptides containing methionine and valine were also toxic. There was usually a clear correlation between the growth-retarding or growth-stimulating effect of the dipeptides and the effects of their amino acid components.     

Nitrogen fixation in soybean as influenced by cultivar and Rhizobium strain

Summary The¹⁵N-substratum labeling technique and other indirect methods were used to compare nitrogen (N₂) fixation in soybean varieties grown in the field in Greece and Romania. Significant variation in the amount (Ndfa) and proportion of N derived from fixation (% Ndfa) was found in different varieties. With 20 kg N/ha applied to soil, N₂ fixed ranged from 22 to 236 kg N/ha in Greece and from 17 to 132 kg N/ha in Romania. In general, varieties or treatments with higher dry matter yield supported greater fixation. Also, varieties with high Ndfa had high % Ndfa andvice versa. Breeding N₂-fixing legumes for high yields at low soil N levels therefore appears to be a reasonable strategy for enhancing N₂ fixation. Heavy applications of inorganic N fertilizer severely depressed N₂ fixation in two out of the three varieties used in Romania. One variety, F 74–412, however, derived slightly higher amounts of N₂ from fixation at 100 kg N/ha rate than when fertilized with 20 kg N/ha. In Greece, Chippewa, Williams and Amsoy-71 inoculated with a Nitragin inoculant fixed similar amounts of N₂ at both 20 and 100 kg N/ha fertilizer rates. However, when Chippewa and Williams were inoculated with amother, locally-isolated Rhizobium strain, N₂ fixation was substantially depressed at the higher N rate.     

Nitrogen fixation by Rhizobium in pure cultures.

Strains of Rhizobium trifolii and Rhizobium meliloti were tested for their asymbiotic nitrogen fixation ability. From among ten tested strains two R. trifolii and one R. meliloti expressed nitrogenase activity within the range of 1.3--9.3 nM C2H4/h/mg protein. Asymbiotic nitrogen fixation was affected by the composition of the medium.     

Dipeptides and Amino Acids as Nitrogen Sources for the Callus of Atropa belladonna

Morphactin ester applied as a bark band to stems of Monterey pine seedlings (Pinus radiata D. Don) at first stimulated and then inhibited the rate of stem elongation over a 20-day period. The inhibition was caused by a direct action of morphactin on the shoot rather than by indirect action on the root system. ¹⁴C labelled morphactin applied to the bark accumulated in the shoot apices and the amount accumulated could be related to the inhibition of growth. More than 70% of labelled material isolated from roots and shoot apices was in the form of a metabolite similar to the carboxylic acid derivative of the morphactin ester. A preparation of ¹⁴C labelled acid derivative applied to the bark was translocated to the shoot apex and to the roots and inhibited stem elongation.     

Amino Acids as Nitrogen Sources for the Growth of Euglena gracilis and Astasia longa

SYNOPSIS. Euglena gracilis (bacillaris variety, strain SM-L1, streptomycin-bleached) used the following amino adds (10⁻³ M) as sole nitrogen source for growth on a defined medium: glycine, alanine, valine, leucine, isoleucine, serine, threonine, and glutamic acid. Aspartic acid was used at 10⁻² M. Glutamine and asparagine were used at 10⁻³ M and were better N sources than their parent dicarboxylic amino acids. Not used as sole N source for growth were phenylalanine, tyrosine, tryptophan, cysteine, cystine, methionine, proline, hydroxyproline, histidine, arginine, lysine, and taurine. Astasia longa (Jahn strain) was more restricted than Euglena and used only asparagine and glutamine as N sources for growth.     

Sources of Nitrogen for Spruce Tissue Cultures

The major heat-labile organic ingredient of the nutrient previously developed in this laboratory for spruce cells is glutamine. This cannot be fully replaced by any single stable organic substance or group of substances tested. Its replacement by inorganic ammonium salts results in no significant reduction in growth rate in terms of wet weights but produces qualitative defects which make such sutistitutions undesirable. Of special interest is the production of reticulated xylem cells when cultures are grown in the light on NH₄ supplements.     

Nitrogen Metabolism of Lemna minor. I. Growth, Nitrogen Sources and Amino Acid Inhibition

Lemna minor grown in sterile culture on a minerals-sucrose medium can utilize as nitrogen source, in order of increasing growth rate: ammonia, nitrate, a mixture of glutamic and aspartic acids plus arginine, or a balanced mixture of amino acids (hydrolyzed casein). Maximum growth is found with nitrate plus hydrolyzed casein.Many synthetic mixtures of amino acids are unable to support growth. Many single amino acids are inhibitory, and when added (at 2 mm or less) to cultures, growing in the presence of nitrate, cause a decrease in growth rate or even death of the plants (e.g. with alanine, valine, methionine or leucine). Some of these inhibitory effects are also found when the amino acid is added to cultures growing on ammonia or hydrolyzed casein. Arginine was the only amino acid of those tested which gave a marked stimulation of growth when added to cultures growing with inorganic nitrogen.The rapid rate of growth, sterile nature of tissue, decreased biological variation of samples containing many plants and ability to utilize different culture media make this an attractive organism for studies on higher plant metabolism.     

Postmortem Blood Sugar and Blood Urea Nitrogen Determinations

Glucose and urea nitrogen determinations were made on blood and cerebrospinal fluid samples collected during 160 postmortem examinations in order to determine the usefulness of such tests in diagnosing diabetes and uremia at the time of autopsy. The results indicated that: (1) Blood is unsuitable for postmortem glucose determination, and no postmortem normal can be established. (2) Cerebrospinal fluid gave more uniform but very low glucose values. (3) Diabetics as a group had very high postmortem glucose levels but showed a marked overlap with non-diabetics. (4) Infants less than 3 months of age showed high postmortem glucose values. (5) Postmortem blood urea nitrogen and cerebrospinal fluid urea nitrogen levels were within normal limits in previously healthy persons who died suddenly from accidental causes. (6) Hospital autopsy cases had high urea nitrogen levels. (7) Postmortem urea nitrogen levels higher than 100 mg.% were indicative of uremia.     

Heme compounds as iron sources for nonpathogenic Rhizobium bacteria.

Many animal-pathogenic bacteria can use heme compounds as iron sources. Like these microorganisms, rhizobium strains interact with host organisms where heme compounds are available. Results presented in this paper indicate that the use of hemoglobin as an iron source is not restricted to animal-pathogenic microorganisms. We also demonstrate that heme, hemoglobin, and leghemoglobin can act as iron sources under iron-depleted conditions for Rhizobium meliloti 242. Analysis of iron acquisition mutant strains indicates that siderophore-, heme-, hemoglobin-, and leghemoglobin-mediated iron transport systems expressed by R. meliloti 242 share at least one component.     

Nitrogen fixation (nif) genes and large plasmids of Rhizobium japonicum.

The location of structural nitrogen-fixation genes was determined for the slow- and fast-growing types of Rhizobium japonicum. Slow-growing R. japonicum strains do not harbor structural nif genes, homologous to nifD and nifH, on large plasmids (100 to 200 megadaltons). In contrast, all fast-growing R. japonicum strains, except PRC194, contain structural nif genes on large plasmids.     

Distinguishing between Nitrification and Denitrification as Sources of Gaseous Nitrogen Production in Soil

The source of N₂O produced in soil is often uncertain because denitrification and nitrification can occur simultaneously in the same soil aggregate. A technique which exploits the differential sensitivity of these processes to C₂H₂ inhibition is proposed for distinguishing among gaseous N losses from soils. Denitrification N₂O was estimated from 24-h laboratory incubations in which nitrification was inhibited by 10-Pa C₂H₂. Nitrification N₂O was estimated from the difference between N₂O production under no C₂H₂ and that determined for denitrification. Denitrification N₂ was estimated from the difference between N₂O production under 10-kPa C₂H₂ and that under 10 Pa. Laboratory estimates of N₂O production were significantly correlated with in situ N₂O diffusion measurements made during a 10-month period in two forested watersheds. Nitrous oxide production from nitrification was most important on well-drained sites of a disturbed watershed where ambient NO₃⁻ was high. In contrast, denitrification N₂O was most important on poorly drained sites near the stream of the same watershed. Distinction between N₂O production from nitrification and denitrification was corroborated by correlations between denitrification N₂O and water-filled pore space and between nitrification N₂O and ambient NO₃⁻. This technique permits qualitative study of environmental parameters that regulate gaseous N losses via denitrification and nitrification.     

Free-Living Rhizobium Strain Able To Grow on N2 as the Sole Nitrogen Source

A Rhizobium strain isolated from stem nodules of the legume Sesbania rostrata was shown to grow on atmospheric nitrogen (N₂) as the sole nitrogen source. Non-N₂-fixing mutants isolated directly on agar plates formed nodules that did not fix N₂ when inoculated into the host plant.