Uptake and transport of N,P and K in tomato supplied with nettle water and nutrient solution

Water extract of stinging nettle (Urtica dioica) has a growth stimulating effect on plants. This investigation elucidated effects of nettle water on uptake and transport of N, P and K. Tomato plants (Solanum lycopersicum L. cv. Dansk export) were grown in sand culture 6–8 weeks. Plants were supplied with nettle water and nutrient solution was used as a control medium. Uptake and transport of N, P and K⁺ were determined with isotopes (¹⁵N,³²P and⁸⁶Rb⁺ as a tracer for K⁺) and ion-selective electrodes and in exudation experiments. A 15% higher uptake of nitrogen (¹⁵N assay) was found after nettle water treatment compared with the nutrient solution control. The total amount of nitrogen was also higher in plants cultivated with nettle water. Transport of inorganic and organic nitrogen, measured in exudation experiments, was more than 50% higher for plants supplied with nettle water compared with plants supplied with nutrient solution. In contrast, nettle water had no effect on uptake, transport or total amount of phosphorus and potassium in the plants. Experiments in hydroculture showed that nettle water had a strong pH-elevating effect. Uptake of NH ₄ ⁺ was strongly stimulated by nettle water compared with nutrient solution. By holding pH at a constant level during the uptake period for 6 h, the uptake of NH ₄ ⁺ from nettle water was significantly lower when no adjustment of pH was made. Consequently a good deal of the NH ₄ ⁺ uptake enhancement by nettle water could be explained by pH-stimulation. Assays with the uncoupler/inhibitor 2,4-dinitrophenol (DNP) and dichlorophenyl-dimethyl-urea (DCMU) showed that uptake of nitrogen from nettle water was less metabolically-linked than uptake from a corresponding nutrient solution. All together, nettle water seems to stimulate the uptake of nitrogen, but not phosphorus or potassium.     

Strong Indirect Effects of a Submersed Aquatic Macrophyte, Vallisneria americana, on Bacterioplankton Densities in a Mesotrophic Lake

Phytoplankton and allochthonous matter are important sources of dissolved organic carbon (DOC) for planktonic bacteria in aquatic ecosystems. But in small temperate lakes, aquatic macrophytes may also be an important source of DOC, as well as a source or sink for inorganic nutrients. We conducted micro- and mesocosm studies to investigate the possible effects of an actively growing macrophyte, Vallisneria americana, on bacterial growth and water chemistry in mesotrophic Calder Lake. A first microcosm (1 L) study conducted under high ambient NH ₄ ⁺ levels (NH ₄ ⁺ 10 µM) demonstrated that macrophytes had a positive effect on bacterial densities through release of DOC and P. A second microcosm experiment, conducted under NH ₄ ⁺ -depleted conditions (NH ₄ ⁺ < 10 µM), examined interactive effects of macrophytes and their sediments on bacterial growth and water chemistry. Non-rooted macrophytes had negative effects on bacterial numbers, while rooted macrophytes had no significant effects, despite significant increases in DOC and P. A 70-L mesocosm experiment manipulated macrophytes, as well as N and P supply under surplus NH ₄ ⁺ conditions (NH ₄ ⁺ 10 µM), and measured effects on bacterial growth, Chl a concentrations, and water chemistry. Bacterial growth and Chl a concentrations declined with macrophyte additions, while bacterial densities increased with P addition (with or without N). Results suggest that the submersed macrophyte Vallisneria exerts a strong but indirect effect on bacteria by modifying nutrient conditions and/or suppressing phytoplankton. Effects of living macrophytes differed with ambient nutrient conditions: under NH ₄ ⁺ -surplus conditions, submersed macrophytes stimulated bacterioplankton through release of DOC or P, but in NH ₄ ⁺ -depleted conditions, the influence of Vallisneria was negative or neutral. Effects of living macrophytes on planktonic bacteria were apparently mediated by the macrophytes use and/or release of nutrients, as well as through possible effects on phytoplankton production.     

Nitrate or ammonium nutrition in french bean

Summary Bean Plants were grown in a greenhouse in sand irrigated with nutrient solutions containing either 2 mM NO ₃ ^– or 2 mM NH ₄ ⁺ . After 45 days fresh weight of NH ₄ ⁺ plants was half that of NO ₃ ^– plants. Cation concentration in NH ₄ ⁺ plants was 30% less than in NO ₃ ^– plants. Amino acids (SER, ASN, GLN) accummulated 3 to 10 times more in NH ₄ ⁺ plants. The concentration of organic acids (malic, malonic, citric) was 10 to 30 times higher in NO ₃ ^– plants. The ATP-costings for the synthesis of amino acids and organic acids in NH ₄ ⁺ plants was half that of NO ₃ ^– ones: therefore it could not account for the reduction of growth in the ammonium-fed plants.     

Interspecies differences in the preference of ammonium and nitrate in vascular plants

Three solution experiments were performed to test the importance of NH ₄ ⁺ versus NO ₃ ^- +NH ₄ ⁺ to growth of 23 wild-forest and open-land species, using field-relevant soil solution concentrations at pH 4.5. At N concentrations of 1–200 M growth increased with increasing N supply in Carex pilulifera, Deschampsia flexuosa, Elymus caninus and Bromus benekenii. Geum urbanum was the most N demanding species and had little growth below 200 M. The preference for NH ₄ ⁺ or NO ₃ ^- +NH ₄ ⁺ was tested also at pH 4.0; no antagonism was found between NH ₄ ⁺ and H⁺, as indicated by similar relative growth in both of the N treatments at both pH levels. Growth in solution with NH ₄ ⁺ relative to NO ₃ ^- +NH ₄ ⁺ , 200 M, was negatively related to the mean pH of the field occurrence of the species tested; acid-tolerant species grew equally well with only NH ₄ ⁺ as with NO ₃ ^- +NH ₄ ⁺ (Oxalis acetosella, Carex pilulifera, Festuca gigantea, Poa nemoralis, Deschampsia flexuosa, Stellaria holostea, Rumex acetosella), while species of less acid soils were favoured by NO ₃ ^- +NH ₄ ⁺ (Urtica dioica, Ficaria verna, Melandrium rubrum, Aegopodium podagraria, Geum urbanum, Bromus benekenii, Sanguisorba minor, Melica ciliata, Silene rupestris, Viscaria vulgaris, Plantago lanceolata). Intermediate species were Convallaria majalis, Elymus caninus, Hordelymus europaeus and Milium effusum. No antagonism between NH ₄ ⁺ and Ca²⁺, Mg²⁺ and K⁺ was indicated by the total uptake of the elements during the experiment.     

Acid-base regulation during ammonium assimilation in Hydrodictyon africanum

Abstract The acid-base balance during ammonium (used to mean NH ₄⁺ and/or NH₃) assimilation in Hydrodictyon africanum has been measured on cells growing with about 1 mol m^?3 ammonium at an external pH of about 6.5. Measurements made included (1) ash alkalinity (corrected for intracellular ammonium) which yields net organic negative charge, (2) the accumulation of organic N in the cells and (3) the change in extracellular H⁺ (from the pH change and the buffer capacity). These measurements showed that some 0.25 excess organic negative charge (half in the cell wall, half inside the plasmalemma) accumulates per organic N synthesized, while some 1.25H⁺ accumulate in the medium per organic N synthesized. Granted a permeability (PNH₃) of some 10^?3 cm s^?1, and a finite [NH₃] in the cytoplasm of these N-assimilating cells it is likely that most of the ammonium entering these growing cells is as NH ₄⁺. This means that most of the H ⁺ appearing in the medium must have originated from inside the cell and have been subjected to active efflux at the plasmalemma: H⁺ accumulates in the medium equivalent to any NH₃ entry by requilibration from exogenous NH ₄⁺. The cell composition (net organic negative charge, organic N content) is very similar in these ammonium-grown cells to that of NO₃⁺grown cells, suggesting that there is no action of a ‘biochemical pH stat’ during longterm assimilation of NO₃⁺in H. africanum. Short-term experiments were carried out at an external pH of 7.2 in which ammonium at various concentrations were supplied to NO₃⁺-grown cells. There was in all cases a rapid influx followed by a slower uptake; at least at the lower concentrations (less than 100 μmol dm^?3) the net influx was all attributable to NH₄⁺influx via a uniporter, probably partly short-circuited by a passive NH₃ efflux due to intrinsic membrane permeability to NH₃. The net ammonium influx was in all cases associated with H⁺ accumulation in the medium. (1.3-1.7 H ⁺ per ammonium taken up); as in the growth experiments, most of the ammonium taken up was assimilated. Determinations of cytoplasmic pH showed either no effect on, or a slight decrease in, pH during ammonium assimilation; the changes that occurred were in the direction expected for actuating a ‘pH-regulating’ change in H⁺ fluxes.     

Evidence for an ammonium transport system in free-living and symbiotic cyanobacteria

The free-living cyanobacterium Anabaena variabilis showed a biphasic pattern of ¹⁴CH₃NH ₃ ⁺ uptake. Initial accumulation (up to 60 s) was independent of CH₃NH ₃ ⁺ metabolism, but long-term uptake was dependent on its metabolism via glutamine synthetase (GS). The CH₃NH ₃ ⁺ was converted into methylglutamine which was not further metabolised. The addition of l-methionine-dl-sulphoximine (MSX), to inhibit GS, inhibited CH₃NH ₃ ⁺ metabolism, but did not affect the CH₃NH ₃ ⁺ transport system.NH ₄ ⁺ , when added after the addition of ¹⁴CH₃NH ₃ ⁺ , caused the efflux of free CH₃NH ₃ ⁺ ; when added before ¹⁴CH₃NH ₃ ⁺ , NH ₄ ⁺ inhibited its uptake indicating that both NH ₄ ⁺ and CH₃NH ₃ ⁺ share a common transport system. Carbonylcyanide m-chlorophenylhydrazone and triphenyl-methylphosphonium both inhibited CH₃NH ₃ ⁺ accumulation indicating that the transport system was -dependent. At pH 7 and at an external CH₃NH ₃ ⁺ concentration of 30 mol dm^-3, A. variabilis showed a 40-fold intracellular accumulation of CH₃NH ₃ ⁺ (internal concentration 1.4 mmol dm^-3). Packets of the symbiotic cyanobacterium Anabaena azollae, directly isolated from the water fern Azolla caroliniana, also showed a -dependent NH ₄ ⁺ transport system suggesting that the reduced inhibitory effect of NH ₄ ⁺ on nitrogenase cannot be attributed to the absence of an NH ₄ ⁺ transport system but is probably related to the reduced GS activity of the cyanobiont.Abbreviations CCCP carbonylcyanide m-chlorophenylhydrazone - GS glutamine synthetase - HEPES 4-(2-hydroxyethyl)-1-piperazine ethanesulphonic acid - MSX l-methionine-dl-sulphoximine - membrane potential - pH transmembrane pH difference - TPMP⁺ triphenylmethylphosphonium     

Ureolytic and nitrifying bacterial heterotrophs from acid soils

Nitrification following ureolysis in soil samples from tea growing soils (pH 4.5–5.5) was found to be chiefly due to the activity of heterotrophic bacteria belonging to generaBacillus, Arthrobacter, Sporosarcina, Micrococcus, Clostridium, Pseudomonas andProteus. A correlation between the intensity of ureolytic activity of organisms in a given soil sample and the yield levels of tea was observed. In culture media the increase in the quantity of NH ₄ ⁺ -N indicating ureolysis was not accompanied by proportional increase in biomass. Ureolysis and nitrification in sterile soil sample inoculated with the isolates improved through amendment of organic carbon to the soil.     

The uptake and metabolism of methylamine by N2-fixing cyanobacteria

The cyanobacteria Anabaena variabilis and Nostoc CAN showed a biphasic pattern of ¹⁴CH₃NH ₃ ⁺ uptake at external pH values of 7.0 and 9.0. The initial phase of uptake, which was independent of metabolism of ¹⁴CH₃NH ₃ ⁺ , was attributed to uptake via a CH₃NH ₃ ⁺ (NH ₄ ⁺ ) transport system at pH 7.0 and probably to passive diffusion of uncharged CH₃NH₂ and trapping by protonation at pH 9.0. The second slower phase of uptake was attributed to metabolism of CH₃NH ₃ ⁺ via glutamine synthetase to form -methylglutamine which accumulates. Anabaena cylindrica showed an initial rapid uptake at pH 7.0 and pH 9.0 but metabolism of ¹⁴CH₃NH ₃ ⁺ was undetectable at pH 7.0 and was barely detectable at pH 9.0. Pretreatment of A. variabilis with l-methionine-d,l-sulphoximine to inactivate glutamine synthetase, inhibited the second phase of ¹⁴CH₃NH ₃ ⁺ uptake at both pH 7.0 and pH 9.0 and the accumulation of -methylglutamine but had no effect on the first phase of uptake. Following transfer of A. variabilis to darkness the initial phase of ¹⁴CH₃NH ₃ ⁺ uptake at pH 7.0 and 9.0 was unaffected but the subsequent metabolism via glutamine synthetase was inhibited.Abbreviations MSX l-methionine-d,l-sulphoximine - GS glutamine synthetase     

氨基酸添加对亚热带森林红壤氮素转化的影响

为探究氨基酸氮形态对亚热带土壤氮素含量及转化的影响,选择建瓯市万木林保护区的山地红壤为对象,采用室内培养实验法,通过设计60%和90%WHC两种土壤含水量并添加不同性质氨基酸,测定了土壤中铵态氮、硝态氮、可溶性有机氮的含量和氧化亚氮的释放量,分析了可溶性有机碳、土壤p H值的大小变化及其与氮素的相互关系。结果表明:与对照处理相比,氨基酸添加显著增加了土壤NH_4~+-N含量并使土壤p H值升高,且在一定程度上解除了高含水量(90%WHC)对NH_4~+-N产生的抑制,其中甲硫氨基酸的效果最为明显。酸性、碱性、中性氨基酸对土壤NO_3~--N含量和N_2O释放影响不显著,但甲硫氨基酸可显著抑制土壤硝化从而导致NH_4~+-N的积累,并在培养前期抑制土壤N_2O产生而在培养后期促进N_2O释放,总体上促进N_2O释放。60%WHC的氨基酸添加处理较90%WHC条件下降低土壤可溶性有机氮的幅度更大。氨基酸对土壤氮素转化的影响与带电性关系较小,而可能与其分解产物密切相关。可见,不同性质氨基酸处理对森林土壤氮素含量及转化存在不同程度的影响,且甲硫氨基酸对土壤氮素转化的影响机理值得深入研究。     

Ammonium uptake in Rhodopseudomonas capsulata

Investigations of the uptake of ammonium (NH ₄ ⁺ ) by Rhodopseudomonas capsulata B100 supported the presence of an NH ₄ ⁺ transport system. Experimentally NH ₄ ⁺ was determined by electrode or indophenol assay and saturation kinetics were observed with two apparent K _m's of 1.7 M and 11.1 M (pH 6.8, 30°) and a V _max at saturation of 50–60 nmol/min·mg protein. The optimum pH and temperature were 7.0 and 33° C, respectively. The Q₁₀ quotient was calculated to be 1.9 at 100 M NH ₄ ⁺ , indicating enzymatic involvement. In contrast to the wild type, B100, excretion of NH ₄ ⁺ , not uptake, was observed in a glutamine auxotroph, R. capsulata G29, which is derepressed for nitrogenase and lacks glutamine synthetase activity. G29R1, a revertant of G29, also took up NH ₄ ⁺ at the same rate as wild type and had fully restored glutamine synthetase activity. Partially restored derivatives, G29R5 and G29R6, grew more slowly than wild type on NH ₄ ⁺ as the nitrogen source, remained derepressed for nitrogenase in the presence of NH ₄ ⁺ , and displayed rates of NH ₄ ⁺ uptake in proportion to their glutamine synthetase activity. Ammonium uptake and glutamine synthetase activity were also restored in R. capsulata G29 exconjugants which had received the plasmid pPS25, containing the R. capsulata glutamine synthetase structural gene. These data suggest that NH ₄ ⁺ transport is tightly coupled to assimilation.Abbreviations used CHES cyclohexylaminoethanesulfonic acid - GS glutamine synthetase - SDS sodium dodecylsulfate     

Nitrogen nutrition influences some biochemical responses to iron deficiency in tolerant and sensitive genotypes of Vitis

The effects of nitrogen source on iron deficiency responses were investigated in two Vitis genotypes, one tolerant to limestone chlorosis Cabernet Sauvignon (Vitis vinifera cv.) and the other susceptible Gloire de Montpellier (Vitis riparia cv.). Plants were grown with or without Fe(III)-EDTA, and with NO₃⁻ alone or a mixture of NO₃⁻ and NH₄⁺. Changes in pH of the nutrient solution and root ferric chelate reductase (FC-R) activity were monitored over one week. We carried out quantitative metabolic profiling (¹H-NMR) and determined the activity of enzymes involved in organic acid metabolism in root tips. In iron free-solutions, with NO₃⁻ as the sole nitrogen source, the typical Fe-deficiency response reactions as acidification of the growth medium and enhanced FC-R activity in the roots were observed only in the tolerant genotype. Under the same nutritional conditions, organic acid accumulation (mainly citrate and malate) was found for both genotypes. In the presence of NH4⁺, the sensitive genotype displayed some decrease in pH of the growth medium and an increase in FC-R activity. For both genotypes, the presence of NH₄⁺ ions decreased significantly the organic acid content of roots. Both Vitis genotypes were able to take up NH₄⁺ from the nutrient solution, regardless of their sensitivity to iron deficiency. The presence of N-NH₄⁺ modified typical Fe stress responses in tolerant and sensitive Vitis genotypes.     

Response of Salvinia spp. to different nitrogen sources: the acid-base regulation approach

This paper shows the importance of acid-base analyses and ¹³C measurements in the evaluation of the responses of Salvinia species' responses to different N sources. It also highlights the importance of these methodologies as potential tools in the study of differences between habitats and nutrient acquisition, particularly N. This study used three different species of Salvinia cultured in the absence of combined N or in the presence of either NO _inf3 ^sup– or NH _inf4 ^sup+ as N sources. The interaction between NO _inf3 ^sup– or NH _inf4 ^sup+ as N source and organic acid metabolism, and the information on diazotrophy from the organic acid measurements, were also examined. Nevertheless, the results presented may not be used per se to assign diazotrophy. Carboxylate (C-A) levels in the different Salvinia species are much lower than the norm for bryophytes and tracheophytes, consistent with previously published work on Azolla. This might be related to the aquatic life form of these plants, since they seem to have no potential to increase the availability of Fe or P by the acidification of their rooting medium (water) that a larger net synthesis of organic acids, with cation-H⁺ exchange, could achieve.     

Evidence that the low-affinity K+ transport system of Rhodobacter capsulatus is a uniporter. The effects of ammonium on the transporter

Comparison of the rate of accumulation of ⁸⁶Rb⁺ by intact cells of Rhodobacter capsulatus during short periods of illumination with the Rb⁺-dependent membrane ionic current measured by electrochromism supports the view that both activities reflect the operation of a low-affinity K⁺ transport system. In experiments performed under similar conditions the ratio of ⁸⁶Rb⁺ uptake to charge uptake was approx. 1.0, suggesting that the transport system operates as a uniporter. The addition of NH _inf4 ^sup+ to a cell suspension led to an increase in membrane ionic current but failed to inhibit the accumulation of ⁸⁶Rb⁺ during illumination. The presence of K⁺ and NH _inf4 ^sup+ inhibited the increase in cellular ATP levels at the onset of illumination. This effect was prevented by Cs⁺. The results are considered within the context of the hypothesis (Golby et al. Eur J Biochem 187: 589–597) that NH _inf4 ^sup+ can be transported by the K⁺ carrier and in the context of an alternative hypothesis that NH _inf4 ^sup+ increases the affinity of the K⁺ transport system for its natural substrate and for Rb⁺.Abbreviations pH pH difference across the cytoplasmic membrane - electrical potential difference across the cytoplasmic membrane     

Photoproduction of ammonium ion from N2 inRhodospirillum rubrum

NH ₄ ⁺ excretion was undetectable in N₂-fixing cultures ofRhodospirillum rubrum (S-1) and nitrogenase activity in these cultures was repressed by the addition of 10 mM NH ₄ ⁺ to the medium. The glutamate analog,l-methionine-dl-sulfoximine (MSX), derepressed N₂ fixation even in the presence of 10 mM extracellular NH ₄ ⁺ . When 10 mg MSX/ml was added to cultures just prior to nitrogenase induction they developed nitrogenase activity (20% of the control activities) and excreted most of their fixed N₂ as NH ₄ ⁺ . Nitrogenase activities and NH ₄ ⁺ production from fixed N₂ were increased considerably when a combined nitrogen source, NH ₄ ⁺ (>40 moles NH ₄ ⁺ /mg cell protein in 6 days) orl-glutamate (>60 moles NH ₄ ⁺ /mg cell protein in 6 days) was added to the cultures together with MSX.Biochemical analysis revealed thatR. rubrum produced glutamine synthetase and glutamate synthase (NADP-dependent) but no detectable NADP-dependent glutamate dehydrogenase. The specific activity of glutamine synthetase was observed to be maximal when nitrogenase activity was also maximal. Nitrogenase and glutamine synthetase activities were repressed by NH ₄ ⁺ as well as by glutamate.The results demonstrate that utilization of solar energy to photoproduce large quantities of NH ₄ ⁺ from N₂ is possible with photosynthetic bacteria by interfering with their regulatory control of N₂ fixation.     

Nitrogen metabolism inRhodopseudomonas globiformis

Rhodopseudomonas globiformis strain 7950 grew with a variety of amino acids, urea, or N₂ as sole nitrogen sources. Cultures grown on N₂ reduced acetylene to ethylene; this activity was absent from cells grown on nonlimiting NH ₄ ⁺ . Glutamate dehydrogenase could not be detected in extracts of cells of strain 7950, although low levels of an alanine dehydrogenase were present. Growth ofR. globiformis on NH ₄ ⁺ was severely inhibited by the glutamate analogue and glutamine synthetase inhibitor, methionine sulfoximine. High levels of glutamine synthetase (as measured in the -glutamyl transferase assay) were observed in cell extracts of strain 7950 regardless of the nitrogen source, although N₂ and amino acid grown cells contained somewhat higher glutamine synthetase contents than cells grown on excess NH ₄ ⁺ . Levels of glutamate synthase inR. globiformis were consistent with that reported from other phototrophic bacteria. Both glutamate synthase and alanine dehydrogenase were linked to NADH as coenzyme. We conclude thatR. globiformis is capable of fixing N₂, and assimilates NH ₄ ⁺ primarily via the glutamine synthetase/glutamate synthase pathway.Abbreviations GS glutamine synthetase - GOGAT Glutamineoxoglutarate aminotransferase - GDH Glutamate dehydrogenase - ADH Alanine dehydrogenase - MSO Methionine sulfoximine     

Ammonium and nitrate uptake rates and rhizosphere pH in non-mycorrhizal roots of Norway spruce [Picea abies (L.) Karst.]

Summary Relationships between root zone temperature, concentrations and uptake rates of NH ₄ ⁺ and NO ₃ ^– were studied in non-mycorrhizal roots of 4-year-old Norway spruce under controlled environmental conditions. Additionally, in a forest stand NH ₄ ⁺ and NO ₃ ^– uptake rates along the root axis and changes in the rhizosphere pH were measured. In the concentration (C_min) range of 100–150 M uptake rates of NH ₄ ⁺ were 3–4 times higher than those of NO ₃ ^– The preference for NH ₄ ⁺ uptake was also reflected in the minimum concentration (C_min) values. Supplying NH₄NO₃, the rate of NO ₃ ^– uptake was very low until the NH ₄ ⁺ concentrations had fallen below about 100 M. The shift from NH ₄ ⁺ to NO ₃ ^– uptake was correlated with a corresponding shift from net H⁺ production to net H⁺ consumption in the external solution. The uptake rates of NH ₄ ⁺ were correlated with equimolar net production of H⁺. With NO ₃ ^– nutrition net consumption of H⁺ was approximately twice as high as uptake rates of NO ₃ ^– In the forest stand the NO ₃ ^– concentration in the soil solution was more than 10 times higher than the NH ₄ ⁺ concentration (<100 M), and the rhizosphere pH of non-mycorrhizal roots considerably higher than the bulk soil pH. The rhizosphere pH increase was particularly evident in apical root zones where the rates of water and NO ₃ ^– uptake and nitrate reductase activity were also higher. The results are summarized in a model of water and nutrient transport to, and uptake by, non-mycorrhizal roots of Norway spruce in a forest stand. Model calculations indicate that delivery to the roots by mass flow may meet most of the plant demand of nitrogen and calcium, and that non-mycorrhizal root tips have the potential to take up most of the delivered nitrate and calcium.     

Replacement of potassium ions by ammonium ions in different micro-organisms grown in potassium-limited chemostat culture

The biomass concentration extant in potassiumlimited cultures of either Klebsiella pneumoniae or Bacillus stearothermophilus (when growing at a fixed temperature and dilution rate in a glucose/ammonium salts medium) increased progressively as the medium pH value was raised step-wise from 7.0 to 8.5. Because the macromolecular composition of the organisms did not vary significantly, this increase in biomass could not be attributed to an accumulation of storage-type polymers but appeared to reflect a pH-dependent decrease in the cells' minimum K⁺ requirement. Significantly, this effect of pH was not eviden with cultures in which no ammonium salts were present and in which either glutamate or nitrate was added as the sole nitrogen source; however, it was again manifest when various concentrations of NH₄Cl were added to the glutamate-containing medium. This suggested a functional replacement of K⁺ by NH ₄ ⁺ , a proposition consistent with the close similarity of the ionic radii of the potassium ion (1.33 Å) and the ammonium ion (1.43 Å). At pH 8.0, and with a medium containing both glutamate (30 mM) and NH₄Cl (100 mM), cultures of B. stearothermophilus would grow without added potassium at a maximum rate of 0.7 h^-1. Under these conditions the cells contained maximally 0.1% (w/w) potassium (derived from contaminating amounts of this element in the medium constituents), a value which should be compared with one of 1.4% (w/w) for cells growing in a potassiumlimited medium containing initially 0.5 mM K⁺. Qualitatively similar findings were made with cultures of K. pneumoniae; and whereas one may not conclude that NH ₄ ⁺ can totally replace K⁺ in the growth of these bacteria, it can clearly do so very extensively.     

Distribution and activity of bacteria in deep granitic groundwaters of southeastern sweden

This study investigated the distribution of bacteria in groundwater from 16 different levels in five boreholes in granite bedrock down to a maximum of 860 m. Enrichment cultures were used to assay the groups of bacteria present. Autoradiographic studies with¹⁴C- or³H-labeled formate, methanol, acetate, lactate, glucose, sodium bicarbonate, leucine, glutamine, thymidine, orN-acetyl-glucosamine were used to obtain information about bacteria active in substrate uptake. The biofilm formation potential was studied in one borehole. The chemical environment in the groundwater was anaerobic with an E_h between −112 and −383 mV, a pH usually around 8, and a temperature range of 10.2 to 20.5°C, depending on the depth. The organic content ranged between <0.5 and 9.5 mg total organic carbon liter⁻¹. Carbon dioxide, hydrogen, hydrogen sulfide, and methane were present in the water. The nitrate, nitrite, and phosphate concentrations were close to, or below, the detection limits, while there were detectable amounts of NH ₄ ⁺ in the range of 4 to 330 μg liter⁻¹. The average total number of bacteria was 2.6×10⁵ bacteria ml⁻¹, as determined with an acridine organge direct-count (AODC) technique. The average number of bacteria that grew on a medium with 1.5 g liter⁻¹ of organic substrate was 7.7×10³ colony-forming units (CFU) ml⁻¹. The majority of these were facultatively anaerobic, gram-negative, nonfermenting heterotrophs. Enrichment cultures indicated the presence of anaerobic bacteria capable of growth on C-1 compounds and hydrogen, presumably methanogenic bacteria. Most probable number assays with sulfate and lactate revealed up to 5.6×10⁴ viable sulfate-reducing bacteria per ml. A biofilm development experiment indicated an active attached microbial population. Active substrate uptake could not be registered with the bulk water populations, except for an uptake of leucine not associated with growth. The bulk water microbial cells in deep groundwater may be inactive cells detached from active biofilms on the rock surface.     

Comparative effects of salt-stress and alkali-stress on the growth,photosynthesis, solute accumulation,and ion balance of barley plants

We compared the effects of salt-stresses (SS, 1: 1 molar ratio of NaCl to Na₂SO₄) and alkali-stresses (AS, 1: 1 molar ratio of NaHCO₃ to Na₂CO₃) on the growth, photosynthesis, solute accumulation, and ion balance of barley seedlings, to elucidate the mechanism of AS (high-pH) damage to plants and the physiological adaptive mechanism of plants to AS. The effects of SS on the water content, root system activity, membrane permeability, and the content of photosynthetic pigments were much less than those of AS. However, AS damaged root function, photosynthetic pigments, and the membrane system, led to the severe reductions in water content, root system activity, content of photosynthetic pigments, and net photosynthetic rate, and a sharp increase in electrolyte leakage rate. Moreover, with salinity higher than 60 mM, Na⁺ content increased slowly under SS and sharply under AS. This indicates that high-pH caused by AS might interfere with control of Na⁺ uptake in roots and increase intracellular Na⁺ to a toxic level, which may be the main cause of some damage emerging under higher AS. Under SS, barley accumulated organic acids, Cl⁻, SO₄ ²⁻, and NO₃ ⁻ to balance the massive influx of cations, the contribution of inorganic ions to ion balance was greater than that of organic acids. However, AS might inhibit absorptions of NO₃ ⁻ and Cl⁻, enhance organic acid synthesis, and SO₄ ²⁻ absorption to maintain intracellular ion balance and stable pH.     

Production of Alkaline Lipase by Corynebacterium paurometabolum, MTCC 6841 Isolated from Lake Naukuchiatal, Uttaranchal State, India

A moderately psychrophilic bacterium Corynebacterium paurometabolum MTCC 6841 (gram positive, short rod type) producing extracellular alkaline lipase was isolated from Lake Naukuchiatal, Uttaranchal, India. The bacterium was able to grow within a broad range of pH (5–10). Soyabean oil and olive oil served as the best carbon sources for lipase production. The bacterium preferred inorganic nitrogenous compounds, NaNO₃ and KNO₃, over organic nitrogenous compound for its growth. Maximum lipase production occurred at 25°C and 8.5 pH. The enzyme activity was found to be maximum at the same values of temperature and pH. The enzyme was reasonably stable in the presence of various organic solvents. No significant effect of Ca⁺, Cu⁺⁺, Fe⁺⁺, Na⁺, K⁺, Mg⁺⁺, Mn⁺, NH4⁺, Co⁺⁺ ions over enzyme activity was detected. Treatment with EDTA reduced the activity to nearly one half.