Uptake and assimilation of nitrogen from solutions containing multiple N sources

We assessed the extent to which plants can acquire amino acids when supplied as single N-sources or when plants have access to a mixture of amino- and inorganic N sources. Because the uptake of different N-sources is temperature-dependent, the effects of temperature on amino-N uptake were also tested. Lolium perenne (perennial rye-grass) was grown hydroponically at 11 °C or 21 °C. Uptake of N was determined using ¹⁵N tracers at the growth temperature from solutions containing either nitrate, ammonium or glycine as single N sources and from a mixture containing all three N-forms. Estimates of the relative importance of amino acids such as glycine to the total N budget of plants will have been underestimated in studies where uptake was determined in single source solutions compared with those from solutions containing a mixture of N-forms. The proportion of total N acquired from the mixed N source as ammonium increased as temperature was reduced. Regarding the uptake and initial metabolism of glycine, uptake was probably the rate limiting step at 11 °C whilst it was the metabolism of glycine to serine at 21 °C. Although ¹⁵N incorporation into the plant amino-N pool was generally in proportion to the abundance of individual amino acids, its incorporation into the glycine pool was sometimes significantly less than predicted.     

Electrochemical identification of microbially mediated hydrogen sulfide oxidation

The process of H₂S oxidation by the phototrophic bacteriaThiocapsa roseopersicina andChlorobium phaeobacteroides, respectively, was monitored using a Pt-glass-Ag⁰, Ag₂S electrode combination without liquid junction. Due to the resulting pe(pH) and pH₂S plottings three steps can be distinguished: oxidation of H₂S to an S(0) state, oxidation of S (0) to SO₄ ^2–, and oxidation of the remaining H₂S directly to SO₄ ^2–. Differences between the investigated bacteria exist with respect to their individual oxidation strategies.Thiocapsa apparently stops oxidizing H₂S at pH₂S 7.5 (e.g. 10^–7.5M H₂S) and shifts to the utilization of the intracellularly stored S (0). In contrastChlorobium utilizes its extracellularly stored sulfur parallel to the extracellular H₂S fraction. The corresponding Pt-sensor responses (pe₇ values) were found to be similar to the corresponding partial redox equilibria (p₇ values) of H₂S oxidation stoichiometries as proposed by Van Niel (1931) and Trüper (1964). It is concluded that the recording of pe enables investigators to understand (and control) in situ redox processes, independent of their thermodynamic equilibration, only bound to changes of electroactivity vs. sensor.     

Cloning in Escherichia coli of genes involved in the synthesis of proline and leucine in Desulfovibrio desulfuricans Norway

Summary A library of Deusulfovibrio desulfuricans Norway genomic DNA was constructed in Escherichia coli with pBR322 as vector and plasmids able to complement the proA and leuB mutations of the host were screened. It was observed that all the plasmids studied were highly unstable, the insert DNA being rapidely lost under non-selective growth conditions. A 2.75 kb DNA fragment of D. desulfuricans Norway was found to complement E. coli ProA, ProB and ProC deficiencies. From the results of restriction analysis and Southern hybridization, it is proposed that the genes involved in proline and leucine biosynthesis are clustered on the chromosome of D. desulfuricans Norway.     

Ammonium and nitrate nutrition inPlantago lanceolata andPlantago major L. ssp.major

P. lanceolata andP. major were grown in culture solutions with nitrate or ammonium as the nitrogen source. Dry matter accumulation in the shoot was faster with nitrate than with ammonium, whilst that of the roots was not affected by the nitrogen source. As a consequence, the shoot-to-root ratio was lower with ammonium than with nitrate. InP. lanceolata, dry matter percentage of shoot and root tissue was lower with nitrate nutrition, suggesting better elongation growth than with ammonium. However, in shoot tissue ofP. major the opposite was found. The rate of root respiration declined with time, and this was almost completely due to a declining activity of the alternative path, which amounted to about 30–60% of total root respiration. Respiration via the cytochrome path was for a part of time slightly increased by ammonium, whereas the activity of the alternative path was strongly enhanced. The concentration of ethanol-soluble carbohydrates (SC) in the roots of both species was higher when nitrate was used, but no difference in the concentration of starch was found. When the plants were transferred from one nitrogen source to the other, many parameters, including the concentration of nitrate and chloride, and the shoot to root ratio, adjusted to the new situation in both species. Grassland Species Research Group, Publication no. 116.     

Micronutrients and nitrogen metabolism

A sand-culture experiment was conducted to study the influence of a deficiency of and an excess of micronutrients on the uptake and assimilation of NH ₄ ⁺ and NO ₃ ⁻ ions by maize. By studying the fate of¹⁵N supplied as¹⁵NH₄NO₃ or NH₄ ¹⁵NO₃, it was demonstrated that in maize plants NH₄−N was absorbed in preference to NO ₃ ⁻ −N. The uptake and distribution of N originating from both NH ₄ ⁺ and NO ₃ ⁻ was considerably modified by deficiency of, or an excess of, micronutrients in the growth medium. The translocation of NH ₄ ⁺ −N from roots to shoots was relatively less than that of NO ₃ ⁻ −N. Deficiency as well as excessive amounts of micronutrients, in the growth medium, substantially reduced the translocation of absorbed N into protein. This effect was more pronounced in the case of N supplied as NO ₃ ⁻ . Amino-N was the predominant non-protein fraction in which N from both NH ₄ ⁺ and NO ₃ ⁻ tended to accumulate. The next important non-protein fractions were NO ₃ ⁻ −N when N was supplied as NO ₃ ⁻ and amide-N when NH ₄ ⁺ was the source. The relative accumulation of¹⁵N into different protein fractions was also a function of imposed micronutrient levels.     

Ionic balance,proton efflux,nitrate reductase activity and growth ofHippophaë rhamnoides L. ssp.rhamnoides as influenced by combined-N nutrition or N2 fixation

Growth of 2-month-old nonnodulatedHippophaë rhamnoides seedlings supplied with combined N was compared with that of nodulated seedlings grown on zero N. Plant growth was significantly better with combined N than with N₂ fixation and, although not statistically significant for individual harvests, tended to be highest in the presence of NH ₄ ⁺ , a mixture of NH ₄ ⁺ and NO ₃ ^? producing the highest yields. Growth was severely reduced when solely dependent on N₂ fixation and, unlike the combined-N plants, shoot to root ratios had only slightly increased after an initial decrease. An apparently insufficient nodule mass (nodule weight ratio <5 per cent) during the greater part of the experimental period is suggested as the main cause of the growth reduction in N₂-fixing plants. Thein vivo nitrate reductase activity (NRA) of NO ₃ ^? dependent plants was almost entirely located in the roots. However, when grown with a combination of NO ₃ ^? and NH ₄ ⁺ , root NRA was decreased by approximately 85 per cent.H. rhamnoides demonstrated in the mixed supply a strong preference for uptake of N as NH ₄ ⁺ , NO ₃ ^? contributing only for approximately 20 per cent to the total N assimilation. Specific rates of N acquisition and ion uptake were generally highest in NO ₃ ^? +NH ₄ ⁺ plants. The generation of organic anions per unit total plant dry weight was approximately 40 per cent less in the NH ₄ ⁺ plants than in the NO ₃ ^? plants. Measured extrusions of H⁺ or OH^? (HCO ₃ ^? ) were generally in good agreement with calculated values on the basis of plant composition, and the acidity generated with N₂ fixation amounted to 0.45–0.55 meq H⁺. (mmol N_org)^?1. Without acidity control and in the presence of NH ₄ ⁺ , specific rates of ion uptake and carboxylate generation were strongly depressed and growth was reduced by 30–35 per cent. Growth of nonnodulatedH. rhamnoides plants ceased at the lower pH limit of 3.1–3.2 and deterioration set in; in the case of N₂-fixing plants the nutrient solution pH stabilized at a value of 3.8–3.9 without any apparent adverse effects upon plant performance. The chemical composition of experimental and field-growing plants is being compared and some comments are made on the nitrogen supply characteristics of their natural sites.     

Growth measurements of terrestrial microbial species by a continuous-flow technique

A continuous nutrient flow system has been developed to measure microbial activity in soil with various concentrations of added substrate. The system consists of a thin soil layer through which substrate was added continuously over periods up to 4.5 days. Substrate utilization was determined by effluent analysis. Respiration was measured manually by injecting a sample into a gas chromatograph or automatically by coupling the growth chamber to a computer-controlled gas sampling valve. This permitted respiratory CO₂ to be measured by the gas chromatograph at intervals selected by the investigator. Software controlling the valve and gas chromatograph not only automated gas phase sampling, but also provided a scan of CO₂ evolution and a preliminary data summary. This included the date and time of sample, peak height, and percent CO₂ in the gas phase. Data for growth on glucose using a microbial population native to a California annual grassland soil demonstrated that the direct cell count and respiratory techniques for biomass estimation give comparable results. This procedure provides the potential for detailed analyses of substrate utilization in studies of the growth and maintenance of soil microorganisms.     

A comparative study of ammonium toxicity at different constatn pH of the nutrient solution

Seedlings of 14 species were grown for 14–28 days on nutrient solution with 6 mmol.l⁻¹ NH₄ as the sole nitrogen source. Solutions acidity was were kept constant at pH 4.0, 5.0, 6.0 and 7.0 by continuous titration with diluted KOH. The following species were used: barley, maize, oats sorghum, yellow and white lupin, pea, soybean, carrot, flax, castor-oil, spinach, sugarbeet and sunflower. Most plant species grew optimally at pH 6.0 with slight reductions at pH 5.0. Growth of many species was severely inhibited at pH 4.0, but this inhibition was not observed with the legume and cereal species. Yield depressions at pH 4.0 relative to pH 6.0 were well correlated with the respective relative decreases of the K concentration in their roots (P<0.002). In the roots of two species (sunflower and flax) total N concentrations were also strongly reduced at pH 4.0. apparently, the interactions between uptake of K, NH₄ and H ions become the prevalent problem at suboptimal pH. At pH 7.0, yields were also considerably decreased, with the exception of the lupines. At this pH, the roots of the growth inhibited plants were characterized by increased levels of total N and free NH₄. It is thought that the binding capacity of the roots for NH₄ is an important factor in preventing NH₄/NH₃ toxicity at supraoptimal pH.     

Microbial communities colonising nutrient-enriched marine sediment

Sediment cores were set up to study microbial colonisation and interactions on marine sand grains under enrichment conditions. Cores were enriched with photosynthetic media in the light and dark (PL, PD) and heterotrophic media in the light and dark (HL, HD), and were incubated for 25 days. Sediment chlorophylls were then measured by acetone extraction, viable heterotrophic bacteria by plate counts, and numbers of cells mm^–2 sand grain surface by s.e.m. Chlorophyll a occurred in all sediments but was highest in the PL sediment. Bacteriochlorophyll a was only observed in the HL sediment. Heterotrophic viable counts were high in the HL and HD sediments. Dense growth of diatoms and blue-green algae, and a marine fungal Thraustochytrid sp. occurred on PL grains. The blue-green alga Schizothrix was often associated with the diatom Amphora on PL grains. Many different bacteria grew on HL and HD grains and some unusual colony and cell morphologies were recorded (Caulobacter, Flexibacter, polymer strands). Characteristic flakey material sometimes occurred in hollows on grains. The results are discussed in relation to microbial communities in low energy sedimentary environments.     

Effect of rainstorms on heterotrophic bacterial activity in a hypertrophic African lake

Hartbeespoort Dam is a hypertrophic man-made lake which is located in the Transvaal Province of South Africa. This region has recently experienced its most severe drought of the century. However, on three occasions in the summer rainy seasons of 1984 and 1985, major rainfalls (> 50 mm) occurred which caused large inflows to the lake. Inflowing river water entered as a density current causing marked silting of the water. Within the epilimnion (0–10 m) prior to these rainfalls there was usually no variation of bacterial numbers with depth, but heterotrophic bacterial activity (glucose uptake) decreased with depth concomitant with primary production. With the increased river inflow bacterial numbers did not increase but bacterial activity at the bottom of the epilimnion (10 m) increased to as high as 2.7 µg C l^–1 h^–1 in January 1985, reversing the depth profile of bacterial activity within the epilimnion. This resulted in decreased glucose concentrations (K_t + S_n) and turnover times. Heterotrophic activity per cell increased by between 2.5 and 5 times. These data demonstrate that storm events are important phenomena causing short-term changes in the metabolic activity of planktonic heterotrophic bacteria in lakes.