Nitrogen nutrition inAtriplex hastata L.

Summary The investigation was carried out in order to ascertain whyAtriplex hastata, previously recorded as a nitrophilous plant, can colonise fly ash which has a low nitrogen content.Sinapis arvensis, a weed which does not grow on fly ash, was selected as a control plant. Solution-culture experiments showed that the two species differed in their responses to high and low levels of nitrogen, the growth rate ofS. arvensis being much more severely affected by low levels of 0 to 50 ppm.A. hastata was found to make very efficient use of its nitrogen supply. Both species grew better with nitrate than with ammonium nitrogen. InA. hastata, uptake of nitrogen was not influenced by pH when only nitrate was present. Where both nitrate and ammonium were available, nitrate was absorbed preferentially at low pH and ammonium at high pH. Some properties of fly ash which render it a suitable medium for growth ofA. hastata are discussed.     

Kinetic control of ethanol production by Zymomonas mobilis

Summary Zymomonas mobilis UQM 2716 was grown anaerobically in continuous culture (D = 0.1/h; 30° C) 3nder glucose or nitrogen limitation at pH 6.5 or 4.0. The rates of glucose consumption and ethanol production were lowest during glucose-limited growth at pH 6.5, but increased during growth at pH 4.0 or under nitrogen limitation, and were highest during nitrogen-limited growth at pH 4.0. The uncoupling agent CCCP substantially increased the rate of glucose consumption by glucose-limited cultures at pH 6.5, but had much less effect at pH 4.0. Washed cells also metabolised glucose rapidly, irrespective of the conditions under which the original cultures were grown, and the rates were variably increased by low pH and CCCP. Broken cells exhibited substantial ATPase activity, which was increased by growth at low pH. It was concluded that the fermentation rates of cultures growing under glucose or nitrogen limitation at pH 6.5, or under glucose limitation at pH 4.0, are determined by the rate at which energy is dissipated by various cellular activities (including growth, ATP-dependent proton extrusion for maintenance of the protonmotive force and the intracellular pH, and an essentially constitutive ATP-wasting reaction that only operates in the presence of excess glucose). During growth under nitrogen limitation at pH 4.0 the rate of energy dissipation is sufficiently high for the fermentation rate to be determined by the inherent catalytic activity of the catabolic pathway.Abbreviations CCCP carbonyl cyanide p-trifluoromethoxyphenylhydrazone - qG rate of glucose consumption (g glucose/g dry wt cells/h) - qE rate of ethanol production (g ethanol/g dry wt cells/h) - Y growth yield (g dry wt cells/g glucose) - D dilution rate Offprint requests to: C. W. Jones     

Evaluation of a new fungus <Emphasis Type="Italic">Ceriporia lacerate</Emphasis> strain P2—its ability to decolorize Alizarin Red and Methyl Orange

A new fungus Ceriporia lacerate P2 which belongs to family Polyporaceae was evaluated for its ability to decolorize two different dyes Alizarin Red and Methyl Orange. Different parameters such as incubation time, pH, carbon source, nitrogen source and carbon/nitrogen regimes were used to find out the optimum medium for Ceriporia lacerate P2 on its ability of decolorization. The results show that the fungus had different ability to decolorize the two tested dyes. For Alizarin Red, the most suitable medium was at pH 3 and the best carbon and nitrogen source were sucrose and ammonium nitrate. While for Methyl Orange, the optimum medium was at pH 7–9 and the best carbon and nitrogen source were sucrose and urea.     

Ecology of Dictyosphaerium pulchellum Wood (Chlorophyta, Chlorococcales) in a shallow, acid, forest lake

This study relates to the ecology of Dictyosphaerium pulchellum Wood in Delamere Lake in Cheshire, UK. Dictyosphaerium pulchellum is a cosmopolitan, green colonial phytoplankton species that occasionally forms dense, monospecific populations in lakes. Delamere Lake is a small, shallow, acid lake (mean pH, 4.5) with very high phytoplankton biomass (annual mean chlorophyll a, 290 μg l⁻¹) and devoid of any significant cladoceran population, the efficient grazers of phytoplankton. A predominantly unicellular form of D. pulchellum was the dominant species in Lake Delamere, and it comprised on average ca. 80% (maximum >99%) of the lake phytoplankton biovolume. Laboratory and lake experiments were conducted on this species showed that its pH tolerance varied between 2.4 and 10.7, and its optimum tolerance range between 3.3 and 8.5 depending on other environmental variables. Low pH was not responsible for the unicellular habit of this alga, but a very high nutrient regime could be an important factor. Bioassays revealed that in Delamere Lake this species was limited by nitrogen, but nitrogen did not hamper high growth in the lake. Dictyosphaerium pulchellum can persist at low light levels, tolerate CO₂-deficiency and can grow in polyhumic water with water colour around 300 mg Pt l⁻¹, but probably not in darker waters. The dominance of D. pulchellum in Delamere Lake is apparently due to a combination of several factors: its ability to tolerate both low pH and high turbidity, exploit high nutrient conditions, absence of effective grazing pressure by zooplankton and being a superior competitor.     

Medium pH and nitrate concentration effects on accumulation of triacylglycerol in two members of the chlorophyta

Algal-derived biodiesel is of particular interest because of several factors including: the potential for a near-carbon-neutral life cycle, the prospective ability for algae to capture carbon dioxide generated from coal, and algae’s high per acre yield potential. Our group and others have shown that in nitrogen limitation, and for a single species of Chlorella, a rise in culture medium pH yields triacylglycerol (TAG) accumulation. To solidify and expand on these triggers, the influence and interaction of pH and nitrogen concentration on lipid production was further investigated on Chlorophyceae Scenedesmus sp. and Coelastrella sp. Growth was monitored optically and TAG accumulation was monitored by Nile red fluorescence and confirmed by gas chromatography. Both organisms grew in all treatments and TAG accumulation was observed by two distinct conditions: high pH and nitrogen limitation. The Scenedesmus sp. was shown to grow and produce lipids to a larger degree in alkaliphilic conditions (pH >9) and was used to further investigate the interplay between TAG accumulation from high pH and/or nitrate depletion. Results given here indicate that TAG accumulation per cell, monitored by Nile red fluorescence, correlates with pH at the time of nitrate depletion.     

Evidence for two transport systems for nitrate in the acidophilic thermophilic alga Cyanidium caldarium

In the unicellular non-vacuolate red alga Cyanidium caldarium nitrate uptake occurs through two specific permease systems which, on the basis of kinetic constants can be defined as low affinity system and high affinity system. The high affinity system is saturated at very low nitrate concentrations (<1 M), whereas the low affinity system is saturated only at high nitrate concentrations (K _m=0.45±0.10 mM). The low affinity system is present in cells growing under conditions of nitrogen limitation as well as in cells growing in excess nitrate. In contrast, the high affinity system is present only in cells growing under conditions of nitrogen limitation. The high affinity system works only at acid pH and is inactive at neutral pH. The low affinity system is active both at acid and at neutral pH.     

Studies on the production of extracellular protease by Alcaligenes faecalis

Alcaligenes faecalis produced extracellular protease when incubated in media containing protein substrates. Enzyme production was found to be influenced by various culture conditions. Enzyme production was growth-associated, expressed linearity with growth and reached a maximum at the end of the growth phase. Carbohydrates and inorganic nitrogen sources could not be utilized by the bacterium for its growth, and organic nitrogen appeared to be a primary determinant in protease production. Enzyme production reached its maximum level of 171.2 U/ml when the culture was incubated at 30 °C at pH 8.0. Ca²⁺ and Mg²⁺ enhanced the enzyme production. The crude enzyme powder was stable at high alkaline pH and stable upto 6 months at the storage temperature of 0–4 °C. This revised version was published online in July 2006 with corrections to the Cover Date.     

2′‐Deoxymugineic acid promotes growth of rice (Oryza sativa L.) by orchestrating iron and nitrate uptake processes under high pH conditions

Poaceae plants release 2′‐deoxymugineic acid (DMA) and related phytosiderophores to chelate iron (Fe), which often exists as insoluble Fe(III) in the rhizosphere, especially under high pH conditions. Although the molecular mechanisms behind the biosynthesis and secretion of DMA have been studied extensively, little information is known about whether DMA has biological roles other than chelating Fe in vivo. Here, we demonstrate that hydroponic cultures of rice (Oryza sativa) seedlings show almost complete restoration in shoot height and soil‐plant analysis development (SPAD) values after treatment with 3–30 μm DMA at high pH (pH 8.0), compared with untreated control seedlings at normal pH (pH 5.8). These changes were accompanied by selective accumulation of Fe over other metals. While this enhanced growth was evident under high pH conditions, DMA application also enhanced seedling growth under normal pH conditions in which Fe was fairly accessible. Microarray and qRT‐PCR analyses revealed that exogenous DMA application attenuated the increased expression levels of various genes related to Fe transport and accumulation. Surprisingly, despite the preferential utilization of ammonium over nitrate as a nitrogen source by rice, DMA application also increased nitrate reductase activity and the expression of genes encoding high‐affinity nitrate transporters and nitrate reductases, all of which were otherwise considerably lower under high pH conditions. These data suggest that exogenous DMA not only plays an important role in facilitating the uptake of environmental Fe, but also orchestrates Fe and nitrate assimilation for optimal growth under high pH conditions.     

Purification and characterization of a neutral serine protease with nematicidal activity from <Emphasis Type="Italic">Hirsutella rhossiliensis</Emphasis>

Serine protease plays an important role in fungal infection to invertebrate hosts. An extracellular protease (Hnsp) was detected in liquid culture of Hirsutella rhossiliensis OWVT-1 with nematodes (Panagrellus redivivus) as the unique nitrogen source and purified to homogeneity by ammonium sulphate precipitation, anion exchange chromatography and gel filtration. Its molecular mass was about 32 kDa, and the optimal reaction pH value and temperature were pH 7 and 40°C, respectively. The Hnsp activity was stable at pH 6–8 and decreased radically at 50°C for 10 min. Hnsp was highly sensitive to inhibitor of PMSF and well decomposed the substrate N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide, suggesting that it belonged to the chymotrypsin/subtilisin of serine proteases. The N-terminal amino acid sequence of Hnsp was SVTDQQGADCGLARISHRE, which showed high homology with other serine proteases from nematophagous fungi. Ability to kill nematode and degrade its cuticle in vitro indicated that Hnsp could be involved in the infection of nematode.     

Effects of plastic composite support and pH profiles on pullulan production in a biofilm reactor

Pullulan is a linear homopolysaccharide which is composed of glucose units and often described as α-1, 6-linked maltotriose. The applications of pullulan range from usage as blood plasma substitutes to environmental pollution control agents. In this study, a biofilm reactor with plastic composite support (PCS) was evaluated for pullulan production using Aureobasidium pullulans. In test tube fermentations, PCS with soybean hulls, defatted soy bean flour, yeast extract, dried bovine red blood cells, and mineral salts was selected for biofilm reactor fermentation (due to its high nitrogen content, moderate nitrogen leaching rate, and high biomass attachment). Three pH profiles were later applied to evaluate their effects on pullulan production in a PCS biofilm reactor. The results demonstrated that when a constant pH at 5.0 was applied, the time course of pullulan production was advanced and the concentration of pullulan reached 32.9 g/L after 7-day cultivation, which is 1.8-fold higher than its respective suspension culture. The quality analysis demonstrated that the purity of produced pullulan was 95.8% and its viscosity was 2.4 centipoise. Fourier transform infrared spectroscopy spectra also supported the supposition that the produced exopolysaccharide was mostly pullulan. Overall, this study demonstrated that a biofilm reactor can be successfully implemented to enhance pullulan production and maintain its high purity.     

Fungal remediation of Amarula distillery wastewater

Amarula Cream is an alcoholic beverage derived from the distillation of fermented marula fruit and to date there is no scientific data as to the characteristics of the distillery wastewater generated from its production. The wastewater was found to have a chemical oxygen demand (COD) of 27 g/l, a pH of 3.8, a high concentration of phenolic compounds (866 mg/l) and a high suspended solids content (10.5 g/l), all of which could adversely affect biological treatment. Full-strength wastewater was treated using shake-flask monocultures of four white rot fungi (Trametes pubescens MB 89, Ceriporiopsis subvermispora, Pycnoporus cinnabarinus or Phanerochaete chrysosporium) at pH 5.0 with no additional carbon or nitrogen supplements. Trametes pubescens performed the best with regards to degrading phenolic compounds, COD and colour, while P. cinnabarinus improved the pH to the greatest extent. Laccase synthesis was only detected in the T. pubescens and C. subvermispora cultures. Six wastewater concentrations (100, 80, 60, 40, 20 and 10%) were assessed at pH 4.5 to establish an optimum concentration for remediation and laccase production by T. pubescens. Similar COD removal efficiencies (71–77%) and phenolic removal efficiencies (87–92%) were achieved at all concentrations. The phenolic removal efficiencies improved by approximately 5% compared to the screening experiment at pH 5.0, indicating that the laccase was more efficient at pH 4.5. The pH became more basic as a result of treatment and the colour decreased for samples below 60% wastewater concentration. The maximum laccase activity (1063 ± 26 units/l) was obtained in the 80% wastewater concentration. This study has resulted in the first characterization of Amarula distillery wastewater and showed that it has a high phenolic compound concentration, COD and suspended solids content. It was possible to biologically treat the wastewater at full strength using a number of white-rot fungi just by raising the pH.     

An obligate methylotrophic, methane-oxidizing Methylomicrobium species from a highly alkaline environment

A new, obligately methylotrophic, methane-oxidizing bacterium, strain AMO 1, was isolated from a mixed sample of sediments from five highly alkaline soda lakes (Kenya). Based on its cell ultrastructure and high activity of the hexulose-6-phosphate synthase, the new isolate belongs to the type I methanotrophs. It differed, however, from the known neutrophilic methanotrophs by the ability to grow and oxidize methane at high pH values. The bacterium grew optimally with methane at pH 9–10. The oxidation of methane, methanol, and formaldehyde was optimal at pH 10, and cells were still active up to pH 11. AMO 1 was able to oxidize ammonia to nitrite at high pH. A maximal production of nitrite from ammonia in batch cultures at pH 10 was observed with 10% of CH₄ in the gas phase when nitrate was present as nitrogen source. Washed cells of AMO 1 oxidized ammonia most actively at pH 10–10.5 in the presence of limiting amounts of methanol or CH₄. The bacterium was also capable of oxidizing organic sulfur compounds at high pH. Washed cells grown with methane exhibited high activity of CS₂ oxidation and low, but detectable, levels of DMS and DMDS oxidation. The GC content of AMO 1 was 50.9 mol%. It showed only weak DNA homology with the previously described alkaliphilic methanotroph "Methylobacter alcaliphilus" strain 20 Z and with the neutrophilic species of the genera Methylobacter and Methylomonas. According to the 16S rRNA gene sequence analysis, strain AMO 1 was most closely related to a neutrophilic methanotroph, Methylomicrobium pelagicum (98.2% sequence similarity), within the gamma-Proteobacteria. Received: July 26, 1999 / Accepted: January 4, 2000     

Photosynthetic rate,needle longevity,and nutrient contents in <Emphasis Type="Italic">Picea glehnii</Emphasis> growing on strongly acidic volcanic ash soil in northern Japan

Picea glehnii Masters can grow in strongly acidic volcanic ash soil (pH 3.6) in northern Japan. We compared needle longevity, photosynthetic rate, and concentrations of elements in needles, in mature trees of P. glehnii growing in volcanic ash soil and in brown forest soil (pH 5.4). P. glehnii growing in volcanic ash soil showed suppressed photosynthetic rate and growth by the deficiency in nitrogen compared with its growth in brown forest soil. However, the younger needles of P. glehnii growing in volcanic ash soil maintained a high photosynthetic rate, as a result of large amounts of remobilized nitrogen from senesced needles. Needles of P. glehnii growing in volcanic ash soil did not show deficiencies in Ca, Mg, or K. Moreover, Al was at low levels in the needles, suggesting that P. glehnii was able to avoid Al toxicity by Al exclusion. P. glehnii thus exhibits great ability to adapt to an acidic environment.     

Nitrogen accumulation in Kaolin wastes in Cornwall

Summary Nitrogen accumulation and nitrogen mineralisation rates were measured in a series of waste heaps, produced by the china clay mining industry, which had been reclaimed at different times with a sward ofAgrostis tenuis, Festuca rubra, andTrifolium repens. The best swards tended to have high ammonification rates and rapid N turnover (which is represented by a nitrogen turnover index) —nitrification rates or nitrogen accumulation were not such good predictors of sward quality. Ammonification increased with pH and with organic nitrogen accumulation whereas N turnover was not related to these factors. Nitrification levels were generally low and it was concluded that nitrification was not important to sward health. Organic nitrogen increased with age in all swards, ammonification in certain types only and nitrification not at all. Levels of all are well short of those in adjacent grazing land. Rates of turnover had however a tendency to decline towards those in the grazings owing probably to the build up of resistant humus. The proportion of the total nitrogen which is in the biomass (30%) is also higher than in adjacent grazings (6%). Rapid nitrogen cycling is thus needed to maintain productivity and greenness, and the disadvantages of this are discussed. The adequacy of nitrogen cycle development to date is considered, and possible future strategies outlined.     

Cyanuric acid as nitrogen source for micro-organisms

Summary Several fungi, among which species of Penicillium and Hormodendrum (?), were found to utilize cyanuric acid as a nitrogen source, assimilating 66 to 75% of its nitrogen in cell substance. The growth with cyanurate was essentially as good as with ammonia and urea nitrogen. The optimum reaction for growth was around pH 5 to 6, but pH 7 to 8 was suboptimal. No decrease in growth was seen at a cyanurate concentration of 9.3 mM. The triazine ring as such is thus not biologically very stable, and the strong persistance of the triazine herbicides simazine and atrazine must be ascribed to the external substituents and not to the central ring.     

Production of peroxidase by Inonotus weirii

Summary The white rot fungus Inonotus weirii produced an extracellular peroxidase which was excreted in association with cell growth and in the absence of an inducer. Production of peroxidase was greatly influenced by the carbon and nitrogen source. The highest activities were obtained on glucose-and xylose-based media containing a combination of ammonium nitrate, yeast extract and distiller's spent grain as nitrogen source. The enzyme produced had a molecular weight of 42000, was stable in the pH range 3–8 at room temperature and had optimal activity at pH 3. The fungus Inonotus weirii could be a potential producer of peroxidase for industrial applications in spite of its rather slow production rate.     

Amino acids as a nitrogen source for Chlorella symbiotic with green hydra

Supply of amino acids may be important in controlling cell division of Chlorella symbiotic with green hydra. Freshly isolated symbionts display characteristics of N-limited algae, and low pH in perialgal vacuoles and high levels of host glutamine synthetase (GS) limit uptake of ammonium. Movement of tritiated amino acids from host to algal pools suggests that symbiotic algae utilize amino acids derived from host digestion of prey. Amounts are significant in relation to host and algal amino acids pools. During host starvation, glutamine produced by host GS may be important as a nitrogen supply to the algae, which take up this amino acid at high rates at low pH.     

The effects of lime-pelleting and lime-superphosphate fertilizer on the growth of three annual legumes in an acid sandy soil

Summary Three pioneer pasture legume species,Trifolium subterraneum, Trifolium glomeratum andOrnithopus compressus, were grown in a sandy soil of pH 5.0. The growth and nodulation of each species was examined in the presence and absence of lime pelleting and with superphosphate and lime-superphosphate fertilizer. In each species tested, plant yield, nodule number and nitrogen recovery as maximal where lime pelleting and lime-super were applied together. Plant calcium indicated that the response to lime was due to changes in soil pH rather than to a calcium response. T. glomeratum was the species most responsive to lime application but its growth was at best only half that ofT. subterraneum andO. compressus, which produced equally under both favourable (pH 5.9) and unfavourable (pH 4.4) conditions created by fertilizer applications.     

Nodulation and nitrogen fixation in extreme environments

Biological nitrogen fixation is a phenomenon occurring in all known ecosystems. Symbiotic nitrogen fixation is dependent on the host plant genotype, theRhizobium strain, and the interaction of these symbionts with the pedoclimatic factors and the environmental conditions. Extremes of pH affect nodulation by reducing the colonization of soil and the legume rhizosphere by rhizobia. Highly acidic soils (pH<4.0) frequently have low levels of phosphorus, calcium, and molybdenum and high concentrations of aluminium and manganese which are often toxic for both partners; nodulation is more affected than host-plant growth and nitrogen fixation. Highly alkaline soils (pH>8.0) tend to be high in sodium chloride, bicarbonate, and borate, and are often associated with high salinity which reduce nitrogen fixation. Nodulation and N-fixation are observed under a wide range of temperatures with optima between 20–30°C. Elevated temperatures may delay nodule initiation and development, and interfere with nodule structure and functioning in temperate Iegumes, whereas in tropical legumes nitrogen fixation efficiency is mainly affected. Furthermore, temperature changes affect the competitive ability ofRhizobium strains. Low temperatures reduce nodule formation and nitrogen fixation in temperate legumes; however, in the extreme environment of the high arctic, native legumes can nodulate and fix nitrogen at rates comparable to those observed with legumes in temperate climates, indicating that both the plants and their rhizobia have successfully adapted to arctic conditions. In addition to low temperatures, arctic legumes are exposed to a short growing season, a long photoperiod, low precipitation and low soil nitrogen levels. In this review, we present results on a number of structural and physiological characteristics which allow arctic legumes to function in extreme environments.     

The effects of pH and aluminium toxicity on the growth and symbiotic development of cowpeas (Vigna unguiculata (L.) Walp)

Summary The interaction of pH (4 or 6), aluminium (0 or 16 ppm at pH 4) and N source (symbiotic or combined) on the growth and nutrient status of cowpea (Vigna unguiculata) was studied in a glasshouse experiment.Low pH significantly decreased the growth of the plants dependent on symbiotic nitrogen fixation but at pH 4 the addition of 16 ppm Al further depressed growth in both nitrogen regimes. Al-ions appear to exert their effect primarily on the root system, as shown by the reduction in total length and fresh weight. The symbiotic development of the plants was affected by low pH but more markedly by the Al treatment.Shoot nitrogen concentrations were reduced from ca. 2.6% at pH 6 to 1.8% and 0.9% at pH 4 without and with aluminium respectively. Calcium concentration was decreased by low pH and further by Al in both nitrogen regimes.In all Al-treated plants, the aluminium was mainly accumulated in the roots and was associated with an increase in their phosphorus concentration.