Xanthine dehydrogenase expression inNeurospora crassa does not require a functionalnit-2 regulatory gene

Xanthine dehydrogenase (XDH) is the initial enzyme in the purine catabolic pathway ofN. crassa. Secondary nitrogen sources such as purines are metabolized when preferred sources of reduced nitrogen (ammonium or glutamine) are unavailable. XDH synthesis is regulated by glutamine repression and uric acid induction. Thenit-2 locus is believed to encode atrans-acting positive regulator essential for the expression of genes encoding enzymes involved in secondary pathways of nitrogen acquisition, such as XDH and nitrate reductase. However, immunoblot analyses and enzyme assays reveal that XDH protein is synthesized and XDH activity is expressed innit-2 mutants. Nevertheless, XDH responds to nitrogen metabolite repression. The generality thatnit-2 is an obligate control element in nitrogen metabolite repression is questioned. Additionally, mutants defective in XDH activity, namely,xdh-1 and the molybdenum cofactor mutantsnit-1, -7, -8 and -9, are observed to grow on xanthine but not hypoxanthine.This research was supported in part by National Science Foundation Grant DMB 8516203.     

Hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) degradation by <Emphasis Type="Italic">Acetobacterium paludosum</Emphasis>

Substrates and nutrients are often added to contaminated soil or groundwater to enhance bioremediation. Nevertheless, this practice may be counterproductive in some cases where nutrient addition might relieve selective pressure for pollutant biodegradation. Batch experiments with a homoacetogenic pure culture of Acetobacterium paludosum showed that anaerobic RDX degradation is the fastest when auxiliary growth substrates (yeast extract plus fructose) and nitrogen sources (ammonium) are not added. This bacterium degraded RDX faster under autotrophic (H₂-fed) than under heterotrophic conditions, even though heterotrophic growth was faster. The inhibitory effect of ammonium is postulated to be due to the repression of enzymes that initiate RDX degradation by reducing its nitro groups, based on the known fact that ammonia represses nitrate and nitrite reductases. This observation suggests that the absence of easily assimilated nitrogen sources, such as ammonium, enhances RDX degradation. Although specific end products of RDX degradation were not determined, the production of nitrous oxide (N₂O) suggests that A. paludosum cleaved the triazine ring.     

Absorption and Assimilation of Inorganic Nitrogen from Different Sources by Storage Root-Tissue

1. The rate of ammonium and nitrate absorption and assimilationby storage tissues of sweet potato tubers, radish, and carrotroots cultured in aerated dilute solutions (5 mg. eq./1.) ofNH₄Cl, KNO₃ or NH₄NO₂ at 25° C. and the effect of the sourceof nitrogen on the nitrogenous composition of the tissues werestudied.
2. Both ions were absorbed at more or less equal ratesunder theprevailing experimental conditions by cells of thedifferentplant materials used.
3. The rate of nitrate assimilationwas lower than absorption,leading to the accumulation of nitrate-Nin the cells of alltissues examined. Ammonium-N, on the otherhand, was assimilatedalmost as soon as it entered the cellsof sweet potato tubersand carrot roots, and there was onlya small increase in thelevel of arnmonium-N; but in radishroot cells ammonium assimilationlagged very much behind absorption,resulting in a great increasein the level of arnmonium-N.
4. Sweetpotato tubers and carrot tissues were shown to be adaptedtothe utilization of more ammonium than nitrate nitrogen, butradish root tissues are adapted to the utilization of more nitratenitrogen.
5. Ammonium or nitrate-N caused similar increases inthe complexorganic-N level (protein-+rest-N) in radish root,but the rateof synthesis was higher with ammonium in sweetpotato tubersand with nitrate in carrot roots.

Furthermore, with both sources changes in the content of amideand amino-N were negligible in radish tissues of both series.In contrast to radish, carrot tissues showed marked increasesin the two fractions. The increase of these fractions in sweetpotato tuber tissues was much lower. The marked contrast in nitrogenous composition in the threedifferent plant materials fed with ammonium and nitrate saltsseparately indicated that the actual course of protein metabolismis most probably dependent on the source of nitrogen utilized.     

Changes in the Morphology of Roots and Leaves of Carob Seedlings Induced by Nitrogen Source and Atmospheric Carbon Dioxide

Carob seedlings were grown hydroponically for 9 weeks under360 and 800 µl l^-1CO₂. One of two nitrogen sources, nitrateor ammonium, was added to the nutrient medium at concentrationsof 3 mol m^-3. Root systems of the developing plants suppliedwith nitrate compared to those supplied with ammonium were characterizedby:(a)more biomass on the lower part of the root;(b)fewer lateralroots of first and second order;(c)longer roots;(d)higher specificroot length;(e)a smaller root diameter. The morphology of theroot systems of nitrate-fed plants changed in the presence ofelevated carbon dioxide concentrations, resembling, more closely,that of ammonium-fed plants. Total leaf area was higher in ammonium-than in nitrate-fed plants. Nitrate-fed plants had greater totalleaf area in the presence of high carbon dioxide than in normalCO₂, due to an increase in epidermal cell size that led to developmentof larger leaflets with lower stomatal frequency. The observedchanges in the morphology of roots and shoots agreed with theresults observed for total biomass production. Nitrate-fed plantsincreased their biomass production by 100% in the presence ofelevated CO₂compared to 15% in ammonium-fed plants, indicatingthat the response of carob to high CO₂concentrations is verydependent on the nitrogen source. Under elevated CO₂, nitrate-grownplants had a larger content of sucrose in both roots and shoots,while no significant difference was observed in the contentof sucrose in ammonium-grown plants, whether in ambient or enrichedcarbon dioxide. Hence, the differences in soluble carbohydratecontents can, at least partly, account for differences in rootand shoot morphology.Copyright 1997 Annals of Botany Company Ceratonia siliquaL.; carob; ammonium; carbohydrate; carbon dioxide; nitrate; morphology; sucrose     

Role of Polyamines in SO2-Polluted Pea Plants

The effect of SO₂ fumigation on free and bound putrescine andspermidine has been investigated in pea plants grown in nitrate-basedand ammonium-containing nutrient solutions. Both amines increasesignificantly more in response to SO₂ fumigation when 50% ofthe nitrate nitrogen is substituted by ammonium. Amine levelsare also increased in the unfumigated, ammonium-supplied plantsrelative to the exclusively nitrate-supplied ones. Since bothSO₂ pollution and ammonium nutrition increase the H⁺ ion concentrationof the cells and cause a shift in the cation/anion ratio, itis concluded that with both treatments amines are synthesizedto bind these H⁺ ions and to compensate the relative cationdeficit. The importance of this mode of metabolic bufferingis discussed and its effectiveness calculated.     

The Uptake and Flow of C, N and lons between Roots and Shoots in Ricinus communis L.

Seedlings of Ricinus communis L. cultivated in quartz sand weresupplied with a nutrient solution containing either 1 mol m^–3NO^–₃ or 1 mol m^–3 NH⁺₄ as the nitrogen source. Duringthe period between 41 and 51 d after sowing, the flows of N,C and inorganic ions between root and shoot were modelled andexpressed on a fresh weight basis. Plant growth was clearlyinhibited in the presence of NH⁺₄. In the xylem sap the majornitrogenous solutes were nitrate (74%) or glutamine (78%) innitrate or ammonium-fed plants, respectively. The pattern ofamino acids was not markedly influenced by nitrogen nutrition;glutamine was the dominant compound in both cases. NH⁺₄ wasnot transported in significant amounts in both treatments. Inthe phloem, nitrogen was transported almost exclusively in organicform, glutamine being the dominant nitrogenous solute, but theN-source affected the amino acids transported. Uptake of nitrogenand carbon per unit fresh weight was only slightly decreasedby ammonium. The partitioning of nitrogen was independent ofthe form of N-nutrition, although the flow of nitrogen and carbonin the phloem was enhanced in ammonium-fed plants. Cation uptakerates were halved in the presence of ammonium and lower quantitiesof K⁺, Na⁺ and Ca²⁺ but not of Mg²⁺ were transported to theshoot. As NH⁺₄ was balanced by a 30-fold increase in chloride in thesolution, chloride uptake was increased 6-fold under ammoniumnutrition. We concluded that ammonium was predominantly assimilated inthe root. Nitrate reduction and assimilation occurred in bothshoot and root. The assimilation of ammonium in roots of ammonium-fedplants was associated with a higher respiration rate. Key words: Ricinus communis, nitrogen nutrition (nitrate/ammonium), phloem, xylem, transport, partitioning, nitrogen, carbon, potassium, sodium, magnesium, calcium, chloride     

Chemical Changes In Young Tea Plant (Camellia sinensis L.) Tissues Following Application of Fertilizer Nitrogen

The uptake of ammonium sulphate by 14-month-old potted tea plantsgrown in a glasshouse was studied over the 11-week period followingapplication. Concurrent changes in the starch of root-wood,amino acids of xylem sap, and total nitrogen of leaves, stems,root-wood, and feeder roots were determined. Depletion of nitrogenfrom the soil at different depths and transformation of NH₄⁺to NO₃^– was also followed. The results show that the uptake of nitrogen commences within2 days of application as indicated by a marked increase in theamino-acid content of the xylem sap. Glutamine and, to a lesserextent, theanine were quantitatively the most important aminoacids in the sap. The amino-acid content of the sap was a maximumat about the time rapid depletion of the ammonium of the soiltook place. An interesting feature of the work is the reciprocalrelationship between the changes in the starch of root-woodand amino acids in the sap a few days after fertilizer application.Studies on the ammonium and nitrate levels of the soil at differentdepths showed that transformation of NH₄⁺ to NO₃^– occurredin the soil. The response of the various tissues to applied fertilizer nitrogenand increase in the fresh weight of the shoot system showedsimilar trends and may be correlated with the depletion of ammonium-nitrogenfrom the soil.     

Glutamine Involvement in Nitrogen Control of Gibberellic Acid Production in Gibberella fujikuroi

When the fungus Gibberella fujikuroi ATCC 12616 was grown in fermentor cultures, both intracellular kaurene biosynthetic activities and extracellular GA₃ accumulation reached high levels when exogenous nitrogen was depleted in the culture. Similar patterns were exhibited by several nonrelated enzymatic activities, such as formamidase and urease, suggesting that all are subject to nitrogen regulation. The behavior of the enzymes involved in nitrogen assimilation (glutamine synthetase, glutamate dehydrogenase, and glutamate synthase) during fungal growth in different nitrogen sources suggests that glutamine is the final product of nitrogen assimilation in G. fujikuroi. When ammonium or glutamine was added to hormone-producing cultures, extracellular GA₃ did not accumulate. However, when the conversion of ammonium into glutamine was inhibited by L-methionine-DL-sulfoximine, only glutamine maintained this effect. These results suggest that glutamine may well be the metabolite effector in nitrogen repression of GA₃ synthesis, as well as in other nonrelated enzymatic activities in G. fujikuroi.     

Studies on the growth and nitrogen assimilation of the bloom dinoflagellate Gyrodinium aureolum Hulburt

Growth experiments were carried out with laboratory culturesof Gyrodinium aureolum. Physiological studies on CO₂ fixationand nitrogen nutrition were undertaken with a natural populationdominated by the same organism. These studies suggested thatthe population was in a late stage of bloom development andsuffering from a degree of nitrogen limitation. The relevanceof the ammonium enhancement of dark CO₂ fixation as a techniquefor determining the nitrogen status of phytoplankton is discussedas are the difficulties involved in extrapolating results fromlaboratory studies to field situations.     

Suppression of long-day flowering by nitrogenous compounds in Lemna perpusilla 6746

The long-day flowering of Lemna perpusilla 6746 on an SH inhibitor-containingmedium was inhibited by the application of ammonium ion to themedium. Ammonium ion not only suppressed long-day flowering,but relieved the inhibition of vegetative growth caused by theinhibitors. Nitrite, casamino acids, glutamine and asparaginehad a similar effect, suggesting that the inhibition of long-dayflowering by ammonium ion is not a direct effect of the ion.Most amino acids, with the exception of glutamate and aspartate,also prevented long-day flowering, but their effects on vegetativegrowth varied. No qualitative differences in amino acid compositionwere observed among plants cultured on media containing nitrate,nitrite or NH4₄NO₃as the sole nitrogen source. However, theamounts of free and total amino acids werehigher in plants fedwith nitrite or NH₄NO₃ than in those fed with nitrate. Thissuggests that the inhibition of long-day flowering by ammoniumand nitrite can be ascribed to increased nitrogen metabolism. Though decreased activity by SH inhibitors of nitrate reductase(SH enzyme) is assumed to result in long-day flowering by loweringthe nitrogen metabolism, lowering the nitrogen level in M mediumdid not bring about floral initiation in the absence of SH inhibitors. (Received January 7, 1975; )     

Effects of Different Nitrogen Sources and Concentrations on CAM Photosynthesis in Kalanchoe blossfeldiana

When Kalanchoë blossfeldiana Poelln. cv. Hikan plants werecultured in solutions containing 0.2, 1.0, 5.0 or 10 mM of nitrateor ammonium under a long-day photoperiod, some criteria of CAM(Crassulacean acid metabolism) photosynthesis (diurnal changesof CO₂ uptake, titratable acidity and malate content in leaves)were examined. The plants absorbed 90 to 100% of CO₂ duringthe light phase regardless of the supplied nitrogen. Nitrate-grownplants absorbed about 10% of CO₂ during the dark phase regardlessof the supplied concentration, whereas in ammonium-grown plantsthe nocturnal CO₂ uptake occurred at 0.2 mM, at which the plantsdepleted nitrogen and no uptake was observed at the higher concentrations.Changes of nocturnal increase in titratable acidity and malatecontent almost corresponded with the changes in the amount ofnocturnal CO₂ uptake. Also K. daigremontiana plants suppliedwith 10 mM of ammonium had a less CAM-like pattern of diurnalCO₂ uptake than the plants supplied with 10 mM of nitrate. Theseresults suggest that a sufficient supply of ammonium depressesCAM photosynthesis.     

氮素添加对内蒙古羊草草原净氮矿化的影响

为了更好地了解天然草原氮素矿化对全球氮沉降背景和草原施肥管理模式的响应, 从2000年起对内蒙古典型草原羊草 (Leymu schinensis) 群落开展了长期的氮素添加实验, 分别设置对照 (N0), 添加5gNH4NO3·m-2 (N1.75) 、30gNH4NO3·m-2 (N10.5) 和80gNH4NO3·m-2 (N28) 4个氮素添加梯度。2002年, 从相邻的同时进行施肥的两个生态系统类型, 即1979年围封的样地A和1999年围封的样地B进行土壤取样, 在最佳温度 (25℃) 和最适土壤湿度 (即60%田间持水量) 下进行5周的室内培养, 并用阶段性淋溶方法研究了氮素添加对土壤氮矿化动态的影响。在A和B两个样地内, 氮素添加都显著改变了土壤的累积氮矿化量。最高氮素处理N28对应于最低的累积氮矿化量, 而低氮素处理N1.75使得累积氮矿化量达到最高。在N0和N1.75处理中, 硝态氮的含量高于铵态氮;在N28处理中, 却表现出相反的趋势。氮素添加显著降低了土壤的pH值, 但累积氮矿化量与土壤pH值、有机碳和全氮均没有显著的相关性。大多数氮素添加处理水平在样地A具有比样地B更高的土壤累积氮矿化量。     

Compensatory Roles of Nitrogen Uptake and Photosynthetic N-use Efficiency in Determining Plant Growth Response to Elevated CO2: Evaluation Using a Functional Balance Model

We used a modified functional balance (FB) model to predictgrowth response of Helianthus annuus L. to elevated CO₂. Modelpredictions were evaluated against measurements obtained twiceduring the experiment. There was a good agreement between modelpredictions of relative growth rate (RGR) responses to elevatedCO₂and observations, particularly at the second harvest. Themodel was then used to compare the relative effects of biomassallocation to roots, nitrogen (N) uptake and photosyntheticN-use efficiency (PNUE) in determining plant growth responseto elevated CO₂. The model predicted that a rather substantialincrease in biomass allocation to root growth had little effecton whole plant growth response to elevated CO₂, suggesting thatplasticity in root allocation is relatively unimportant in determininggrowth response. Average N uptake rate at elevated comparedto ambient CO₂was decreased by 21–29%. In contrast, elevatedCO₂increased PNUE by approx. 50% due to a corresponding risein the CO₂-saturation factor for carboxylation at elevated CO₂.The model predicted that the decreased N uptake rate at elevatedCO₂lowered RGR modestly, but this effect was counterbalancedby an increase in PNUE resulting in a positive CO₂effect ongrowth. Increased PNUE may also explain why in many experimentselevated CO₂enhances biomass accumulation despite a significantdrop in tissue nitrogen concentration. The formulation of theFB model as presented here successfully predicted plant growthresponses to elevated CO₂. It also proved effective in resolvingwhich plant properties had the greatest leverage on such responses.Copyright 2000 Annals of Botany Company Elevated CO₂, functional balance model, Helianthus annuus L., N uptake, photosynthetic nitrogen use efficiency, root:shoot ratio     

Uptake of 13C and 15N (ammonium, nitrate and urea) by Microcystis in Lake Kasumigaura

The uptake rate of carbon and nitrogen (ammonium, nitrate andurea) by the Microcystis predominating among phytoplankton wasinvestigated in the summer of 1984 in Takahamaira Bay of LakeKasumigaura. The V_max values of Microcystis for nitrate (0.025–0.046h^–1) and ammonium (0.15–0.17 h^–1) were considerablyhigher than other natural phytoplankton. The ammonium, nitrateand urea uptake by Microcystis was light dependent and was notinhibited with nigh light intensity. The K₁ values were farlower than the I_k values. The carbon uptake was not influencedby nitrogen enrichment. Microcystis accelerated the uptake rateby changing V_max/K _s value when nitrogen versus carbon contentin cells declined. Nitrate was scarcely existent in TakahamairiBay during the summer, when Microcystis usually used ammoniumas the nitrogen source. However, the standing stock of ammoniumin the water was far lower than the daily ammonium uptake rates.Therefore, the ammonium in this water had to be supplied becauseof its rapid turn-over time (–0.7–2.6 h).     

鼎湖山主要森林土壤N2O排放及其对模拟N沉降的响应

研究了鼎湖山生物圈保护区马尾松(Pinus massoniana)林、混交林和季风常绿阔叶林(季风林)土壤N₂O排放特征及其对氮沉降增加的响应。在1999~2002年期间,3种森林土壤N₂O排放速率均表现明显的季节性变化特点,但这种季节性变化因年份和森林类型不同而异,总的来说,3种森林土壤N₂O排放速率呈现夏秋季较高而冬春季较低的变化。土壤N₂O排放速率在3年观测期间的平均值分别为(g·hm^-2·d^-1):14.2±3.1(季风林),5.8±0.9(混交林)和5.1±0.9(马尾松林)。土壤N₂O排放速率与土壤温度之间在季风林呈现显著的指数正相关关系,但在混交林和马尾松林中它们之间的关系则均不明显。经3个月的模拟氮沉降试验后,氮沉降增加对季风林和马尾松林土壤N₂O的排放均具有明显的促进作用,且这种促进作用随氮沉降水平的升高而增强,但对混交林土壤N₂O排放的影响则不明显。     

Control of l-amino acid oxidase in Neurospora crassa by different regulatory circuits

l-Amino acid oxidase is synthesized in Neurospora crassa in response to three different physiological stimuli: (i) starvation in phosphate buffer, (ii) mating, and (iii) nitrogen derepression in the presence of amino acids. During starvation in phosphate buffer, or after mating, l-amino acid oxidase synthesis occurred in parallel with that of tyrosinase. Exogenous sulfate repressed the formation of the two enzymes in starved cultures, but not in mated cultures. Sulfate repression was relieved by protein synthesis inhibitors, suggesting that the effect of sulfate required the synthesis of a metabolically unstable protein repressor. With amino acids as the sole nitrogen source only l-amino acid oxidase was produced. Under these conditions enzyme synthesis was repressed by ammonium and was insensitive to sulfate. Biochemical evidence suggested that the l-amino acid oxidase formed under the three different conditions was the same protein. Therefore, the expression of l-amino acid oxidase appeared to be under the control of least two regulatory circuits. One, also controlling tyrosinase, seems to respond to developmental signals related to sexual morphogenesis. The other, controlling other enzymes of the nitrogen catabolic system, is used by the organism to obtain nitrogen from alternative sources such as proteins and amino acids.     

Inhibition of Nodule Development in Soybean by Nitrate or Reduced Nitrogen

Imsande, J. 1986. Inhibition of nodule development in soybeanby nitrate or reduced nitrogen.—J. exp. Bot. 37: 348–355. Nodulation of hydroponically grown soybean plants [Glycine max(L.) Merr.] is inhibited by continuous growth in the presenceof 4· mol m^–3 KNO₃ The presence of 4·0 molm^–3 ‘starter nitrate’ for 3-6 d during noduledevelopment, however, subsequently stimulates nodule dry weightaccumulation and nitrogenase activity. These stimulations occureven though 4· mol m^–3 nitrate temporarily delaysnodule development, i.e. the late steps of nodule developmentare reversibly inhibited by a short-term exposure to 4·0mol m^–3 nitrate. On the other hand, treatment with 4·0mol m^–3 nitrate in excess of 14 d significantly reducesnodule dry weight Thus, extended growth in the presence of 4·0mol m^–3 KNO₃ seems to block both early and late stepsof nodule development. Nodulation of hydroponically grown soybeansis also inhibited by continuous growth in the presence of 2·0mol m^–3 (NH₄)₂SO₄ This inhibition is not caused by acidityof the growth medium. On the other hand, nodule development6 d after inoculation with Rhizoblum japonicum is not delayedby a 7-d exposure to 2·0 mol m^–3 (NH₄)₂SO₄ butis partially inhibited by a prolonged exposure to (NH₄)₂SO₄Because repression of nodulation by 4·0 mol m^–3KNO₃ is more severe than that by 2·0 mol m^–3 (NH₄)₂SO₄and because ammonium taken up by the soybean plant is not activelyoxidized to nitrate, it is suggested that there are at leasttwo mechanisms by which nitrate utilization represses noduleformation in soybean. Key words: Glycine max, nitrogen, nitrogen fixation, nodulation     

Ammonia emission from young barley plants: influence of N source, light/dark cycles and inhibition of glutamine synthetase

Barley (Hordeum vulgare L. cv. Golf) plants were grown at twodifferent relative addition rates; 0.1 and 0.2 d^–1 ofnitrate. Three to five days before measurements started theplants were transferred to a nutrient solution with 2 mM nitrateor ammonium. The ammonium-grown plants showed increased ammoniumlevels in both shoots and roots and also increased ammoniumconcentrations in xylem sap. Ammonia emission measured in cuvettes connected to an automaticNH₃ monitor was close to zero for nitrate-grown plants but increasedto 0.59 and 0.88 nmol NH₃ m^–2 S^–1 for plants transferredto ammonium after growing at RA=0.2 and 0.1 d^–1, respectively.In darkness, NH₃ emission decreased together with photosynthesisand transpiration, but increased rapidly when the light wasturned on again. Addition of 0.5 mM methionine sulphoximine (MSO) to the plantscaused an almost complete inhibition of both root and shootglutamine synthetase (GS) activity after 24 h. Ammonia emissionincreased dramatically and photosynthesis and transpirationdecreased in both nitrate- and ammonium-grown plants as a resultof the GS inhibition. At the same time plant tissue and xylemsap ammonium concentrations increased, indicating the importanceof GS in controlling plant ammonium levels and thereby NH₃ emissionfrom the leaves. Key words: Hordeum vulgare, ammonia emission, ammonium, glutamine synthetase, nitrogen nutrition, photosynthesis, transpiration