Reassessing the nitrogen relations of Arctic plants: a mini-review

The Arctic is often assumed to be an NH₄⁺-dominated ecosystem. This review assesses the validity of this assumption. It also addresses the question of whether Arctic plant growth is limited by the ability to use the forms of nitrogen that are available. The review demonstrates that several sources of soil nitrogen are available to Arctic plants, including soluble organic nitrogen (e.g. glycine, aspartic acid and glutamic acid), NH₄⁺ and NO^?₃. In mesic Arctic soils, soluble organic nitrogen is potentially more important than either NH⁺₄ or NO^?₃. Many Arctic species are capable of taking up soluble organic nitrogen (either directly and/or in association with ectomycorrhizae), with the greatest potential for soluble organic nitrogen uptake being exhibited by deciduous species. The ability to take up soluble organic nitrogen may enable some Arctic plants to avoid nitrogen limitations imposed by the slow rate of organic matter decomposition. NO^?₃ is also present in many Arctic soils, especially calcareous soils and soils near flowing water, animal burrows and bird cliffs. Arctic species characteristic of mesic and xeric habitats are capable of taking up and assimilating NO^?₃. Even when present in lower concentrations in soils than NH⁺₄, NO^?₃ is still an important source of nitrogen for some Arctic plants. Arctic-plants therefore have a variety of nitrogen sources available to them, and are capable of using those nitrogen sources. Taken together, these findings demonstrate that the Arctic is not an NH⁺₄dominated ecosystem. Symbiotic fixation of atmospheric N₂ does not appear to be an important source of nitrogen for Arctic plants. The reliance of Arctic plants on internal recycling of nitrogen substantially reduces their dependence on soil nitrogen uptake (this is particularly the case for slow-growing evergreens). Despite the high level of internal nitrogen recycling, Arctic plant growth remains limited by the low levels of available soil nitrogen. However, Arctic plant growth is not limited by an inability to utilize any of the available forms of nitrogen. The potential effects of climatic warming on nitrogen availability and use are discussed. The question of whether the Arctic ecosystem is uniquely different from temperate nitrogen-deficient ecosystems is also assessed.     

Regulation of mineral nitrogen uptake in plants

In the biosphere plants are exposed to different forms of N, which comprise mineral and organic N forms in soils as well as gaseous NH3, NOx, and molecular N2 in the atmosphere. The form of N uptake is mainly determined by its abundance and accessibility, which make and the most important N forms for plant nutrition under agricultural conditions. With minor importance, the form of N uptake is also subject to plant preferences, by which plants maintain their cation/anion balance during uptake. However, some species seem to have an obligatory preference which even prevents their growth on certain other N sources. In general, uptake of a certain N form closely matches the growth-related demand of the plant, at least when N transport to the root surface is not limiting. In addition, many plants accumulate large pools of N during vegetative growth which are remobilized in the generative stage. As a consequence, systems responsible for N transport need to be tightly regulated in their expression and activity upon sensing N availability and plant demand. Employing the tools of molecular genetics, the first plant genes encoding transporters for inorganic N have recently been isolated and characterized. These data can now complete the wealth of physiological and nutritional studies on N uptake. The present article will focus on the uptake of and into root cells and tries to link data derived from physiological, genetic and molecular studies.     

Transporters for uptake and allocation of organic nitrogen compounds in plants

Nitrogen is an essential macronutrient for plant growth. Following uptake from the soil or assimilation within the plant, organic nitrogen compounds are transported between organelles, from cell to cell and over long distances in support of plant metabolism and development. These translocation processes require the function of integral membrane transporters. The review summarizes our current understanding of the molecular mechanisms of organic nitrogen transport processes, with a focus on amino acid, ureide and peptide transporters.     

Photoheterotrophy and light-dependent uptake of organic and organic nitrogenous compounds by Planktothrix rubescens under low irradiance

1. Planktothrix rubescens is the dominant photoautotrophic organism in Lake Zürich, a prealpine, deep, mesotrophic freshwater lake with an oxic hypolimnion. Over long periods of the year, P. rubescens accumulates at the metalimnion and growth occurs in situ at irradiance near the photosynthesis compensation point. Experiments were conducted to evaluate the contribution of photoheterotrophy, heterotrophy and light‐dependent uptake of nitrogenous organic compounds to the carbon and nitrogen budget of this cyanobacterium under conditions of restricted availability of light quanta. 2. We used both purified natural populations of P. rubescens from the depth of 9 m and an axenic culture grown under low irradiance at 11 μmol m^?2 s^?1 on a light : dark cycle (10 : 14 h) to determine the uptake rates of various amino acids, urea, glucose, fructose, acetate and inorganic carbon. The components were added to artificial lake water in low amounts that simulated the naturally occurring potential concentrations. 3. The uptake rates of acetate and amino acids (glycine, serine, glutamate and aspartate) were strongly enhanced at low irradiance as compared with the dark. However, no difference was observed in the uptake of arginine, which was taken up at high rates under both treatments. The uptake rates of glucose, fructose and urea were very low under all conditions. Similar results were obtained for both axenic P. rubescens and for purified natural populations of P. rubescens that were separated from bacterioplankton and other phytoplankton. 4. Metalimnetic P. rubescens that was stratified at low irradiance for weeks exhibited much higher uptake rates than filaments that were entrained in the deepening surface mixed layer and experienced higher irradiance. The added organic compounds contributed up to 62% to the total carbon uptake of metalimnetic P. rubescens. On the basis of a molar C : N ratio of 4.9, the nitrogen uptake as organic compounds satisfied up to 84% of the nitrogen demand. 5. The experiments indicate that photoheterotrophy and light‐dependent uptake of nitrogenous organic compounds may contribute significantly to the carbon and nitrogen budget of filaments at low irradiance typical for growth of P. rubescens in the metalimnion and at the bottom of the surface mixed layer.     

氨基酸转运载体研究进展

氨基酸转运载体是介导氨基酸跨膜转运的膜蛋白,在氨基酸营养机体细胞和神经调节过程中起着重要作用;而且,其功能异常会导致严重的氨基酸吸收和代谢障碍性疾病,也具有重要的病理学意义。本文就近年来关于中性氨基酸、酸性氨基酸和碱性氨基酸转运载体家族成员及其组织分布、分子生物学特征、生理功能和病理学意义等研究进展进行了综述。     

The nitrogen balance of Raphanus sativus X raphanistrum plants. I. Daily nitrogen use under high nitrate supply

Abstract Growth-chamber cultivated Raphanus plants accumulate nitrate during their vegetative growth. After 25 days of growth at a constant supply to the roots of 1 mol m^?3 (NO^?₃) in a balanced nutrient solution, the oldest leaves (eight-leaf stage) accumulated 2.5% NO^?₃-nitrogen (NO₃-N) in their lamina, and almost 5% NO₃-N in their petioles on a dry weight basis. This is equivalent to approximately 190 and 400 mol^?3 m^?3 concentration of NO^?₃ in the lamina and the petiole, respectively, as calculated on a total tissue water content basis. Measurements were made of root NO^?₃ uptake, NO^?₃ fluxes in the xylem, nitrate uptake by the mesophyll cells, and nitrate reduction as measured by an in vivo test. NO^?₃ uptake by roots and mesophyll cells was greater in the light than in the dark. The NO^?₃ concentration in the xylem fluid was constant with leaf age, but showed a distinct daily variation as a result of the independent fluxes of root uptake, transpiration and mesophyll uptake. NO^?₃ was reduced in the leaf at a higher rate in the light than in the dark. The reduction was inhibited at the high concentrations calculated to exist in the mesophyll vacuoles, but reduction continued at a low rate, even when there was no supply from the incubation medium. Sixty-four per cent of the NO^?₃ influx was turned into organic nitrogen, with the remaining NO^?₃ accumulating in both the light and the dark.     

Characteristics of inorganic nitrogen assimilation of plants in fire-prone Mediterranean-type vegetation

A range of approaches was used to investigate how species within a fire-prone Banksia woodland in South West Australia exploited inorganic soil nitrogen sources and how this changes through the development of the fire chronosequence. Nitrate and ammonium were present in soil solution throughout the chronosequence but nitrate predominated in recently burnt sites. Mean shoot nitrate reductase activities were high for all species in recently burnt sites and showed little increase when nitrate was supplied via the transpiration stream. Nitrate reductase of shoots of most species was low at sites not burnt for several years, but following transpirational induction with nitrate, developed activities similar to those at recently burnt sites. The principal amino compounds transported in the xylem were species specific, including asparagine, glutamine and citrulline-dominated species, and changed little in relative composition across the chronosequence. Species most active in leaf nitrate reduction transported the largest amounts of nitrate in their xylem sap and proportional amounts of nitrate in xylem tended to be greatest in recently burnt sites. Most of the species examined appeared to be shoot rather than root nitrate assimilators, but marked differences were recorded in potential of leafy shoots of different species to reduce nitrate. As a general rule, shallow-rooted herbaceous, non-mycorrhizal or VAM-positive species had the highest capacity to reduce nitrate, whereas woody species with ericoid mycorrhizae or combined vesicular arbuscular/ectomycorrhizal associations exhibited little capacity to reduce nitrate in roots or shoots. It seems likely that this latter group utilize ammonium or even organic forms of nitrogen rather than nitrate. Some putative nitrogen-fixing species were active in reducing and transporting nitrate, others were virtually inactive in these respects.     

Effect of amino compounds on nitrate utilization by roots of dwarf bean

Amino compounds (1 mM, pH 5) were given prior to, together with, or after the addition of nitrate to study their effect on nitrate uptake and in vivo nitrate reductase activity (NRA) in roots of Phaseolus vulgaris. The effect of amino compounds varied with the amino species, the nitrate status of the plant (induced vs uninduced) and the aspect of nitrate utilization. Cysteine inhibited the nitrate uptake rate and root NRA under all conditions tested. NRA in uninduced roots was stimulated by tryptophan, and arginine inhibited NRA under all conditions tested. Uptake was inhibited by aspartate and glutamate and stimulated by leucine when these amino compounds were given prior to or after completion of the apparent induction of nitrate uptake. In the presence of β-alanine and tryptophan, induction of uptake was accelerated.     

Nitrate accumulation in leaves of vegetation of a forested ecosystem receiving high amounts of atmospheric ammonium sulfate

Vegetation of an acid woodland, receiving an atmospheric ammonium input of about 3 kmol (40 kg N) per hectare per year, was analyzed on the content of organic nitrogen, ammonium and nitrate. A high nitrate content (50–320 μmol g⁻¹ dry weight) was found in bird-cherry, black-berry and bracken, whereas only low amounts (up to 2 μmol g⁻¹ dry weight) of nitrate were present in mountain-ash, hazel and the two dominant tree species oak and birch. The impact of this nitrate uptake and nitrate accumulation on soil pH and autotrophic nitrification is discussed.     

Carbohydrate and free amino acid contents in tomato plants grown in media with bicarbonate and nitrate or ammonium

Tomato plants were cultivated (from 2 to 23 days after germination) in media with NO ₃ ⁻ , NH ₄ ⁺ , or a mixture of both forms in different proportions used as the N source given with or without 5 mol dm⁻³ HCO ₃ ⁻ . The accumulation of soluble sugars (reducing sugars and sucrose) and free amino acids was higher in the roots and leaves of NH ₄ ⁺ -fed plants than in NO ₃ ⁻ -fed plants. Starch accumulation in NH ₄ ⁺ -fed plants was higher in leaves (about 28%) and lower in roots (about 37%) in comparison with that of NO ₃ ⁻ -fed plants. Plants cultivated in media containing a mixture of NO ₃ ⁻ /NH ₄ ⁺ were characterized by a lower content of sugars and amino acids accumulation in comparison with that in plants fed with NO ₃ ⁻ or NH ₄ ⁺ . An elevated HCO ₃ ⁻ concentration in the rhizosphere stimulated the accumulation of soluble sugars and free amino acids in all the experimental variants. There were only small differences in the starch content.     

氨基酸转运体在肿瘤代谢中的研究进展

代谢重编程是肿瘤的重要特征,是指肿瘤细胞为满足其快速增殖的生物合成与能量需求,对其糖代谢、脂代谢以及氨基酸代谢等代谢路径进行的重编程,以维持增长速度以及补偿能量代谢所造成的氧化还原压力。虽然不同的癌症代谢变化不同,但有些特征是所有癌症共有的,氨基酸代谢重编程是其中一个重要的特征。氨基酸进出细胞需要氨基酸转运体的协助,因而在肿瘤细胞中多种特定的氨基酸转运体均过表达。靶向氨基酸转运体通过影响肿瘤细胞的氨基酸代谢从而达到抗肿瘤的目的,是目前抗肿瘤药物的研究热点之一。主要介绍了几种在肿瘤代谢中发挥重要作用的氨基酸转运体以及靶向氨基酸转运体抗肿瘤治疗的研究进展及相关作用机制,旨在了解氨基酸转运体在抗肿瘤研究中的作用,以期促进靶向氨基酸转运体抗肿瘤药物的发展。     

Qualitative and quantitative changes in nitrogenous compounds in senescing leaf and bark tissues of the apple

Quantitative and qualitative changes in proteins and ethanol-soluble nitrogen were followed in senescing leaf and bark tissues of ‘Golden Delicious’ apple trees (Malus domestica Borkh.). While senescing leaves lost 46% of their proteins, total bark protein increased 240% during senescence. However, the protein nitrogen gain in bark tissue was about the same as the protein nitrogen loss in leaf tissue per unit fresh weight of tissues. The pattern of bark protein accumulation appears to be gradual from early August to November and sequential from lower to higher molecular weight species of proteins. The final electrophoretic profile of total bark proteins was established at the later stages of senescence. By late November, 89% of the nitrogen in the bark tissue was found in proteins with 11% in the ethanol-soluble fractions. The total protein content of dormant bark tissue was 3.5% per gram dry tissue. Fractionation of the total bark proteins by DEAE-cellulose chromatography indicated that the final upsurge of bark proteins observed in November was associated primarily with one group of proteins (Peak III).     

A Na+-DEPENDENT SYSTEM A AND ASC-INDEPENDENT AMINO ACID TRANSPORT SYSTEM STIMULATED BY GLUCAGON IN RAT HEPATOCYTES

The effects of glucagon on amino acid transport in rat hepatocytes are not fully understood. We examined the effect of this hormone on alanine, serine and cysteine preferring system (system ASC)-mediated amino acid transport in rat hepatocyte monolayers using 2-aminoisobutyric acid (AIB) and L -cysteine. Glucagon induced a time and protein synthesis-dependent stimulation of Na⁺-dependent alanine preferring system (system A)-independent AIB transport. The glucagon-induced increase in transport activity was not modified by substrate starvation and not related to changes in the intracellular pool of amino acids. Glucagon did not modify system ASC activity measured by L -cysteine. Therefore the transport activity of AIB independent of system A stimulated by glucagon cannot be attributed to system ASC. This suggests a Na⁺-dependent transport system in rat hepatocytes not identified until now.     

Nitrate uptake and reduction in higher and lower plants

The nitrogen compounds nitrate and ammonium are the minerals that plants need in large quantities and which limit their growth in temperate zones. The nitrate assimilation pathway starts with nitrate uptake followed by nitrate reduction resulting in ammonium which is fixed into the amino acids glutamine and glutamate in most plants. This review concentrates on nitrate uptake and nitrate reduction with respect to higher and lower plants. The physiology and the progress in molecular approaches of both processes are considered. For nitrate uptake the well‐established uptake systems are discussed and special attention is drawn to nitrate sensing and the nitrate carrier. Knowledge, particularly on nitrate sensing is rare, but it seems to be the first step in a signal transduction chain triggered by nitrate. Therefore further work should consider this topic more frequently. For nitrate reductase the focus is on the post‐translational modification as a regulatory tool for nitrate assimilation, on the intersections of carbon and nitrogen metabolism and on the molecular approaches. A few remarks on how environmental conditions affect nitrate assimilation are also included. Further progress is needed to understand the transduction of positive and negative signals from the environment affecting the expression of genes coding for the nitrate assimilating pathway.     

Alpine plants show species-level differences in the uptake of organic and inorganic nitrogen

As an estimate of species-level differences in the capacity to take up different forms of N, we measured plant uptake of ¹⁵N-NH₄ ⁺, ¹⁵N-NO₃ ^– and ¹⁵N, [1]-¹³C glycine within a set of herbaceous species collected from three alpine community types. Plants grown from cuttings in the greenhouse showed similar growth responses to the three forms of N but varied in the capacity to take up NH₄ ⁺, NO₃ ^– and glycine. Glycine uptake ranged from approximately 42% to greater than 100% of NH₄ ⁺ uptake; however, four out of nine species showed significantly greater uptake of either NH₄ ⁺ or NO₃ ^– than of glycine. Relative concentrations of exchangeable N at the sites of plant collection did not correspond with patterns of N uptake among species; instead, species from the same community varied widely in the capacity to take up NH₄ ⁺, NO₃ ^–, and glycine, suggesting the potential for differentiation among species in resource (N) use.     

The uptake of nitrate and ammonium nitrogen in Chara hispida L.: the contribution of the rhizoid

Abstract The uptake of ammonium and nitrate nitrogen by cultured plants of the green freshwater alga Chara hispida L. has been compared quantitatively with the contribution of its rhizoidal tissue. In the short-term, the rhizoid takes up 7–20% of the ammonium nitrogen, and about 15% of the nitrate that is taken up by whole plants under similar conditions. The uptake was studied over a range of both temperatures and external concentrations. The apparent activation energy for the uptake of NH₄⁺ and NO₃^? by the whole plant was found to be 50 kJ mol^?1 and 30 kJ mol^?1, respectively. For the rhizoid, the values were similar for both nitrogenous ions, 106 kJ mol^?1 and 70–100 kJ mol^?1. The rhizoidal uptake mechanism for ammonium nitrogen operates more efficiently compared to that in the whole plant. Nitrate is taken up by the rhizoid by a mechanism with a substrate affinity higher than in the plant taken as a whole. The possible ecological significance of the results is discussed.     

Transport of osmoprotective compounds in hybridoma cells exposed to hyperosmotic stress

Addition of osmoprotective compounds has a positive effect on growth and monoclonal antibody production in hyperosmotic hybridoma cell cultures. In order to better understand the processes involved in the osmoprotective response, uptake of the osmoprotective compounds glycine betaine, proline, sarcosine and glycine in mouse hybridoma cell line 6H11 during exposure to hyperosmotic stress was studied. Hyperosmotic stress (510 mOsmol/kg) was introduced through the addition of NaCl (100 mM) to the growth medium, and amino acid transport activity was measured immediately after transfer of the cells to the hyperosmotic medium. The osmoprotective capability of the four osmoprotectants tested was negatively affected if methylaminosobutyric acid (MeAiB), a specific substrate for amino acid transport system A, was simultaneously included in the hyperosmotic medium in equimolar amounts with one of the osmoprotective compounds. This was due to accumulation of MeAiB in the stressed cells, giving a significant reduction in the concentration of the osmoprotective compound inside the cells. Furthermore, addition of excess meAiB gave approx. 905 reduction in the initial rate of uptake of glycine betaine, while 40–50% reduction in the initial rate of uptake of proline, glycine and sarcosine. Similarly, addition of proline, glycine or sarcosine also gave a significant reduction in the initial rate of glycine betaine uptake. These results suggest that the four osmoprotective compounds share, at least in part, a common, MeAiB inhibitable carrier for transport into osmotically stressed hybridoma cells. This carrier is probably equal to amino acid transport system A.