The pathway of nitrogen assimilation in plants

The major route of nitrogen assimilation has been considered for many years to occur via the reductive amination of α-oxoglutarate, catalysed by glutamate dehydrogenase. However, recent work has shown that in most bacteria an alternative route via glutamine synthetase and glutamine: 2-oxoglutarate aminotransferase (glutamate synthase) operates under conditions of ammonia limitation. Subsequently the presence of a ferredoxin-dependent glutamate synthase in green leaves and green and blue-green algae, and a NAD(P)H and ferredoxin-dependent enzyme in roots and other non-green plant tissues, has suggested that this route may also function in most members of the plant kingdom. The only exceptions are probably the majority of the fungi, where so far most organisms studied do not appear to contain glutamate synthase. Besides the presence of the necessary enzymes there is other evidence to support the contention that the assimilation of ammonia into amino acids occurs via glutamine synthetase and glutamate synthase, and that it is unlikely that glutamate dehydrogenase plays a major role in nitrogen assimilation in bacteria or higher plants except in circumstances of ammonia excess.     

Response of Zea mays to the Inoculation with Azospirillum on Nitrogen Metabolism under Greenhouse Conditions

The maize (Zea mays L.) plants inoculated by N₂-fixing bacterium Azospirillum showed increased activity of glutamate dehydrogenase (GDH) and glutamine synthetase (GS) in root cells free extracts over uninoculated control plants. Maximum differences in NADH-GDH activity were observed during the second and third weeks after sowing. The specific activity of GS showed a greater increase at the end of the assay. The percentage of nitrogen in leaves, root and foliage length, total fresh mass and nitrogenase activity were higher in inoculated plants than in the control ones.     

Effect of Azotobacter strains on sugar beet callus proliferation and nitrogen metabolism enzymes

The effect of five Azotobacter chroococcum strains and nitrogen content in nutrient media on callus growth of two Beta vulgaris L. cultivars were investigated, as well as the activity of nitrate reductase (NR), glutamine synthetase (GS) and glutamate dehydrogenase (GDH) in inoculated callus tissue. On medium with full nitrogen content (1 N) the inoculation with A. chroococcum strain A2 resulted in the highest calli mass, while strains A8 and A14 maximally increased NR activity. On media with 1/8 N the highest effect on calli growth, GS and GDH activity had the strain A8. The strain A2/1 significantly increased callus proliferation on medium without N. Asymbiotic association between sugar beet calli and Azotobacter depended on genotype/strain interaction and was realised in presence of different nitrogen levels. This revised version was published online in July 2006 with corrections to the Cover Date.     

施钾对甘薯氮素转移分配及氮代谢酶活性的影响

利用¹⁵N示踪技术,研究了施钾对甘薯发根结薯期、薯块膨大期地上和地下部氮素转移分配、光合特性及氮代谢酶活性的影响.结果表明: 在发根结薯期,施钾显著提高¹⁵N向地上部的转移分配,其中K₃(K₂O, 300 mg·kg^-1)处理与对照相比¹⁵N向叶片转移速率提高了76.2%,¹⁵N积累量提高了92.1%.在薯块膨大期,随施钾量增加地上部叶片¹⁵N总分配率由33.7%降低至24.4%,块根¹⁵N分配率由5.8%升高至17%,其中K₃处理块根¹⁵N积累量是对照的3倍.两个关键生长期硝酸还原酶、谷氨酸脱氢酶、谷氨酰胺合酶、谷氨酸合酶和净光合速率(P_n)均随施钾量的增加而提高.逐步回归分析表明,氮代谢酶活性和P_n是影响甘薯¹⁵N转移和分配的主要因素(R分别为0.965和0.942),通径分析表明,在发根结薯期主要通过促进硝酸还原酶和谷氨酸脱氢酶介导的氮素催化能力促进氮素向地上部分配;在薯块膨大期主要通过提高谷氨酰胺合酶/谷氨酸合酶循环介导的氮素同化能力促进氮素向地下部分配.     

Effect of nitrogen starvation on ammonium assimilation by Chlamydomonas reinhardtii

In nitrogen-starved Chlamydomonas reinhardtii , wild type, strain 21 gr cells, consumption of nitrate, nitrite and ammonium may occur in the dark in the absence of an added carbon source. Consumption of ammonium in the dark was about 25% higher than in the light, while consumption of nitrate or nitrite in the dark was lower than in the light.
N starvation produced a linear increase with time in the intracellular level of glutamine synthetase (GS, EC 6.3.2.1) and glutamate synthase (NADH-GOGAT, EC 1.4.1.14 and ferredoxin-GOGAT, EC 1.4.7.1) activities in C. reinhardtii . The effect on GS₁ (3-fold) and NADH-GOGAT (4.5-fold) was higher than that on GS₂ (1.5-fold) and ferredoxin-GOGAT (1.5-fold).
Experiments with methylammonium, L-methionine-D, L-sulfoximine (MSX) and azaserine suggest that: 1) Ammonium itself decreases the intracellular levels of glutamine synthetase and ferredoxin-glutamate synthase activities; and 2) a metabolite resulting from ammonium assimilation by the alga may be a negative modulator of NADH-glutamate synthase activity.     

陆生植物生物量分配对模拟氮沉降响应的Meta分析

分析了陆生植物地上、地下各组织中生物量分配对氮沉降的响应,为研究大气氮沉降背景下陆地生态系统的碳、氮循环过程及植物生物量分配、立木收获、定向培育等相关研究和实践提供参考依据。共收集整理了国内外63篇论文的原始数据资料进行Meta分析(Meta-analysis),用以定量评估氮沉降对植物生物量分配的影响,并通过亚组分析进一步探讨了不同生态系统类型、植物种类、氮肥形式、施氮水平和持续时间对生物量分配的影响。结果表明,总体来看施氮会显著促进植物地上部分生物量分配,植物叶生物量和茎生物量在施氮条件下均显著增加;然而地下生物量所受促进作用要低于地上部分,表现为植物细根生物量和粗根生物量在氮输入下并没有显著变化;植物根冠比在氮沉降下显著降低;叶重比、茎重比和根重比在氮沉降下没有显著变化。此外,亚组分析结果表明生态系统类型和植物类型会显著影响植物总生物量和根冠比对氮沉降的响应,草本植物在氮沉降下的生物量累积明显优于木本,这说明短期氮沉降可能会增加草本的覆盖面积;施肥形式对根冠比的影响存在明显差异,相比于尿素,硝酸铵对植物根冠比的作用更显著;不同施氮水平显著影响地上生物量分配,中氮水平(本研究为60—120 kg hm-2a-1)促进作用最大,高氮水平(本研究为≥120 kg hm-2a-1)促进作用明显减弱,这与总生物量的变化一致,表明过高的氮沉降量将抑制植物生长;氮沉降处理时间长短对植物地上生物量的影响也存在显著差异,当施氮时间高于3年,氮沉降对地上生物量的促进作用几乎消失。总之,短期氮沉降会使植物分配更多生物量给地上部分,且氮沉降对草本植物生物量的累积作用明显优于木本,这些发现可为未来大气氮沉降背景下植物地上、地下部分碳存储、植物群落结构、植被动态等相关研究提供科学依据。     

氮沉降对森林植物的影响

综述了氮沉降对森林植物的影响。氮沉降对森林植物的影响主要表现在以下6个方面：(1)在一定量范围内的氮沉降有利于植物的光合作用,但过量后则会引起植物的光合速率下降;(2)当植物生长受氮限制时,在一定程度上的氮沉降增加植物生产力,但当氮过量后,氮沉降则使植物的生产力下降;(3)过量的氮沉降导致植物体各种营养元素含量的比例失衡;(4)氮沉降会改变植物的形态结构,集中表现为根／冠比减小;(5)氮沉降会增加植物对天然胁迫如干旱、病虫害和风的敏感性,减少其抵御能力;(6)氮沉降会改变植物组成和降低森林植物的多样性。     

Effect of atmospheric ammonia on the nitrogen metabolism of Scots pine (Pinus sylvestris) needles

Four-year-old seedlings of Scots pine ( Pinus sylvestris L.) were exposed to filtered air (FA), and to FA supplemented with NH₃ (60 and 240 μg m⁻³) in controlled-environment chambers for 14 weeks. Exposure to the higher NH₃ concentration resulted in an increased activity of glutamine synthetase (GS, EC 6.3.1.2), and an increase in the concentrations of soluble proteins, total nitrogen, free amino acids and leaf pigments in the needles. The GS activity (μmol g⁻¹ fresh weight h⁻¹) in the needle extract increased to levels 69% higher than in FA and the soluble protein concentration to levels 22% higher. Total nitrogen concentration in the needles was 42% higher than in FA, while the free amino acid concentration was 300% higher, which was caused by an increase in arginine, glutamate, aspartate and glutamine. Chlorophyll a , chlorophyll b and carotenoid concentrations were 29, 38 and 11% higher, respectively. Neither the glutamate dehydrogenase (GDH, EC 1.4.1.2) activity nor the concentrations of free NH₄⁺ and glucose in the needles were affected by exposure to NH₃. After NH₃ fumigation at 240 μg m⁻³ the starch concentration decreased by 39% relative to the FA. The results indicate that the metabolism of Scots pine acclimates to concentrations of NH₃ which are 3 to 10 times higher than the average concentration in areas with intensive stock farming. The possible mechanisms underlying acclimation to NH₃ are discussed.     

Atmospheric nitrogen deposition explains patterns of plant species loss

Atmospheric nitrogen (N) deposition across Europe increased substantially from the 1950s to the 1990s. Targeted surveys suggest a negative correlation between N deposition and species richness within quadrats in sensitive habitats. However, it remains unclear whether plant species losses at national recording scales are correlated with nitrogen deposition. We relate plant species losses before 1987 in Great Britain to reduced and oxidized N deposition, land use change and climate change. The mean Ellenberg fertility (N) indices of plant species lost in each 100 km² cell before 1987 was compared with those of species that were recorded between 1987 and 1999. In 45% of squares, indices of species lost were significantly lower than those for species present after 1986. For 17%, primarily upland, squares, the opposite effect was found. A generalized least squares regression model, with difference in the mean Ellenberg N index between samples as the dependent variable, showed that higher deposition of reduced N was significantly associated with selective loss of species with a lower index. Arable land use and change in arable land use also demonstrated this positive relationship. Rough grazing, change in rough grazing, change in pasture and change in annual precipitation showed negative effects. Difference in Ellenberg R index was highly correlated with difference in Ellenberg N and was negatively correlated with oxidized N deposition, suggesting that the lack of a significant effect of oxidized N deposition on Ellenberg N was because it had effects through both acidification and eutrophication, while the effect of reduced N deposition was primarily through eutrophication. Our results suggest that N deposition, along with land use and precipitation changes, has been a significant driver of local plant extinctions. With N deposition increasing in many parts of the world, local extinctions of plant species may be experienced in other regions.     

马衔山不同海拔土壤碳、氮、磷含量及生态化学计量特征

研究半干旱地区土壤碳、氮、磷化学计量特征,了解其空间变化规律,有助于揭示半干旱地区C、N、P循环对全球气候变化的响应。本研究以半干旱区的马衔山为对象,选择5个海拔的7个样地,采集0～15、15～30 cm层的土壤,测定其有机碳(SOC)、全氮(TN)、全磷(TP)、pH、含水率等理化性质,分析其SOC、TN、TP化学计量与土壤理化因子之间的关系。结果表明:(1) 0～15 cm土壤SOC、TN、TP含量高于15～30 cm土壤。表层土壤SOC、TN含量随海拔升高呈增加趋势,TP含量随海拔升高变化较小。(2) C∶N随海拔增加呈先增加后降低趋势,C∶P、N∶P随海拔升高均呈增加趋势。(3)在0～15 cm土壤中,pH与SOC、TN含量及C∶P呈显著负相关,在15～30 cm土层中,pH与SOC、TN、TP含量及化学计量特征关系不显著;土壤含水率与0～15、15～30 cm层土壤中SOC、TN含量均呈极显著正相关。本研究显示,在半干旱区的马衔山地区,土壤含水率随海拔增加而增加,而SOC、TN含量及C∶P、N∶P也呈增加趋势,土壤养分含量及化学计量均受土壤含水率影响。     

The influence of leaf‐atmosphere NH3(g) exchange on the isotopic composition of nitrogen in plants and the atmosphere

The distribution of nitrogen isotopes in the biosphere has the potential to offer insights into the past, present and future of the nitrogen cycle, but it is challenging to unravel the processes controlling patterns of mixing and fractionation. We present a mathematical model describing a previously overlooked process: nitrogen isotope fractionation during leaf‐atmosphere NH_3(g) exchange. The model predicts that when leaf‐atmosphere exchange of NH_3(g) occurs in a closed system, the atmospheric reservoir of NH_3(g) equilibrates at a concentration equal to the ammonia compensation point and an isotopic composition 8.1‰ lighter than nitrogen in protein. In an open system, when atmospheric concentrations of NH_3(g) fall below or rise above the compensation point, protein can be isotopically enriched by net efflux of NH_3(g) or depleted by net uptake. Comparison of model output with existing measurements in the literature suggests that this process contributes to variation in the isotopic composition of nitrogen in plants as well as NH_3(g) in the atmosphere, and should be considered in future analyses of nitrogen isotope circulation. The matrix‐based modelling approach that is introduced may be useful for quantifying isotope dynamics in other complex systems that can be described by first‐order kinetics.     

大气NO2影响植物生长与代谢的研究进展

二氧化氮(NO₂)是大气氮氧化物之一,是大气气溶胶颗粒形成的主要成分,降低大气NO₂浓度可减轻空气中的雾霾.大气NO₂通过干沉降和湿沉降两种方式降落到植物叶片.植物吸收NO₂后主要通过两种代谢途径来降低空气中NO₂浓度: 一是主要在细胞质和叶绿体中利用还原酶的氮代谢途径,二是在质外体和细胞质中的歧化反应.植物吸收NO₂干扰了植物正常的生长和生理代谢,包括: 植物营养和生殖生长,植物体内硝酸还原酶(NaR)活性、亚硝酸还原酶(NiR)活性、氮素吸收、光合等生理代谢过程.对目前国内外有关大气NO₂影响植物生长与代谢的研究进展进行了综述,并对植物吸收NO₂的生理及分子机制的未来研究方向进行了展望.     

The effect of increased deposition of atmospheric nitrogen on Calluna vulgaris in upland Britain

Regular (monthly) additions of NH₄NO₃ (4–12 g N m⁻² yr⁻¹) were made over a period of 8 yr (1989–98) to areas of moorland in North Wales dominated by the ericaceous shrub Calluna vulgaris . Results from the early stages of the experiment (1990–94) have shown marked and dose-related increases in shoot extension and canopy height in response to the nitrogen treatments, with significantly higher shoot nitrogen contents. The nitrogen-related stimulation in the growth of the C. vulgaris canopy over this period has resulted in large accumulations of litter on the high-nitrogen-treated plots (6.6 kg m⁻² in plots treated with 12 g N m⁻² yr⁻¹, compared with 3.8 kg m⁻² for the water controls). Litter nitrogen concentrations were also significantly increased at the higher rates of nitrogen addition, leading to a doubling of the total return of nitrogen to the litter layer over the experimental period. These changes in vegetation structure were associated with large reductions in the abundance of the bryophyte and lichen species normally present under the untreated canopy. Results since 1994, however, show little increase in shoot extension in response to the nitrogen treatments, with no clear dose response to increasing levels of addition. These findings are associated with a dose-related increase in the susceptibility of the nitrogen-treated areas of the C. vulgaris canopy to late winter injury, characterized as browning of the shoot tips in early to late spring. These results indicate that deleterious effects are now accumulating as a result of the long-term addition of nitrogen to these moorland plots.     

Changes in the activity of enzymes involved with primary nitrogen metabolism due to ectomycorrhizal symbiosis on jack pine seedlings

Jack pine (Pinus banksiana Lamb.) seedlings were inoculated with either one of the ectomycorrhizal (ECM) fungi, Laccaria bicolor (Maire) Orton or Pisolithus tinctorius (Pers.) Coker and Couch, and grown for 16 weeks in a growth chamber along with non-ECM controls. Five enzymes involved with the assimilation of nitrogen or the synthesis of amino acids were measured in the 3 jack pine root systems as well as in the pure fungal cultures. Pisolithus tinctorius in pure culture had no detectable activity of nitrate reductase (NR. EC 1.6.6.1), glutamate dehydrogenase (GDH. EC 1.4.1.2), glutamate decarboxylase (GDCO. EC 4.1.1.15) or glutamate oxoglutarate aminotransferase (GOGAT, EC 1.4.1.13) but did have some glutamine synthetase (GS, EC 6.3.1.2) activity. Laccaria bicolor in pure culture had no NR activity, small levels of GDCO activity, and high GS, GDH and GOGAT activity. The high levels of enzymatic activity present in L. bicolor indicate that it may play a greater role in the nitrogen metabolism of its host plant than P. tinctorius. ECM infection clearly altered the enzymatic activity in jack pine roots but the nature of these changes depended on the fungal associate. Non-ECM root systems had higher specific activities than ECM root systems for NR, GS, GDH and GDCO but GOGAT activites were the same for both the ECM and non-ECM roots. Root systems infected with L. bicolor had significantly greater NR and GDCO activity than those infected with P. tinctorius. Differences in the GS activity of the two fungi in pure culture corresponded to the GS activity of jack pine roots in symbiotic association with these fungi. While the free amino acid profiles in roots were significantly affected by ECM infection, the profile of free amino acids exported to the stem was the same for all treatments. High asparagine and low glutamine in roots infected with P. tinctorius indicates that asparagine synthetase (EC x.x.x.x) activity should be higher within this symbiotic association than in the L. bicolor association or in the non-mycorrhizal roots.     

氮沉降对森林生态系统土壤碳库的影响

森林土壤碳库是陆地生态系统碳库的重要组成部分,对维持全球碳平衡具有重要意义。不断加剧的全球氮沉降有可能改变森林生态系统中碳元素的地球化学循环过程,从而引起森林土壤碳储量的变化。本文从森林土壤碳收支的角度,将氮沉降对森林生态系统土壤碳库影响的复杂过程划分为凋落物分解、细根周转、土壤呼吸和土壤可溶性有机碳淋失4个相对独立的过程。综合国内外研究现状,对其进行了简要评述,指出了目前研究的不足,并探讨了这一研究领域的发展方向。     

不同形态氮素对专用型小麦花后氮代谢关键酶活性及籽粒蛋白质含量的影响

采用盆栽方法研究了氮素形态对不同专用型小麦开花后氮素同化关键酶活性及籽粒蛋白质含量的影响。结果表明:不同专用型小麦氮素同化关键酶硝酸还原酶、谷氨酰胺合成酶和谷氨酸合酶对氮素形态的反应不同。强筋小麦豫麦34施用酰胺态氮对旗叶硝酸还原酶和谷氨酰胺合成酶活性、籽粒谷氨酰胺合成酶和谷氨酸合酶活性具有明显的促进作用,最终籽粒蛋白质含量较高;中筋小麦豫麦4 9在施用铵态氮时,3种氮素同化关键酶活性均有较大增强,籽粒蛋白质含量最高;弱筋小麦豫麦5 0硝酸还原酶活性以铵态氮处理最高,而籽粒和旗叶谷氨酰胺合成酶和谷氨酸合酶活性在酰胺态氮处理下明显增强,酰胺态氮对籽粒中蛋白质含量的增加具有明显的促进作用。相关性分析表明,籽粒蛋白质含量与旗叶GS活性和籽粒GOGAT活性呈显著或极显著正相关,与旗叶NR活性和GS活性、籽粒GOGAT活性相关性不显著     

Effects of nitrogen deposition on the interaction between an aphid and its host plant

Abstract 1. Anthropogenic increases in nitrogen deposition are impacting terrestrial ecosystems worldwide. While some of the direct ecosystem‐level effects of nitrogen deposition are understood, the effects of nitrogen deposition on plant–insect interactions and on herbivore population dynamics have received less attention. 2. Nitrogen deposition will potentially influence both plant resource availability and herbivore population growth. If increases in herbivore population growth outstrip increases in resource availability, then increases in the strength of density dependence expressed within the herbivore population would be predicted. Alternatively, if plant resources respond more vigorously to nitrogen deposition than do herbivore populations, a decline in the strength of density dependence would be expected. No change in the strength of density dependence acting upon the herbivore population would suggest equivalent responses by herbivores and plants. 3. A density manipulation experiment was performed to examine the effect of nitrogen deposition on the interaction between a host plant, Asclepias tuberosa, and its herbivore, Aphis nerii. Aphid maximum per capita growth rate (R_max), carrying capacity (K), and the strength of density dependence were measured under three nitrogen deposition treatments. The effect of nitrogen deposition on the relationship among these three measures of insect population dynamics was explored. 4. Simulated nitrogen deposition increased aphid per capita population growth, plant foliar nitrogen concentrations, and plant biomass. Nitrogen deposition caused R_max and K to increase proportionally, leading to no overall change in the strength of density dependence. In this system, potential changes in the negative feedback processes operating on herbivore populations following nitrogen deposition appear to be buffered by concomitant changes in resource availability.