15N自然丰度法在陆地生态系统氮循环研究中的应用

随着氮沉降的不断增加以及人们对全球变化问题的日益关注, 稳定同位素技术在全球变化研究中得到广泛的应用。因为植物和土壤的氮同位素组成记录了氮循环影响因子的综合作用, 并且具有测量简单以及不受取样时间和空间限制的优点, 所以氮同位素自然丰度法被用于氮循环的研究中。该文从氮循环过程中植物和土壤的氮分馏入手, 总结国内外相关文献, 阐述了植物和土壤氮自然丰度在预测生态系统氮饱和和氮循环长期变化趋势中的应用; 总结了利用树轮δ ¹⁵N法研究氮循环过程中应该注意的事项以及目前尚未解决的问题。     

利用15N示踪技术研究木荷与马尾松幼苗叶片对NO2的吸收与分配

大气氮氧化物(NO_x=NO+NO₂)随着干沉降进入森林生态系统时,会首先接触森林冠层。森林乔木能通过叶片吸收多少NO₂以及对吸收的NO₂是如何分配的,目前尚不清楚。该研究利用¹⁵N稳定同位素示踪技术,对中国南方常见乔木树种木荷(Schima superba)和马尾松(Pinus massoniana)幼苗在黑暗和光照两种条件下进行了¹⁵NO₂静态箱熏蒸实验,检测并分析了两种植物的¹⁵N回收率以及吸收的NO₂在植物各组织中的分配结果。结果显示:植物主要通过气孔吸收NO₂,木荷和马尾松在黑暗条件下整体分别能回收10.3%±5.9%和20.4%±7.0%¹⁵NO₂,在光照条件下整体分别能回收35.9%±5.4%和68.2%±7.6%¹⁵NO₂。两种植物各组织中的平均干质量¹⁵     

15N自然丰度法在生态系统氮素循环研究中的应用

稳定性同位素技术是现代生态学研究中的一门新兴技术,在生态学研究的诸多领域都展示了广阔的应用前景,其中,稳定性同位素＾１５Ｎ自然丰度法近年来在生态系统氮素循环研究中发挥了正在发挥着极为重要的作用,首先简述了自然生态系统氮素循环诸过程中的＾１５Ｎ同位素分馏机制,然后,在此基础上,综述了＾１５Ｎ自然丰度法的基本原理与方法,列举了近年 来此法在生物固氮及氮素转化过程研究中的一些应用实例,并预测了该方法在国     

土壤-植物-大气连续体系中氮的研究进展

氮素是植物最需要的重要养分元素之一.近年来,土壤-植物-大气这一连续体系(SPAC)中的氮循环成为研究的热点之一.大气中的氮素可以通过生物固定和N沉降等作用进入土壤和植物内,同时土壤和植物内的氮素又会以氨挥发和氮氧化物等方式排放到大气中.氮素通过生物固持和植物吸收等方式进入植物体内,植物器官脱落使植物损失一部分的氮素,另外雨水的淋洗和植物溢出液也会造成植物的N损失.植物氮素在植物体内的积累和分布随着生长时期和各营养器官而有所不同.另外,植物吸收氮素的过程又受到大气状况和土壤状况的制约.土壤中氮素经过矿化作用、硝化作用和反硝化作用进行转化,一部分把氮素转化成植物能吸收的营养形态,另一部分则从土壤中损失.凋落物的分解和N沉降能补充土壤中的氮素,而植物吸收、微生物固持、水文流失和N溢出等方式使氮素从土壤中损失出去.另外,凋落物的分解和根际土壤、CO2浓度和臭氧对氮素循环有着重要的作用.N污染、N沉降、碳氮循环的耦合作用是今后研究的热点问题.     

稻田土壤－作物－家畜系统中氮的循环研究

本文用Ｎ￣（１５）标记水稻和绿肥研究了稻田土壤－作物－家畜系统中氮的循环。Ｎ￣（１５）标记稻草喂羊,羊体回收饲料稻草Ｎ３１．１６％,羊粪２８．２６％,羊尿５．７２％,总回收６５．１４％,损失３４．８６％。将羊粪尿单施,稻谷回收饲料稻草Ｎ３．１９％,水稻全株回收４．８２％,土壤残留１９．００％,损失１０．１４％。故羊体、水稻及土壤残留共回收饲料稻草Ｎ５４．９８％．将羊粪与尿素配施,则饲料稻草Ｎ的总回收率为５５．８８％。Ｎ￣（１５）标记绿肥喂猪;猪体回收饲料绿肥Ｎ２３．５１％．猪粪２３．８５％,猪尿２８．７６％,总回收率７６．１２％,损失率２３．８８％。将猪粪、尿还田,稻谷回收饲料绿肥Ｎ６．６９％,水稻全株回收１０．０５％,土壤残留１９．１７％,故猪体、水稻和土壤残留共回收饲料绿肥Ｎ５２．７３％,将猪粪与尿素配施,则饲料绿肥Ｎ的总回收率为５２．７５％。     

等量分次施氮对冬枣15N和13C利用与分配特性的影响

以4年生盆栽冬枣为试材,采用¹³C、¹⁵N双标记示踪技术,在果实发育期研究了等氮量分次追施氮肥对冬枣植株¹⁵N和¹³C吸收、利用、积累和分配的影响.结果表明: 至果实采收期,冬枣各器官Ndff值(植株器官从肥料中吸收分配到的¹⁵N量对该器官全氮量的贡献率)随追氮次数的增多而显著增大.生殖器官(果实)和营养器官(叶片、枣吊、新生枣头枝和细根)的¹⁵N分配率以4次追氮处理最高,1次追氮处理最低,贮藏器官(主干、多年生枝和粗根)¹⁵N分配率的趋势相反;4次追氮处理¹⁵N利用率分别比1次和2次追氮处理高27.4%和15.5%.追氮次数越多,植株总氮量和¹⁵N吸收量越大;随时间的推移,1次追氮处理土壤¹⁵N丰度和总氮含量持续降低,2次追氮处理呈先升高后降低的趋势,4次追氮处理变化相对最为平稳,至处理后期显著高于其他处理;果实白熟至采收期,叶片叶绿素、氮含量和净光合速率均表现为4次追氮>2次追氮>1次追氮.不同处理¹³C同化物积累与分配不同.4次追氮处理¹³C固定总量分别是1次和2次追氮处理的1.1和1.2倍.增加追氮次数,促进了¹³C同化物向果实和贮藏器官的转移,而减少了向当年生营养器官的分配.综上,果实发育期4次追氮通过保证根层稳定、充足的氮素供应,提高了对氮素的吸收和利用,进而维持了较高的净光合速率,促进并优化了光合同化物的积累和分配,最有利于冬枣树体的生长及产量和品质的提高.     

羊草草地植被—土壤系统氮循环研究

研究表明,0－30cm土层7月氮（N）总储量为479．2g.m^-2,其中主要为有机N,占总N量的98．5％,土壤中的无机N年度变化很大,在2．55－11．3g.m^-2之间,7月无机N储量为7．3g.m^-2,与其它类型草地不同。该类型草地土壤铵态N与硝态N含量有些季节相差不大,有些季节硝态N的含量超过铵态N的含量,铵态N的峰值出现的时间早于硝态N。植物根系吸收利用的无机N约为3.48g.m^-2.a^-1,植物根系向地上每年输送的N量为2．97g.m^-2.a^-1,地上活体向地下转移的N量为1.54g.m^-2.a^-1,植物地上部分每年转为立估凋落物的N量为1.43g.m^-2.a^-1,由立枯凋落物转为土壤有机N的量大于1.08g.m^-2.a^-1,植物根系每年转为土壤有机N的量为1.51g.m^-2.a^-1。     

15N交叉标记有机与无机肥料氮的转化与残留

有机无机肥配施能够培肥土壤,改善土壤氮素供给,但目前有机无机肥配施主要集中在化肥氮的研究,忽略秸秆氮对化肥氮转化的影响。为了解秸秆还田对不同氮源转化和残留的影响,采用¹⁵N对尿素和水稻秸秆进行交叉标记,在两种不同肥力水稻土 (粘土矿物类型为1 ∶ 1型红黄泥和2 ∶ 1型紫潮泥) 进行水稻盆栽试验。设置对照(CK),单施尿素(¹⁵NU)、标记尿素与稻草配施(¹⁵NU-S) 和标记稻草与尿素配施(¹⁵NS-U)4个处理。结果表明,水稻吸收的氮素60%以上来自土壤氮,土壤氮素肥力相对较低的红黄泥较之紫潮泥对肥料氮的依赖更强;水稻生长期间微生物同化的尿素氮占标记底物的百分数红黄泥为1.8%-8.3%,紫潮泥为1.8%-19.2%;微生物同化的秸杆氮占标记底物的百分数红黄泥为1.7%-5.0%,紫潮泥为2.0%-6.2%。而粘土矿物固持的尿素氮占标记底物的百分数,红黄泥为0.3%-2.1%,紫潮泥为3.5%-18.7%;粘土矿物固持的秸杆氮红黄泥为0.2%-0.9%,紫潮泥为1.7%-5.0%。水稻成熟期尿素氮的残留率,红黄泥¹⁵NU处理、¹⁵NU+S分别为14.5%和17.0%,紫潮泥分别为16.9%和17.1%。秸秆氮的残留率分别为红黄泥38.8%、紫潮泥41.5%;有机无机肥配施提高了微生物同化化肥氮的能力,降低了粘土矿物晶格固持化肥氮的水平。有机无机配施提高了化肥氮利用率同时,提高了有机形态氮残留,降低了无机形态氮(矿质氮+固定态铵)的残留。     

15N示踪控释氮肥的氮肥利用率及去向研究

选用¹⁵N同位素标记的新型回收塑料包膜控释肥和大颗粒尿素,采用池栽试验研究夏玉米-冬小麦轮作体系中肥料氮的去向及利用率。结果表明,整个轮作体系中,控释肥处理（PCU）作物吸收的肥料氮为241.03 kg/hm,高于尿素处理（Urea)的211.02 kg/hm。控释肥处理施用的肥料氮主要残留在0~40 cm土层,而尿素处理则残留在0~60 cm土层,控释肥延缓了肥料氮向土壤深层迁移的趋势。在夏玉米和冬小麦轮作体系中,控释肥处理的氮肥利用率（32.86%,32.47%）高于尿素处理（28.23%,30.16%）。在冬小麦季,控释肥处理损失率相比尿素处理从36.07% 降至28.75%,而夏玉米季,控释肥处理损失率相比尿素处理从37.17%降至29.50%。玉米季控释肥处理与尿素处理差异不显著,但在冬小麦季控释肥处理的产量显著高于尿素处理。因此,在玉米和小麦整个生长季,新型回收塑料包膜控释肥的养分释放与作物养分需求吻合,既提高氮肥利用率,也降低了肥料氮的损失。     

入侵植物与菌根真菌互作及其对土壤氮循环的影响研究进展

在全球气候变化背景下,植物入侵现象日益加剧,入侵植物通过降低本地生物多样性,改变土壤微生物群落结构与组成,影响生态系统结构与功能,显著改变土壤氮循环等生态过程。土壤氮循环是生态系统物质循环的重要环节,影响生态系统中氮供应与分配,而全球气候变化和植物入侵正在改变土壤氮循环的效率和途径。菌根作为真菌和植物根系的重要共生体,在土壤氮循环过程中起着至关重要的作用。目前,对于菌根真菌与入侵植物互作对土壤氮循环的影响,仍缺乏系统研究和深入理解。该文综述了近年来关于入侵植物与菌根真菌互作对土壤氮循环影响的相关研究进展,重点分析了入侵植物与菌根真菌互作通过调控土壤微生物群落,影响土壤硝化、反硝化作用及相关土壤酶活性,改变土壤理化性质等机制影响土壤氮循环,并对未来研究方向提出展望。该研究为理解入侵植物在全球土壤氮循环中的作用提供新的视角,并为入侵植物管理与植物入侵影响下氮循环响应评估提供理论依据。     

Net and gross incorporation of nitrogen by marine copepods fed on N-labelled diatoms: Methodology and trophic studies

The stable isotope of nitrogen (¹⁵N) and an appropriate three-compartment model were used in two 24-h lasting feeding experiments to trace the flow of N through the copepod Acartia discaudata and Calanus helgolandicus fed on ¹⁵N-labelled Skeletonema costatum and Thalassiosira weissflogii, respectively. Details of the labelling technique and principles of the computation of N transport rates are given. At the end of a single 24-h feeding period only about one third of the total amount of N ingested by A. discaudata was incorporated into the copepod's body N; we refer to this rate as net incorporation. Most of the N ingested was lost as ammonium (48% of total N ingested), followed by losses in the form of eggs + fecal pellets (13%) and dissolved organic N (DON, 9%). The sum of net incorporation and the latter losses is defined as gross incorporation. Net incorporation by C. helgolandicus and N losses did not vary over time during a 24 h lasting time-series feeding experiment. On average, 79% of total N ingested was actually incorporated by the copepod whereas mean N losses as ammonium, eggs + fecal pellets represented only 12 and 9%, respectively. After a 24-h feeding period only 2% of N ingested was lost as DON. Inspection of individual DON pathways showed that both A. discaudata and C. helgolandicus highly contributed to total DON production via direct excretion (79 and 64%, respectively). The remaining DON appearing in the DON pool was derived from phytoplankton via direct release and/or indirect release (copepod ‘sloppy feeding’).     

Forests losing large quantities of nitrogen have elevated 15N:14N ratios

Summary Urea (U) and ammonium nitrate (AN) had been applied to a Scots pine (Pinus sylvestris L.) forest in northern Sweden for 18 consecutive years at four doses resulting in total N applications ranging from 0 to 1980 kg ha^–1. The ¹⁵N abundance ( ¹⁵N) of the grass Deschampsia flexuosa (L.) Trin. increased linearly (from –0.7 to 11.0) with application rate in the case of U. The response to AN was in the same direction but smaller. While others have shown that the initial response of nitrogen-limited systems to additions of N is a change of ¹⁵N abundance towards that of added N, this study shows that further and excessive additions leads to a retention of ¹⁵N. Monitoring ¹⁵N abundance over time in dose-response trials of this type thus opens new possibilities to estimate critical loads of N and the point of nitrogen saturation.     

Contributions of benthic and planktonic primary producers to nitrate and ammonium uptake fluxes in a nutrient-poor shallow coastal area (Corsica, NW Mediterranean)

By using the stable isotope ¹⁵N, we have measured in situ the uptake of nitrate and ammonium by the seagrass Posidonia oceanica, its leaf epiphyte community, the brown macroalgae Halopteris scoparia and the suspended particulate organic matter (SPOM). In Revellata Bay (Gulf of Calvi, Western Corsica), which is a very nutrient-poor region, the specific uptake rates (V) (μg N g N⁻¹ h⁻¹) of SPOM measured at ambient concentrations are 10-1000 higher than those of benthic primary producers. Macroalgae have intermediary V, between the seagrass leaf and leaf epiphytes. V are quite variable and the reasons for this variability remain unclear.Despite the difference of specific uptake rates found between benthic and pelagic primary producers, when integrating the uptake fluxes for a water column of 10 m depth, the contribution of benthic primary producers to N uptake fluxes (g N m⁻² h⁻¹) is significant, corresponding on average to 40% of total uptake flux. This results from the dominance in terms of N biomass of benthic primary producers in this shallow nutrient-poor area. When reported for the entire volume of the Revellata Bay, the contribution of benthic primary producers is reduced to 5-10% of total N uptake flux.Although this contribution could appear relatively low, it results in a significant direct transfer of inorganic nitrogen from the water column to the benthic compartment. By this transfer, the benthic plants act as a biological pump incorporating the pelagic N into the benthic compartment for a time longer than the characteristic time of phytoplankton dynamics (month-years vs. day-week).     

Flow of Deposited Inorganic N in Two Gleysol-dominated Mountain Catchments Traced with <Superscript>15</Superscript>NO<Subscript>3</Subscript><Superscript>−</Superscript> and <Superscript>15</Superscript>NH<Subscript>4</Subscript><Superscript>+</Superscript>

In two mountain ecosystems at the Alptal research site in central Switzerland, pulses of ¹⁵NO₃ and ¹⁵NH₄ were separately applied to trace deposited inorganic N. One forested and one litter meadow catchment, each approximately 1600 m², were delimited by trenches in the Gleysols. K¹⁵NO₃ was applied weekly or fortnightly over one year with a backpack sprayer, thus labelling the atmospheric nitrate deposition. After the sampling and a one-year break, ¹⁵NH₄Cl was applied as a second one-year pulse, followed by a second sampling campaign. Trees (needles, branches and bole wood), ground vegetation, litter layer and soil (LF, A and B horizon) were sampled at the end of each labelling period. Extractable inorganic N, microbial N, and immobilised soil N were analysed in the LF and A horizons. During the whole labelling period, the runoff water was sampled as well. Most of the added tracer remained in both ecosystems. More NO₃⁻ than NH₄⁺ tracer was retained, especially in the forest. The highest recovery was in the soil, mainly in the organic horizon, and in the ground vegetation, especially in the mosses. Event-based runoff analyses showed an immediate response of ¹⁵NO₃⁻ in runoff, with sharp ¹⁵N peaks corresponding to discharge peaks. NO₃⁻ leaching showed a clear seasonal pattern, being highest in spring during snowmelt. The high capacity of N retention in these ecosystems leads to the assumption that deposited N accumulates in the soil organic matter, causing a progressive decline of its C:N ratio.     

Abiotic immobilization of nitrate in two soils of relic <Emphasis Type="Italic">Abies pinsapo</Emphasis>-fir forests under Mediterranean climate

Evidence for abiotic immobilization of nitrogen (N) in soil is accumulating, but remains controversial. Identifying the fate of N from atmospheric deposition is important for understanding the N cycle of forest ecosystems. We studied soils of two Abies pinsapo fir forests under Mediterranean climate seasonality in southern Spain—one with low N availability and the other with symptoms of N saturation. We hypothesized that biotic and abiotic immobilization of nitrate (NO₃ ⁻) would be lower in soils under these forests compared to more mesic temperate forests, and that the N saturated stand would have the lowest rates of NO₃ ⁻ immobilization. Live and autoclaved soils were incubated with added ¹⁵NO₃ ⁻ (10 μg N g⁻¹ dry soil; 99% enriched) for 24 h, and the label was recovered as total dissolved-N, NO₃ ⁻, ammonium (NH₄ ⁺), or dissolved organic-N (DON). To evaluate concerns about possible iron interference in analysis of NO₃ ⁻ concentrations, both flow injection analysis (FIA) and ion chromatography (IC) were applied to water extracts, soluble iron was measured in both water and salt extracts, and standard additions of NO₃ ⁻ to salt extracts were analyzed. Good agreement between FIA and IC analysis, low concentrations of soluble Fe, and 100% (±3%) recovery of NO₃ ⁻ standard additions all pointed to absence of an interference problem for NO₃ ⁻ quantification. On average, 85% of the added ¹⁵NO₃ ⁻ label was recovered as ¹⁵NO₃ ⁻, which supports our hypothesis that rates of immobilization were generally low in these soils. A small amount (mean = 0.06 μg N g⁻¹ dry soil) was recovered as ¹⁵NH₄ ⁺ in live soils and none in sterilized soils. Mean recovery as DO¹⁵N ranged from 0.6 to 1.5 μg N g⁻¹ dry soil, with no statistically significant effect of sterilization or soil type, indicating that this was an abiotic process that occurred at similar rates in both soils. These results demonstrate a detectable, but modest rate of abiotic immobilization of NO₃ ⁻ to DON, supporting our first hypothesis. These mineral soils may not have adequate carbon availability to support the regeneration of reducing microsites needed for high rates of NO₃ ⁻ reduction. Our second hypothesis regarding lower expected abiotic immobilization in soils from the N-saturated site was not supported. The rates of N deposition in this region may not be high enough to have swamped the capacity for soil NO₃ ⁻ immobilization, even in the stand showing some symptoms of N saturation. A growing body of evidence suggests that soil abiotic NO₃ ⁻ immobilization is common, but that rates are influenced by a combination of factors, including the presence of plentiful available carbon, reduced minerals in anaerobic microsites and adequate NO₃ ⁻ supply.     

A further extension of the method for independently measuring the amount of nitrogen fixed by a legume crop

Summary The combination of using¹⁵N for determining the amount of nitrogen fixed by a legume crop in field experiments and the labelling of only one treatment at a time in each treatment combination is shown to be conceptually and experimentally valid for determining the effect of cultural practices on the amount of nitrogen fixed by a legume crop.     

A15N tracer study to compare nitrogen supply by Azolla and ammonium sulphate to IR8 rice plants grown under flooded conditions

Summary A pot experiment was carried out using a Bangladesh sandy loam paddy soil of pH 6.9 to compare the rates at which nitrogen from Azolla and ammonium sulphate was available to a high yielding rice variety, IR8, grown for 60 days in pots with 4 cm standing flood water.¹⁵N tracer studies confirm that nitrogen from ammonium sulphate was more available to the rice plants than from Azolla. An application of 6, 9 and 18 mg N of Azolla pot^–1 (each pot contained 250 g soil) increased shoot dry matter yields by 13, 29 and 49% for an uptake of 19, 36 and 85% more nitrogen; the corresponding increases on using ammonium sulphate were 33, 54 and 114% for an increased uptake of 57, 90 and 177% more nitrogen, respectively. About 34% of applied¹⁵N of Azolla was taken up by the rice plants in 60 days but 61% of¹⁵N of the ammonium sulphate was absorbed during this period. About 45% of the Azolla-N was released in 60 days, 55% remained in the soils as undecomposed material and 11% was lost as gas. The gaseous loss of¹⁵N from ammonium sulphate was 14%; 25% remained in the soils.     

Leaf vs. epiphyte nitrogen uptake in a nutrient enriched Mediterranean seagrass (Posidonia oceanica) meadow

In situ nitrogen uptake by leaves and epiphytes was studied in a Mediterranean seagrass (Posidonia oceanica) meadow impacted from a fish farm and a pristine meadow, using ¹⁵NH₄ and ¹⁵NO₃ as tracers. In the impacted meadow both leaves and epiphytes yielded higher N concentrations and showed higher specific N uptake, suggesting a linkage between N uptake and its accumulation. Epiphytes took up N faster than leaves in relation to their corresponding biomass, but when assessed per unit area, N uptake was higher in leaves. Leaf N uptake was negatively correlated with epiphyte N uptake. With increasing epiphyte load on leaves, N leaf uptake decreased while N epiphyte uptake increased, indicating that epiphyte overgrowth hinders N uptake by P. oceanica leaves. Epiphyte contribution to total N uptake increased, while that of leaves decreased at the impacted meadow. However, 2-3 times less N was transferred daily from the water column to the benthic compartment, through seagrass and epiphyte uptake on total, at the impacted meadow. Therefore, it is probably still the loss of the key species - the seagrass - which plays the most important role in N cycling in seagrass ecosystems.