15N标记的土壤估测某些豆科植物N2固定的温室实验

Pot experiments were carried out to estimate N2 fixation by vetch,milk vetch,sickle alfalfa and broadbean in pure stand using a ^15N-labelled soil.Winter wheat was used as the non-fixing control.The 15N-labelled soil used was prepared by growing corn-wheat-corn successively on a nearly organic-matter-free Xiashu loess supplemented with adequate amounts of (15NH4)2SO4,P,K and micronutrients,then incorporating these 15N-labelled plant materials into the soil after each havest,and allowing the plant materials to be decomposed aerobically for 410d after incorporation of the plant material of the thire crop.The 15N enrichment of wheat plant-N varied slightly with organs,with a maximum difference of 9.8%,Based on 15N enrichment of soil N inferred from the mean value of the 15N enrichment in different organs of wheat 79%-91% of total N in the tops and 67%-74% of total N in the roots of legumes studied were derived from atmosphere .Estimate by isotope dilution method was in good agreement with that by the conventional difference method provided values obtained by the latter were corrected for seed N,and also with that from the measurement of N accumulated in the tops of the legumes.     

Influence of mycorrhiza vs. soluble phosphate on growth,nodulation, and N2 fixation (15N) in alfalfa under different levels of water potential

Summary The legume Medicago sativa (+Rhizobium melilott) was grown under controlled conditions to study the interactions between soluble P in soil (four levels), or a mycorrhizal inoculum, and the degree of water potential (four levels) in relation to plant development and N₂ fixation. ¹⁵N-labelled ammonium sulphate was added to each pot for a qualitative estimate of N₂ fixation, in order to rank the effects of the different treatments.Dry-matter yield, nutrient content and nodulation increased with the amount of plant-available P in the soil, and decreased as the water stress increased, for each P-level. The mycorrhizal effect on dry matter, N yield, and on nodulation was little affected by the water potential. Since P uptake was affected by the water content in mycorrhizal plants, additional mechanisms, other than those mediated by P, must be involved in the mycorrhizal activity.There was a positive correlation between N yield and nodulation for the different P levels and the mycorrhizal treatment at all water levels. A high correlation between plant unlabelled N content and atom% ¹⁵N excess was also found for all levels of P. In mycorrhizal plants, however, the correlation between unlabelled N yield and ¹⁵N was lower. This suggests that mycorrhiza supply plants with other N sources in addition to those derived from the improvement on N₂ fixation.     

Influence of arbuscular mycorrhizae and phosphorus fertilization on growth, nodulation and N2 fixation (15N) inMedicago sativa at four salinity levels

The rose of an isolate of the arbuscular mycorrhizal (AM) fungusGlomus mosseae in the protection ofMedicago sativa (+Rhizobium meliloti) against salt stress induced by the addition of increasing levels of soluble salts was studied. The interactions between soluble P in soil (four levels), mycorrhizal inoculum and degree of salinity in relation to plant growth, nutrition and infective parameters were evaluated. Salt stress was induced by sequential irrigation with saline water having four concentrations of three salts (NaCl, CaCl₂, and MgCl₂).¹⁵N-labelled ammonium sulphate was added to provide a quantitative estimate of N₂ fixation under moderate to high salinity levels. N and P concentration and nodule formation increased with the amount of plant-available P or mycorrhizal inoculum in the soil and generally declined as the salinity in the solution culture increased from a moderate to a high level. The mycorrhizal inoculation protected the plants from salt stress more efficiently than any amount of plant-available P in soil, particularly at the highest salinity level applied (43.5 dS m^?1). Mycorrhizal inoculation matched the effect on dry matter and nutrition of the addition in the soil of 150 mg P kg^?1. Nevertheless the highest saline solution assayed (43.5 dS m^?1) affected more severely plants supplemented with phosphorus than those with the addition of mycorrhizal inoculum. Such a saline-depressing effect was 1.5 (biomass), 1.4 (N) and 1.5 (P) times higher in plants supplied with soluble phosphate than with AM inoculum. Mechanisms beyond those mediated by P must be involved in the AM-protectioe effect against salinity. The¹⁵N methodology used allowed the determination of N₂ fixation as influenced by different P applications compared to mycorrhizal inoculation. A lack of correlation between nodule formation and function (N₂ fixation) was evidenced in mycorrhizal-inoculated plants. In spite of the reduced activity per nodule in mycorrhizal-inoculated In spite of the reduced activity per nodule in mycorrhizal-inoculated plants, the N contents determined indicated the highest acquisition of N occurred in plants with the symbiotic status. Moreover, N and P uptake increased while Ca and Mg decreased in AM-inoculated plants. Thus P/Ca ratios and cation/anion balance in general were altered in mycorrhizal treatments. This study therefore confirms previous findings that AM-colonized plants have optional and alternative mechanisms available to satisfy their nutritive requirements and to maintain their physiological status in stress situations and in disturbed ecosystems.     

Paddy System with a Hybrid Rice Enhances Cyanobacteria Nostoc and Increases N2 Fixation

Biological nitrogen(N) fixation(BNF) plays a significant role in maintaining soil fertility in paddy field ecosystems. Rice variety influences BNF, but how different rice varieties regulate BNF and associated diazotroph communities has not been quantified. Airtight,field-based ~(15)N_2-labelling growth chamber experiments were used to assess the BNF capacity of different rice varieties. In addition,both the 16 S rRNA and nifH genes were sequenced to assess the influence of different rice varieties on bacterial and diazotrophic communities in paddy soils. After subjecting a rice-soil system to 74 d of continuous airtight, field-based ~(15)N_2 labelling in pots in a growth chamber, the amounts of fixed N were 22.3 and 38.9 kg ha~(-1) in inbred japonica(W23) and hybrid indica(IIY) rice cultivars planted in the rice-soil systems, respectively, and only 1%–2.5% of the fixed N was allocated to the rice plants and weeds. A greater abundance of diazotrophs was found in the surface soil(0–1 cm) under IIY than under W23. Sequencing of the 16 S rRNA gene showed significantly greater abundances of the cyanobacterial genera Nostoc, Anabaena, and Cylindrospermum under IIY than under W23.Sequencing of the nifH gene also showed a significantly greater abundance of Nostoc under IIY than under W23. These results indicate that the hybrid rice cultivar(IIY) promoted BNF to a greater extent than the inbred rice cultivar(W23) and that the increase in BNF might have been due to the enhanced heterocystous cyanobacteria Nostoc.     

Yield,nodulation, and N2 fixation by cowpea cultivars at different phosphorus levels

A greenhouse experiment was conducted to investigate the effect of a P application (0 vs. 50 mg P kg^-1) on yield, nodulation, and N₂ fixation by three cowpea cultivars (Soronko, Amantin, and IT81D-1137) using the ¹⁵N isotope-dilution method. When P was not applied the inoculated cowpea genotypes showed significant differences (Soronko>Amantin> IT81D-1137) in N accumulation, in contrast to the uninoculated cowpea cultivars, which accumulated similar amounts of N. The differences in shoot N in inoculated plants were thus caused by differences in N₂ fixation. The average values of N fixed (for both P levels) were 74% in Soronko, 59% in Amantin, and 42% in IT81D-1137, corresponding to 80, 51, and 24 mg N plant^-1, respectively. Inoculation increased the total shoot-N accumulation in cv. Soronko by 270% without P and by 204% with P, cv. Amantin by 152 and 104%, and cv. IT81D-1137 by 74 and 58%, respectively. With P, the % N derived from atmosphere (%Ndfa) was 42% for IT81D-1137, 62% for Amantin, and 76% for Soronko. The high value for Soronko indicates that in a soil of medium fertility, certain cowpea cultivars are capable of satisfying their total N requirement through N₂ fixation. The P effect on N₂ fixation was mainly in the total amount of N fixed rather than on the percentage derived from the atmosphere.     

Biomass,nodulation and N2 fixing response by subclover and pink serradela to phosphorus fertilization

In Mediterranean areas, sustainable production of rainfed pastures is associated with plant species which withstand low soil fertility and variable annual drought, phosphorus (P) being the primary limiting nutrient for legume growth and N₂ fixation. A study was undertaken under environmentally controlled conditions to compare the response of subclover and pink serradela to P supply using a moderately acid soil, very poor in available P, in terms of biomass, nodulation pattern, N₂ fixation and P use efficiency. A P dose in the range of 30–60 kg P ha^?1 is recommended as a starter for pastures under the present conditions. A higher P rate reduced the fixing nodule number and biomass, and the N₂ fixation rate. Fixed N₂ per unit of nodule P by pink serradela as monocrop was about 80% greater than the sole subclover, demonstrating its higher efficiency on the use of nodule P. Thus, pink serradela needs a lower P rate to produce the same amount of biomass per unit area as sole subclover. Long-term studies in field conditions, using these and other new pasture legume species are needed to consolidate the present findings.     

利用15N2直接标记法研究水稻种植对稻田固氮量和固氮活性的影响

稻田固氮对土壤维持肥力有着重要的作用,但水稻种植与固氮菌及其活性之间的关系尚不清楚。本试验利用¹⁵N₂直接标记法测定了下位砂姜土发育的简育水耕人为土在种水稻和不种水稻条件下的生物固氮量,及其在土壤不同层次(0～1、1～5、5～15 cm)和水稻中的分配,并通过实时荧光定量PCR技术测定了土壤中固氮菌nifH DNA及RNA基因数量。结果表明:种水稻处理显著提高了土壤各层固氮量,尤其提高了1～5 cm和5～15 cm土层土壤固氮量对总固氮量的贡献;种水稻处理的总固氮量是不种水稻处理的10.3倍;水稻植株中生物固定的氮占总固氮量的31.48%;在0～1 cm土层,种水稻处理显著提高了nifH RNA基因数量,而对nifH DNA基因数量的增加不显著。可见,水稻种植没有增加固氮菌的数量,稻田固氮量的增加是因为水稻种植极大地促进了固氮菌nifH基因的表达,提高了固氮菌的固氮活性。     

15N tracers and microbial analyses reveal in situ N2O sources in contrasting water regimes of a drained peatland forest

Managed peatlands are a significant source of nitrous oxide (N₂O), a powerful greenhouse gas and stratospheric ozone depleter. Due to the complexity and diversity of microbial N₂O processes, different methods such as tracer, isotopomer, and microbiological technologies are required to understand these processes. The combined application of different methods helps to precisely estimate these processes, which is crucial for the future management of drained peatlands, and to mitigate soil degradation and negative atmospheric impact. In this study, we investigated N₂O sources by combining tracer, isotopomer, and microbial analysis in a drained peatland forest under flooded and drained treatments. On average, the nitrification genes showed higher abundances in the drained treatment, and the denitrification genes showed higher abundances in the flooded treatment. This is consistent with the underlying chemistry, as nitrification requires oxygen while denitrification is anaerobic. We observed significant differences in labelled N₂O fluxes between the drained and flooded treatments. The emissions of N₂O from the flooded treatment were nearly negligible, whereas the N₂O evolved from the nitrogen-15 (¹⁵N)-labelled ammonium (¹⁵NH⁺₄) in the drained treatment peaked at 147 μg ¹⁵N m^-2 h^-1. This initially suggested nitrification as the driving mechanism behind N₂O fluxes in drained peatlands, but based on the genetic data, isotopic analysis, and N₂O mass enrichment, we conclude that hybrid N₂O formation involving ammonia oxidation was the main source of N₂O emissions in the drained treatment. Based on the ¹⁵N-labelled nitrate (¹⁵NO-₃) tracer addition and gene copy numbers, the low N₂O emissions in the flooded treatment came possibly from complete denitrification producing inert dinitrogen. At atomic level, we observed selective enrichment of mass 45 of N₂O molecule under ¹⁵NH⁺₄ amendment in the drained treatment and enrichment of both masses 45 and 46 under ¹⁵NO-₃ amendment in the flooded treatment. The selective enrichment of mass 45 in the drained treatment indicated the presence of hybrid N₂O formation, which was also supported by the high abundances of archaeal genes.     

Emission of N2O, N2 and CO2 from soil fertilized with nitrate: effect of compaction, soil moisture and rewetting

Soil compaction and soil moisture are important factors influencing denitrification and N₂O emission from fertilized soils. We analyzed the combined effects of these factors on the emission of N₂O, N₂ and CO₂ from undisturbed soil cores fertilized with (150 kg N ha⁻¹) in a laboratory experiment. The soil cores were collected from differently compacted areas in a potato field, i.e. the ridges (ρ_D=1.03 g cm⁻³), the interrow area (ρ_D=1.24 g cm⁻³), and the tractor compacted interrow area (ρ_D=1.64 g cm⁻³), and adjusted to constant soil moisture levels between 40 and 98% water-filled pore space (WFPS).High N₂O emissions were a result of denitrification and occurred at a WFPS≥70% in all compaction treatments. N₂ production occurred only at the highest soil moisture level (≥90% WFPS) but it was considerably smaller than the N₂O-N emission in most cases. There was no soil moisture effect on CO₂ emission from the differently compacted soils with the exception of the highest soil moisture level (98% WFPS) of the tractor-compacted soil in which soil respiration was significantly reduced. The maximum N₂O emission rates from all treatments occurred after rewetting of dry soil. This rewetting effect increased with the amount of water added. The results show the importance of increased carbon availability and associated respiratory O₂ consumption induced by soil drying and rewetting for the emissions of N₂O.     

N2O产生法测定土壤无机态氮15N丰度

用化学方法分别将土壤中微量的铵、硝酸盐和亚硝酸盐转化为N2O气体,然后用带自动预浓缩装置的同位素比值质谱仪测定N2O中的15N丰度.N2O中的15N丰度测量值完全符合铵、硝酸盐和亚硝酸盐的15N参考值.方法快速、简单和准确,不受空气氮的污染.特别是方法的检测限很低,每批次样品中只需含5～ 20μg N.它将有助于土壤氮素的矿化作用、硝化作用和反硝化作用的研究.     

N2 fixation and growth of legumes as affected by sulphur fertilization

The influence of three sulphur application rates in combination with two nitrogen application rates on N₂ fixation and growth of different legumes was investigated. N was applied as N-labelled ¹⁵NH₄ ¹⁵NO₃. The ¹⁵N isotope dilution technique was used to estimate N₂ fixation. At both N increments dry matter yield was highest with high S supply. Independently of the N supply, the high S application rate resulted in a significantly higher N accumulation, which was mainly caused by a higher N₂ fixation rate. With the grain legumes the weight of nodules was increased by the high S application rate. The higher number of nodules per pot with optimum S supply was the result of a better root growth. Rates of acetylene reduction correlated significantly with S supply.     

CO2浓度升高条件下不同程度豌豆蚜危害对紫花苜蓿叶片营养物质和次生代谢物质的影响

由于人类大量开采使用石油、煤炭、天然气等化石燃料,使大气CO₂浓度升高,这不但加速全球变暖,还将影响地球上动植物的生存和分布,从而对整个生态系统产生深远影响。为探明CO₂浓度升高与豌豆蚜（Acyrthosiphon pisum）虫口密度对紫花苜蓿（Medicago sativa）叶片内化学物质的影响,明确CO₂浓度升高和蚜虫密度在紫花苜蓿生理生化中的作用,试验在CO₂光照培养箱内设置380 μL·L^-1（对照）、550 μL·L^-1和750 μL·L^-1 3个CO₂浓度培育苜蓿幼苗并接入10日龄成蚜10头·株^-1、20头·株^-1、30头·株^-1,并以0头·株^-1作为空白对照,1周后测定植物体内营养物质和次生代谢物质含量。结果表明,随CO₂浓度升高,蚜虫密度为30头·株^-1时紫花苜蓿可溶性蛋白、可溶性糖以及淀粉含量均上升,在750 μL·L^-1 CO₂浓度下分别比CK上升11.62倍、0.49倍和0.24倍;黄酮、总酚和简单酚含量也显著上升。随蚜虫危害程度加重,同一CO₂浓度下紫花苜蓿淀粉、简单酚含量先上升后下降,高CO₂浓度蚜虫密度为30头·株^-1时比0头·株^-1时可溶性糖、总酚以及单宁含量上升1.66倍、1.49 mg·g^-1和1.09 mg·g^-1,差异均显著（P<0.05）。说明具有固氮作用的豆科植物更易于适应CO₂浓度升高的变化,从而在受到刺吸胁迫后增强自身诱导抗虫性以抵御害虫为害。     

三氯生和三氯卡班对水稻土好氧氮转化及N_2O排放的影响

三氯生(Triclosan, TCS)和三氯卡班(Triclocarban, TCC)是典型的药品与个人护理用品,在土壤生态系统中被广泛检出,且存在增加土壤微生物抗药性及抑制土壤呼吸的潜在风险,但目前有关TCS和TCC对土壤氮转化过程及氧化亚氮(N_2O)排放的影响尚不清楚。基于此,采用室内培养实验和15N稀释-富集法,结合氮转化数值模型,研究了不同浓度梯度下TCS(2和5mg·kg~(-1))和TCC(1和2 mg·kg~(-1))的单独及联合存在对水稻土氮初级转化速率以及N_2O排放的影响。结果表明,1mg·kg~(-1)TCC及5mg·kg~(-1)TCS+2mg·kg~(-1)TCC处理对水稻土氮素的矿化-同化无显著影响,其余TCS和TCC处理均显著促进了氮的矿化-同化循环。此外,TCS和TCC处理显著降低了自养硝化速率、硝态氮的微生物固定速率以及硝酸盐异化还原成铵(Dissimilatory nitrate reduction to ammonium, DNRA)速率(2 mg·kg~(-1)TCS处理及5mg·kg~(-1)TCS+2mg·kg~(-1)TCC对DNRA速率无显著影响)。值得关注的是,TCS和TCC单一和联合处理均显著增加了N_2O的累积排放量,其累积排放量为对照的1.13倍～1.44倍。本研究表明,TCS和TCC改变了水稻土好氧氮转化过程,可能对稻田生态系统氮循环产生不利影响;TCC和TCS对水稻土N_2O排放的促进作用也增加了稻田生态系统对温室效应和臭氧层破坏的潜在贡献,因此,未来评价TCS和TCC土壤生态风险时,应考虑其对氮转化过程和N_2O排放的潜在影响。     

Estimating N2 fixation by field-grown lupins (Lupinus angustifolius L.) using soil and plant 15N enrichment

Summary Biological N₂ fixation was estimated in a field experiment following the addition of NH₄Cl or KNO₃ to unconfined microplots (1.5 m²) at 2.5 g N m^-2 (10 atom% ¹⁵N). A model of total N and ¹⁵N accumulation in lupins and decreasing ¹⁵N enrichment in the KCl-extractable soil-N pool (0–0.15 m depth) was used to estimate the proportion of N in lupins derived from biological N₂ fixation. Estimates of N₂ fixation derived from the model were compared with ¹⁵N isotope-dilution estimates obtained using canola, annual ryegrass, and wheat as nonfixing reference plants. Biomass, total N accumulation, or ¹⁵N enrichment in the lupin and reference crops did not differ whether NH _inf4 ^sup+ or NO _inf3 ^sup- was added as the labelled inorganic-N source. The decrease in soil ¹⁵N enrichment was described by first-order kinetics, whereas total N and ¹⁵N accumulation in the lupins were described by logistical equations. Using these equations, the uptake of soil N by lupins was estimated and was then used to calculate fixed N₂. Estimates of N₂ fixation derived from the model increased from 0 at 50 days after sowing to a maximum of 0.79 at 190 days after sowing. Those based on the ¹⁵N enrichment of the NO _inf3 ^sup- pool were 10% higher than those based on the mineral-N pool. ¹⁵N isotope-dilution estimates of N₂ fixation ranged from 0.37 to 0.55 at 68 days after sowing and from 0.71 to 0.77 at 190 days after sowing. Reference plant-derived values of N₂ fixation were all higher than modelled estimates during the early states of growth, but were similar to modelled estimates at physiological maturity. The use of the model to estimate N₂ derived from the atmosphere has the intrinsic advantage that the need for a non-fixing reference plant is avoided.     

Comparative growth and symbiotic performance of four Acacia mangium provenances from Papua New Guinea in response to the supply of phosphorus at various concentrations

Plant growth performance, the P content in root and nodule tissues, and nodulation and N₂-fixing ability were studied in four provenances of Acacia mangium from Papua New Guinea following different levels of P fertilizer application. A. mangium did not seem to need high levels of P for growth and N₂ fixation. The response by this leguminous tree to the P supply varied significantly according to provenance and to P concentrations in the culture solution. The provenances of A. mangium were classified into three types according to their P response: (1) Growth performance, nodulation, and N₂ fixation of plants were stimulated as concentrations of P increased (provenance PH 482); (2) the maximal effect of P on plant growth was found only at P concentrations higher than 500 M (provenance PH 484); and (3) the plant response to P fertilization was low, even with nutrient solutions containing P concentratins higher than 500 M (provenances PH 483 and PH 485). Provenance PH 483 was distinguished by its low nodulating ability. However, this provenance grew well, probably because of its high N₂ fixation efficiency as expressed by specific acetylene reduction activity and its high P content in nodule tissues. Therefore, in certain cases, these two parameters may be useful criteria in selecting leguminous plants for field use. Statistical analyses of the study results showed that the effect of the factor P supply on N₂ fixation efficiency and nodule development was only significant at P concentrations lower than 250 M whereas the effect of the factor plant provenance was significant regardless of the P concentration used. This observation emphasizes the value of provenance screening in the identification of plants for use in a wide range of soil types.     

庐山毛竹扩张及模拟氮沉降对土壤N_2O和CO_2排放的影响

毛竹是我国南方广泛分布的一种典型的森林资源,其扩张已引发了多方面的生态问题,但是目前关于氮沉降背景下毛竹扩张引起的土壤N_2O和CO_2气体排放变化的研究甚少,且无原位观测数据。采用静态箱-气相色谱法,分析江西庐山毛竹纯林、毛竹扩张形成的毛竹-日本柳杉混交林及日本柳杉纯林3种林分土壤的N_2O和CO_2排放速率和累积排放量及其对模拟氮沉降的响应。结果表明:(1)混交林土壤的NH_4~+-N含量、NO_3~--N含量及pH分别为14.39mg·kg~(-1)、8.65mg·kg~(-1)、4.88,显著高于日本柳杉纯林的9.75 mg·kg~(-1)、5.58 mg·kg~(-1)、4.05,但是混交林土壤DOC含量(236.5 mg·kg~(-1))却显著低于日本柳杉纯林(382.0mg·kg~(-1))。(2)混交林土壤N_2O累积排放量(393.6mg·m~(-2))显著高于毛竹纯林(202.5mg·m~(-2))和日本柳杉纯林(192.8mg·m~(-2)),混交林土壤CO_2累积排放量(4 655 g·m~(-2))显著高于日本柳杉纯林(2 815 g·m~(-2))。(3)模拟氮沉降未对3种林分类型土壤的CO_2排放速率和累积排放量产生显著影响,但明显增加了混交林和日本柳杉纯林的N_2O累积排放量。本研究表明:毛竹扩张不同阶段土壤的理化性质、N_2O及CO_2排放表现出不同特征。毛竹扩张过程中一定程度上增大了土壤N_2O和CO_2的排放量,但是完全扩张后N_2O排放出现明显下降趋势,而CO_2的排放未发生显著变化。同时,氮沉降促进了毛竹未扩张和扩张初期土壤的N_2O排放,而对CO_2排放未产生显著影响。表明在未来气候变化条件下管理亚热带毛竹扩张时,必须明确考虑这些生态系统组成、结构和影响因子之间的影响。     

Field measurements of nitrogen fixation in leguminous trees used in agroforestry systems: Influence of 15N-labeling approaches and reference trees

Appropriate ¹⁵N-labeling methods are crucial for estimating N₂-fixation in trees used in agroforestry systems. A 4-year field experiment was conducted on an Alfisol in Southwestern Nigeria to compare the estimates of N₂ fixed in Leucaena leucocephala, using two non-N₂-fixing leguminous trees, Senna siamea and S. spectabilis, as reference plants and three different methods of introducing ¹⁵N into soil. The atom % ¹⁵N uptake pattern (as reflected in the leaves) was identical in both N₂- and non-N₂-fixing tree species irrespective of the ¹⁵N-application method. There was a significant decline in atom % ¹⁵N excess in the leaves of L. leucocephala (from 0.266 to 0.039), S. siamea (0.625 to 0.121), and S. spectabilis (from 0.683 to 0.118) from the first sampling 12 months after planting and the second sampling 18 months after sampling. From the second harvest in 1991 until the end of the experiment (fifth) harvest in 1993, however, the atom ¹⁵N % excess decline in leaves of the three species was less pronounced and depended on the method of ¹⁵N application. In those plants to which the tracer was applied once at planting, the ¹⁵N decline was steady between the second and the last prunings. In the split-application treatment, the atom ¹⁵N % excess increased slightly at the third pruning and decreased during the subsequent two prunings. The reference tree and the method of ¹⁵N application influenced the estimated proportion of N derived from atmospheric N₂ by L. leucocephala, calculated as 73 and 64%, corresponding to 119 and 98 kg N ha^-1 of N₂ fixed per 6 months, when S. spectabilis and S. siamea were used as reference trees, respectively. The approach by which ¹⁵N-labeled fertilizer was applied to the soil in three splits gave slightly higher estimates of N derived from the atmosphere but this was of little agronomic significance because total N₂ fixed was similar for all methods.     

N2O and NO fluxes between a Norway spruce forest soil and atmosphere as affected by prolonged summer drought

Global change scenarios predict an increasing frequency and duration of summer drought periods in Central Europe especially for higher elevation areas. Our current knowledge about the effects of soil drought on nitrogen trace gas fluxes from temperate forest soils is scarce. In this study, the effects of experimentally induced drought on soil N₂O and NO emissions were investigated in a mature Norway spruce forest in the Fichtelgebirge (northeastern Bavaria, Germany) in two consecutive years. Drought was induced by roof constructions over a period of 46 days. The experiment was run in three replicates and three non-manipulated plots served as controls. Additionally to the N₂O and NO flux measurements in weekly to monthly intervals, soil gas samples from six different soil depths were analysed in time series for N₂O concentration as well as isotope abundances to investigate N₂O dynamics within the soil. N₂O fluxes from soil to the atmosphere at the experimental plots decreased gradually during the drought period from 0.2 to −0.0 μmol m⁻² h⁻¹, respectively, and mean cumulative N₂O emissions from the manipulated plots were reduced by 43% during experimental drought compared to the controls in 2007. N₂O concentration as well as isotope abundance analysis along the soil profiles revealed that a major part of the soil acted as a net sink for N₂O, even during drought. This N₂O sink, together with diminished N₂O production in the organic layers, resulted in successively decreased N₂O fluxes during drought, and may even turn this forest soil into a net sink of atmospheric N₂O as observed in the first year of the experiment. Enhanced N₂O fluxes observed after rewetting up to 0.1 μmol m⁻² h⁻¹ were not able to compensate for the preceding drought effect. During the experiment in 2006, with soil matric potentials in 20 cm depth down to −630 hPa, cumulative NO emissions from the throughfall exclusion plots were reduced by 69% compared to the controls, whereas cumulative NO emissions from the experimental plots in 2007, with minimum soil matric potentials of −210 hPa, were 180% of those of the controls. Following wetting, the soil of the throughfall exclusion plots showed significantly larger NO fluxes compared to the controls (up to 9 μmol m⁻² h⁻¹ versus 2 μmol m⁻² h⁻¹). These fluxes were responsible for 44% of the total emission of NO throughout the whole course of the experiment. NO emissions from this forest soil usually exceeded N₂O emissions by one order of magnitude or more except during wintertime.     

Estimating N2 fixation by Sesbania rostrata and S. cannabina (syn. S. aculeata) in lowland rice soil by the 15N dilution method

Summary A field experiment in concrete-based plots was conducted to estimate the contribution of N derived from air (Ndfa) or biological N₂ fixation in Sesbania rostrata and S. cannabina (syn. S. aculeata), using various references, by the ¹⁵N dilution method. The two Sesbania species as N₂-fixing reference plants and four aquatic weed species as non-N₂-fixing references were grown for 65 days after sowing in two consecutive crops, in the dry and the wet seasons, under flooded conditions. Soil previously labeled with ¹⁵N at 0.26 atom % ¹⁵N excess in mineralizable N was further labeled by ammonium sulfate with 3 and 6 atom % ¹⁵N excess. The results showed that ¹⁵N enrichment of soil NH ₄ ⁺ -N dropped exponentially in the first crop to half the original level in 50 days while in the second crop, it declined gradually to half the level in 130 days. The decline in ¹⁵N enrichment, in both N₂-fixing and non-fixing species, was also steeper in the first crop than in the second crop. Variations in ¹⁵N enrichment among non-fixing species were smaller in the second crop. The ratio of the uptake of soil N to that of fertilizer N in N₂-fixing and non-fixing species was estimated by the technique of varying the ¹⁵N level. In the second crop, this ratio in non-fixing species was higher than that in N₂-fixing species. Comparable estimates of % Ndfa were obtained by using ¹⁵N enrichment of various non-fixing species. There was also good agreement between the estimates obtained by using ¹⁵N enrichment of non-fixing species and those by using soil NH ₄ ⁺ -N, particularly in the second crop. By 25 days after sowing, the first crop of both Sesbania spp. had obtained 50% of total N from the atmosphere and the second crop had obtained 75%. The contribution from air increased with the age of the plant and ranged from 70% to 95% in 45–55 days. S. rostrata fixed substantially higher amounts of N₂ due to its higher biomass production compared with S. cannabina. Mathematical considerations in applying the ¹⁵N dilution method are discussed with reference to these results.