Real-time imaging of nitrogen fixation in an intact soybean plant with nodules using 13N-labeled nitrogen gas

Real-time images of nitrogen fixation in an intact nodule of hydroponically cultured soybean ( Glycine max [L] Merr.) were obtained. In the present study, we developed a rapid method to produce and purify ¹³N-labeled radioactive nitrogen gas (half life: 9.97 min). ¹³N was produced from a ¹⁶O (p, α) ¹³N nuclear reaction. The target chamber was filled with CO₂ and irradiated for 10 min with protons at an energy of 18.3 MeV and an electric current of 5 μA, which was delivered from a cyclotron. All CO₂ in the collected gas was absorbed and removed with powdered soda-lime in a syringe and replaced with helium gas. The resulting gas was injected into gas chromatography and separated and a 35 mL fraction, including the peak of [¹³N]-nitrogen gas, was collected by monitoring the chromatogram. The obtained gas was mixed with 10 mL of O₂ and 5 mL of N₂ and used in the tracer experiment. The tracer gas was fed into the underground part of intact nodulated soybean plants and serial images of the distribution of ¹³N were obtained non-invasively using a positron-emitting tracer imaging system (PETIS). The rates of nitrogen fixation of the six test plants were estimated to be 0.17 ± 0.10 μmol N₂ h⁻¹ from the PETIS image data. The decreasing rates of assimilated nitrogen were also estimated to be 0.012 ± 0.011 μmol N₂ h⁻¹. In conclusion, we successfully observed nitrogen fixation in soybean plants with nodules non-invasively and quantitatively using [¹³N]N₂ and PETIS.     

Nitrogen uptake by sago palm (Metroxylon sagu Rottb.) in the early growth stages

Previous trials have revealed variable responses of sago palm ( Metroxylon sagu Rottb.) to fertilizer application, particularly nitrogen (N). In the present study, we quantified the fertilizer use efficiency (FUE) of sago palm for the first time using ¹⁵N-labeled fertilizer in pot and field experiments. The pot experiment was conducted in Japan using a 2:1 mixture of sand to Philippine soil. The field experiment was conducted in Leyte in the Philippines. Both experiments consisted of three replicates in each of three treatments: control, ¹⁵N urea at 50 kg N ha⁻¹ and ¹⁵N urea at 100 kg N ha⁻¹. The N uptake of sago palm increased significantly, but inconsistently with increasing N application. The few instances of a significant increase in N uptake did not translate into significant improvements in growth parameters, except for the number of leaflets in the pot experiment. The FUE values for sago seedlings (< 6 months) in the pot experiment treated with 50 and 100 kg N ha⁻¹ were 10.5 and 13.2%, respectively, whereas for the 2-year-old sago palms in the field, the corresponding FUE values were 14.8 and 12.0%. The FUE values were similar at the two levels of N application in both experiments. Sago growth parameters appeared to be insensitive to N application, suggesting that the form of N and the timing of N fertilization are important factors for sago palms. Therefore, the use of N fertilizer in sago production can only be justified after determining and fully understanding the response of sago palm to N application.     

Gaseous nitrogen losses from soil under Sitka spruce following the application of fertilizer 15N urea

Gaseous N loss, through denitrification and NH₃⁻volatilization, was monitored throughout the growing season after spring application of ¹⁵N labelled urea fertilizer to peaty gley soils supporting N-deficient Sitka spruce. From the ¹⁵N data, it was calculated that only about 0.28% of applied N was lost through NH₃-volatilization, almost all within the first few days after fertilizer application. Approximately 0.05% of applied N was calculated to be lost through denitrification. Denitrification decreased slowly over a 4-month period after fertilizer application. Rates of NH₃-volatilization correlated with available NH₄⁺ in the litter layer, while for the early part of the study when N-losses were highest, denitrification rates correlated with available NO₃⁻ in the litter layer. Observations of gaseous N-loss are also discussed in relation to data from lysimetry, changes in soil pH, and the soil moisture regime.     

The fate at several time intervals of 15N-labelled ammonium nitrate applied to an established grass sward

¹⁵N-labelled ammonium nitrate solution was applied in late April to circular, enclosed micro-plots prepared by pressing open-ended polypropylene cylinders into an established sward of perennial ryegrass. Cylinders were removed from the ground at intervals between 2 and 370 days after the application and assessments made of the distribution of ¹⁵N in plant and soil components. Of the added labelled N, 54.7% was recovered in the herbage which was cut four times during the growing season and again at the final sampling date. After two days, 37% of the labelled N was recovered in the soil microbial biomass. Large fluctuations occurred in the amount of ¹⁵N recovered in the soil microbial biomass over the next 14 days suggesting that rapid cycling of ¹⁵N occurred between this fraction and the mineral N fraction. After the first cut in late May, translocation of¹⁵N occurred more slowly from the roots into the stubble than from stubble into new herbage, so that the amount in the stubble declined more rapidly than did that in the roots. During the winter, there was no net transfer of N from the roots to above-ground components of the sward. By the end of the growing season, half the ¹⁵N remaining in the sward was immobilized in the humified fraction of the soil organic matter; some of this was mineralized in the following spring.     

猪粪比例对烟草废弃物高温堆肥腐熟进程的影响

以烟草废弃物为基本原料进行堆肥试验,研究了不同比例猪粪与烟草废弃物混合堆肥体系中温度、pH、全氮、C/N比、水溶性NH＋4-N和种子发芽指数（GI,germination index）的动态变化规律,探讨了不同猪粪比例对烟草废弃物高温堆肥腐熟进程的影响。结果表明,在烟草废弃物中加入猪粪能缩短进入高温分解阶段的时间,增加高温分解持续时间,增加全氮含量,加快物料NH＋4-N和C/N比降低的速率,加快烟草废弃物堆肥腐熟化进程。添加一定比例猪粪的处理（烟草废弃物∶猪粪=7∶3、烟草废弃物∶猪粪=8∶2和烟草废弃物∶猪粪=9∶1）,分别在堆肥第3、4、5 d进入高温分解阶段（〉50 ℃）,高温持续时间分别为11、10、8 d;而纯烟草废弃物处理,最高温度为43 ℃,未进入高温分解阶段;至堆肥26 d,NH＋4-N含量分别比纯烟草废弃物对照降低47.7%、61.9%和25.6%;GI分别达到81.4%、84.1%和83.7%。     

Residual Effects of Compost and Green Manure of Pea with Other Organic Wastes on Nutrient-Use Efficiency of Successive Rice after Wheat

Adoption of legumes in cereal-based cropping systems and improvement of organic fertilizers are important to reduce chemical fertilizer use. We supplied compost and green manure (GM) of green pea plant residue (GP), without supplementation or supplemented with dried chicken manure (CM) and/or rapeseed oil residue (RR), for wheat and examined the effects of compost and GM (remaining fertilizer) on the rice of pea–wheat–rice system in two factorial pot experiments. Additions of pea residue resulted in an increase of 30% grains of rice compared to without addition, and compost increased 25% grains against GM. Supplementation increased 63–70% of rice grains as compared to the control. Nitrogen (N) and potassium (K) recovery efficiencies were correlated with yield, which suggested that composting and supplementation enabled effective nutrients accumulation in plant, resulting in better yield. Therefore, it is recommended that composting of GP with CM plus RR improves effectiveness of pea on rice of a pea–wheat–rice system.     

沼液堆肥化与牛粪堆肥化的发酵特性及腐熟进程

为评价沼液作为堆肥含氮添加剂的应用效果,开发沼液的处理应用技术,以牛粪树叶堆肥为对照,将沼液和树叶混合堆制发酵,探讨其发酵特性与腐熟进程。研究结果表明,环境温度一直在10℃以下,沼液堆肥化和牛粪堆肥化均能经历35d以上的堆温超过50℃的高温发酵;而沼液堆肥化超过50℃的高温期持续时间比牛粪堆肥化少8d;经60d的发酵沼液堆肥化的半纤维素含量从发酵初期的12.14%下降到6.53%,纤维素含量由20.5%下降到9.8%;而牛粪堆肥化的半纤维素含量从12.8%下降到9.56%,纤维素含量由21.5%下降到15.9%。可见沼液堆肥化的分解更彻底。从C/N、温度、可溶性糖含量、含水量、种子发芽指数综合评价两种堆肥的腐熟度,沼液堆肥化进入腐熟状态约经30d,而牛粪堆肥化进入腐熟约需45d。     

Role of soil redox in growth and fertilizer nitrogen uptake by rice in Thailand paddy soils

Abstract. In a laboratory study, ¹⁵N ammonium fertilizer uptake and rice growth was determined in a non-acid sulphate marine soil (Typic Tropaquept) and an acid sulphate soil (Sulfic Tropaquept). Acid sulphate sensitive (IR 26) and acid sulphate tolerant (IR 46) rice varieties were grown in soil suspensions incubated at four Eh levels (+500, +250, +50, and -150 mV) in microcosms for three weeks. The results showed that rice grown in non-acid sulphate marine soils gave slightly better dry matter weight of 1.8g/pot, greater ¹⁵N uptake of 12.8 mg N/pot, and higher total N uptake of 38.4 mg N/pot than under acid sulphate soil conditions indicating the non-acid marine soil is more favourable to rice culture. Growth as measured by weight of dry matter was significantly reduced from 2.1g/pot under oxidized condition (+500 mV) to 0.8g/pot under highly reduced condition (-150 mV). N uptake by rice was significantly reduced from 16.9 mg/pot at + 500 mV to 4.5 mg N/pot at -150 mV Total N uptake also decreased with decreasing Eh. Growth, ¹⁵N uptake and total N uptake by acid sulphate tolerant rice, IR 46 were significantly higher than the acid sulphate sensitive rice variety, IR 26. Under highly reduced soil conditions (-150 mV), growing rice in acid sulphate soil would require additions of lime, intermittent irrigation and/or mid season drainage in order to increase soil redox potential and remove toxic substances.     

Microbial Immobilization and Plant Uptake of Different N Forms in Three Forest Types in Shikoku District, Southern Japan

Soil microbial immobilization and plant uptake of N were evaluated for three forest types in Kochi, Shikoku district. During 196-d laboratory incubation, soil NO₃-N production in the Hinoki cypress forest was negligible for the initial 40 d and then rapidly increased, whereas NO₃-N production was rapid from the beginning in Japanese cedar and deciduous hardwood forests. Microbial immobilization of the labeled ¹⁵N decreased in the order of NH₄-N>glycine-N>NO₃-N. The ¹⁵N immobilization was higher for soil in the Hinoki cypress forest than other two soils. The delayed NO₃-N production in the Hinoki cypress forest was likely related with low availability of NH₄-N due to NH₄-N immobilization and substantial NO₃-N immobilization. In the field experiment, ¹⁵N uptake by roots decreased in the order of NH₄-N>NO₃-N>glycine-N. The absorption of the labeled ¹³C suggested direct uptake of organic N. The preference of N forms by root uptake was not different among forest types. Trees in three forest types can absorb inorganic and organic forms of N, suggesting trees absorb the N form that is the most abundant in the soil.     

Recovery of 15N-labelled fertilizer in crops, drainage water and soil using monolith lysimeters in south-east Nigeria

Labelled urea was applied to monolith lysimeters in the 1st year of a 2-year experiment at Onne in south-east Nigeria. On eight lysimeters maize and rice were grown in each of the 2 years. Four lysimeters were similarly cropped in the 2nd year after being uncropped in the 1st year. Measurements were made over the 2-year period of labelled and unlabelled mineral nitrogen in the drainage water, and labelled and unlabelled nitrogen in the crops. At the end of the experiment, weeds and the soil were also analysed for labelled and unlabelled nitrogen.
The total recovery of ¹⁵N in crop, soil and leachate varied between 70 and 93%. It was lowest when applied to the second season rice crop, which recovered only 15%, and highest when it was leached in the 1st year or was taken up by the maize crop. The highest crop uptake was 31%. The results indicate that, depending on the treatment, between 10 and 30% of the ¹⁵N was immobilized in the soil, lysimeters cropped in the 1st year lost between 22 and 29% of the ¹⁵N in drainage water, and between 7 and 30% was lost by denitrification. The accuracy of these figures is discussed.     

Short-term kinetics of residual wheat straw C and N under field conditions: characterization by 13C15N tracing and soil particle size fractionation

Summary A clear understanding of the short-term decomposition and fate of crop residues is necessary to predict the availability of mineral N in soil. The fate of ¹³ C¹⁵N-labelled wheat straw in a silty soil (Typic Hapludalf) was studied using particle size fractionation and in situ incubation in which the equivalent of 8 t dry matter per ha of straw was incorporated into the soil over 574 days. Soil samples were separated into five particle-size fractions by wet sieving after disruption of aggregates. The weight, C and N contents, and ¹³C and ¹⁵N atom excess of each fraction were determined. Straw-derived C disappeared rapidly from the > 2000-μm fraction with an estimated half-life of 53 'normalized' days (equivalent of 10°C and −0−01 MPA water potential). Straw-derived C appeared to be only temporarily stored in the intermediate fractions (1000–2000 and 200–1000 pm). The maximum net ¹³C accumulation in the 50–200-μm fraction was 4·4% of added ¹³C. Straw-derived C accumulated most rapidly and preferentially in the 50-μm fraction, which stabilized after 265 days and accounted for 70% of the residual ¹³C on day 574. Although there was more residual ¹⁵N than ¹³C, the distributions and kinetics of the two isotopes in the fractions were similar.     

Effect of nitrite accumulation on nitrous oxide emission and total nitrogen loss during swine manure composting

The present study investigated the nitrogen balance in swine manure composting to evaluate the effect of nitrite (     ) accumulation, which induces nitrogenous emissions, such as N₂O, during compost maturation. During active composting, most N losses result from NH₃ emission, which was 9.5% of the initial total nitrogen (TN_initial), after which,     began to accumulate as only ammonia-oxidizing bacteria proliferated. After active composting, the addition of mature swine compost (MSC), including nitrite-oxidizing bacteria (NOB), could prevent     accumulation and reduce N₂O emission by 70% compared with the control in which     accumulated as a result of delayed growth of indigenous NOB. Total N₂O emissions in the control and in the treatment of MSC addition (MA) were 9.3% and 3.0% of TN_initial, respectively, whereas N losses as the sum total of NH₃ and N₂O over the whole period were 19.0% (control) and 12.8% (MA) of TN_initial, respectively. However, the difference in total N losses was markedly greater than that measured as NH₃ and N₂O, which were 27.8% (control) and 13.3% (MA) of TN_initial, respectively. These results demonstrated that the magnitude of nitrogen losses induced by     accumulation is too large to ignore in the composting of swine manure.     

穗肥施用时期对优质稻产量、氮素积累及品质的影响

为了探明优质晚籼稻(Oryza sativa L. subsp. xian)产量和品质协同提升的适宜的穗肥施用时期,采用大田微区¹⁵N示踪技术,以优质晚籼稻Y两优911和野香优莉丝为材料,设置3个穗肥施用时期(D₁：倒4叶期施肥;D₂：倒3叶期施肥;D₃：倒2叶期施肥),研究穗肥施用时期对优质稻产量、籽粒氮素积累及食味品质的影响。结果表明,随着穗肥施用时期的推迟,两个优质晚籼稻品种的产量先升高后降低,在D₂达到最高产量,较D₁、D₃显著增产4.30%～6.39%;籽粒总氮积累量、¹⁵N标记肥料氮素积累量及其占比、¹⁵N标记肥料氮素回收利用率、¹⁵N标记肥料氮素收获指数表现为增加趋势,D₂和D₃较D₁显著增加19.16%～21.26%、32.54%～50.75%、0.91～2.49、12.42～23.34和...     

Measurement of ammonia volatilization loss using a dynamic chamber technique: A case study of surface-incorporated manure and ammonium sulfate in an upland field of light-colored Andosol

The present study aimed to elucidate ammonia (NH₃) volatilization loss following surface incorporation (0–15 cm mixing depth) of nitrogen (N) fertilizer in an upland field of light-colored Andosol in central Japan. A dynamic chamber technique was used to measure the NH₃ effluxes. Poultry manure, pelleted poultry manure, cattle manure, pelleted cattle manure and ammonium sulfate were used as N fertilizers for basal fertilization to a bare soil with surface incorporation. All three experiments in summer and autumn 2007 and in summer 2008 showed negligible NH₃ volatilization losses following the application of all N fertilizers with the same application rate of 120 kg N ha⁻¹ as total N; these negligible losses were primarily ascribed to chemical properties of the soil, that is, its high cation exchange capacity (283 mmol_c kg⁻¹ dry soil) and relatively low pH(H₂O) (5.9). In addition, the surface incorporation, the very small ratio of ammoniacal N to total N for the manure, and the decrease in soil pH to ≤5.5 following applications of ammonium sulfate were also advantageous to the inhibition of NH₃ volatilization loss from the field-applied N fertilizers.     

Organic matter dynamics in a compost-amended anthropogenic landfill capping-soil

The application of municipal waste compost and other organic materials may serve to improve fertility and organic carbon (C) stocks of soils used in land reclamation activities, particularly in the recovery of degraded areas at exhausted quarries, mines and landfill sites amongst others. We investigated long-term organic matter dynamics in such anthropogenic soils by collecting samples at different depths over a 10-year chronosequence subsequent to compost application to the top layer of a landfill capping-soil. Variations in the stable isotope composition (¹³C and ¹⁵N) of the soil samples showed that, even after 10 years, amended topsoils were enriched in compost-derived organic matter. The addition of compost to the superficial layer also resulted in an input of soluble organic compounds that was subject to leaching along the soil profile. Sorption isotherms for compost-derived water-extractable organic matter onto mineral materials used for landfill covering suggest that sorptive preservation was primarily responsible for the increase in C content and the shift in the C isotopic signature to values similar to that of the applied compost, in the deeper soil horizons over the 10-year period. This was also confirmed by the accumulation of lignin-derived phenolic compounds in the deeper horizons. Nevertheless, analysis for non-cellulosic carbohydrates in soil samples and their respective water-extractable fractions suggest that a proportion of the compost-derived, labile organic matter fraction is leached through the profile and potentially lost from the soil system.     

Increase in the Nitrogen Content of Soil by the Introduction of Earthworms into Soil

The contribution of an earthworm species ( Amynthas vittatus ) to the increase of the nitrogen content of soil was examined. Three specimens of adult earthworms were introduced into 300 g of soil (Gray Lowland soil, silty clay) supplemented with 1% carboxymethyl cellulose in a container and incubated for 32 d at 22°C in the dark. The contents of total-N, NH₄-N and NO₃-N, and the population of aerobic nitrogen-fixing bacteria in soil significantly increased after incubation with the earthworms, while the natural abundance of ¹⁵N (δ¹⁵N) in soil decreased. The amount of nitrogen in the earthworms did not decrease during the incubation in the microcosm. Both acetylene reduction activity of the microcosm and incorporation of ¹⁵N to soil from atmospheric ¹⁵N₂ were significantly enhanced by the introduction of the earthworms into soil, though the observed increment of nitrogen in soil was much higher than the estimated one based on the nitrogen-fixing activity. The results obtained in the present study indicated that the earthworms increased the nitrogen content of soil, presumably due to the enhancement of the nitrogen-fixing activity of the soil from the microcosm by the earthworms.     

Fate of nitrogen derived from 15N-labeled plant residues and composts in rice-planted paddy soil

Pot experiments that lasted for 3 y were conducted to investigate the dynamics of nitrogen derived from plant residues (rice root, hull, straw, corn root, and rapeseed pod-wall), and composts (rice straw compost, cattle manure compost, and cattle manure sawdust compost), which were labeled with ¹⁵N. The rates of nitrogen uptake by rice (=N efficiency), denitrification, and immobilization derived from the organic materials incorporated before the first year of cultivation were investigated throughout 3 y of cultivation. At the end of the first year of cultivation, relatively high rates of N efficiency were obtained for rapeseed pod-wall (24.6%), rice straw (19.1%), and rice hull (18.6%), while corn root and cattle manure sawdust compost displayed a noticeably high denitrification rate. Corn root, cattle manure sawdust compost, rice hull, and rapeseed pod-wall exhibited remarkably high N mineralization rates ranging from 60 to 75% of the organic materials N applied. Cumulative rates of N efficiencies from the organic materials applied before the first year of cultivation fitted well to a first-order kinetic model and their asymptotes were compared among the organic materials. The asymptotic rates of N efficiency tended to depend on the rates at the end of the first year of cultivation.     

Changes in natural 15N abundance in paddy soils under different,long-term soil management regimes in the Tohoku region of Japan

Abstract

Long-term temporal changes in natural ¹⁵N abundance (δ¹⁵N value) in paddy soils from long-term field experiments with livestock manure and rice straw composts, and in the composts used for the experiments, were investigated. These field experiments using livestock manure and rice straw composts had been conducted since 1973 and 1968, respectively. In both experiments, control plots to which no compost had been applied were also maintained. The δ¹⁵N values of livestock manure compost reflected the composting method. Composting period had no significant effect on the δ¹⁵N value of rice straw compost. The δ¹⁵N values increased in soils to which livestock manure compost was successively applied, and tended to decrease in soils without compost. In soils to which rice straw compost was successively applied, the δ¹⁵N values of the soils remained constant. Conversely, δ¹⁵N values in soils without rice straw compost decreased. The downward trend in δ¹⁵N values observed in soils to which compost and chemical N fertilizer were not applied could be attributed to the natural input of N, which had a lower δ¹⁵N value than the soils. Thus, the transition of the δ¹⁵N values in soils observed in long-term paddy field experiments indicated that the δ¹⁵N values of paddy soils could be affected by natural N input in addition to extraneous N that was applied in the form of chemical N fertilizers and organic materials.     

Effects of Inoculation of Phototrophic Purple Bacteria on Grain Yield of Rice and Nitrogenase Activity of Paddy Soil in a Pot Experiment

Phototrophic purple bacteria (PPB) are one group of dinitrogen (N₂)-fixing bacteria often found in the anoxic and photic zone of paddy soil. In the present study, the effects of inoculation of PPB on grain yield of rice and changes in their populations and nitrogenase activity were investigated with and without surface application of rice straw in a pot experiment. As an inoculant, Rhodopseudomonas palustris strain KN122 isolated from a paddy soil was selected, and the cell suspension was inoculated into the floodwater once or three times during the cultivation. As a result, the inoculation of PPB increased the grain yield of rice. Compared to the control, the grain yield was 9% higher in the inoculated plot without rice straw application. The treatment was more effective in combination with rice straw application. In the plots where PPB were inoculated with rice straw, grain yields were 21% (single inoculation) and 29% (triple inoculation) higher. Populations of PPB in the plots without rice straw application significantly increased by the inoculation, unlike those in the plots with rice straw application, except for the bulk soil. Phototrophic and heterotrophic nitrogenase activities (acetylene-reducing activity) associated with the soils or residues of the rice straw were not affected by PPB inoculation throughout the experiment. This indicates that the inoculation of PPB into floodwater may not be effective for enhancing N₂ fixation in paddy soils and that the beneficial effect of PPB inoculation on the grain yield of rice may be due to unidentified functions of PPB other than biological N₂ fixation.     

Studies on the rate of decomposition of plant residues in soil by following the changes in sugar components

Uniformly ¹⁴C labelled straw and ryegrass leaf were incubated in soil in the proportions 0.5 and 1% w/w, respectively at 12°C for various periods up to 5 years.
Samples were hydrolysed with 2.5 m H₂SO₄ under reflux for 20 min to release non-cellulosic sugars and subsequently with 12 M/0.5 m H₂SO₄ to release glucose from cellulose-like polymers. Total radioactivity was determined in whole soil and hydrolysates and in individual sugars.
Fitting the data to simple exponential equations, total and 2.5 m H₂SO₄ hydrolysate radioactivity were best expressed by three part exponential functions, whereas radioactivity in 12M/0.5M H₂SO₄ hydrolysates and sugars were best expressed by two part functions, except for arabinose from the straw, for which a three-part function was better.
A comparison of the fit was made with that using the single-term exponential expressions devised by Janssen, 1984 and Weibull, 1951 and with a modification of the two part exponential expression by the substitution of one term by the Weibull expression. Apart from the Weibull expression on its own, there was no significant difference between the expressions.
It is suggested that, even for well defined carbohydrate components of the substrate, the decomposability changes with time because of their increasing inaccessibility to microorganisms, rather than that several components are present each with a different constant rate of decomposition.