Nitrogen fixation in paddy soils

Several important features of the N. fixation in paddy fields which were reported previously were confirmed and some new additional results regarding the evaluation of the N₂ fixation in the rhizosphere were obtained by reinvestigation in the fields. In addition, rice plants were cultivated in the submerged soil in pots and various parts of the soil were analyzed for the N₂-fixing activity as well as several other properties. The results of the pot experiments were found to be fairly similar to those observed in the field investigations, indicating the validity of the submerged soil in a pot as a rather simulated model for the actual paddy field. By using this model system, the following facts were ascertained: (1) Water-percolation had almost no effect on the N₂-fixing activities of both the rhizosphere and the non-rhizosphere soils. (2) Suppressing effect of washing the root of rice plant on the N₂-fixing activity was slight in the seedling stage and marked in the tillering and flowering stages. (3) The N₂-fixing activity of a single rice root varied from tip to base.     

Free-living nitrogen-fixing bacteria in temperate cropping systems: Influence of nitrogen source

Sustainable cropping systems rely on a minimum of external inputs. In these systems N is largely acquired in animal manures and leguminous green manures. Little is known of how these organic forms of N fertilizer influence the presence and activity of free-living N₂-fixing bacteria. High concentrations of inorganic N in soil inhibit N₂-fixation in cyanobacteria and Azotobacter spp. It is likely that manure and fertilizer applications would result in concentrations of inorganic N capable of inhibiting N₂ fixation and, ultimately, the presence of these organisms. We investigated the effect of synthetic and organic N fertilizer sources on the populations and N₂-fixation potential of free-living N₂-fixing bacteria in the Farming Systems Trial at the Rodale Research Institute. Field plots received the following N treatments prior to corn (Zea mays L.) production: (1) Legume rotations and green manures supplying about 165 kg N ha^-1; (2) beef cattle manure applied at a rate of 220 kg N ha^-1 (plus 60 kg N ha^-1 from 1994 hay plow-down); or (3) fertilizer N (urea and NH₄NO₃) applied at a rate of 145 kg N ha^-1. Soil samples were collected at two depths from corn plots four times during the growing season, and analyzed for soil moisture, soil pH, numbers of N₂-fixing cyanobacteria and Azotobacter spp., extractable NH _inf4 ^sup+ and NO _inf3 ^sup- , and potentially mineralizable N. Soil samples collected in mid-July were analyzed for nitrogenase activity (by C₂H₂ reduction) and total C and N. Populations of Azotobacter spp. and cyanobacteria were influenced only slightly by treatment; however, cyanobacteria species composition was notably influenced by treatment. Nitrogenase activity in surface soils was greatest in legume-N plots and in subsurface plots levels were greatest in fertilizer-N plots. Populations and activity of free-living N-fixing bacteria appeared to be somewhat reduced in all plots as a result of low soil pH levels and high concentrations of inorganic N across all treatments. Annual applications of N to all plots resulted in high levels of potentially mineralizable N that in turn may have reduced non-symbiotic N₂-fixation in all plots.     

太湖地区长期不同施肥水稻土N2和CO2固定细菌群落结构的特征与差异

土壤N2固定细菌和CO2固定细菌是土壤碳氮循环重要的微生物群落,不同施肥下土壤有机碳积累与这两类微生物群落结构及活性的关系有助于了解施肥对农田碳氮循环的影响特点.本文研究了太湖地区自1987年开始的水稻土长期施肥试验,选取不施肥(NF)、氮磷钾(NPK)肥(CF)、NPK肥与猪粪配施(CFM)和NPK肥与秸秆还田配施(CFS)小区,采集(0-20 cm)表层样本,以nifH和cbbLR基因分别作为N2和CO2固定细菌的指示基因,用PCR-DGGE和荧光定量PCR的方法研究二者的群落结构,并用平板菌落计数法测定土壤自生固氮细菌数量和用乙炔还原法测定土壤固氮酶活性.结果表明,与CF处理相比,CF、CFM和CFS处理下的自生固氮细菌数量分别提高了58％、66％和106％;CF、CFM和CFS处理下的nifH基因丰度分别提高了213％、1079％和344％.CF与CFM处理的土壤固氮酶活性显著高于NF和CFS处理.因此,施肥提高了cbbLR基因的多样性.与NF处理相比,CF、CFM和CFS处理下的cbbLR基因丰度分别提高了465％、1827％和758％.相关性分析表明,土壤自生固氮菌数量与土壤有机碳含量呈显著正相关,cbbLR和nifH基因丰度均与归一化的土壤养分呈极其显著正相关.研究表明,有机/无机肥配施下土壤养分平衡对维持N2和CO2固定细菌较高的丰度具有重要的作用.     

Assessment of slow release fertilizers and nitrification inhibitors in flooded rice

This work evaluates the effect of different slow-release fertilizers and nitrification inhibitors (NI) on N-use efficiency, grain yield and N₂ fixation in rice fields of Valencia (Spain) during three consecutive crop seasons (1998–2000). Eight N sources [ammonium sulphate, urea, polymer-coated urea (PCU 32% and 40% N), sulphur coated urea, isobutylidene diurea (IBDU), ammonium sulphate nitrate (ASN) plus dicyandiamide and ASN plus dimethyl pyrazole phosphate, were applied at 120 kg N ha^–1 and at 2 or 15 days before flooding (DBF) during 1998. Another experiment was based on the use of urea blended with PCU (40% N) at four ratios (1:0, 3:1, 1:1, 1:3) and applied at 15 DBF and at four rates (70, 95, 120 and 145 kg N ha-1) during 1999 and at only one rate (120 kg N ha^–1) during 2000. Both experiments also included a control (no N). The results showed that, when applied shortly before flooding, PCU (32% and 40% N) and IBDU application improved biological N₂ fixation compared to the conventional fertilizer application, with or without NI, reaching similar values to those observed in the absence of N fertilizer. Slow release fertilizers, particularly PCU 40% N applied basally before flooding, were the best N source for grain yield and improved recovery efficiency. Differences among N sources were only significant when the flooding was delayed for 15 days after fertilizer application.     

Growth and Nitrogen Fixation in Soybean as Affected by Phosphorus Fertilizer and Sheep Manure using 15N Isotopic Dilution

A field experiment was conducted to study the effect of adding different phosphorus (P) fertilizer levels [0, 40, and 80 kg phosphorus pentoxide (P₂O₅) ha^?1 (abbreviated as P0, P1, and P2, respectively)] and rates of sheep manure (M) [0, 20, and 40 ton ha^?1 (abbreviated as M0, M1, and M2, respectively)] on growth and nitrogen (N₂) fixation of soybean (Glycine max L.). Sorghum bicolor L. was employed as a reference crop to evaluate N₂ fixation using the ¹⁵N-isotpic dilution technique. Results showed that addition of P fertilizer or sheep manure had positive effects on dry-matter production, N accumulation, and seed yield. Such effects were more pronounced when adding sheep manure and P together than adding separately. Solely P fertilizer had a small impact on N₂ fixation. A tangible increase in the amounts of N₂ fixed due to manure addition occurred. The efficient use of N fertilizer (%NUE) increased significantly as the result of adding a high level of P fertilizer. However, a drastic decrease in %NUE was observed when sheep manure was added solely or in combination with P fertilizer. From productivity and ecological standpoints, P2M1 and P2M2 surpassed the other treatments in showing greater grain yield and greater N₂ fixation. However, considering the high cost of sheep manure, P2M1 was the optimal treatment for improving growth and N₂ fixation in soybean plants with minimal manure consumption. In conclusion, the integrated use of manure and P fertilizer could be considered a useful agricultural practice for improving the performance of soybean plants grown in an Aridisol. Their beneficial effects were mainly attributed to the enhancement of N₂ fixation through root growth and soil property improvements besides being a source of P and other nutrients that are essential for N₂-fixation process.     

The effects on N2 fixation (C2H2 reduction), bacterial population and rice plant growth of two modes of straw application to a wetland rice field

Summary The effects of incorporation and surface application of straw to a wetland rice field on nitrogen fixation (C₂H₂ reduction), bacterial population and rice plant growth were studied. Rice straw (5 t ha^–1) was chopped (10- to 15-cm pieces) and applied to the field 2 weeks before transplanting IR42, a long-duration variety, and IR50, a short-duration variety. The acetylene-reducing activity (ARA) of IR42 and IR50 measured at heading stage for 3 consecutive days showed significantly higher ARA in IR42 as a result of the 2 straw application methods. Mostly up to 20 days after straw surface application and incorporation, the dark ARA in the soil, total and N₂-fixing heterotrophs, and photoorganotrophic purple nonsulphur bacteria (POPNS) in the soil and in association with degrading straw were stimulated. Higher bacterial populations were associated with straw on the surface than with straw incorporated. The POPNS counts, in particular, were increased hundreds fold in the surface-applied straw treatment. Straw applications also increased the root, shoot and total plant biomass at heading stage and the total dry matter yield at harvest in both varieties. The data show the potentials of straw as a source of substrate for the production of microbial biomass and for the non-symbiotic N₂ fixation to improve soil fertility and plant nutrition.     

Short-term measurements of uptake of nitrogen fixed in the rhizospheres of sorghum (Sorghum bicolor) and millet (Pennisetum americanum)

Summary In a series of short-term experiments root systems of young sorghum and millet plants inoculated with N₂-fixing bacteria were exposed to ¹⁵N₂-enriched atmospheres for 72 h. The plants were grown in a normal atmosphere for up to 22 days after the end of the exposure to allow them to take up the fixed N₂. Environmental conditions and genotypes of sorghum and millet were selected to maximise N₂-fixation in the rhizosphere. Detectable amounts of fixed N (> 16 g/plant) were rapidly incorporated into sorghum plants grown in a sand/farmyard manure medium, but measurable fixation was found on only one occasion in plants grown in soil. N₂ fixation was detectable in some experiments with soil-grown millet plants but the amounts were small (2–4 g/plant) and represented less than 1 % of plant N accumulated over the same period. In many cases there was no detectable ¹⁵N₂ incorporation despite measurable increases in ethylene concentration found during an acetylene reduction assay.Published as ICRISAT Journal Article No. JA 740     

Heterotrophy-coordinated diazotrophy is associated with significant changes of rare taxa in soil microbiome

Soil heterotrophic respiration during decomposition of carbon (C)-rich organic matter plays a vital role in sustaining soil fertility. However, it remains poorly understood whether dinitrogen (N₂) fixation occurs in support of soil heterotrophic respiration. In this study, ¹⁵N₂-tracing indicated that strong N₂ fixation occurred during heterotrophic respiration of carbon-rich glucose. Soil organic ¹⁵N increased from 0.37 atom% to 2.50 atom% under aerobic conditions and to 4.23 atom% under anaerobic conditions, while the concomitant CO₂ flux increased by 12.0-fold under aerobic conditions and 5.18-fold under anaerobic conditions. Soil N₂ fixation was completely absent in soils replete with inorganic N, although soil N bioavailability did not alter soil respiration. High-throughput sequencing of the 16S rRNA gene further indicated that: i) under aerobic conditions, only 15.2% of soil microbiome responded positively to glucose addition, and these responses were significantly associated with soil respiration and N₂ fixation and ii) under anaerobic conditions, the percentage of responses was even lower at 5.70%. Intriguingly, more than 95% of these responses were originally rare with < 0.5% relative abundance in background soils, including typical N₂-fixing heterotrophs such as Azotobacter and Clostridium and well-recognized non-N₂-fixing heterotrophs such as Sporosarcina, Agromyces, and Sedimentibacter. These results suggest that only a small portion of the soil microbiome could respond quickly to the amendment of readily accessible organic C in a fluvo-aquic soil and highlighted that rare phylotypes might have played more important roles than previously appreciated in catalyzing soil C and nitrogen turnovers. Our study indicates that N₂ fixation could be closely associated with microbial turnover of soil organic C when available in excess.     

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.     

Changes in acetylene reduction activities and effects of inoculated rhizosphere nitrogen-fixing bacteria on rice

Acetylene reduction activities (ARAs) of soils and rice plants during rice-growing season were monitored in temperate region in northeast China. This activity was significantly higher in rhizosphere soil than that in inter-row soil after rice seedlings were transplanted. The ARA was high for most of growing season, suggesting that the native N₂-fixing bacteria responded to rice roots very quickly. Sixteen strains of free-living N₂-fixing bacteria were isolated from three different soils. The ARAs of these strains were correlated with the averaged soil ARAs, suggesting that the isolated strains were likely the active flora responsive to rice roots. The strains were inoculated by soaking seedling roots into the liquid culture for 2 h, and the seedlings were transplanted into pots. Most strains tested did not show any growth-promoting effects except Azotobacter armeniacus and Azotobacter nigricans, which showed growth-promoting effects only at late rice growth stage and only when inoculated in combination but not separately. Present data indicated the promising future applications of these two strains in combination in the region, but further research is needed to understand the underlying mechanisms.     

Mitigating nitrous oxide emissions from a maize-cropping black soil in northeast China by a combination of reducing chemical N fertilizer application and applying manure in autumn

Abstract

Nitrous oxide (N₂O) emissions from agricultural soils, mainly caused by chemical nitrogen (N) fertilizer inputs, are major sources of N₂O in Chinese terrestrial ecosystems. Thus, attempts to reduce N₂O emissions from agricultural soils by optimizing N applications are receiving increasing attention. Further, organic fertilizers are being increasingly used in China to improve crop production/quality and prevent or reduce soil degradation. However, organic and chemical fertilizers are often both applied in spring in northeast China, which promotes N₂O emissions and may be sub-optimal. Therefore, we hypothesized that reducing applications of chemical fertilizer N and applying manure in autumn could be an effective strategy for mitigating N₂O emissions from cropped soils in the region. To test this hypothesis, we established a field trial to investigate the effects of different combinations of chemical N fertilizer applications and animal manure in autumn on both N₂O emissions and maize (Zea mays L.) grain yields in northeast China. The treatments, expressed as NxMy (where Nx and My denote the total amounts of chemical fertilizer nitrogen (N) and manure (M) applied in kg N ha^?1 and m³ M ha^?1, respectively), were N₀M₀, N₂₃₀M₀, N₂₇₀M₁₂, N₂₃₀M₁₅, N₃₂₀M₁₈ in 2010 and N₀M₀, N₂₃₀M₀, N₂₀₀M₁₂, N₂₀₀M₁₅, N₂₈₀M₁₈ in 2011. Measurements of the resulting N₂O emissions showed that pulse fluxes occurred after each chemical N fertilizer application, but not after manure inputs in autumn or during soil-thawing periods in the following spring. Emission factors for the chemical fertilizer N were on average 1.07% (1.00?1.10%) and 1.14% (0.49?1.83%) in 2010 and 2011, respectively. Furthermore, by comparing the nine pairs of fertilization treatments, the relative increase in cumulative nitrous oxide-nitrogen (N₂O-N) emissions was found to be proportional to the relative increase in urea application, but independent of the amount of autumn-applied manure. These findings imply that N₂O emissions from fertilized agricultural soils in northeast China could be mitigated by supplying manure in the autumn and reducing the total amount of chemical N fertilizer applied in the following year. Although no significant difference in maize grain yield was found among the fertilization treatments, the grain yield-scaled N₂O emissions for the treatments with a lower chemical N application (e.g., N₂₃₀M₁₅ and N₂₀₀M₁₅ treatments) were significantly lower than those with a higher chemical N application (e.g., N₃₂₀M₁₈ and N₂₈₀M₁₈ treatments). Meanwhile, under the condition of the same application amount of chemical fertilizer N, the grain yield-scaled N₂O emission decreased with the increase of manure application rate. Thus, the results support the hypothesis that combining reductions in chemical N fertilizer and applying manure in autumn could be an effective strategy for mitigating N₂O emissions from N-fertilized soils in northeast China.     

N2O emission mitigation and microbial activity after Biochar and Cao application in a flooded nitrate-rich vegetable soil

Adding easily decomposable organic materials into flooded nitrate-rich soils can effectively decrease the soil nitrate concentration and repair nitrate-rich soil. However, nitrate reduction is usually accompanied with an increase in N₂O emission. This study was conducted to reduce N₂O emission in a nitrate-rich vegetable soil flooded for remediation and amended with biochar. Nitrate-rich vegetable soil was placed in five treatment groups: flooding (F); flooding with rice straw (F?+?RS); flooding with rice straw and 1% biochar (F?+?RS?+?1% biochar); flooding with rice straw and 3% biochar (F?+?RS?+?3% biochar); flooding with rice straw and CaO (F?+?RS?+?CaO). Biochar and CaO reduced the N₂O emission levels relative to the F?+?RS group, with the former being more effective than the latter, achieving reduction of 40.70% (3% biochar) and 17.35% (CaO) of cumulative N₂O emission. The 3% biochar was more effective than the 1% biochar. Regression analysis showed a positive correlation between the abundance of NO reductase gene (norB) and soil N₂O emission flux. In general, biochar and CaO could effectively reduce N₂O emissions from a nitrate-rich vegetable soil during flooding remediation, duo to elevating soil pH and altering denitrifying activity. The norB gene was the most important denitrifying gene driving soil N₂O emission in the remediation.     

Response of Determinate and Indeterminate Common Bean Genotypes to Rhizobium Inoculant in a Short Season Rainfed Production System in the Canadian Prairie

ABSTRACT

Common bean (Phaseolus vulgaris L.) has been shown to be a poor di-nitrogen (N₂) fixer and nitrogen (N) fertilizers are usually recommended in bean production. Recent research results suggest that the success of the bean/Rhizobium symbiosis may depend, in part, on the specific bean genotype. Twelve dry bean genotypes differing in growth habit, commercial class, and maturity were evaluated for N₂ fixation in field experiments. Response to inoculant application was highly influenced by environmental conditions. Genotypes differed in nodule dry mass, seed yield, seed N yield, and in amount of N₂ fixed. Growth habit alone was not adequate in classifying bean genotypes for N₂ fixation. The actual amount of N₂ fixed was low ranging from 16 kg ha^?1 to 27 kg ha^?1, suggesting that the symbiotic process alone may not provide adequate N for optimum seed yield in dry environments.     

Influences of organic debris and rice root on the nitrogen fixation in the submerged soil

Abstract

A series of experiments has been conducted on the N2 fixation in the paddy soils by the authors (1–4). The amount of organic substrates for microorganisms and the degree of reduction of the soil are found to be two major factors affecting the N₂-fixing activity of the heterotrophic microorganisms in the submerged soil. Organic debris, rice root and their neighboring soils are identified to be the important micro-sites for the heterotrophic N₂ fixers. The organic debris and the rice root are considered to play dual roles by supplying the organic substances; (1) increase of the population of the heterotrophic N₂ fixers—the amount of nitrogenase, (2) preparation of the reduced conditions favorable for the nitrogenase activity.

However, it is not yet clearly known which of these two roles of the organic substrates is more essential to the N₂-fixing activity in the paddy soil. In addition, it is expected that there must be some differences between the organic debris and rice root in their contribution to the N₂ fixation in the paddy soil.

An experiment was carried out to clarify these problems. Moist soil sample was collected from the plough layer of the paddy field at Central Agric. Exp. Sta. in Konosu City, Saitama Pref., passed through a 5 mm sieve and placed in pots (3 kg moist soil/pot). Ammonium sulfate, calcium superphosphate, and potassium chloride at the rate of 0.4-0.4-0.4 (N-P₂O₅-K₂0) g/pot were incorporated into the soil 7 days before transplanting. Split application of ammonium sulfate at the rates of 0.2 and 0.4 g N/pot were also incorporated at 30 and 41 days after transplanting respectively. These pots were divided into three series; planted (P-series), non-planted (N-series), and non-planted and applied with organic manure (O-series). In case of O-series, 60 g of fairly rotted organic manure was applied to each pot. Each pot of P-series was planted with two 4O-day-old seedlings of rice plant at 7 days after submergence. The Nseries was regarded as a control. Each series was not replicated in this preliminary experiment.     

Phosphorus Deficiency Delays the Onset of Nodule Function in Soybean

Effects of phosphorus (P) deficiency on nodulation were examined in soybean grown in nutrient solution for 7 weeks. Increasing P supply increased shoot growth of nitrogen (N₂)-fixing plants from week 5 and that of nitrate-fed plant from week 4 after treatment. Nitrogen (N₂)-fixing plants had a greater P requirement for maximum growth at week 5. Increasing P supply from 1 to 16 μ M increased N concentration in N₂-fixing plants at week 4 but did not affect it from week 5. By contrast, P deficiency increased N concentration in nitrate-fed plants. Increasing P supply improved nodule formation from week 3. Nodule mass was affected more by P supply than nodule number, which, in turn, was affected more than plant growth. However, P supply did not decrease nodule specific N₂ fixation from week 5. The results suggest that P deficiency impaired symbiotic N₂ fixation through delaying onset of nodule function and decreasing nodule development.     

Nitrous oxide emissions from wetland soil amended with inorganic and organic fertilizers

Agricultural soils are a primary source of anthropogenic trace gas emissions, and the subtropics contribute greatly, particularly since 51% of world soils are in these climate zones. A field experiment was carried out in an ephemeral wetland in central Zimbabwe in order to determine the effect of cattle manure (1.36% N) and mineral N fertilizer (ammonium nitrate, 34.5% N) application on N₂O fluxes from soil. Combined applications of 0 kg N fertilizer + 0 Mg cattle manure ha^?1 (control), 100 kg N fertilizer + 15 Mg manure ha^?1 and 200 kg N fertilizer + 30 Mg manure ha^?1 constituted the three treatments arranged in a randomized complete block design with four replications. Tomato and rape crops were grown in rotation over a period of two seasons. Emissions of N₂O were sampled using the static chamber technique. Increasing N fertilizer and manure application rates from low to high rates increased the N₂O fluxes by 37–106%. When low and high rates were applied to the tomato and rape crops, 0.51%, 0.40%, and 0.93%, 0.64% of applied N was lost as N₂O, respectively. This implies that rape production has a greater N₂O emitting potential than the production of tomatoes in wetlands.     

Effect of deep placement of calcium cyanamide,coated urea,and urea on soybean (Glycine max (L.) Merr.) seed yield in relation to different inoculation methods

Abstract

The main objective of this study was to increase the productivity of soybean [Glycine max (L.) Merr. cv. Enrei] seed by deep placement of controlled release nitrogen fertilizers and by the application of different methods of inoculation of bradyrhizobia. Ten days old seedlings in an inoculated paper pot (IPP), in a non-inoculated paper pot (NIPP), and those grown in a vermiculite bed without paper pot (DT) were transplanted to an upland field converted from a drained paddy field in Nagaoka. In addition to the basal application of 16 kg N ha^?1 in the surface layer (Control), deep placement of 100 kg N ha^?1 of urea (Urea), 100-day type coated urea (CU-100), and calcium cyanamide (CaCN₂) treatments were applied at the depth of 20 cm. In the IPP method, a significantly higher seed yield was obtained with the deep placement of CaCN₂ and CU-100 compared with the Urea and Control treatments. A similar tendency was observed for the DT and NIPP methods. Among the same N fertilizer treatments, the seed yield for IPP and DT tended to exceed that for NIPP, although the NIPP roots also showed nodulation probably due to infection with indigenous bradyrhizobia. The percentage of nitrogen derived from atmospheric N₂ estimated by the simple relative ureide method was higher in the plants with CU-100 and CaCN₂ compared with those with the Urea and Control treatments at the RI stage, suggesting that the basal deep placement of CaCN₂ or CU-100 for soybean cultivation enabled the supply of N without concomitant depression of N₂ fixation. Thus the deep placement of cheaper CaCN₂ was found to be as effective as that of CU-100 for enhancing the soybean seed yield.     

Greenhouse gas emissions in response to straw incorporation,water management and their interaction in a paddy field in subtropical central China

A field experiment was conducted to study the effects of combination of straw incorporation and water management on fluxes of CH₄, N₂O and soil heterotrophic respiration (Rh) in a paddy field in subtropical central China by using a static opaque chamber/gas chromatography method. Four treatments were set up: two rice straw incorporation rates at 0 (S1) and 6 (S2) t ha^?1 combined with two water managements of intermittent irrigation (W1, with mid-season drainage) and continuous flooding (W2, without mid-season drainage). The cumulative seasonal CH₄ emissions for the treatments of S1W2, S2W1 and S2W2 increased significantly by 1.84, 5.47 and 6.63 times, respectively, while seasonal N₂O emissions decreased by 0.67, 0.29 and 1.21 times, respectively, as compared to S1W1 treatment. The significant increase in the cumulative Rh for the treatments S1W1, S2W1 and S2W2 were 0.54, 1.35 and 0.52 times, respectively, in comparison with S1W2. On a seasonal basis, both the CO₂-equivalents (CO₂e) and yield-scaled CO₂e (GHGI) of CH₄ and N₂O emissions increased with straw incorporation and continuous flooding, following the order: S2W2>S2W1>S1W2>S1W1. Thus, the practices of in season straw incorporation should be discouraged, while mid-season drainage is recommended in paddy rice production from a point view of reducing greenhouse gas emissions.     

Use of 15N isotope dilution for quantification of N2 fixation associated with roots of kallar grass (Leptochloa fusca (L.))

Summary Leptochloa fusca (L.) Kunth (kallar grass) has previously been found to exhibit high rates of nitrogen fixation. A series of experiments to determine the level of biological nitrogen fixation using ¹⁵N isotopic dilution were carried out in nutrient solution and saline soil. In the nutrient solution, E. coli inoculated plants were taken as non-nitrogen-fixing control. It was observed that nearly 60%–80% of the plant N was derived from atmospheric fixation. Estimations based on the N difference method gave much lower values (18%–35%). In experiments with saline soil which was initially sterilized with chloroform fumigation, a mixed culture of N₂-fixing rhizospheric isolates from kallar grass roots was inoculated and planted to kallar grass. Uninoculated treatments were regarded as controls. The soil was previously labelled with ¹⁵N by adding cellulose and (¹⁵NH₄)₂SO₄. The results of these studies showed fixation values of 6%–32% when estimated by ¹⁵N dilution, whereas by the N difference method 54% of the plant N was estimated to be derived from fixation. This discrepancy is due to the increase in root proliferation due to inoculation, which results in greater uptake of soil N. The distribution of ¹⁵N in different fractions of the soil-N indicted isotopic dilution due to bacterial fixation of atmospheric N₂.     

Effects of water content and N addition on potential greenhouse gas production from two differently textured soils under laboratory conditions

GHGs production and emission may vary depending on soil physical properties, water management and fertilization. Two paddy soils characterized by different texture were incubated to evaluate the impact of flooding (permanent or intermittent) and N addition on potential N₂O, CH₄ and CO₂ production and release into atmosphere and soil solution. Relationships with volumetric water content (VWC) and water filled pore space (WFPS) were evaluated. Overall, the finer clayey soil (CL) produced 58% more CH₄ than the coarser sandy soil (SA) and showed an earlier sink to source transition; the difference was lower with N addition. Permanent flooding favoured the amount of dissolved CH₄. SA produced more N₂O emissions than CL under permanent flooding (31.0 vs. 3.7%); an opposite pattern was observed for dissolved N₂O (16.4 vs. 52.7%). Fertilization increased N₂O emissions under dry conditions in CL and under flooding in SA.

Our findings showed that i) VWC had a larger influence on N₂O and CH₄ emissions than WFPS, ii) soil type influenced the gas release into atmosphere or soil solution and the timing of sink to source transition in CH₄ emissions. Further investigation on timing of fertilization and drainage are needed to improve climate change mitigation strategies.