Bacterial and fungal contributions to soil nitrogen cycling under Douglas fir and red alder at two sites in Oregon

A study was conducted at two experimental tree plantations in the Pacific Northwest to assess the roles of bacteria and fungi in nitrogen (N) cycling. Soils from red alder (Alnus rubra) and Douglas-fir (Pseudotsuga menziesii) plots in low- (H.J. Andrews) and high- (Cascade Head) productivity stands were sampled in 2005 and 2006. Fungal:bacterial ratios were determined using phospholipid fatty acid (PLFA) profiles and quantitative (Q)-PCR. Ratios from these two molecular methods were highly correlated and showed that microbial biomass varied significantly between the two experimental sites and to a lesser extent between tree types with fungal:bacterial biomass ratios lower in more N-rich plots. ¹⁵N isotope dilution experiments, with ammonium (NH₄⁺) and nitrate (NO₃^?), were paired with antibiotics that blocked bacterial (bronopol) and fungal (cycloheximide) protein synthesis. This modified isotope dilution technique was used to determine the relative contribution of bacteria and fungi to net N mineralization and gross rates of ammonification and nitrification. When bacterial protein synthesis was blocked NH₄⁺ consumption and nitrification rates decreased in all treatments except for NH₄⁺ consumption in the Douglas-fir plots at H.J. Andrews, suggesting that prokaryotic nitrifiers are a major sink for mineral NH₄⁺ in forest soils with higher N availability. Cycloheximide consistently increased NH₄⁺ consumption, however the trend was not statistically significant. Both antibiotics additions also significantly increased gross ammonification, which may have been due to continued activity of extra- and intracellular enzymes involved in producing NH₄⁺ combined with the inhibition of NH₄⁺ assimilation into proteins. The implication of this result is that microorganisms are likely a major sink for soil dissolved organic N (DON) in soils.     

Stimulated activity of the soil nitrifying community accelerates community adaptation to Zn stress

Field data have shown that soil nitrifying communities gradually adapt to zinc (Zn) after a single contamination event with reported adaptation times exceeding 1 year. It was hypothesized that this relatively slow adaptation relates to the restricted microbial diversity and low growth rate of the soil nitrifying community. This hypothesis was tested experimentally by recording adaptation rates under varying nitrification activities (assumed to affect growth rates) and by monitoring shifts in community composition. Soils were spiked at various Zn concentrations (0-4000 mg Zn kg⁻¹) and two NH₄⁺-N doses (N1, N2) were applied to stimulate growth. A control series receiving no extra NH₄⁺-N was also included. Soils were incubated in pots under field conditions with free drainage. The pore water Zn concentration at which nitrification was halved (EC50, mg Zn l⁻¹) did not change significantly during 12 months in the control series (without NH₄⁺-N applications), although nitrification recovered after 12 months at the highest Zn dose only. The EC50 after 12 months incubation increased by more than a factor 10 with increasing NH₄⁺-N dose (p < 0.05) illustrating that increased activity accelerates adaptation to Zn. Zinc tolerance tests confirmed the role of Zn exposure, time and NH₄⁺-N dose on adaptation. Zinc tolerance development was ascribed to the AOB community since the AOB/AOA ratio (AOB = ammonia oxidizing bacteria; AOA = ammonia oxidizing archaea) increased from 0.4 in the control to 1.4 in the most tolerant community. Moreover, the AOB amoA DGGE profile changed during Zn adaptation whereas the AOA amoA DGGE profile remained unaffected. These data confirm the slow but pronounced adaptation of nitrifiers to Zn contamination. We showed that adaptation to Zn was accelerated at higher activity and was associated with a shift in soil AOB community that gradually dominated the nitrifying community.     

Inhibition of ammonia-oxidizing bacteria promotes the growth of ammonia-oxidizing archaea in ammonium-rich alkaline soils

Disparities in the substrate affinity and tolerance threshold for ammonia have been believed to play a key role in driving niche differentiation between ammonia-oxidizing archaea (AOA) and bacteria (AOB); however, recent surveys argue that direct competition between AOA and AOB is also important in this phenomenon. Accordingly, it is reasonable to predict that diverse AOA lineages would grow in ammonium (NH₄⁺)-rich alkaline arable soils if AOB growth is suppressed. To test this hypothesis, a microcosm study was established using three different types of alkaline arable soils, in which a high NH₄⁺ concentration (200 μg N g^-1 dry soil) was maintained by routinely replenishing urea and the activities of AOB were selectively inhibited by 1-octyne or 3,4-dimethylpyrazole phosphate (DMPP). Compared with amendment with urea alone, 1-octyne partially retarded AOB growth, while DMPP completely inhibited AOB. Both inhibitors accelerated the growth of AOA, with significantly higher ratios of abundance of AOA to AOB observed with DMPP amendment across soils. Nonmetric multidimensional scaling analysis (NMDS) indicated that different treatments significantly altered the community structures of both AOA and AOB and AOA OTUs enriched by high-NH₄⁺ amendment were taxonomically constrained across the soils tested and closely related to Nitrososphaera viennensis EN76 and N. garnensis. Given that these representative strains have been demonstrated to be sensitive to high ammonia concentrations, our results suggest that it is the competitiveness for ammonia, rather than disparities in substrate affinity and tolerance threshold for ammonia, that drives niche differentiation between these phylotypes and AOB in NH₄⁺-rich alkaline soils.     

Soil particle surface electrochemical property effects on abundance of ammonia-oxidizing bacteria and ammonia-oxidizing archaea, NH4 activity, and net nitrification in an acid soil

Soil surface electrochemical properties may have a strong influence on nitrifying microorganisms, H⁺ and NH₄⁺ activities, and therefore on the nitrification process. A gradient of surface electrochemical parameters was obtained by amendment of a subtropical acid pine soil (Oxisol) with 0% (control), 3%, 5%, 8%, 10% and 12% pure Ca-Montmorillonite by weight. The H⁺ and NH₄⁺ activities, the abundance of the ammonia-oxidizing bacterial (AOB) and archaeal (AOA) amoA gene copies, and time-dependent kinetics of net nitrification were investigated. Soil particle surface specific area ranged from 53 to 103 m² g⁻¹ and increased with increasing montmorillonite application rate. Similar to specific area, surface charge quantity, surface charge density, electric field strength and surface potential increased after montmorillonite amendment. The H⁺ and NH₄⁺ activities decreased linearly after montmorillonite addition. AOB amoA gene copy number was 1.82 × 10⁵ copies g⁻¹ for unamended soil, and the highest AOB amoA gene copy numbers were found for the 10% montmorillonite amendment (3.11 × 10⁷ g⁻¹ soil), which was more than 150 times higher than unamended soil. AOA amoA gene copy numbers were 9.19 × 10³ copies g⁻¹ dry unamended soil, and the highest AOA amoA gene copy numbers were found in the 8% montmorillonite amendment (1.22 × 10⁵ g⁻¹ soil). Although pH significantly decreased during the first three weeks of incubation, no significant difference was observed between the unamended control and different rates of montmorillonite addition treatments during the whole incubation. The largest net nitrification (103 mg N kg⁻¹) was observed in the 10% montmorillonite amendment and the lowest in unamended soil (62 mg N kg⁻¹). While montmorillonite did not change the kinetic patterns of net nitrification, the highest nitrification potential (275 mg N kg⁻¹) for the 10% montmorillonite treatment was more than 3 times higher than unamended soil from simulation of time-dependent kinetics. Nitrification was significantly stimulated after montmorillonite amendment in acid soil mainly due to an increase in the quantity and activity of AOB and AOA. We concluded that soil particle surface parameters can significantly influence nitrification, especially in acid soils.     

Ammonia-oxidizing archaea are more important than ammonia-oxidizing bacteria in nitrification and NO3 −-N loss in acidic soil of sloped land

As part of a long-term sloped land use experiment established in 1995 at Taoyuan Agro-ecosystem Research Station (111°26′ E, 28°55′ N) in China, soil samples were collected from three land use types, including cropland (CL), natural forest, and tea plantation. Quantitative polymerase chain reaction and terminal restriction fragment length polymorphism were used to determine the abundance and community composition of amoA-containing bacteria (AOB) and archaea (AOA). The results indicate that land use type induced significant changes in soil potential nitrification rate and community composition, diversity, and abundance of AOB and AOA. Both AOB and AOA community compositions were generally similar between upper and lower slope positions (UP and LP), except within CL. The LP soils had significantly (p?<?0.05) higher diversity and abundance of both AOB and AOA than in the UP. Potential nitrification rate was significantly correlated (p?<?0.05) with diversity and abundance of AOA, but not with AOB. Among land use types, the NO₃ ^? and amoA-containing AOA runoff loss was greatest in CL. Nitrate-N runoff loss was significantly correlated (p?<?0.05) with the loss of AOA amoA copies in the runoff water. Furthermore, relationships between NO₃ ^?-N runoff loss and abundance of AOA but not of AOB at both slope positions were significantly correlated (p?<?0.05). These findings suggest that AOA are more important than AOB in nitrification and NO₃ ^?-N runoff loss in acidic soils across sloped land use types.     

Potato cultivar type affects the structure of ammonia oxidizer communities in field soil under potato beyond the rhizosphere

The effects of plants on the microbiota involved in the oxidation of ammonia in soils have been controversial. Here, we investigated the dynamics in the abundances and community structures of the bacterial and archaeal ammonia oxidizers (AOB and AOA, respectively) in two fields that were cropped with potato. Six different potato cultivars were used, including a genetically-modified one, in a fourfold replicated experimental set-up. On the basis of bulk and rhizosphere soil extracted microbial community DNA, AOB and AOA quantitative PCR as well as PCR-DGGE were performed. In addition, samples were used for the production and analysis of amoA gene fragment based clone libraries. Regardless of sample type (bulk versus rhizosphere soil) and across soils, the population sizes of AOA (of the order 10⁴–10⁸ amoA gene copies g⁻¹ dry soil), were generally higher than those of AOB in the same samples (about 10⁴–10⁵ g⁻¹ dry soil), resulting in ratio's of log-transformed values > 1.0. Whereas the AOB numbers were generally raised in the rhizosphere versus bulk soils in both soils, the opposite was true for the AOA numbers. Moreover, significant effects of cultivar type on both the AOB and AOA community structures were found in both soils, and these extended to beyond the rhizospheres. The effects were found across the whole growth season. Soil type did not significantly affect the community structures of AOA, but had a small effect on the community structure of AOB. Analysis of the structures of the AOB communities revealed a prevalence of AOB subgroups 2, 3a, 3b and 4 in one field soil and of 2 and 4 in the other one. With respect to the AOA, soil/sediment clusters (SS) I, II, III and IV were found to prevail.     

The relative contribution of nitrifiers to autotrophic nitrification across a pH-gradient in a vegetable cropped soil

Purpose

Microbial nitrification plays an important role in nitrogen cycling in ecosystems. Nitrification is performed by ammonia-oxidizing archaea (AOA), ammonia-oxidizing bacteria (AOB), and nitrite-oxidizing bacteria (NOB) including complete ammonia oxidizers. However, the relative importance of nitrifiers in autotrophic nitrification in relation to soil pH is still unclear.

Materials and methods

Combining DNA-based stable isotope probing (SIP) and molecular biological techniques, we investigated the abundance, structure, and activity of AOA, AOB, and NOB along a pH-gradient (3.97–7.04) in a vegetable cropped soil.

Results and discussion

We found that AOA abundance outnumbered AOB abundance and had a significantly negative relationship with soil pH. The abundances of NOB Nitrospira 16S rRNA, nxrB gene, and Nitrobacter nxrA gene were affected by soil pH. Incubation of soil with ¹³CO₂ and DNA-SIP analysis demonstrated that significant ¹³CO₂ assimilation by AOA rather than by AOB occurred in the acidic soils, whereas the labeled ¹³C level of AOA was much less in the neutral soil than in the acidic soils. There was no evidence of ¹³CO₂ assimilation by NOB except for Nitrobacter with NxrB gene at pH 3.97. Phylogenetic analysis of AOA amoA gene in the ¹³C- and ¹²C-labeled treatments showed that the active AOA mainly belonged to Nitrososphaera in the acidic soils.

Conclusions

These results suggested that the main performer of nitrification was AOA in the acidic soils, but both AOA and AOB participated in nitrification in the neutral soil with low nitrification activity. NOB Nitrospira and Nitrobacter did not grow in the soils with pH 4.82–7.04 and other populations of NOB were probably involved in nitrite oxidation in the vegetable cropped soil.

     

Effects of NH4+ and NO3− on litter and soil organic carbon decomposition in a Chinese fir plantation forest in South China

Soil organic carbon (SOC) dynamics and nutrient availability determine the soil quality and fertility in a Chinese fir plantation forest in subtropical China. Uniformly ¹³C-labeled Chinese fir (Cunninghamia lanceolata) and alder (Alnus cremastogyne) leaf litter with or without 100 mg NH₄⁺ or NO₃⁻ were added to the soil. The purpose was to investigate the influence of N availability on the decomposition of the litter and native SOC. The production of CO₂, the natural abundance of ¹³C–CO₂, and the inorganic N dynamics were monitored. The results showed that Chinese fir (with a high C:N ratio) and alder (with a low C:N ratio) leaf litter caused significant positive priming effects (PEs) of 24% and 42%, respectively, at the end of the experiment (235 d). The PE dynamics showed that positive PE can last for at least 87 d. However, the possible occurrence of a significant negative PE with a sufficient incubation period is difficult to confirm. The application of both NH₄⁺ and NO₃⁻ was found to have a stimulating effect on the decomposition of Chinese fir and alder leaf litter in the early stage (0–15 d) of incubation, but an adverse effect in the late stage. Compared with NO₃⁻, NH₄⁺ caused a greater decrease in the PE induced by both Chinese fir and alder leaf litter. The effects of NH₄⁺ and NO₃⁻ on the PE dynamics had different patterns for different incubation stages. This result may indicate that the stability or recalcitrance of SOC, especially in such plantation forest soils, strongly depends on available leaf litter and application of N to the soil.     

Variation in the relationship between nitrification and acidification of subtropical soils as affected by the addition of urea or ammonium sulfate

The effects on nitrification and acidification in three subtropical soils to which (NH₄)₂SO₄ or urea had been added at rate of 250 mg N kg⁻¹ was studied using laboratory-based incubations. The results indicated that NH₄⁺ input did not stimulate nitrification in a red forest soil, nor was there any soil acidification. Unlike red forest soil, (NH₄)₂SO₄ enhanced nitrification of an upland soil, whilst urea was more effective in stimulating nitrification, and here the soil was slightly acidified. For another upland soil, NH₄⁺ input greatly enhanced nitrification and as a result, this soil was significantly acidified. We conclude that the effects of NH₄⁺ addition on nitrification and acidification in cultivated soils would be quite different from in forest soils. During the incubation, N isotope fractionation was closely related to the nitrifying capacity of the soils.     

Influence of bacterial-feeding nematodes on nitrification and the ammonia-oxidizing bacteria (AOB) community composition

The effects of bacterial-feeding nematodes on nitrification and the ammonia-oxidizing bacteria (AOB) community composition were studied in soil microcosms. Sterilized soils were inoculated with mixed soil bacteria (obtained by filtering) or with bacteria and bacterial-feeding nematodes, after which the dynamic inorganic nitrogen concentration was measured weekly. After 28 days of incubation, denaturing gradient gel electrophoresis (DGGE) based on PCR amplification of the amoA gene was used to analyze the AOB community composition. In addition, a clone library from the amoA gene fragments was established using clones randomly selected and sequenced from the two treatments. The results showed that the presence of bacterial-feeding nematodes led to significantly greater NH₄⁺ and NO₃⁻ contents over the entire incubation period, indicating that bacterial-feeding nematodes promoted both N mineralization and nitrification. The results of DGGE showed that the AOB community composition was significantly changed in the presence of bacterial-feeding nematodes. Furthermore, the sequencing results suggested that Nitrosospira sp. was the dominant species in the treatment without nematodes, while Nitrosomonas sp. and Nitrosospira sp. were the dominant species in the treatment with nematodes. Such changes in the AOB community may be one of explanation of the important role that nematodes play in promoting nitrification.     

Contrasting response of nitrification capacity in three agricultural soils to N addition during short-term incubation

Purpose

Human disturbance is a major culprit driving imbalances in the biological transformation of nitrogen from the nonreactive to the reactive pool and is therefore one of the greatest concerns for nitrogen (N) cycling. The objective of this study was to compare potential nitrification rates and the abundance of ammonia oxidizers responsible for nitrification, with the amendment of external N in different agricultural soils.

Materials and methods

Three typical Chinese agricultural soils, QiYang (QY) acid soil, ShenYang (SY) neutral soil, and FengQiu (FQ) alkaline soil, were amended with 0, 20, 150, and 300 μg NH₄ ⁺-N g^?1 soil and incubated for 40 days. The abundance of ammonia oxidizing bacteria (AOB) and archaea (AOA) at the end of incubation in the soil microcosms was determined using the real-time PCR.

Results and discussion

There was a significant decrease in ammonium concentration in the QY soil from the highest to the lowest N-loading treatments, while no significant difference in ammonium concentrations was detected among the different N-loading treatments for the SY and FQ soils. A significantly higher potential nitrification rate (PNR) was observed in the FQ soil while lowest PNR was found in the QY soil. Quantitative PCR analysis of AOB amoA genes demonstrated that AOB abundance was significantly higher in the high N-loading treatments than in the control for the QY soil only, while no significant difference among treatments in the SY and FQ soils. A significant positive correlation between PNR and AOB amoA abundance, however, was found for the SY and FQ soils, but not for the QY soil. Little difference in AOA amoA abundance between different N-loading treatments was observed for all the soils.

Conclusions

This study suggested that ammonia oxidation capacity in the FQ and SY soils was higher than those in the QY soil with the addition of ammonium fertilizer for a short-term. These findings indicated that understanding the differential responses of biological nitrification to varying input levels of ammonium fertilizer is important for maximizing N use efficiency and thereby improving agricultural fertilization management.     

冬种绿肥对水稻土硝化作用的影响

冬闲田种植绿肥是传统的水稻土培肥增产措施,但绿肥-水稻种植系统中,不同绿肥种类对硝化作用的影响规律及调控机制尚不明确.采用盆栽试验,研究了冬种紫云英、油菜、黑麦草对土壤性状及硝化作用的影响,并通过特异性细菌抑制剂(卡那霉素和大观霉素)研究了氨氧化细菌(AOB)和氨氧化古菌(AOA)对硝化作用的相对贡献.结果表明,冬种三...     

Short-term population dynamics of ammonia oxidizing bacteria in an agricultural soil

Ammonia oxidizing bacteria (AOB) control the rate limiting step of nitrification, the conversion of ammonia (NH₄⁺) to nitrite (NO₂⁻). The AOB therefore have an important role to play in regulating soil nitrogen cycling. Tillage aerates the soil, stimulating rapid changes in soil N cycling and microbial communities. Here we report results of a study of the short term responses of AOB and net nitrification to simulated tillage and NH₄⁺ addition to soil. The intensively farmed vegetable soils of the Salinas Valley, California, provide the context for this study. These soils are cultivated frequently, receive large N fertilizer inputs and there are regional concerns about groundwater N concentrations. An understanding of N dynamics in these systems is therefore important. AOB population sizes were quantified using a real-time PCR approach. In a 15 day experiment AOB populations, increased rapidly following tillage and NH₄⁺ addition and persisted after the depletion of soil NH₄⁺. AOB population sizes increased to a similar degree, over a 1.5-day period, irrespective of the amount of NH₄⁺ supplied. These data suggest selection of an AOB community in this intensively farmed and C-limited soil, that rapidly uses NH₄⁺ that becomes available. These data also suggest that mineralization may play an especially important role in regulating AOB populations where NH₄⁺ pool sizes are very low. Methodological considerations in the study of soil AOB communities are also discussed.     

Responses of bacterial and archaeal ammonia oxidisers of an acidic luvisols soil to different nitrogen fertilization rates after 9?years

It is still not clear which group of ammonia-oxidizing microorganisms plays the most important roles in nitrification in soils. Change in abundances and community compositions of ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA) under long-term different nitrogen (N) fertilization rates were investigated in an acidic luvisols soil using real-time polymerase chain reaction and denaturing gradient gel electrophoresis, respectively, based on the ammonia monooxygenase a-subunit gene. The experimental plan included the following treatments: control without N fertilization (N_CK), low N fertilization rate, middle N fertilization rate, and high N fertilization rate as 0, 100, 150, and 250?kg urea-N?ha^?1, respectively. Long-term different N fertilization rates did not significantly alter the total C and N contents of soil while it significantly decreased soil pH, which ranged from 5.60 to 5.20. The AOB abundance was more abundant in the N fertilization treatments than the N_CK treatment; the AOA abundance decreased by the increasing N fertilization rates, as did the ratios of AOA/AOB. The large differences in the potential nitrification rates among four treatments depended on the changes in AOA abundance but not to changes in AOB abundance. Phylogenetic analysis showed that the AOB communities were dominated by Nitrosospira clusters 1, 3, and 9 while all AOA sequences were grouped into soil/sediment cluster except for one sequence. Taken together, these results indicated that AOB and AOA preferred different soil N conditions and AOA were functionally more important in the nitrification than AOB in the acidic luvisols soil.     

Effect of application rate of a nitrification inhibitor,dicyandiamide (DCD), on nitrification rate,and ammonia-oxidizing bacteria and archaea growth in a grazed pasture soil: An incubation study

Purpose

Dicyandiamide (DCD) has been used commercially in New Zealand to reduce nitrate leaching and N₂O emissions in grazed pastures. However, there is a lack of information in the literature on the optimum rate of DCD to achieve the environmental benefits while at the same time reducing the cost of the technology. The objective of this study was to determine the effect of DCD application rate on its effectiveness to inhibit ammonia oxidizer growth and nitrification rate in a grazed pasture soil.

Materials and methods

The soil was a Templeton silt loam (Immature Pallic Soil; Udic Haplustepts) collected from Lincoln University Research Dairy Farm with a mixed pasture consisting of perennial ryegrass (Lolium perenne L.) and white clover (Trifolium repens L.) and was incubated alone (control) or with cow urine at 700 kg N/ha with 6 rates of DCD [0, 2.5, 5, 7.5, 10 (applied twice), 15 and 20 kg/ha] in incubation vessels. The incubation vessels were placed randomly in an incubator with a constant temperature of 12 °C. During 112 days of incubation, soil subsamples were taken at different time intervals to measure the concentrations of NO₃ ^?-N and NH₄ ⁺-N and the amoA gene copy numbers of ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA).

Results and discussion

DCD applied at all the different rates inhibited nitrification in urine-treated soils, but the effectiveness increased with DCD application rate. In addition, AOB growth and the amounts of nitrate-N in the soil were significantly related to the application rate of DCD. However, AOA population abundance showed no relationship to the application rate of DCD. The DCD rate at which the AOB growth rate and nitrate-N concentration were halved (effective dosage that causes 50 % reduction in nitrification rate, or ED₅₀) was about 10 kg DCD/ha.

Conclusions

These results suggest that DCD applied at relatively low rates still slowed down the nitrification rate, and the current recommended rate of 10 kg DCD/ha for DCD use in New Zealand grazed pastures would result in a 50 % reduction in nitrification rate in this soil. The actual rate of DCD application used would depend on the cost of the product and the environmental and agronomic benefits that would result from its use.     

The effects of different irrigation regimes on nitrous oxide emissions and influencing factors in greenhouse tomato fields

Purpose

Intensive agricultural practices have enhanced problems associated with the competing use of limited water resources. Nitrous oxide (N₂O) is a major contributor to global warming. It is important for researchers to ascertain the relationship between irrigation and soil N₂O emissions in order to identify mitigation strategies to reduce nitrous oxide emissions. Different irrigation amounts affect soil water dynamics and nitrogen turnover. The effect of three lower limits of irrigation on soil N₂O emissions, influencing factors, and abundance of genes involved in nitrification and denitrification were investigated in tomato irrigated in a greenhouse.

Materials and methods

Observations were performed between April and August 2015 in a long-term irrigated field subjected to different lower limits of irrigation: 20 kPa (D₂₀), 30 kPa (D₃₀), and 40 kPa (D₄₀) from greenhouse soil during the tomato crop season. Soil N₂O fluxes were monitored using the static chamber-gas chromatograph method. Copy numbers of genes were determined using the real-time quantitative polymerase chain reaction (real-time PCR) technique. Characteristics of soil N₂O emissions were analyzed, and differences between irrigation regimes were determined. The effects of influencing factors on soil N₂O emissions were analyzed, including soil temperature, soil moisture, soil pH, and soil mineral nitrogen, as well as changes in the abundance of soil ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA) based on amoA genes and denitrifier genes (nosZ, nirK, and cnorB).

Results and discussion

Our results showed that peaks in N₂O emissions occurred 1–5 days after each irrigation. During the whole tomato growth period, soil N₂O fluxes were lowest under D₃₀ treatment compared with those under D₂₀ and D₄₀ treatments. Soil NO₃ ^?-N concentrations were significantly higher than NH₄ ⁺-N concentrations. Soil N₂O fluxes were significantly related to soil moisture, NH₄ ⁺-N concentrations (P < 0.01), soil pH, and AOA copy numbers (P < 0.05). There was no consistent correlation between soil N₂O emissions, soil temperature, and soil NO₃ ^?-N concentrations. Different irrigation regimes significantly affected AOA copy numbers but did not affect the expression of other genes. AOA copy numbers were higher than those of AOB. Soil N₂O fluxes significantly affected the AOA copy numbers and potential nitrification rates (P < 0.05).

Conclusions

Soil moisture, pH, and NH₄ ⁺-N concentration were important factors affecting soil N₂O emissions. Compared with other genes associated with nitrification and denitrification, AOA plays an important role in N₂O emissions from greenhouse soils. Selecting a lower limit of irrigation of 30 kPa could effectively reduce N₂O emissions from vegetable soils.

     

Conversion of grassland to coniferous woodland has limited effects on soil nitrogen cycle processes

In the last century, conversion of native North American grasslands to Juniperus virginiana forests or woodlands has dramatically altered ecosystem structure and significantly increased ecosystem carbon (C) stocks. We compared soils under recently established J. virginiana forests and adjacent native C₄-dominated grassland to assess changes in potential soil nitrogen (N) transformations and plant available N. Over a 2-year period, concentrations of extractable inorganic N were measured in soils from forest and grassland sites. Potential gross N ammonification, nitrification, and consumption rates were determined using ¹⁵N isotope-dilution under laboratory conditions, controlling for soil temperature and moisture content. Potential nitrification rates (V_max) and microbial biomass, as well as soil physical and chemical properties were also assessed. Extractable NH₄⁺ concentrations were significantly greater in grassland soils across the study period (P ≤ 0.01), but analysis by date indicated that differences in extractable inorganic N occurred more frequently in fall and winter, when grasses were senescent but J. virginiana was still active. Laboratory-based rates of gross N mineralization (ammonification) and nitrification were greater in grassland soils (P ≤ 0.05), but only on one of four dates. Potential nitrification rates (V_max) were an order of magnitude greater than gross nitrification rates in both ecosystems, suggesting that nitrification is highly constrained by NH₄⁺ availability. Differences in plant uptake of N, C inputs, and soil microclimate as forests replace grasslands may influence plant available N in the field, as evidenced by seasonal differences in soil extractable NH₄⁺, and total soil C and N accumulation. However, we found few differences in potential soil N transformations under laboratory conditions, suggesting that this grassland-to-forest conversion caused little change in mineralizable organic N pools or potential microbial activity.     

Effects of 3,4-dimethylpyrazole phosphate (DMPP) on the abundance of ammonia oxidizers and denitrifiers in two different intensive vegetable cultivation soils

Purpose

Nitrification and denitrification in the N cycle are affected by various ammonia oxidizers and denitrifying microbes in intensive vegetable cultivation soils, but our current understanding of the effect these microbes have on N₂O emissions is limited. The nitrification inhibitor, 3,4-dimethylpyrazole phosphate (DMPP), acts by slowing nitrification and is used to improve fertilizer use efficiency and reduce N losses from agricultural systems; however, its effects on nitrifier and denitrifier activities in intensive vegetable cultivation soils are unknown.

Materials and methods

In this study, we measured the impacts of DMPP on N₂O emissions, ammonia oxidizers, and denitrifying microbes in two intensive vegetable cultivation soils: one that had been cultivated for a short term (1 year) and one that had been cultivated over a longer term (29 years). The quantitative PCR technique was used in this study. Three treatments, including control (no fertilizer), urea alone, and urea with DMPP, were included for each soil. The application rates of urea and DMPP were 1800 kg ha^?1 and 0.5% of the urea-N application rate.

Results and discussion

The application of N significantly increased N₂O emissions in both soils. The abundance of ammonia-oxidizing bacteria (AOB) increased significantly with high rate of N fertilizer application in both soils. Conversely, there was no change in the growth rate of ammonia-oxidizing archaea (AOA) in response to the applied urea despite the presence of larger numbers of AOA in these soils. This suggests AOB may play a greater role than AOA in the nitrification process, and N₂O emission in intensive vegetable cultivation soils. The application of DMPP significantly reduced soil NO₃^?-N content and N₂O emission, and delayed ammonia oxidation. It greatly reduced AOB abundance, but not AOA abundance. Moreover, the presence of DMPP was correlated with a significant decrease in the abundance of nitrite reductase (nirS and nirK) genes.

Conclusions

Long-term intensive vegetable cultivation with heavy N fertilization altered AOB and nirS abundance. In vegetable cultivation soils with high N levels, DMPP can be effective in mitigating N₂O emissions by directly inhibiting both ammonia oxidizing and denitrifying microbes.

     

High contribution of ammonia-oxidizing archaea (AOA) to ammonia oxidation related to a potential active AOA species in various arable land soils

Purpose

Ammonia oxidation is the limiting step in soil nitrification and critical in the global nitrogen cycle. The discovery of ammonia-oxidizing archaea (AOA) has improved our knowledge of microbial mechanisms for ammonia oxidation in complex soil environments. However, the relative contributions of AOA and ammonia-oxidizing bacteria (AOB) to ammonia oxidation remain unclear.

Materials and methods

In this study, through large geographical scale sampling in China, totally nine samples representing various types of arable land soils were selected for analyzing the ammonia oxidation activity. The AOA and AOB activities were separately determined by using the dicyandiamide and 1-octyne inhibition method. High-throughput pyrosequencing and DNA stable-isotope probing (DNA-SIP) analysis were applied to investigate the distribution and activity of Candidatus Nitrosocosmicus franklandus in the arable land soils.

Results and discussion

In this study, AOA abundance (3.2?×?10⁷–3.4?×?10⁹ copies g^?1) and activity (0.01–1.33 mg N kg^?1 dry soil day^?1) were evaluated for nine selected arable land soils and accounted for 4–100% of ammonia oxidation. By separately determining AOA and AOB rates, we observed that archaeal ammonia oxidation dominated the ammonia oxidation process in six soils, revealing a considerable contribution of AOA in ammonia oxidation in arable land soils. Based on high-throughput pyrosequencing analysis, the AOA species Ca. N. franklandus with relatively low abundance (0.6–13.5% in AOA) was ubiquitously distributed in all the tested samples. Moreover, according to the DNA-SIP analysis for Urumqi sample, the high activity and efficiency of Ca. N. franklandus in using CO₂ suggests that this species plays an important role in archaeal ammonia oxidation in arable land soils.

Conclusions

Through determining the AOA activity and analyzing the potential predominant functional AOA species, this study greatly improves our understanding of ammonia oxidation in arable land soils.

     

亚热带松林中强酸性土壤自养和异养硝化作用的研究

The occurrence of nitrification in some acidic forest soils is still a subject of debate. Identification of main nitrification pathways in acidic forest soils is still largely unknown. Acidic yellow soil (Oxisol) samples were selected to test whether nitrification can occur or not in acidic subtropical pine forest ecosystems. Relative contributions of autotrophs and heterotrophs to nitrification were studied by adding selective nitrification inhibitor nitrapyrin. Soil NH₄⁺-N concentrations decreased, but NO₃^--N concentrations increased significantly for the no-nitrapyrin control during the first week of incubation, indicating that nitrification did occur in the acidic subtropical soil. The calculated net nitrification rate was 0.49 mg N kg^-1 d^-1 for the no-nitrapyrin control during the first week of incubation. Nitrapyrin amendment resulted in a significant reduction of NO₃^--N concentration. Autotrophic nitrification rate averaged 0.28 mg N kg^-1 d^-1 and the heterotrophic nitrification rate was 0.21 mg N kg^-1 d^-1 in the first week. Ammonia-oxidizing bacteria (AOB) abundance increased slightly during incubation, but nitrapyrin amendment significantly decreased AOB amoA gene copy numbers by about 80%. However, the ammonia-oxidizing archaea (AOA) abundance showed significant increases only in the last 2 weeks of incubation and it was also decreased by nitrapyrin amendment. Our results indicated that nitrification did occur in the present acidic subtropical pine forest soil, and autotrophic nitrification was the main nitrification pathway. Both AOA and AOB were the active biotic agents responsible for autotrophic nitrification in the acidic subtropical pine forest soil.