Responses of soil microbial community to nitrogen fertilizer and precipitation regimes in a semi-arid steppe

Purpose

Changes of nitrogen (N) cycle caused by N fertilization and precipitation regimes have affected the key ecosystem structure and functions in temperate steppe, which may modify the structure of soil microbial communities involved in N transformation. This paper was designated to examine the response of soil ammonia oxidizers and denitrifiers to the N fertilization and precipitation regimes in a semi-arid steppe where N and water contents are major limiting factors of the grassland productivity.

Materials and methods

This study was based on a long-term N fertilization and precipitation regimes experiment in Inner Mongolia (116° 17′ 20″ E, 42° 2′ 29″ N). The treatments including CK (control), R (reduced precipitation), W (30% increase in precipitation), N (10 g N m^?2 y^?1), RN (reduced precipitation and 10 g N m^?2 y^?1), and WN (30% increase in precipitation and 10 g N m^?2 y^?1). Soil basic chemical properties and microbial activities were analyzed. Molecular methods were applied to determine the abundance, structure and diversity of ammonia oxidizers and denitrifiers. Statistical analysis detected the main and interactive effect of treatments on soil microbial communities and revealed the relationship between soil microbial community structures and environmental factors.

Results and discussion

N fertilization significantly increased ammonia-oxidizing bacteria (AOB) abundance. Ammonia-oxidizing archaea (AOA) community structure was markedly changed in N fertilizer treatment and strongly affected by soil pH, while soil nitrate and water content correlated with AOB community structure. Soil nitrate was the key factor influencing nirK gene community structure, while soil pH and water content explained much of the variations of nosZ gene community. AOB-amoA and nosZ gene community diversities were influenced by precipitation regimes and interaction of N fertilization and precipitation regimes, respectively.

Conclusions

N fertilization and precipitation regimes had significant influences on the changes of soil properties and microbial functional communities. Soil nitrification was mainly driven by AOB in the semi-arid grassland. Changes of substrate content and soil pH were the key factors in shifting functional microbial communities. The non-synergistic effects of N fertilization and precipitation regimes on the microbial functional groups indicated that the negative effect of lower pH induced by N fertilization would be alleviated by precipitation regimes, which should be well considered in grassland restoration.

     

Responses of soil nitrous oxide production and abundances and composition of associated microbial communities to nitrogen and water amendment

Soil moisture and nitrogen (N) are two important factors influencing N₂O emissions and the growth of microorganisms. Here, we carried out a microcosm experiment to evaluate effects of soil moisture level and N fertilizer type on N₂O emissions and abundances and composition of associated microbial communities in the two typical arable soils. The abundances and community composition of functional microbes involved in nitrification and denitrification were determined via quantitative PCR (qPCR) and terminal restriction length fragment polymorphism (T-RFLP), respectively. Results showed that N₂O production was higher at 90% water-filled pore (WFPS) than at 50% WFPS. The N₂O emissions in the two soils amended with ammonium were higher than those amended with nitrate, especially at relatively high moisture level. In both soils, increased soil moisture stimulated the growth of ammonia-oxidizing bacteria (AOB) and nitrite reducer (nirK). Ammonium fertilizer treatment increased the population size of AOB and nirK genes in the alluvial soil, while reduced the abundances of ammonia-oxidizing archaea (AOA) and denitrifiers (nirK and nosZ) in the red soil. Nitrate addition had a negative effect on AOA abundance in the red soil. Total N₂O emissions were positively correlated to AOB abundance, but not to other functional genes in the two soils. Changed soil moisture significantly affected AOA rather than AOB community composition in both soils. The way and extent of N fertilizers impacted on nitrifier and denitrifier community composition varied with N form and soil type. These results indicate that N₂O emissions and the succession of nitrifying and denitrifying communities are selectively affected by soil moisture and N fertilizer form in the two contrasting types of soil.     

Nitrogen loading levels affect abundance and composition of soil ammonia oxidizing prokaryotes in semiarid temperate grassland

Purpose  

Global nitrogen deposition has profound impact on the terrestrial ecosystem including the semiarid temperate grassland, causing vegetation community shifts and soil acidification. Little is known regarding the effect of nitrogen (N) deposition on the belowground microbial communities. This study aimed to examine the response of ammonia-oxidizing bacteria (AOB) and archaea (AOA) to added N in semiarid temperate grassland.     

Methanotroph abundance not affected by applications of animal urine and a nitrification inhibitor,dicyandiamide, in six grazed grassland soils

Purpose  

Methanotrophs are an important group of methane (CH₄)-oxidizing bacteria in the soil, which act as a major sink for the greenhouse gas, CH₄. In grazed grassland, one of the ecologically most sensitive areas is the animal urine patch soil, which is a major source of both nitrate (NO₃ ⁻) leaching and nitrous oxide (N₂O) emissions. Nitrification inhibitors, such as dicyandiamide (DCD), have been used to mitigate NO₃ ⁻ leaching and N₂O emissions in grazed pastures. However, it is not clear if the high nitrogen loading rate in the animal urine patch soil and the use of nitrification inhibitors would have an impact on the abundance of methanotrophs in grazed grassland soils. The purpose of this study was to determine the effect of animal urine and DCD on methanotroph abundance in grazed grassland soils.     

Frontiers in the microbial processes of ammonia oxidation in soils and sediments

Purpose

Two recent discoveries in nitrogen (N) cycling processes, i.e., archaeal ammonia oxidizers and anaerobic ammonia (ammonium) oxidation (anammox), have triggered great interest in studying microbial ammonia oxidation processes. The purpose of this review is to highlight recent progress in ammonia oxidation processes in soils and sediments and to propose future research activities in this topic.

Results and discussion

Aerobic ammonia oxidation and anammox processes are linked through the production and consumption of nitrite, respectively, thereby removing the reactive N (NH₄ ⁺, NO₂ ^?, NO₃ ^?) from soil and sediment ecosystems. Ammonia-oxidizing microorganisms are widely distributed in soils and sediments, and increasing evidence suggests that ammonia-oxidizing archaea and bacteria are functionally dominant in the ammonia oxidation of acid soils and other soils, respectively. The widespread occurrence and great variation in the abundance of anammox bacteria indicate their heterogeneous distribution and niche differentiation. Therefore, the worldwide distribution of both microbial groups in nature has stimulated researchers to investigate the physiology and metabolism of related groups, as well as appraising their contribution to N cycling.

Conclusions

We summarized the current progress and provided future perspectives in the microbiology of aerobic and anaerobic ammonia oxidation in soils and sediments. With increasing concern and interest in soil and sediment ammonia oxidation processes, studies in the microbial mechanisms underlying nitrification and anammox, as well as their interactions, are essential for understanding their contribution to the loss of N either through nitrate leaching or N-related gas emissions.     

DNA stable isotope probing revealed no incorporation of 13CO2 into comammox Nitrospira but ammonia-oxidizing archaea in a subtropical acid soil

Journal of Soils and Sediments - The recently discovered complete ammonia oxidizers (comammox Nitrospira) challenged our concept of the whole nitrification process, i.e., ammonia oxidation...     

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.     

Soil type determines the abundance and community structure of ammonia-oxidizing bacteria and archaea in flooded paddy soils

Purpose  

Ammonia oxidation plays an important role in global nitrogen cycle. However, little information is available on ammonia oxidizers in paddy soils. This study aimed to understand the controlling factors of ammonia oxidizers in the paddy soils.     

Dynamics of sulfate reduction and sulfate-reducing prokaryotes in anaerobic paddy soil amended with rice straw

Incorporation of rice straw to soil is a common agricultural practice in rice cultivation. In anaerobic paddy soil, the complete mineralization of organic matter to CH₄ and CO₂ is accomplished by the sequential reduction of nitrate, ferric iron, sulfate, and methanogenesis. In order to estimate the temporal changes of sulfate-reducing prokaryotes (SRP) as decomposers of organic matters, the effects of rice straw amendment on the dynamics of sulfate reduction and SRP were investigated by combining the monitoring of CH₄, sulfate, and organic acids with molecular tools such as soil DNA extraction, real-time PCR, cloning, sequencing, and phylogenetic analysis. The incorporation of rice straw into paddy soil significantly increased concentrations of sulfate, formate, acetate, propionate, and lactate and CH₄ production. The rate of sulfate reduction in the straw-amended slurries was significantly higher than that in the unamended slurries. The dsrAB gene copy numbers of SRP in the straw-amended soil slurries ranged from 4.26 × 10⁶ to 1.96 × 10⁸ per gram of dry soil, which were significantly higher than those in the unamended control ranging from 1.99 × 10⁶ to 7.90 × 10⁷ per gram of dry soil. Significant correlations were observed between SRP dsrAB gene copy numbers and the concentrations of sulfate and acetate. Cloning and sequencing analyses showed a clear shift of SRP community structure between treatments and time. In the straw-amended slurries, Clostridia-like SRP significantly increased, while Deltaproterobacteria-like SRP (Sytrophobacter, Desulfobacterium, Desulfovibrio, and Desulfomonile) decreased during the incubation period. Novel uncultured SRP were abundant in the straw-amended slurries and changed during the incubation period.     

Contrasting response of two grassland soils to N addition and moisture levels: N<Subscript>2</Subscript>O emission and functional gene abundance

Purpose

Nitrification and denitrification processes dominate nitrous oxide (N₂O) emission in grassland ecosystems, but their relative contribution as well as the abiotic factors are still not well understood.

Materials and methods

Two grassland soils from Duolun in Inner Mongolia, China, and Canterbury in New Zealand were used to quantitatively compare N₂O production and the abundance of bacterial and archaeal amoA, denitrifying nirK and nirS genes in response to N additions (0 and 100 μg NH₄ ⁺–N g^?1 dry soil) and two soil moisture levels (40 and 80 % water holding capacity) using microcosms.

Results and discussion

Soil moisture rather than N availability significantly increased the nitrification rate in the Duolun soil but not in the Canterbury soil. Moreover, N addition promoted denitrification enzyme activities in the Canterbury soil but not in the Duolun soil. The abundance of bacterial and archaeal amoA genes significantly increased as soil moisture increased in the Duolun soil, whereas in the Canterbury soil, only the abundance of bacterial amoA gene increased. The increase in N₂O flux induced by N addition was significantly greater in the Duolun soil than in the Canterbury soil, suggesting that nitrification may have a dominant role in N₂O emission for the Duolun soil, while denitrification for the Canterbury soil.

Conclusions

Microbial processes controlling N₂O emission differed in grassland soils, thus providing important baseline data in terms of global change.