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.     

Chronic Nitrogen Fertilization Modulates Competitive Interactions Among Microbial Ammonia Oxidizers in a Loess Soil

Nitrogen(N) application may lead to niche segregation of soil ammonia-oxidizing archaea(AOA) and bacteria(AOB), thereby reducing the competitive interactions between AOA and AOB due to higher ammonium substrate availability. However, the adaptive mechanisms of AOA and AOB under N enrichment remain poorly understood. Stable isotope probing(SIP) microcosm incubation was employed to reveal community changes of active AOA and AOB in a loess soil from a field experiment growing potatoes that received no N(control, CK), low N(LN, 75 kg N ha~(-1)), and high N(HN, 375 kg N ha~(-1)). The results showed that the soil potential nitrification rate(PNR) was measured by culturing of the soil samples from the field experiment. Soil PNR was significantly increased in HN by87.5% and 67.5% compared with CK and LN, respectively. Compared with CK, the~(13)C-amoA genes of soil AOA and AOB in HN had 2.58 × 10~4 and 1.55 × 10~6 copies, representing 1.6-and 16.2-fold increase respectively. It was indicated that AOB dominated soil ammonia oxidation. A phylogenetic analysis of the~(13)C-amoA gene showed that N application significantly increased the proportion of54 d9-like AOA up to 90% in HN, while the Nitrososphaera gargensis-like and Nitrososphaera viennensis-like AOA were inhibited and completely disappeared. Nitrogen application also resulted in the community shift of active AOB-dominant group from Nitrosospira briensis-like to Nitrosospira sp. TCH711-like. Our study provides compelling evidence for the emergence and maintenance of active nitrifying communities under the intensified N input to an agricultural ecosystem.     

水稻根际和周围土壤中微生物功能基因丰度与碳、氮状况及其通量之间的关系

Rapid nitrogen（N） transformations and losses occur in the rice rhizosphere through root uptake and microbial activities. However,the relationships between rice roots and rhizosphere microbes for N utilization are still unclear. We analyzed different N forms（NH＋4,NO-3, and dissolved organic N）, microbial biomass N and C, dissolved organic C, CH4 and N2O emissions, and abundance of microbial functional genes in both rhizosphere and bulk soils after 37-d rice growth in a greenhouse pot experiment. Results showed that the dissolved organic C was significantly higher in the rhizosphere soil than in the non-rhizosphere bulk soil, but microbial biomass C showed no significant difference. The concentrations of NH＋4, dissolved organic N, and microbial biomass N in the rhizosphere soil were significantly lower than those of the bulk soil, whereas NO-3in the rhizosphere soil was comparable to that in the bulk soil. The CH4 and N2O fluxes from the rhizosphere soil were much higher than those from the bulk soil. Real-time polymerase chain reaction analysis showed that the abundance of seven selected genes, bacterial and archaeal 16 S rRNA genes, amoA genes of ammonia-oxidizing archaea and ammonia-oxidizing bacteria, nosZ gene, mcrA gene, and pmoA gene, was lower in the rhizosphere soil than in the bulk soil, which is contrary to the results of previous studies. The lower concentration of N in the rhizosphere soil indicated that the competition for N in the rhizosphere soil was very strong, thus having a negative effect on the numbers of microbes. We concluded that when N was limiting, the growth of rhizosphere microorganisms depended on their competitive abilities with rice roots for N.     

Leaching and Transformation of Nitrogen Fertilizers in Soil After Application of N with Irrigation： A Soil Column Method

A soil column method was used to compare the effect of drip fertigation （the application of fertilizer through drip irrigation systems, DFI） on the leaching loss and transformation of urea-N in soil with that of surface fertilization combined with flood irrigation （SFI）, and to study the leaching loss and transformation of three kinds of nitrogen fertilizers （nitrate fertilizer, ammonium fertilizer, and urea fertilizer） in two contrasting soils after the fertigation. In comparison to SFI, DFI decreased leaching loss of urea-N from the soil and increased the mineral N （NH4^＋-N ＋ NO3^--N） in the soil. The N leached from a clay loam soil ranged from 5.7% to 9.6% of the total N added as fertilizer, whereas for a sandy loam soil they ranged between 16.2% and 30.4%. Leaching losses of mineral N were higher when nitrate fertilizer was used compared to urea or ammonium fertilizer. Compared to the control （without urea addition）, on the first day when soils were fertigated with urea, there were increases in NH4^＋-N in the soils. This confirmed the rapid hydrolysis of urea in soil during fertigation. NH4^＋-N in soils reached a peak about 5 days after fertigation, and due to nitrification it began to decrease at day 10. After applying NH4^＋-N fertilizer and urea and during the incubation period, the mineral nitrogen in the soil decreased. This may be related to the occurrence of NH4^＋-N fixation or volatilization in the soil during the fertigation process.     

钙盐诱导下土壤锰和铁的释放及其对胡椒的生物有效性

Releases of manganese and iron ions from an albic soil (Albic-Udic Luvisol), a yellow-red soil (Hap-Udic Ferrisol) and a yellow-brown soil (Arp-Udic Luvisol) induced by calcium salt addition and their bioavailability to pepper (Capsicum frutescens L.) were studied in a pot experiment. Addition of Ca(NO₃)₂ decreased soil pH and increased both exchangeable and DTPA (diethylenetriamine pentaacetic acid)-extractable Mn and Fe in soils. Meanwhile, total Mn accumulation in the shoots of Capsicum frutescens L. on the salt-treated soils increased significantly (P< 0.01) compared with the control, suggesting that salt addition to soil induced Mn toxicity in Capsicum frutescens L. Although exchangeable and DTPA-extractable Fe increased also in the salt-treated soils, Fe uptake by the shoots of Capsicum frutescens L. decreased. The effect of added salts in soils on dry matter weight of pepper varied with the soil characteristics, showing different buffer capacities of the soils for salt toxicity in an order of yellow-brown soil > albic soil > yellow-red soil. Fe/Mn ratio in shoots of Capsicum frutescens L. decreased with increasing salt addition for all the soils, which was ascribed to the antagonistic effect of Mn on Fe accumulation. The ratio of Fe/Mn in the tissue was a better indicator of the appearance of Mn toxicity symptoms than Mn concentration alone.     

红壤中La的生物富集及其对玉米幼苗生长影响研究

Through a pot culture lanthanum nitrate was applied to maize seedlings grown in a red loamy soil to investigate the physiological and toxic effects of added La on the growth of crop seedlings and La bioaccumulation to help understand the environmental chemistry behaviors of rare earth element as fertilizers in soils. Compared to the control, La concentrations in shoots and especially in roots of maize seedlings increased with an increase of La in the soil. Also, with added concentrations of La 〉 0.75 g La kg-1 soil and ≥ 0.05 g La kg^-1 soil, the dry weight of shoots and roots of maize seedlings was significantly reduced （P ≤ 0.05）, respectively, compared with the control. Additionally, La ≥0.5 g kg^-1 in the soil significantly inhibited （P ≤ 0.05） primary root elongation. Roots were more sensitive to La stress than shoots and thus could be used as a biomarker to La stress. Overall, in the red loamy soil studied, La had no significant beneficial effects on the growth of maize at the added La levels above 0.1 g kg^-1 soil.     

Evidence for niche differentiation of nitrifying communities in grassland soils after 44 years of different field fertilization scenarios

Long-term nitrogen(N)fertilization imposes strong selection on nitrifying communities in agricultural soil,but how a progressively changing niche affects potentially active nitrifiers in the field remains poorly understood.Using a 44-year grassland fertilization experiment,we investigated community shifts of active nitrifiers by DNA-based stable isotope probing(SIP)of field soils that received no fertilization(CK),high levels of organic cattle manure(HC),and chemical N fertilization(CF).Incubation of DNA-SIP microcosms showed significant nitrification activities in CF and HC soils,whereas no activity occurred in CK soils.The 44 years of inorganic N fertilization selected only 13C-ammonia-oxidizing bacteria(AOB),whereas cattle slurry applications created a niche in which both ammonia-oxidizing archaea(AOA)and AOB could be actively13C-labeled.Phylogenetic analysis indicated that Nitrosospira sp.62-like AOB dominated inorganically fertilized CF soils,while Nitrosospira sp.41-like AOB were abundant in organically fertilized HC soils.The 13C-AOA in HC soils were affiliated with the 29i4 lineage.The 13C-nitrite-oxidizing bacteria(NOB)were dominated by both Nitrospira-and Nitrobacter-like communities in CF soils,and the latter was overwhelmingly abundant in HC soils.The 13C-labeled nitrifying communities in SIP microcosms of CF and HC soils were largely similar to those predominant under field conditions.These results provide direct evidence for a strong selection of distinctly active nitrifiers after 44 years of different fertilization regimes in the field.Our findings imply that niche differentiation of nitrifying communities could be assessed as a net result of microbial adaption over 44 years to inorganic and organic N fertilization in the field,where distinct nitrifiers have been shaped by intensified anthropogenic N input.     

DMPP硝化抑制效果受其浓度、施用方式及土壤基质势的影响

The efficacy of nitrification inhibitors depends on soil properties and environmental conditions. The nitrification inhibitor 3.4-dimethylpyrazole phosphate （DMPP） was investigated in a sandy loam and a loamy soil to study its effectiveness as influenced by inhibitor concentration, application form, and soil matric potential. DMPP was applied with concentrations up to 34.6 mg DMPP kg^-1 soil as solution or as ammonium-sulfate/ammonium-nitrate granules formulated with DMPP. DMPP inhibited the oxidation of ammonium in both soils, but this effect was more pronounced in the sandy loam than in the loamy soil. When applied as solution, increasing DMPP concentrations up to 7 mg DMPP kg^-1 soil had no influence on the inhibition. The effectiveness of DMPP formulated as fertilizer granules was superior to the liquid application of DMPP and NH4^＋, particularly in the loamy soil. Without DMPP, a decline in soil matric potential down to -600 kPa decreased nitrification in both soils, but this effect was more pronounced in the sandy loam than in the loamy soil. DMPP was most effective in the sandy loam particularly under conditions of higher soil moisture, i.e., under conditions favorable for nitrate leaching.     

添加生物炭对酸性红壤中玉米生长和氮素利用率的影响

Biochar added to soil can improve crop growth through both direct and indirect effects, particularly in acidic, highly weathered soils in subtropical and tropical regions. However, the mechanisms of biochar improving crop growth are not well understood. The objectives of this study were i） to determine the crop responses to biochar addition and ii） to understand the effect of biochar addition on N use efficiency. Seven acidic red soils varying in texture, p H, and soil nutrient were taken from southern China and subjected to four treatments： zero biochar and fertilizer as a control（CK）, 10 g kg-1biochar（BC）, NPK fertilizers（NPK）, and 10 g kg-1biochar plus NPK fertilizers（BC＋NPK）.15N-labeled fertilizer was used as a tracer to assess N use efficiency. After a 46-d pot experiment,biochar addition increased soil p H and available P, and decreased soil exchangable Al3＋, but did not impact soil availabe N and cation exchange capacity（P 〉 0.05）. The N use efficiency and N retained in the soil were not significantly affected by biochar application except for the soil with the lowest available P（3.81 mg kg-1） and highest exchanageable Al3＋（4.54 cmol kg-1）. Greater maize biomass was observed in all soils amended with biochar compared to soils without biochar（BC vs. CK, BC＋NPK vs. NPK）. This agronomic effect was negatively related to the concentration of soil exchangeable Al3＋（P 〈 0.1）. The results of this study implied that the liming effect of biochar improved plant growth through alleviating Al toxicity and P deficiency, especially in poor acidic red soils.     

喀麦隆南部地区酸性土壤上增加P的施入后土壤质量和玉米生长的关系

A large array of soil properties influences plant growth response to phosphorus （P） fertilizer input in acid soils. We carried out a pot experiment using three contrasted acid soils from southern Cameroon with the following main objectives： i） to assess the main soil causal factors of different maize （Zea mays L.） growth response to applied P and ii） to statistically model soil quality variation across soil types as well as their relationships to dry matter production. The soils used are classified as Typic Kandiudox （TKO）, Rhodic Kandiudult （RKU）, and Typic Kandiudult （TKU）. Analysis of variance, regression, and principal component analyses were used for data analysis and interpretation. Shoot dry matter yield （DMY） was significantly affected by soil type and P rate with no significant interaction. Predicted maximum attainable DMY was lowest in the TKO （26.2 g pot^-1） as compared to 35.6 and 36.7 g pot^-1 for the RKU and TKU, respectively. Properties that positively influenced DMY were the levels of inorganic NaHCO3-extractable P, individual basic cations （Ca, Mg, and K）, and pH. Their effects contrasted with those of exchangeable Al and C/N ratio, which significantly depressed DMY. Principal component analysis yielded similar results, identifying 4 orthogonal components, which accounted for 84.7% of the total system variance （TSV）. Principal component 1 was identified as soil nutrient deficiency explaining 35.9% of TSV. This soil quality varied significantly among the studied soils, emerging as the only soil quality which significantly （P 〈 0.05） correlated with maize growth. The 2nd, 3rd, and 4th components were identified as soil organic matter contents, texture, and HCl-extractable P, respectively.     

硝化抑制剂对稻田土壤N2O排放和硝化、反硝化菌数量的影响

【目的】本研究旨在明确硝化抑制剂对稻田土壤氮素周转的影响,探讨抑制剂提高氮肥利用率及微生物响应机理。【方法】以草甸黑土发育的水稻土为研究对象,进行了两组培养试验 (25℃),培养周期均为150天。共设4个处理:1) 不施肥 (CK);2) 单施尿素 (Urea);3) 尿素 + 双氰胺 (Urea + DCD);4) 尿素 + 3, 4-二甲基吡唑磷酸盐 (Urea + DMPP)。一组试验从培养第1天起,抽取气体样品,用气相色谱法测定N₂O排放量。另一组试验从培养第1天直到结束,取土样测定氨氧化细菌和氨氧化古菌数量,采用荧光定量PCR等技术测定nirK基因和nirS基因拷贝数,用常规方法测定土壤无机氮含量。【结果】施用尿素显著增加了N₂O排放量,其中85%的N₂O排放发生在培养开始后的前两周内。Urea + DMPP处理土壤NH₄+浓度在前23天稳定在较高水平,与Urea处理相比,N₂O减排率为78.3%,Urea + DCD处理为21.6%。Urea + DMPP处理排放系数为0.05%,Urea + DCD为0.18%,Urea + DMPP处理显著低于Urea + DCD处理。施用尿素培养,土壤氨氧化细菌 (AOB) 数量显著增加而氨氧化古菌 (AOA) 数量则显著减少。添加DCD和DMPP能显著抑制AOB的数量,但对AOA没有影响。培养第3、30和90天,Urea + DMPP处理土壤中的AOB数量显著低于Urea + DCD处理的30%、56%和60%。对于反硝化细菌来说,所有处理中的nirK基因拷贝数均显著高于nirS基因拷贝数。添加DMPP在培养第3和30天显著减少了含nirK和nirS基因的反硝化细菌数量,而添加DCD对两类反硝化细菌数量无明显作用。【结论】东北黑土水稻生产中,硝化抑制剂DMPP降低N₂O排放量和排放系数的效果显著好于DCD,因为DMPP在培养后的30天内,可以显著抑制氨氧化细菌繁衍,降低反硝化细菌数量,从而起到减少N₂O排放、提高肥料利用率的作用。     

基于宏基因组学的酸性森林土壤氨氧化微生物群落特征研究

全球30%以上陆地面积是酸性土壤(pH5.5),而酸性土壤中氨氧化微生物群落特征研究是破译其硝化过程微生物学机理的基础。尤其随着完全硝化微生物(Complete ammonia oxidizer,comammox)的发现,亟需重新认知酸性土壤中氨氧化微生物类群。以酸性马尾松林为研究对象,综合利用荧光定量PCR(qPCR)、凝胶电泳半定量和宏基因组测序等技术研究土壤中氨氧化古菌(Ammonia-oxidizing archaea,AOA)、氨氧化细菌(Ammonia-oxidizing bacteria,AOB)和Comammox的相对丰度以及群落组成特征。研究发现AOA和AOB amoA基因丰度分别为2.61×106 copies·g~(-1)和1.45×106copies·g~(-1);而comammoxamoA基因qPCR结果存在显著的非特异性扩增,导致其丰度被高估,而经凝胶电泳半定量矫正后,约为(1.38～1.47)×106copies·g~(-1),该结果和土壤宏基因测序揭示的comammox相对丰度基本吻合。此外,宏基因组分析发现经典嗜酸group1.1a-associated仅占AOA总类群的12%,而group1.1b则占88%,尽管目前仍未有嗜酸group 1.1b AOA纯菌株的报道。AOB主要类群为Nitrosospira(约64%),而Nitrosomonas约占36%。Comammox主要类群为clade B(约64%),而clade A仅占36%且均隶属于clade A.1亚枝,这暗示clade B与已报道的嗜中性comammox clade A纯菌株有极大的生理代谢差异。总之,本研究提供了综合利用qPCR、半定量和宏基因组分析土壤氨氧化微生物群落的策略,并建议优化comammox的qPCR引物,同时本研究系统分析了酸性马尾松林土壤中氨氧化微生物的相对丰度和群落组成特征。     

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.

     

Nitrapyrin affects the abundance of ammonia oxidizers rather than community structure in a yellow clay paddy soil

Purpose

Yellow clay paddy soil (Oxisols) is a low-yield soil with low nitrogen use efficiency (NUE) in southern China. The nitrification inhibitor nitrapyrin (2-chloro-6- (tricholoromethyl)-pyridine, CP) has been applied to improve NUE and reduce environmental pollution in paddy soil. However, the effects of nitrapyrin combined with nitrogen fertilizers on ammonia oxidizers in yellow clay paddy soil have not been examined.

Materials and methods

A randomized complete block design was set with three treatments: (1) without nitrogen fertilizer (CK), (2) common prilled urea (PU), and (3) prilled urea with nitrapyrin (NPU). Soil samples were collected from three treatments where CK, PU, and NPU had been repeatedly applied over 5 years. Soil samples were analyzed by quantitative PCR and 454 high-throughput pyrosequencing of the amoA gene to investigate the influence of nitrapyrin combined with nitrogen on the abundance and community structure of ammonia oxidizers in yellow clay paddy soil.

Results and discussion

The potential nitrification rate (PNR) of the soil was significantly correlated with the abundances of both ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB). Application of urea significantly stimulated AOA and AOB growth, whereas nitrapyrin exhibited inhibitory effects on AOA. Phylogenetic analysis showed that the most dominant operational taxonomic units (OTUs) of AOA and AOB were affiliated with the Nitrosotalea cluster and Nitrosospira cluster 12, respectively. AOA and AOB community structures were not altered by urea and nitrapyrin application.

Conclusions

Nitrogen fertilization stimulated nitrification and increased the population sizes of AOA and AOB. Nitrapyrin affected the abundance, but not community structure of ammonia oxidizers in yellow clay soil. Our results suggested that nitrapyrin improving NUE and inhibiting PNR was attributable to the inhibition of AOA growth.

     

Responses of ammonia-oxidizing bacteria and archaea to nitrogen fertilization and precipitation increment in a typical temperate steppe in Inner Mongolia

As the first and rate-limiting step of nitrification, ammonia oxidation can be realized either by ammonia-oxidizing bacteria (AOB) or archaea (AOA). However, the key factors driving the abundance, community structure and activity of ammonia oxidizers are still unclear, and the relative importance of AOA and AOB in ammonia oxidation is unresolved. In the present study, we examined the effects of long-term (6 years) nitrogen (N) addition and simulated precipitation increment on the abundance and community composition of AOA and AOB based on a field trial in a typical temperate steppe of northern China. We used combined approaches of quantitative PCR, terminal-restriction fragment length polymorphism (T-RFLP) and clone library analyses of amoA genes. The study objective was to determine (1) AOA and AOB diversity and activity in response to N addition and increased precipitation and (2) the relative contributions of AOA and AOB to soil ammonia oxidation in the typical temperate steppe. The results showed that the potential nitrification rate (PNR) increased with N addition, but decreased with increased precipitation. Both N addition and increased precipitation significantly increased AOB but not AOA abundance, and a significant correlation was only observed between PNR and AOB amoA gene copies. The T-RFLP analysis showed that both N and precipitation were key factors in shaping the composition of AOB, while AOA were only marginally influenced. Phylogenetic analysis indicated that all AOA clones fell within the soil and sediment lineage while all AOB clones fell within the Nitrosospira. The study suggested that AOA and AOB had distinct physiological characteristics and ecological niches. AOB were shown to be more sensitive to N and precipitation than AOA, and the ammonia oxidation process was therefore supposed to be mainly driven by AOB in this temperate steppe.     

石灰和双氰胺对红壤酸化和硝化作用的影响及其机制

施用石灰是改良酸性土壤的重要措施,但其对土壤硝化作用的增强不仅加速土壤酸化,也增加硝态氮流失风险。传统的硝化抑制剂双氰胺(Dicyandiamide,DCD)能否在石灰改变pH的条件下始终有效抑制硝化是当前红壤区生产中亟需解决的问题。采用短期土壤培养试验,探讨了不同用量石灰与DCD配合施用对土壤酸化和硝化作用的影响及其机制。结果表明:施用一定量的石灰(≤4 g·kg^–1)显著提高土壤pH,通过促进氨氧化细菌的生长以促进硝化作用。在不同pH条件下,DCD对红壤硝化过程均有显著抑制效果。在较高pH(pH 7.0~7.8)条件下,DCD主要通过降低氨氧化细菌的丰度以抑制硝化,而在低pH(pH<6.0)条件下,DCD对氨氧化古菌和氨氧化细菌的丰度均有抑制作用。此外,DCD通过抑制土壤硝化,显著提高了土壤pH。上述结果表明,适宜量(2~4 g·kg^–1)的石灰和DCD结合施用不仅能够减缓红壤酸化,而且能够抑制硝化作用,降低硝态氮的潜在环境风险。     

Abundance and community structure of ammonia-oxidizing archaea and bacteria in an acid paddy soil

Nitrification is essential to the nitrogen cycle in paddy soils. However, it is still not clear which group of ammonia-oxidizing microorganisms plays more important roles in nitrification in the paddy soils. The changes in the abundance and composition of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) were investigated by real-time PCR, terminal restriction fragment length polymorphism, and clone library approaches in an acid red paddy soil subjected to long-term fertilization treatments, including treatment without fertilizers (CT); chemical fertilizer nitrogen (N); N and potassium (NK); N and phosphorus (NP); N, P, and K (NPK); and NPK plus recycled crop residues (NPK+C). The AOA population size in NPK+C was higher than those in CT, while minor changes in AOB population sizes were detected among the treatments. There were also some changes in AOA community composition responding to different fertilization treatments. Still few differences were detected in AOB community composition among the treatments. Phylogenetic analysis showed that the AOA sequences fell into two main clusters: cluster A and cluster soil/sediment. The AOB composition in this paddy soil was dominated by Nitrosospira cluster 12. These results suggested that the AOA were more sensitive than AOB to different fertilization treatments in the acid red paddy soil.     

硝化抑制剂对毛竹林土壤N_2O排放和氨氧化微生物的影响

为了探索硝化抑制剂在毛竹生产中的施用技术,通过培养试验研究3,4-二甲基吡唑磷酸盐(DMPP)和双氰胺(DCD)两种硝化抑制剂对毛竹林施用尿素后土壤N2O排放、氮素转化和相关氨氧化细菌(AOB)、氨氧化古菌(AOA)群落结构和丰度的影响。试验设(1)对照(CK)、(2)单施尿素(Urea)、(3)尿素+1%DMPP(DMPP占总N的1%,下同);(4)尿素+1.5%DMPP;(5)尿素+10%DCD;(6)尿素+15%DCD等6个处理,测定N2O的排放动态以及气体排放转折点时的土壤特征指标。结果表明:与单施尿素相比,160 d的时间内两种DMPP用量处理的土壤N2O累积排放减排幅度均为54%,而10%DCD和15%DCD处理的土壤分别减少28%和41%。DMPP和DCD处理50 d和90 d时土壤的NH4+-N含量均显著高于(p0.05)单施尿素处理,而NO3--N含量和表观硝化率则恰好相反,但两种抑制剂间无差异。DMPP处理的AOB群落结构的变化从10 d开始显现,至50 d和90 d时仍保持明显的抑制状态,而DCD处理则至90 d时抑制作用基本消失。单施尿素AOB功能基因(amo A)的丰度均显著高于硝化抑制剂处理(90 d时尿素+10%DCD处理除外);在整个培养期内,尿素和对照土壤的AOA群落结构相似,硝化抑制剂反而增加了AOA功能基因的丰度,表明硝化抑制剂对AOA丰度无明显抑制作用。即两种硝化抑制剂主要通过抑制AOB起作用;调节土壤p H至中性范围,并在1%DMPP施用条件下,硝化抑制剂的抑制效果最显著。