氮肥施加条件下增温对硝化菌活性和丰度的影响

温度在多种生物地球化学过程中起到关键的调节作用,是影响土壤硝化作用和微生物分布的重要因素之一。硝化过程的第1个步骤由氨氧化细菌(AOB)和氨氧化古菌(AOA)催化,然而,不同施氮量下,增温对硝化菌活性和丰度的影响尚不清楚。本研究基于2008年10月起设立于太行山山前平原的长期增温试验平台(高于地表2m的红外加热器使土壤温度升高1.5℃),于2018年5月对不施氮(N0)和施氮[N1,240kg(N)·hm-2·a-1]下增温分别对0～10 cm和10～20 cm土壤硝化潜势(PNR)、AOA和AOB丰度的影响进行了研究。硝态氮(NO3--N和铵态氮(NH4+-N)含量用分光光度法测量,应用缓冲液培养法测定土壤PNR,提取土壤DNA后用实时荧光定量PCR技术测定功能基因AOA和AOB的丰度。结果表明:温度升高显著增加N1条件下PNR和NO3--N含量(P0.05),降低了N0条件下PNR和NO3--N含量,但差异不显著。N1条件下,增温土壤AOB丰度显著提高(P0.05); N0条件下,增温土壤AOA丰度显著降低(P0.05)。与N0相比, N1条件下的AOA/AOB比值明显降低,表明增温加氮肥处理对AOB的生长刺激更强烈。在增温加施氮条件下,细菌(AOB)表现显著的正反应,在增温不施氮条件下,古菌(AOA)和AOB表现显著的负反应。本研究结果可为全球增温背景下进一步了解硝化活性和氨氧化微生物对增温和氮有效性的响应提供科学依据。     

Nitrous oxide production in grassland soils: assessing the contribution of nitrifier denitrification

Nitrifier denitrification is the reduction of NO₂⁻ to N₂ by nitrifiers. It leads to the production of the greenhouse gas nitrous oxide (N₂O) as an intermediate and possible end product. It is not known how important nitrifier denitrification is for the production of N₂O in soils. We explored N₂O production by nitrifier denitrification in relation to other N₂O producing processes such as nitrification and denitrification under different soil conditions. The influence of aeration of the soil, different N sources, and pH were tested in four experiments. To differentiate between sources of N₂O, an incubation method with inhibitors was used [Biol. Fertil. Soils 22 (1996) 331]. Sets of four incubations included controls without addition of inhibitors, incubations with addition of small concentrations of C₂H₂ (0.01-0.1 kPa), large concentrations of O₂ (100 kPa), or a combination of C₂H₂ and O₂. The results indicate that the availability of NO₂⁻ stimulated the apparent N₂O production by nitrifier denitrification. A decreasing O₂ content increased the total N₂O production, but decreased N₂O production by nitrifier denitrification. No significant effect of pH could be found. The study revealed problems concerning the use of the inhibitors C₂H₂ and O₂. Almost one-third of all incubations with inhibitors produced more N₂O than the controls. Possible reasons for the problems are discussed. The inhibitors C₂H₂ and O₂ need to be tested thoroughly for their effects on different N₂O producing processes before further application.     

Nitrous oxide production of heavy metal contaminated soil

Arsenic (As), lead (Pb), copper (Cu) and zinc (Zn) can be found in large concentrations in mine spills of central and northern Mexico. Interest in these heavy metals has increased recently as they contaminate drinking water and aquifers in large parts of the world and severely affect human health, but little is known about how they affect biological functioning of soil. Soils were sampled in seven locations along a gradient of heavy metal contamination with distance from a mine in San Luis Potosí (Mexico), active since about 1800 AD. C mineralization and N₂O production were monitored in an aerobic incubation experiment. Concentrations of As in the top 0-10 cm soil layer ranged from 8 to 22,992 mg kg⁻¹, from 31 to 1845 mg kg⁻¹ for Pb, from 27 to 1620 mg kg⁻¹ for Cu and from 81 to 4218 mg kg⁻¹ for Zn. There was a significant negative correlation between production rates of CO₂ and concentrations of As, Pb, Cu and Zn, and there was a significant positive correlation with pH, water holding capacity (WHC), total N and soil organic C. There was a significant negative correlation (P<0.05) between production rate of nitrous oxide (N₂O) attributed to nitrification by the inhibition method in soil incubated at 50% WHC and total concentrations of Pb and Zn, and there was a significant positive correlation (P<0.05) with pH and total N content. There was a significant negative correlation (P<0.05) between the production rate of N₂O attributed to denitrification by the inhibition method in soil incubated at 100% WHC and total concentrations of Pb, Cu and Zn, and a significant positive correlation (P<0.01) with pH; there was a significant positive correlation (P<0.05) between the production of N₂O attributed to other processes by the inhibition method and WHC, inorganic C and clay content. A negative value for production rate of N₂O attributed to nitrifier denitrification by the inhibition method was obtained at 100% WHC. The large concentrations of heavy metals in soil inhibited microbial activity and the production rate of N₂O attributed to nitrification by the inhibition method when soil was incubated at 50% WHC and denitrification when soil was incubated at 100% WHC. The inhibitor/suppression technique used appeared to be flawed, as negative values for nitrifier denitrification were obtained and as the production rate of N₂O through denitrification increased when soil was incubated with C₂H₂.     

Nitrogen transformations in tropical soils under conventional and sustainable farming systems

Samples of alluvial soil from mixed sandstone shale and slate and of Taiwan clay were collected from two sites, both managed under a similar crop rotation scheme. The fields were further divided into sections which were managed under either conventional farming or sustainable farming practices. When the soil samples were collected in April 1989, after 1 year of operation under conventional or sustainable practices, the nitrification activities of both soils managed under sustainable practices practices. The nitrifying activities in Taiwan clay samples collected in April 1993 which had been managed with chemical or with organic fertilizer were not significantly different. However, nitrifying activity in the alluvial soil was higher under sustainable than under conventional practices. Numbers of NH ₄ ⁺ -oxidizing bacteria were not significantly different in any of the soil samples irrespective of the different management practices. In contrast, higher numbers of NO ₂ ^- -oxidizing bacteria were detected in both soils managed sustainably. The results also indicated that the composition of NH ₄ ⁺ -oxidizing bacteria differed in the alluvial soil when managed with different kinds of fertilizer.     

Nitrous oxide flux from soil amino acid mineralization

Nitrous oxide (N₂O) is a greenhouse gas produced during microbial transformation of soil N that has been implicated in global climate warming. Nitrous oxide efflux from N fertilized soils has been modeled using NO₃⁻ content with a limited success, but predicting N₂O production in non-fertilized soils has proven to be much more complex. The present study investigates the contribution of soil amino acid (AA) mineralization to N₂O flux from semi-arid soils. In laboratory incubations (−34 kPa moisture potential), soil mineralization of eleven AAs (100 μg AA-N g⁻¹ soil) promoted a wide range in the production of N₂O (156.0±79.3 ng N₂O-N g⁻¹ soil) during 12 d incubations. Comparison of the δ¹³C content (‰) of the individual AAs and the δ¹³C signature of the respired AA-CO₂-C determined that, with the exception of TYR, all of the AAs were completely mineralized during incubations, allowing for the calculation of a N₂O-N conversion rate from each AA. Next, soils from three different semi-arid vegetation ecosystems with a wide range in total N content were incubated and monitored for CO₂ and N₂O efflux. A model utilizing CO₂ respired from the three soils as a measure of organic matter C mineralization, a preincubation soil AA composition of each soil, and the N₂O-N conversion rate from the AA incubations effectively predicted the range of N₂O production by all three soils. Nitrous oxide flux did not correspond to factors shown to influence anaerobic denitrification, including soil NO₃⁻ contents, soil moisture, oxygen consumption, and CO₂ respiration, suggesting that nitrification and aerobic nitrifier denitrification could be contributing to N₂O production in these soils. Results indicate that quantification of AA mineralization may be useful for predicting N₂O production in soils.     

Organic manure input and straw cover improved the community structure of nitrogen cycle function microorganism driven by water erosion

Water erosion process induces differences to the nitrogen (N) functional microbial community structure, which is the driving force to key N processes at soil-water interface. However, how the soil N transformations associated with water erosion is affected by microorganisms, and how the microbial respond, are still unclear. The objective of this study is to investigate the changes of microbial diversity and community structure of the N-cycle function microorganisms as affected by water erosion under application of organic manure and straw cover. On the basis of iso-nitrogen substitution, four treatments were set up: 1) only chemical fertilizer with N 150 kg ha^?1, P₂O₅ 60 kg ha^?1 and K₂O 90 kg ha^?1 (CK); the N was substituted 20% by 2) organic manure (OM); 3) straw (SW); and 4) organic manure + straw (1:1) (OMSW). The results showed that applying organic manure and straw to sloping farmland can increase soil N contents, but reduce runoff depth, K_w, sediment yield and N loss, especially in the OMSW. Straw cover and straw + organic manure increased the diversity (Chao1) of nitrifier (AOB), and both diversity and uniformity (Shannon) of denitrifier (nirK/S) were increased in the OMSW. All erosion control measures reduced N-fixing bacteria diversity and increased their uniformity, and the combined application of organic manure and straw cover was a better erosion control measure than the single application of them. Improved soil chemistry and erodibility were the main drives for the changes of N-functional microbial community structure and the appearance of dominant bacteria with different organic materials.     

Oxygen exchange between nitrogen oxides and H2O can occur during nitrifier pathways

Interpretation of the oxygen isotopic signature of soil-derived N₂O may be flawed when it is based on reaction stoichiometry and fractionation alone. In fact, oxygen (O) exchange between H₂O and intermediates of N₂O production pathways may largely determine this O isotopic signature. Although in our previous work we conclusively proved the occurrence of O exchange during N₂O production by denitrification of NO₃⁻, its occurrence in N₂O production pathways by nitrifiers remains unclear. The aim of this study was to examine the likeliness of O exchange during various stages of N₂O production in soil via nitrification, nitrifier denitrification and denitrification. We evaluated a set of scenarios on the presence of such exchange using data from a series of ¹⁸O and ¹⁵N tracing experiments. The measured actual O incorporation from H₂O into N₂O (AOI) was compared with the theoretical maximum O incorporation (MOI) from various scenarios that differed in their assumptions on the presence of O exchange. We found that scenarios where O exchange was assumed to occur exclusively during denitrification could not explain the observed AOI, as it exceeded the MOI for 9 out of 10 soils. This demonstrates that additional O exchange must have occurred in N₂O production through nitrifier pathways. It remains to be determined in which steps of these pathways O exchange can take place. We conclude that O exchange is likely to be mediated by ammonia oxidizers during NO₂⁻ reduction (nitrifier denitrification), and that it could possibly occur during NO₂⁻ oxidation to NO₃⁻ by nitrite oxidizers as well.     

Nitrifier denitrification as a distinct and significant source of nitrous oxide from soil

Soils are the major source of the greenhouse gas nitrous oxide (N₂O) to our atmosphere. A thorough understanding of terrestrial N₂O production is therefore essential. N₂O can be produced by nitrifiers, denitrifiers, and by nitrifiers paradoxically denitrifying. The latter pathway, though well-known in pure culture, has only recently been demonstrated in soils. Moreover, nitrifier denitrification appeared to be much less important than classical nitrate-driven denitrification. Here we studied a poor sandy soil, and show that when moisture conditions are sub-optimal for denitrification, nitrifier denitrification can be a major contributor to N₂O emission from this soil. We conclude that the relative importance of classical and nitrifier denitrification in N₂O emitted from soil is a function of the soil moisture content, and likely of other environmental conditions as well. Accordingly, we suggest that nitrifier denitrification should be routinely considered as a major source of N₂O from soil.