Evolution of gaseous hydrocarbons from soil: Effect of moisture content and nitrate level

The production of gaseous hydrocarbons has been studied in a clayey, loamy and sandy soil. The soil samples were incubated under closed conditions at field capacity and under waterlogged conditions. Additions of 100 or 300 μgg^?1 nitrate were made to some of the soil samples. It was found that hydrocarbons were produced in two phases: at the beginning of the incubation period, coinciding with the high microbial activity after rewetting a dry soil and after obtaining anaerobic conditions in the flasks. Under anaerobic conditions, the concentration of methane was higher than the concentration of ethylene, ethane and propane. Methane production was high in the neutral soil, and of minor importance in the acid soils, while ethylene production was important in the soil with the highest organic matter content. Waterlogged conditions were much more favourable for hydrocarbon production than field capacity conditions. The addition of nitrate, in general, decreased hydrocarbon production, especially at 300μgg^?1 for all treatments. In some treatments and for some gases, the influence of 100μgg^?1 was less significant.     

氮素水平对土壤甲烷氧化和硝化微生物相互作用的影响

甲烷氧化微生物和氨氧化微生物均是既可以氧化甲烷(CH₄)又可以氧化氨(NH₃),氨氧化是硝化作用的限速步骤,也是好氧土壤氧化亚氮(N₂O)排放的主要生物路径。选取内蒙古草原围封禁牧土壤为研究对象,利用稳定同位素核酸探针技术(DNA-SIP)探讨不同氮水平下土壤活性甲烷氧化微生物与硝化微生物及其相互作用机制。结果发现低氮添加促进甲烷氧化活性,而高氮添加抑制甲烷氧化活性;低氮和高氮添加均显著增强硝化活性。基于DNA-SIP的高通量测序结果发现Methylobacter MOB和Nitrosospira AOB/Nitrospira NOB分别是该土壤的主要活性甲烷氧化和硝化微生物。网络结构分析发现Methylobacter MOB和Nitrosospira AOB/Nitrospira NOB存在显著负相关关系,进一步证明活性甲烷氧化和硝化微生物之间存在竞争性相互作用。以上结果表明,氮素水平影响草原土壤甲烷氧化和硝化微生物的相互作用,研究结果为采取措施调控草原土壤CH₄的汇和N₂O...     

Biological nitrification inhibition by Brachiaria humidicola roots varies with soil type and inhibits nitrifying bacteria, but not other major soil microorganisms

The tropical pasture grass Brachiaria humidiola (Rendle) Schweick releases nitrification inhibitory compounds from its roots, a phenomenon termed 'biological nitrification inhibition' (BNI). We investigated the influence of root exudates of B. humidicola on nitrification, major soil microorganisms and plant growth promoting microorganisms using two contrasting soil types, Andosol and Cambisol. The addition of root exudates (containing BNI activity that is expressed in Allylthiourea unit (ATU) was standardized in a bioassay against a synthetic inhibitor of nitrification, allylthiourea, and their function in soil was compared to inhibition caused by the synthetic nitrification inhibitor dicyandiamide. At 30 and 40 ATU g⁻¹soil, root exudates inhibited nitrification by 95% in fresh Cambisol after 60 days. Nitrification was also similarly inhibited in rhizosphere soils of Cambisol where B. humidicola was grown for 6 months. Root exudates did not inhibit other soil microorganisms, including gram-negative bacteria, total cultivable bacteria and fluorescent pseudomonads. Root exudates, when added to pure cultures of Nitrosomonas europaea , inhibited their growth, but did not inhibit the growth of several plant growth promoting microorganisms, Azospirillum lipoferum , Rhizobium leguminosarum and Azotobacter chroococcum. Our results indicate that the nitrification inhibitors released by B. humidicola roots inhibited nitrifying bacteria, but did not negatively affect other major soil microorganisms and the effectiveness of the inhibitory effect varied with soil type.     

Effect of soil solarization on subsequent nitrification activity at elevated temperatures

Soil solarization is a nonchemical method of soil disinfection achieved by covering the soil surface with sheets of vinyl plastic to generate elevated soil temperature, generally over 45°C. Such elevated temperatures may be detrimental to some nitrifying microorganisms and favorable to others. However, little information exists to indicate how nitrification activity in soil is affected after solarization. We performed several experiments to investigate the effects of soil solarization on nitrification activity. We found that: (1) if a soil was subjected to pretreatment of 45 or 50°C for as little as 1 d, nitrification activity in a subsequent incubation at 30°C was less than that of a soil that did not receive any high-temperature pretreatment. However, if a soil received pretreatments of 45 or 50°C for more than 7 d, nitrification activity in a subsequent incubation at 45 or 50°C was greater than that of soil that did not receive high temperature pretreatment. (2) Nitrification activity in three kinds of soil taken from 0–5 cm depth after solarization treatment was greater at 45°C than 30°C. (3) Nitrification activity at 45°C in soil that had received solarization in the preceding year was greater than that in soil that had not been subjected to solarization. This was consistent with the fact that the population densities of ammonia oxidizers were greater in soils that had been subjected to solarization. These results suggest that soil solarization induces nitrifying microorganisms that are more active at 45–50°C than they are at 30°C, and that the effect of solarization on nitrification persists until the next crop season.     

Changes in nitrification and bacterial community structure upon cross-inoculation of Scots pine forest soils with different initial nitrification rates

Nitrification occurs slowly in many acid Scots pine forest soils. We examined if bacterial community structure and interactions between members of the bacterial community in these forest soils prohibit growth of ammonia-oxidising microorganisms and their nitrifying activity. Native and gamma-irradiated Scots pine forest soils known to have low net nitrification rates were augmented with fresh soils or soil slurries from nitrifying Scots pine forest soil, and vice versa. Augmentation of native non-nitrifying soils with nitrifying soils induced net nitrification, although no significant changes in bacterial community structure, as measured by 16S rRNA gene-based denaturing gradient gel electrophoresis (DGGE), were observed. In sterilised soils, the inoculum, i.e. native nitrifying soil or non-nitrifying soil, determined the occurrence of net nitrification and bacterial community structure, and not the origin of the sterilised soils. Our results demonstrate that low net nitrification rates in acid Scots pine forest soils cannot be (solely) explained by unfavourable abiotic soil conditions, but that still uncaptured biotic factors contribute to suppression of nitrification.     

土壤生态系统硝化微生物研究进展

微生物主导的硝化作用是生态系统中氮素循环的关键过程,其不仅与酸雨、温室气体、水体富营养化等环境问题的发生有关,还作用于土壤中氮素营养的转化,与人类生产生活密切相关。土壤生态系统中进行硝化作用的微生物包括细菌、古细菌、真菌等。这些微生物根据自身能量代谢类型的不同,利用不同的生物酶进行着不同机制的硝化作用。本文综述了目前已报道的生态系统中进行自养(经典自养硝化和全程氨氧化)和异养硝化作用的微生物类群、硝化作用关键酶及其编码基因类型、其在生态系统中多样的分布特征,以及其前沿的分子生态学研究方法。同时对不同类型硝化微生物类群今后的研究热点提出了展望,以期为系统地研究土壤生态系统中硝化微生物提供参考。     

Comammox—a newly discovered nitrification process in the terrestrial nitrogen cycle

Purpose

Nitrification, the microbial oxidation of ammonia to nitrate via nitrite, is a pivotal component of the biogeochemical nitrogen cycle. Nitrification was conventionally assumed as a two-step process in which ammonia oxidation was thought to be catalyzed by ammonia-oxidizing archaea (AOA) and bacteria (AOB), as well as nitrite oxidation by nitrite-oxidizing bacteria (NOB). This long-held assumption of labour division between the two functional groups, however, was challenged by the recent unexpected discovery of complete ammonia oxidizers within the Nitrospira genus that are capable of converting ammonia to nitrate in a single organism (comammox). This breakthrough raised fundamental questions on the niche specialization and differentiation of comammox organisms with other canonical nitrifying prokaryotes in terrestrial ecosystems.

Materials and methods

This article provides an overview of the recent insights into the genomic analysis, physiological characterization and environmental investigation of the comammox organisms, which have dramatically changed our perspective on the aerobic nitrification process. By using quantitative PCR analysis, we also compared the abundances of comammox Nitrospira clade A and clade B, AOA, AOB and NOB in 300 forest soil samples from China spanning a wide range of soil pH.

Results and discussion

Comammox Nitrospira are environmentally widespread and numerically abundant in natural and engineered habitats. Physiological data, including ammonia oxidation kinetics and metabolic versatility, and comparative genomic analysis revealed that comammox organisms might functionally outcompete other canonical nitrifiers under highly oligotrophic conditions. These findings highlight the necessity in future studies to re-evaluate the niche differentiation between ammonia oxidizers and their relative contribution to nitrification in various terrestrial ecosystems by including comammox Nitrospira in such comparisons.

Conclusions

The discovery of comammox and their broad environmental distribution added a new dimension to our knowledge of the biochemistry and physiology of nitrification and has far-reaching implications for refined strategies to manipulate nitrification in terrestrial ecosystems and to maximize agricultural productivity and sustainability.

     

Characterization of nitrifying bacteria communities of soils from different ecological regions of China by molecular and conventional methods

Soil nitrification rate is very different among soil types, as a result of differences in physical and chemical properties. Little is known about the composition of the nitrifying bacteria community. In this investigation, three soils (fluvo-aquic soil, permeable paddy soil and red earth) from different geo-ecological regions in China were characterized for their nitrification activities and their nitrifying bacteria communities determined either by molecular approaches or by conventional culture methods. A 28-day long-term soil incubation showed that the maximum nitrification potential was found in the fluvo-aquic soil with almost 100% of inorganic N present as NO₃^–-N, while the minimum nitrification potential was in red earth with only a 4.9% conversion rate from ammonium into nitrate. There was no relationship between nitrification potential and numbers of nitrifiers in the soil. The conventional most probable number (MPN) method could enumerate ammonia oxidizers, but failed in enumerating nitrite oxidizers. Therefore, we used an MPN-PCR procedure which gave a convincing nitrite oxidizer count result, instead of MPN-diphylamine. Soils were characterized by denaturing gradient gel electrophoresis (DGGE) of DNA extracted from soils and amplified using a primer specific for the 16S rRNA gene and/or for the amoA gene. The DGGE columns of the three soils differed from each other. There were two similar bands present in DGGE columns of the fluvo-aquic and permeable paddy soils, but no similar band was found in DGGE columns of the red earth. The sequence of amoA indicated that all ammonia oxidizers in these soils were grouped into Nitrosospira clusters 1 and 3, and each soil had a common band similar to the other soils and a special band which differed from the other soils.     

Einfluß von Wassergehalt und Salzen auf die Nitrifikation in Proben einer podsoligen Braunerde

Influence of watercontent and salts on the nitrification in samples of a Dystric Cambisol Samples of a Dystric Cambisol from a beech site produced nitrate but autotrophic nitrifying microorganisms could not be detected. Net nitrification of the humic layer and the upper 5 cm of the mineral soil during incubation at 22°C was investigated. Nitrification rate increased with increasing water content of the soil. Additions of ammonium or peptone did not increase the nitrification in the humic layer. Supply of (ammonium-)sulphates and chlorides with concentrations higher than 2 mMol per kg soil inhibited nitrification totally. This could not be ascribed to pH-changes. Additions of phosphates, lime or alkali to the soil samples increased nitrification.     

Current insights into the autotrophic thaumarchaeal ammonia oxidation in acidic soils

Recent studies of ammonia-oxidizing archaea (AOA) suggested their significant contributions to global nitrogen cycling, and phylogenetic analysis categorized AOA into a novel archaeal phylum, the Thaumarchaeota. AOA are ubiquitous in terrestrial ecosystems, have unique mechanisms for nitrification, better adaptation to low-pH pressures, and strikingly lower ammonia requirements compared with ammonia-oxidizing bacteria (AOB). Previous perceptions that microbial ammonia oxidation in acidic soils was minimal, and entirely meditated by autotrophic bacteria and occasionally by heterotrophic nitrifiers have been dramatically challenged, and the dominant nitrifying groups urgently called for re-assessment. Controversially, the relative contributions of AOA and AOB to autotrophic ammonia oxidation have been reported to vary in different soils, but ammonia substrate availability, which was largely restricted under acidic conditions, seemed to be the key driver. Theoretically predicted ammonia concentrations in acidic soils below the substrate threshold of AOB and remarkably high ammonia affinity of AOA raised the supposition that thaumarchaea could represent the dominant ammonia-oxidizing group in ammonia-limited acidic environments. Recently, the functional dominance of thaumarchaea over its bacterial counterpart and autotrophic thaumarchaeal ammonia oxidation in acidic soils has been compellingly confirmed by DNA-stable isotope probing (SIP) experiments and the cultivation of an obligate acidophilic thaumarchaeon, Nitrosotalea devanaterra. Here, we review the currently available knowledge concerning the history and progress in our understanding of the ammonia-oxidizing microorganisms (AOB and AOA) and the mechanisms of nitrification in nutrient-depleted acidic soils, present the possible mechanisms shaping the distinct niches of AOA and AOB, and thus strengthen the assumption that AOA dominate over AOB in ammonia oxidation of acidic soils.     

Effect of temperature on nitrification in soils

Nitrification plays an important role in nitrogen transformation in soils. As a practical problem the fact is noteworthy that by nitrification, the loss of nitrogen from soils is caused at higher temperatures in the summer. On the contrary, it is quite slow at the lower temperatures of early spring. Recently much Work has been done on the effect of temperature on nitrification in soils. It was noticed that its effect is not the same according to the difference of soils. Soils which have high nitrifying activities at ordinary temperatures (around 25°C) show fairly good nitrification even at lower temperatures (1, 2, 5). However, in nitrification by usual experimental methods, the amount of nitrate produced can be only estimated while nitrifying organisms are increasing or decreasing in soil. As the growing Process of microbes is extremely influenced by both the properties of the soil and the cultural conditions, it is necessary to examine the effect of temperature on the growing process of microbes rather than biochemical nitrification by a unit cell. In usual methods nitrification can hardly be treated in these separate aspects. The author has succeeded in obtaining soils in which nitrifying organisms increase to the maximum limit, and no increase can be expected in number of these organisms. By using such samples, experiments were made concerning the effect of temperature on nitrification by a unit cell and its direct effect on the growth of organisms. Furthermore, the author prepared favorable cultural conditions for various kinds of soils by the “washing cultivation method” (6), and examined the effect of temperature on the increasing process of nitrifying ability.     

不同生物硝化抑制剂对红壤性水稻土N2O排放的影响及其机制

为揭示不同生物硝化抑制剂(BNIs)对红壤性水稻土N₂O排放的影响差异及作用机制,通过21 d的土柱淹水培养试验,比较了三种BNIs 1,9-癸二醇(1,9-D)、亚麻酸(LN)和3-(4-羟基苯基)丙酸甲酯(MHPP)与化学合成硝化抑制剂双氰胺(DCD)对土壤N₂O排放及相关硝化、反硝化功能基因的影响。结果表明:不同BNIs(1,9-D、LN、MHPP)可以显著平均降低土壤N₂O日排放峰值40.1%;1,9-D和MHPP可分别抑制N₂O排放总量44.5%和43.9%,而DCD和LN对N₂O排放总量没有显著影响。1,9-D和MHPP对AOA(氨氧化古菌)、AOB(氨氧化细菌)硝化菌和nirS、nirK型反硝化菌的调控均有所不同,1,9-D可以同时抑制AOA、AOB和nirS微生物的生长;MHPP仅可以抑制AOA的生长;其中,AOA-amoA和nirS基因丰度与土壤N₂O的排放呈显著正相关关系。同时,1,9-D和MHPP均增加了nosZ基因丰度及其与AOA-...     

Nitrification activity and putative ammonia-oxidizing archaea in acidic red soils

Purpose  

Acidic red soils account for 21% of land area in China and contain low ammonia concentration due to ionization to ammonium. The unusual high affinity for ammonia of marine Nitrosopumilus maritimus and acidophilic soil Nitrosotalea devanaterra has suggested that ammonia-oxidizing archaea (AOA) may have greater selective advantage over ammonia-oxidizing bacteria (AOB) in ammonia-limited environment because ammonia rather than ammonium is thought to be the actual substrate for oxidation. The aim of this study was to assess whether nitrification activity can be attributed to AOA and/or AOB by relating community structures of AOA and AOB to nitrification activity in acidic red soils in southern China.     

Microbial transformation of nitrogen compounds in middle taiga soils

The intensity of mineralization, nitrogen fixation, and denitrification in forest soils of the Karelian middle taiga ecosystems has been evaluated. Podzol-gleyish soil underlying a birch forest with gramineous plants and miscellaneous herbs was shown to have the highest nitrogen-fixing activity. The loss of gaseous nitrogen during denitrification was insignificant due to the low nitrifying activity of the soils named above. N₂O uptake by microorganisms was rather intensive in all the soils analyzed, and in illuvial-humo-ferric podzols underlying pine and spruce forests this process predominated. Podzolic sandy loam gley-like soil of a birch forest with gramineous plants and miscellaneous herbs had the highest potential for the mineralization of organic nitrogen; the rate of ammonification and nitrification in this soil was maximal.     

Conversion of forest to agricultural land affects the relative contribution of bacteria and fungi to nitrification in humid subtropical soils

Land-use and management practices can affect soil nitrification. However, nitrifying microorganisms responsible for specific nitrification process under different land-use soils remains unknown. Thus, we investigated the relative contribution of bacteria and fungi to specific soil nitrification in different land-use soils (coniferous forest, upland fields planted with corn and rice paddy) in humid subtropical region in China. ¹⁵N dilution technique in combination with selective biomass inhibitors and C₂H₂ inhibition method were used to estimate the relative contribution of bacteria and fungi to heterotrophic nitrification and autotrophic nitrification in the different land-use soils in humid subtropical region. The results showed that autotrophic nitrification was the predominant nitrification process in the two agricultural soils (upland and paddy), while the nitrate production was mainly from heterotrophic nitrification in the acid forest soil. In the upland soils, streptomycin reduced autotrophic nitrification by 94%, whereas cycloheximide had no effect on autotrophic nitrification, indicating that autotrophic nitrification was mainly driven by bacteria. However, the opposite was true in another agricultural soil (paddy), indicating that fungi contributed to the oxidation of NH₄⁺ to NO₃^?. In the acid forest soil, cycloheximide, but not streptomycin, inhibited heterotrophic nitrification, demonstrating that fungi controlled the heterotrophic nitrification. The conversion of forest to agricultural soils resulted in a shift from fungi-dominated heterotrophic nitrification to bacteria- or fungi-dominated autotrophic nitrification. Our results suggest that land-use and management practices, such as the application of N fertilizer and lime, the long-term waterflooding during rice growth, straw return after harvest, and cultivation could markedly influence the relative contribution of bacteria and fungi to specific soil nitrification processes.     

Effect of a cold shock on the activity and composition of the communities of ammonium-oxidizing microorganisms in a chestnut soil

The simulation of a cold shock was performed in an incubation experiment with soil microcosms by a sharp decrease of the temperature to negative values and the subsequent analysis of the nitrification rate of the ammonium-oxidizing microorganisms. Three procedures of the cold shock effect were selected: long, short-time, and cyclic. A significant decrease of the nitrifying activity was recorded after the long effect, whereas the 8-, 16-, and 24-hour cold shocks did not affect the intensity of nitrification. A cyclic temperature decrease alternating with periods of incubation under high temperatures also did not affect the nitrifying activity of the microorganisms. We suppose that the domination of mesophilic microorganisms with a resistant enzyme system or of psychrophilic and psychrotolerant microorganisms contributes to the preservation of a high nitrification level in soils with frequent alternations of high and low temperatures.     

Archaea and bacteria respectively dominate nitrification in lightly and heavily grazed soil in a grassland system

To investigate the effects of stocking rates on nitrification activity and active nitrifying communities in a typical steppe grazing system, we conducted a laboratory incubation study using soil from a 10-year-old grassland gradient grazing experiment with sheep. A combination of molecular methods, such as DNA-based stable-isotope probing (DNA-SIP), real-time quantitative PCR, and high-throughput sequencing, was used to identify changes of nitrification activity and active nitrifying communities under different stocking rates (0 (SR0), 3 (SR3), 6 (SR6), and 9 (SR9) sheep per ha). The nitrification activity of soils was significantly increased by light grazing (SR3), while it was significantly decreased by heavy grazing (SR9). Nitrososphaera viennensis lineage of ammonia-oxidizing archaea (AOA) functionally predominated over ammonia-oxidizing bacteria (AOB) in nitrification in the SR3 soil, while the Nitrosospira cluster 3 of AOB was the major player in the SR9 soil. Therefore, stocking rates altered the distribution of active nitrifying communities by affecting soil chemical and physical conditions.     

Effects of organic manure on nitrification in arable soils

Summary The production of nitrate by the process of nitrification is highly dependent on other N-transforming processes in the soil. Hence, changes in the type of N compound applied to enrich agricultural soils may affect the production of nitrate. The size and activity of the chemolithotrophic bacterial community were studied in an integrated farming system, with increased inputs of organic manure and reduced inputs of mineral nitrogenous fertilizer, versus conventional farming. The integrated farming had a positive effect on potential nitrifying activity, but not on the numbers of chemolithotrophic nitrifying bacteria as determined by a most probable number technique or by fluorescence antibody microscopy. Cells of the recently described nitrite-oxidizing species Nitrobacter hamburgensis and Nitrobacter vulgaris were just as common as the cells of the well known species Nitrobacter winogradskyi. It was concluded that nitrification is stimulated by integrated farming, presumably by an increased mineralization of ammonium which is not immediately consumed by the crop or immobilized in the heterotrophic microflora of the soil. Since nitrifying bacteria are involved in the production of NO and N₂O, integrated farming with the application of manure may favour the production of noxious N-oxides.Communication no. 40 of the Dutch Programme on Soil Ecology of Arable Farming Systems     

Fluctuations in the abundance of ammonia oxidizers and the contents of inorganic nitrogen in solarized soil

Solarization makes a great impact on the abundance of ammonia oxidizers and nitrifying activity in soil. To elucidate fluctuations in the abundance of ammonia oxidizers and nitrification in solarized soil, copy numbers of amoA gene of ammonia-oxidizing bacteria (AOB) and archaea (AOA), viable number of ammonia oxidizers and inorganic nitrogen contents were investigated in greenhouse experiments. The copy number of amoA gene and the viable number of ammonia oxidizers were determined by the quantitative polymerase chain reaction and most probable number methods, respectively. Abundance of AOB based on the estimation of amoA gene copy numbers and viable counts of ammonia oxidizers was decreased by the solarization treatment and increased during the tomato (Solanum lycopersicum L.) cultivation period following the solarization. Effect of solarization on the copy number of amoA gene of AOA was less evident than that on AOB. The proportion of nitrate in inorganic nitrogen contents was declined by the solarization and increased during the tomato cultivation period following the solarization. Positive correlations were found between the proportion of nitrate in inorganic nitrogen content and the copy number of bacterial or archaeal amoA gene or the viable number of ammonia oxidizers; the copy number of bacterial amoA gene showed a strong correlation with the viable number of ammonia oxidizers. The present study revealed influences of solarization on the fluctuation in the abundance of ammonia oxidizers and dynamics of inorganic nitrogen contents in soil and the results indicate that the determination of amoA gene of AOB is possibly a quick and useful diagnostic technique for evaluating suppression and restoration of nitrification following solarization.     

长期定位施肥对中性紫色土硝化作用及氨氧化微生物的影响

为了研究长期不同施肥措施对中性紫色土氨氧化微生物及其硝化作用的影响,以国家紫色土肥力与肥料效益监测基地的中性紫色土为研究对象,进行土壤氨氧化细菌和氨氧化古菌amo A基因的Real-time PCR分析,比较长期不同定位施肥对土壤氨氧化潜势和硝化强度的影响,并分析不同施肥制度对功能微生物丰度与功能的作用。数据显示,土壤中氨氧化古菌amo A基因拷贝数(Log值6.21～7.14)远大于氨氧化细菌(Log值3.65～5.73),相对于对氨氧化细菌丰度的影响,施肥对土壤氨氧化古菌丰度影响较小。施用氮肥与磷肥都显著提高了土壤氨氧化细菌丰度,1.5NPK+M处理氨氧化细菌丰度最高(Log值5.73),有机无机肥配施可以显著提高土壤氨氧化微生物丰度;而含氯化肥的施用在一定程度上降低了土壤氨氧化细菌丰度与硝化细菌生长,与施用不含氯的肥料处理相比,含氯肥料处理的土壤氨氧化细菌丰度与硝化细菌数分别降低了3.74%和88.12%。研究表明,长期施肥能影响中性紫色土中氨氧化细菌的丰度,有机无机肥配施能够提高土壤的氨氧化潜力与土壤的硝化能力。