土壤氨氧化细菌对大气CO2浓度增高的响应

摘要：利用FACE(free．aircarbondioxideenrichment，开放式空气CO2浓度增高)试验平台，研究大气CO2浓度增高对土壤氨氧化细菌的数量、优势菌群及其硝化活性的影响。结果表明，大气CO2浓度增高时，土壤氨氧化细菌的数量在常氮水平上趋于而在高氮水平上与对照没有差异。大气CO2浓度增高对土壤氨氧化细菌的优势菌群也产生明显影响。CO2浓度增高条件下，亚硝化球菌(Nitrosococcus sp.)和亚硝化弧菌(Nitrosovibrio sp．)是优势菌属；而在对照条件下，亚硝化单胞菌(Nitrosomonas sp．)和亚硝化球菌(Nitrosococcus sp．)是优势菌属。另外，CO2浓度增高条件下优势菌株的硝化活性也有不同程度的减弱。     

大气CO2浓度增高对农田土壤硝化活性的影响

利用中国唯一的FACE(Free-Air Carbon dioxide Enrichment,开放式空气CO2浓度增高)平台,研究大气CO2浓度增高对农田土壤硝化活性的影响.位于无锡的中国稻麦轮作农田生态系统FACE试验平台于2001年6月开始运行,设有FACE与Ambient(普通空气对照)2个处理,FACE区CO2浓度比Ambient区高200 μmol·mol-1,每个处理含低氮与常氮2个氮肥水平.在轮作水稻和小麦各3季之后,发现大气CO2浓度增高下,常氮水平上土壤的NO3--N质量分数降低,NH4+-N质量分数增高;而低氮水平上土壤的NO3--N质量分数增高,NH4+-N质量分数没有显著差异.然后分别在土壤样品中加入NH4+-N,好气培养42 d后通过测定土壤中的NO3--N、NO2--N总质量分数来研究土壤的硝化活性.结果显示,不管在CO2浓度增高下还是对照条件下,增加氮肥施用量均增强了土壤的硝化活性;且与对照相比,大气CO2浓度增高在常氮水平上降低了土壤的硝化活性,在低氮水平上却增强了土壤的硝化活性,说明大气CO2浓度增高对农田土壤硝化活性的影响与N肥供应水平有关.     

高氨氮废水处理系统中功能微生物的检测

利用16S rDNA分子检测技术对高氨氮废水处理系统中的氨氧化细菌、亚硝酸盐氧化细菌等难分离培养微生物进行了分析检测.从实验室处理高氨氮废水的生物流化床硝化系统生物膜中提取细菌总DNA,以其为模板,利用氨氧化细菌(AOB)和亚硝酸盐氧化细菌(NOB)的16S rDNA特异性引物进行多聚酶链式反应(PCR)扩增,获得AOB和NOB的特异片段.将特异片段与pGEM-T载体连接并转化到E.coli DH5α中获得重组子,对阳性重组子进行酶切分型,AOB可分为8种不同类型.选取AOB的8种类型代表以及随即选取8个NOB克隆进行测序.经GenBank搜索以及同源性分析,发现所获得的8个AOB类型来自不同的菌株,处理系统中以Nitrosomonas sp.和Nitrosococcus sp.为氨氧化细菌的优势菌属;8株NOB重组子中有7株来自不同的菌株,系统中Nitrobacter sp.和Afipia sp.为亚硝酸盐氧化细菌的优势菌属.图6表1参14     

近地层臭氧浓度升高对稻田土壤氨氧化与反硝化细菌活性的影响

利用中国唯一的稻麦轮作臭氧FACE（free-air O3 concentration enrichment,开放式空气臭氧浓度增高）试验平台,研究近地层臭氧浓度升高对稻田不同生育时期植株氮吸收量、土壤氮含量与脲酶活性、土壤氨氧化细菌与反硝化细菌数量以及成熟期土壤硝化与反硝化作用强度的影响。结果发现,臭氧浓度升高条件下,单株水稻（Oryza sativaL.）的氮吸收量趋于升高,土壤全氮含量趋于下降、脲酶活性趋于增强,在成熟期土壤全氮与对照相比平均下降9%,土壤脲酶活性与对照相比平均升高13%。在水稻整个生长季节,土壤氨氧化细菌和反硝化细菌的数量均呈现出先增多后减少的趋势,且均在开花期达到峰值,但臭氧熏蒸土壤与对照相比趋于升高;在水稻成熟期,土壤中单个氨氧化细菌和反硝化细菌的活性均趋于下降,土壤硝化强度和反硝化强度与对照相比也平均下降17%和24%。结果表明,近地层臭氧浓度升高条件下,稻田土壤氮素转化因为水稻氮素吸收增强而加快,土壤氨氧化细菌和反硝化细菌的数量均趋于增多,但它们的生理代谢活性均趋于下降。     

OLAND生物脱氮系统运行及其硝化菌群的分子生物学检测

采用两阶段限氧自养硝化 -反硝化生物脱氮系统 (oxygen limitedautotrophicnitrificationanddenitrificationsystem ,以下简称OLAND)处理高氨氮、低COD的废水 .应用内浸式多聚醚砜中空膜 ,实现了污泥的完全截留 ,阻止了生物量的大量洗脱 ,并通过控制溶氧在 0 .1～ 0 .3mgL-1之间 ,实现了硝化阶段出水中氨氮与亚硝态氮浓度的比例达到最适值〔1 (1.2± 0 .2 )〕 ,从而为第二阶段的厌氧氨氧化提供理想的进水 ,进而获得较高的脱氮率 .同时应用荧光原位杂交技术对硝化阶段不同时期硝化菌群的变化进行分子生物学检测 ,揭示了随溶氧浓度的降低 ,氨氧化菌的数量基本保持恒定、亚硝酸氧化菌的数量略有减少的变化规律 ,并且发现 ,在两阶段限氧自养硝化 -反硝化生物脱氮系统中氨氮的氧化主要是由Nitrosomonassp .完成 ,亚硝酸的氧化主要由Nitrobactersp .完成 .图 4表 2参 2 2     

近地层臭氧浓度升高对麦田土壤氨氧化与反硝化细菌活性的影响

利用臭氧FACE(free-air O3concentration enrichment,开放式空气臭氧浓度增高)试验平台,研究近地层臭氧浓度(臭氧摩尔分数平均为70 nmol.mol-1)升高对小麦不同生育期植株氮吸收量,土壤w(全氮)、w(矿质氮)、脲酶活性、氨氧化细菌数量、反硝化细菌数量以及小麦成熟期土壤硝化与反硝化作用强度的影响。结果表明,与对照(臭氧摩尔分数平均为45 nmol.mol-1)相比,臭氧浓度升高条件下小麦不同生育期氮吸收量总体趋于升高,土壤w(全氮)、w(铵态氮)和w(硝态氮)总体趋于下降,在小麦成熟期土壤w(全氮)和w(铵态氮)分别比对照下降9%和71%(P0.05),而w(硝态氮)比对照下降36%;臭氧浓度升高使小麦不同生育期土壤脲酶活性总体趋于增强,在拔节期、抽穗期和灌浆期均显著高于对照(P0.05);随着小麦的生长,臭氧熏蒸土壤氨氧化细菌和反硝化细菌数量也趋于升高,在小麦成熟期均显著高于对照(P0.05)。在小麦成熟期,尽管臭氧熏蒸土壤单个氨氧化细菌的硝化活性比对照下降57%,但土壤整体硝化强度却比对照提高123%;臭氧熏蒸土壤反硝化作用强度与对照相比无明显差异,但单个反硝化细菌的反硝化活性却比对照降低96%,达显著水平(P0.05)。认为臭氧浓度升高促进了小麦对土壤氮素的吸收,导致土壤氮素库存量降低,进而加快了土壤中氮素的转化,表现为脲酶活性升高,氨氧化细菌和反硝化细菌数量增加,但它们的代谢活性反而下降。     

SMBBR工艺处理生活污水脱氮效能及其微生物多样性

微生物是生物膜法处理技术的核心,微生物多样性的研究对于生物膜法去除污染物的机理探讨具有重要意义.采用特异性流化床生物膜反应器(Special Moving-Bed Biofilm Reactor,SMBBR)处理城市生活用水,并利用IlluminaHiSeq高通量测序技术对各反应器的微生物分布以及菌群与环境因子的相关性进行研究.结果显示,在水温20-30℃、上清液回流比为150%、DO为4 mg/L、水力停留时间为18 h的条件下,SMBBR对氨氮的去除率高达96.7%;SMBBR的好氧反应器和厌氧反应器的生物膜微生物种群结构组成存在不同,但优势微生物种群均为变形菌门(Proteobacteria)和厚壁菌门(Firmicutes).在属的水平检测到好氧反硝化菌红细菌(Rhodobacter)、陶厄氏菌属(Thauera),以及氨氧化细菌(Ammonia-oxidizing bacteria,AOB)亚硝化单胞菌(Nitrosomonadales)等.另外,DO是影响微生物群落结构最显著的环境因子.本研究表明环境因子会影响微生物群落替演,好氧反硝化以及氨氧化反应可能是SMBBR工艺中重要的脱氮机制.     

单级好氧和限氧污水处理系统中总氮损失的微生物学机理

近年来,单级好氧和限氧污水处理系统中总氮损失的现象引起了人们的普遍关注,本文对这种现象的微生物学机理及研究现状进行了阐述,主要是儿类细菌的单独脱氮或者它们之间的协同脱氮,包括自养(亚)硝化菌单独脱氮、好氧反硝化菌单独脱氮、(亚)硝化菌和好氧反硝化菌的协同脱氮以及(亚)硝化菌和厌氧氨氧化菌的协同脱氮.与传统的硝化-反硝化脱氮工艺相比,这些脱氮新途径具有不可比拟的优越性,对于强化污水生物脱氮具有重要意义.图8参53     

大气CO2浓度升高对添加麦秸条件下稻田土壤酶活性的影响

利用中国扬州开放式大气CO2浓度升高(FACE)系统稻麦轮作试验平台,研究大气CO2浓度升高对稻田小麦秸秆降解过程中土壤酶活性的影响.试验平台设置FACE[(580±60)μmol·mol-1]和对照[(380±40)μmol·mol-1]2个CO2浓度,在低氮(150 kg·hm-2)和高氮(250 kg·hm-2)2种氮水平下添加小麦秸秆,分别在不同生长时期采样并分析土壤脱氢酶、脲酶、磷酸酶和蔗糖酶活性.结果表明:同一氮水平下,大气CO2浓度升高显著增强土壤脱氢酶、脲酶和蔗糖酶活性,但显著降低土壤磷酸酶活性;无论是大气CO2浓度升高条件下还是对照条件下,低氮处理土壤脱氢酶与脲酶活性均高于高氮处理,而高氮处理土壤磷酸酶活性均高于低氮处理,氮水平对蔗糖酶活性并无显著影响.     

氮肥对土壤氧化大气甲烷影响的机制

综合评述了氮氧化甲烷的抑制机制。包括：（1）竞争甲烷单氧化酶的竞争抑制机制,（2）代谢产物的毒害抑制机制,（3）外源盐引起的微生物生理缺水抑制机制和（4）氮素周围作用引起的抑制机制。提出了氧化菌竞争利用土壤空气有限O2的竞争抑制机制,即氨氧化菌利用更多的土壤有限氧气→产生优势氨氧化菌→形成优势菌群→限制甲烷氧化菌繁殖和功能发挥的氨长期抑土壤化大气甲烷的机制,并认为这种抑制作用是不可逆的。     

不同C/N值下亚硝酸盐氧化菌和异养菌混合体系的微生物多样性

采用PCR-RFLP技术研究了不同C/N比下亚硝酸盐氧化菌及异养菌混合体系的微牛物多样性,并探讨了微生物菌群结构与其功能(硝化件能)的关系.C/N=0时,混合体系主要由自养菌和寡营养菌(85.1%)组成,包括亚硝酸盐氧化菌(NOB)、拟杆菌门、α-变形菌纲、浮霉菌门和绿色非硫细菌中的一些菌株.C/N=0.44时,混合体系中的自养菌减少,异养菌(主要是γ-变形菌纲的成员)大量出现.C/N=8.82时,γ-变形菌纲的菌株尤其是反硝化菌Pseudomonas sp.占主导(93.8%),与此同时,随着C/N升高,该混合体系的硝化性能也由专一的亚硝酸盐氧化过程转变为同时硝化反硝化过程.微生物菌群结构的转变较好地解释了其硝化性能的改变.本研究揭示了微生物菌群结构与其功能的内在联系,同时表明PCR-RFLP技术与化学分析相结合是研究微生物菌群结构与功能的有力工具.图3表2参13     

Impact of total organic carbon and chlorine to ammonia ratio on nitrification in a bench-scale drinking water distribution system

Nitrification occurs in chloraminated drinking water systems and is affected by water quality parameters. The aim of this study was to investigate the impact of total organic carbon and chlorine to ammonia ratio on nitrification potential in a simulated drinking water distribution system as during chloramination. The occurrence of nitrification and activity of nitrifying bacteria was primarily monitored using four rotating annular bioreactors (RAB) with different chlorine to ammonia ratios and total organic carbon (TOC) levels. The results indicated that nitrification occurred despite at a low influent concentration of ammonia, and a high concentration of nitrite nitrogen was detected in the effluent. The study illustrated that reactors 1(R1) and 3 (R3), with higher TOC levels, produced more nitrite nitrogen, which was consistent with the ammonia-oxidizing bacteria (AOB) counts, and was linked to a relatively more rapid decay of chloramines in comparison to their counterparts (R2 and R4). The AOB and HPC counts were correlated during the biofilm formation with the establishment of nitrification. Biofilm AOB abundance was also higher in the high TOC reactors compared with the low TOC reactors. The chlorine to ammonia ratio did not have a significant impact on the occurrence of nitrification. Bulk water with a high TOC level supported the occurrence of nitrification, and AOB development occurred at all examined chlorine to ammonia dose ratios (3:1 or 5:1).     

Calcium constrains plant control over forest ecosystem nitrogen cycling

Forest ecosystem nitrogen (N) cycling is a critical controller of the ability of forests to prevent the movement of reactive N to receiving waters and the atmosphere and to sequester elevated levels of atmospheric carbon dioxide (CO2). Here we show that calcium (Ca) constrains the ability of northern hardwood forest trees to control the availability and loss of nitrogen. We evaluated soil N-cycling response to Ca additions in the presence and absence of plants and observed that when plants were present, Ca additions "tightened" the ecosystem N cycle, with decreases in inorganic N levels, potential net N mineralization rates, microbial biomass N content, and denitrification potential. In the absence of plants, Ca additions induced marked increases in nitrification (the key process controlling ecosystem N losses) and inorganic N levels. The observed "tightening" of the N cycle when Ca was added in the presence of plants suggests that the capacity of forests to absorb elevated levels of atmospheric N and CO2 is fundamentally constrained by base cations, which have been depleted in many areas of the globe by acid rain and forest harvesting.     

Nitrous oxide nitrification and denitrification 15N enrichment factors from Amazon forest soils.

The isotopic signatures of 15N and 18O in N2O emitted from tropical soils vary both spatially and temporally, leading to large uncertainty in the overall tropical source signature and thereby limiting the utility of isotopes in constraining the global N2O budget. Determining the reasons for spatial and temporal variations in isotope signatures requires that we know the isotope enrichment factors for nitrification and denitrification, the two processes that produce N2O in soils. We have devised a method for measuring these enrichment factors using soil incubation experiments and report results from this method for three rain forest soils collected in the Brazilian Amazon: soil with differing sand and clay content from the Tapajos National Forest (TNF) near Santarém, Pará, and Nova Vida Farm, Rond?nia. The 15N enrichment factors for nitrification and denitrification differ with soil texture and site: -111 per thousand +/- 12 per thousand and -31 per thousand +/- 11 per thousand for a clay-rich Oxisol (TNF), -102 per thousand +/- 5 per thousand and -45 per thousand +/- 5 per thousand for a sandier Ultisol (TNF), and -10.4 per thousand +/- 3.5 per thousand (enrichment factor for denitrification) for another Ultisol (Nova Vida) soil, respectively. We also show that the isotopomer site preference (delta15Nalpha - delta15Nbeta, where alpha indicates the central nitrogen atom and beta the terminal nitrogen atom in N2O) may allow differentiation between processes of production and consumption of N2O and can potentially be used to determine the contributions of nitrification and denitrification. The site preferences for nitrification and denitrification from the TNF-Ultisol incubated soils are: 4.2 per thousand +/- 8.4 per thousand and 31.6 per thousand +/- 8.1 per thousand, respectively. Thus, nitrifying and denitrifying bacteria populations under the conditions of our study exhibit significantly different 15N site preference fingerprints. Our data set strongly suggests that N2O isotopomers can be used in concert with traditional N2O stable isotope measurements as constraints to differentiate microbial N2O processes in soil and will contribute to interpretations of the isotopic site preference N2O values found in the free troposphere.     

Litterfall 15N abundance indicates declining soil nitrogen availability in a free-air CO2 enrichment experiment

Forest productivity increases in response to carbon dioxide (CO2) enrichment of the atmosphere. However, in nitrogen-limited ecosystems, increased productivity may cause a decline in soil nitrogen (N) availability and induce a negative feedback on further enhancement of forest production. In a free-air CO2 enrichment (FACE) experiment, the response of sweetgum (Liquidambar styraciflua L.) productivity to elevated CO2 concentrations [CO2] has declined over time, but documenting an associated change in soil N availability has been difficult. Here we assess the time history of soil N availability through analysis of natural 15N abundance in archived samples of freshly fallen leaf litterfall. Litterfall delta15N declined from 1998 to 2005, and the rate of decline was significantly faster in elevated [CO2]. Declining leaf litterfall delta15N is indicative of a tighter ecosystem N cycle and more limited soil N availability. By integrating N availability over time and throughout the soil profile, temporal dynamics in leaf litterfall delta15N provide a powerful tool for documenting changes in N availability and the critical feedbacks between C and N cycles that will control forest response to elevated atmospheric CO2 concentrations.     

Effects of hydraulic retention time on nitrification activities and population dynamics of a conventional activated sludge system

The effects of hydraulic retention time (HRT) on the nitrification activities and population dynamics of a conventional activated sludge system fed with synthetic inorganic wastewater were investigated over a period of 260 days. When the HRT was gradually decreased from 30 to 5 h, the specific ammonium-oxidizing rates (SAOR) varied between 0.32 and 0.45 kg NH4+-N (kg mixed liquor suspended solids (MLSS)·d)^-1, and the specific nitrate-forming rates (SNFR) increased from 0.11 to 0.50 kg NO3--N (kg MLSS·d)^-1, showing that the decrease in HRT led to a significant increase in the nitrite oxidation activity. According to fluorescence in situ hybridization (FISH) analysis results, the proportion of ammonia-oxidizing bacteria (AOBs) among the total bacteria decreased from 33% to 15% with the decrease in HRT, whereas the fraction of nitrite-oxidizing bacteria (NOBs), particularly the fast-growing Nitrobacter sp., increased significantly (from 4% to 15% for NOBs and from 1.5% to 10.6% for Nitrobacter sp.) with the decrease in HRT, which was in accordance with the changes in SNFR. A short HRT favored the relative growth of NOBs, particularly the fast-growing Nitrobacter sp., in the conventional activated sludge system.     

Dynamics of nitrogen and dissolved organic carbon at the Hubbard brook experimental forest

The factors controlling spatial and temporal patterns in soil solution and streamwater chemistry are highly uncertain in northern hardwood forest ecosystems in the northeastern United States, where concentrations of reactive nitrogen (Nr) in streams have surprisingly declined over recent decades in the face of persistent high rates of atmospheric Nr deposition and aging forests. Reactive nitrogen includes inorganic species (e.g., ammonium [NH4+], nitrate [NO3-]) and some organic forms (e.g., amino acids) available to support the growth of plants and microbes. The objective of this study was to examine controls on the spatial and temporal patterns in the concentrations and fluxes of nitrogen (N) species and dissolved organic carbon (DOC) in a 12-year record of soil solutions and streamwater along an elevational gradient (540-800 m) of a forested watershed at the Hubbard Brook Experimental Forest (HBEF) in the White Mountains of New Hampshire, USA. Dissolved organic N and DOC concentrations were elevated in the high-elevation spruce-fir-white birch (SFB) zone of the watershed, while NO3- was the dominant N species in the lower elevation hardwood portion of the watershed. Within the soil profile, N retention was centered in the mineral horizon, and significant amounts of N were retained between the lower mineral soil and the stream, supporting the idea that near- and in-stream processes are significant sinks for N at the HBEF. Temporal analysis suggested that hydrologic flow paths can override both abiotic and biotic retention mechanisms (i.e., during the non-growing season when most hydrologic export occurs, or during years with high rainfall), there appears to be direct flushing of N from the organic horizons into the stream via horizontal flow. Significant correlations between soil NO3- concentrations, nitrification rates and streamwater NO3- exports show the importance of biological production as a regulator of inorganic N export. The lack of internal production response (e.g., mineralization, nitrification) to a severe ice storm in 1998 reinforces the idea that plant uptake is the dominant regulator of export response to disturbance.     

Production of N2O in two biologic nitrogen removal processes: a comparison between conventional and short-cut nitrogen removal processes

The N2O production in two nitrogen removal processes treating domestic wastewater was investigated in laboratory-scale aerobic-anoxic sequencing batch reactors （SBRs）. Results showed that N2O emission happened in the aerobic phase rather than in the anoxic phase. During the aerobic phase, the nitrogen conversion to N2O gas was 27.7% and 36.8% of NH＋-N loss for conventional biologic N-removal process and short-cut biologic N-removal process. The dissolved N2O was reduced to N2 in the anoxic denitrification phase. The N2O production rate increased with the increasing of nitrite concentration and ceased when NH＋-N oxidation was terminated. Higher nitrite accumulation resulted in higher NEO emission in the short-cut nitrogen removal process. Pulse-wise addition of 20 mg NO2 -N. L- 1 gave rise to 3-fold of N2O emission in the conventional N-removal process, while little change happened with 20 mg NOS-N L-1 was added to SBR1.     

菌根真菌对大气CO_2浓度升高的响应研究进展

大气CO2浓度升高对植物的光合作用、呼吸作用等产生直接影响,进而影响到运送到根系中碳的量,菌根真菌也随之受到影响.本文对全球CO2浓度升高对菌根真菌的影响、菌根真菌在植物对大气CO2增加响应中的作用、菌根真菌在大气CO2浓度增加条件下对整个生态系统的作用等进行了综述,同时对当前存在的问题和未来的发展做了探讨.图1参37