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。     

秸秆还田种类对稻田N2O排放及硝化反硝化微生物的影响

以太湖流域典型单季稻田的原状土柱为研究对象,通过设置温室土柱试验,同步监测3种秸秆(水稻秸秆RS、小麦秸秆WS、玉米秸秆MS)施用下水稻各生长期N₂O排放、水稻产量和土壤理化因子,同时定量化分析多个N₂O排放相关菌群及功能基因的丰度,以阐明N₂O排放对不同种类秸秆施用引发的微生物响应机制,筛选控制单季稻田N₂O减排增效最佳的秸秆种类。结果表明：与对照相比,RS、WS和MS处理下水稻生长期N₂O排放量分别增加162.32%、107.11%和9.48%,其中RS处理显著高于MS处理。水稻生育期内,土壤氨氧化菌(AOA、AOB)和反硝化菌群落(nir S、nos Z)丰度均呈现先上升后下降的变化趋势。与对照相比,拔节期RS处理显著增加AOA、AOB、nirS和nosZ拷贝数,MS和WS处理对上述功能基因丰度均无显著影响。各生育期土壤NH₄⁺-N含量整体高于NO₃^–-N含量,二者均在水稻分蘖期达到峰...     

生物质炭对酸性菜地土壤N2O排放及相关功能基因丰度的影响

【目的】 生物质炭显著影响土壤氧化亚氮 (N₂O) 排放,但关于其相关微生物机理的研究相对匮乏,尤其是生物质炭对酸性菜地土壤N₂O排放的微生物作用机理。本文通过研究氮肥配施生物质炭对酸性菜地土壤N₂O排放以及硝化和反硝化过程相关功能基因丰度的影响,探讨酸性菜地土壤N₂O排放与功能基因丰度的关系,阐释生物质炭对酸性菜地土壤试验N₂O排放的微生物作用机理。 【方法】 在田间一次性施入生物质炭 40 t/hm2,试验连续进行了3年,共9茬蔬菜。设置4个处理:对照 (CK)、氮肥 (N)、生物质炭 (Bc) 和氮肥 + 生物质炭 (N + Bc)。在施用后第三年,采集土壤样品进行室内培养,应用荧光定量PCR技术检测硝化过程氨氧化古菌 (AOA)、氨氧化细菌 (AOB) 功能基因amoA和反硝化过程亚硝酸还原酶基因 (nirK、nirS) 以及N₂O还原酶基因 (nosZ) 等相关功能基因丰度,同时监测土壤pH值、无机氮 (铵态氮、硝态氮) 含量及N₂O排放。 【结果】 与CK相比,生物质炭 (Bc) 处理的土壤有机碳 (SOC) 提高了27.1%,总氮 (TN) 提高了8.2%,amoA-AOB基因丰度显著降低了11.0%,nosZ基因丰度增加了21.2% (P < 0.05),N ₂O排放没有显著变化 (P > 0.05)。与CK相比,施用氮肥 (N) 显著降低土壤pH ( P < 0.05),显著增加土壤无机氮含量、 nirK、nirS和nosZ功能基因丰度以及土壤N₂O累积排放量 (P < 0.05)。与N处理相比,生物质炭与氮肥联合施用 (N + Bc) 处理显著增加 amoA-AOA、amoA-AOB、nirK、nirS和nosZ基因丰度,增幅分别为68.1%、39.3%、21.1%、19.8%、48.4% (P < 0.05),但 ( nirK + nirS)/nosZ的比值降低,同时N₂O累积排放量显著降低33.3% (P < 0.05)。室内培养期间N ₂O排放峰出现在1～5 d,N和N+Bc处理排放速率分别为 N 1.70 × 103和1.76 × 103 ng/(kg·h)。相关分析结果显示,N₂O排放速率与氧化亚氮还原酶的标记基因nosZ基因拷贝数 (P < 0.05)、NH ₄+-N含量 (P < 0.01) 呈显著正相关,与pH呈显著负相关 ( P < 0.01)。 【结论】 在菜地生态系统中氮肥和生物质炭联合施用可以有效缓解菜地土壤酸化,减少菜地土壤N₂O排放,主要归因于反硝化作用nosZ基因丰度增加,(nirK + nirS)/nosZ比值降低。      

生物硝化抑制剂对黔西南黄壤硝化作用及N2O排放的影响

为探索生物硝化抑制剂对贵州黔西南地区黄壤硝化作用及氧化亚氮(N2O)排放的影响,通过三周的室内培养试验,研究两种生物硝化抑制剂对羟基苯丙酸甲酯(MHPP)和丁香酸(SA)对黄壤中的无机态氮素含量、氨氧微生物功能基因以及N2O排放量的影响。结果表明,与对照CK相比,MHPP和SA在黄壤上均能明显抑制硝化作用,对土壤硝化速率的抑制率分别为6%~43%和5%~51%。MHPP和SA均抑制了黄壤氨氧化古菌AOA(12%~22%,27%~41%)与氨氧化细菌AOB(6%~19%,26%~46%)amoA基因的丰度。整个培养期内,黄壤的硝态氮含量与AOB的amoA基因丰度显著正相关,而与AOA的amoA基因丰度无显著相关,表明AOB对黄壤硝化作用起了主导作用。在N2O排放方面,MHPP和SA分别显著抑制了黄壤51%和21%的N2O排放积累量,MHPP的减排效果优于SA。MHPP降低了黄壤N2O排放的峰值,而SA主要延缓了黄壤N2O产生高峰的出现。总之,生物硝化抑制剂MHPP和SA在贵州黔西南黄壤上具有氮肥减施增效的潜力,这为今后烤烟新型绿色专用肥的开发提供了理论依据。     

有机和无机肥配比对黄褐土硝化和反硝化微生物丰度及功能的影响

【目的】 土壤硝化与反硝化作用是氮循环的两个关键环节,本文研究不同比例的有机、无机肥配施对硝化和反硝化进程产生的影响,为高效施肥提供理论基础。 【方法】 在安徽农业大学农翠园试验基地的黄褐土上进行了小麦–玉米轮作田间试验。试验以不施氮肥为对照 (CK),在小麦、玉米总施氮量相同的条件下,设置5个处理,分别为单施无机肥 (T1)、无机肥∶有机肥 = 2∶1 (T2)、无机肥∶有机肥 = 1∶1 (T3)、无机肥∶有机肥 = 1∶2 (T4)、单施有机肥 (T5)。在小麦拔节期,取0—20 cm土壤样品,利用荧光定量PCR技术测定反硝化和氨氧化微生物丰度,并结合反硝化能力、N₂O/(N₂O+N₂) 产物比、土壤呼吸、硝化势和氨氧化细菌 (AOB) 与古菌 (AOA) 对硝化势相对贡献率的测定,分析江淮地区长期有机和无机肥配施对黄褐土硝化、反硝化微生物丰度及其功能的影响。 【结果】 单施无机肥或有机肥处理的硝化势均高于不同配比处理。与添加有机肥相比,增施无机肥会显著增加AOA的丰度和硝化贡献率。在反硝化方面,反硝化能力和土壤呼吸随着有机肥投入量的增加而增加,单施有机肥处理显著高于其它处理。nirS和nosZ型反硝化菌丰度随着有机肥的增加而增加,而nirK型反硝化菌丰度呈减少趋势。相关分析表明,反硝化能力与nirS型、nosZ型反硝化菌丰度、有机质和可溶性有机碳含量极显著正相关,与nirK相关性不强。 【结论】 与单施无机肥或有机肥处理相比,有机和无机肥适当配施可降低土壤硝化势,并能调控AOA和AOB在硝化过程中的作用,有效地降低土壤反硝化损失。      

硝化抑制剂与尿素配施对旱地土壤温室气体排放及硝化微生物的影响

为明确施肥措施对旱地土壤温室气体排放的综合效应及微生物机理,采集典型麦田土壤进行室内微宇宙培养,研究了双氰胺(DCD)和烯丙基硫脲(ATU)分别与尿素配施对旱地土壤氮素转化及N₂O、CO₂和CH₄排放的影响,同时监测了不同类型微生物群落的动态变化。结果表明氨氧化细菌(AOB)主导了施氮麦田土壤硝化过程及N₂O排放。单施尿素促进AOB迅速繁殖,使N₂O排放总量提高235%,同时促进了细菌生长,CO₂排放量增加18.5%。DCD与尿素配施极大程度抑制了AOB的生长,显著降低了N₂O排放(59.4%),但促进了细菌的生长并提高了CO₂的排放总量(50.6%)。而ATU与尿素配施同时抑制了真菌、细菌和AOB的生长,对反硝化细菌的影响则相反,使CO₂和N2O排放分别下降28.4%和35.2%。与不施肥相比,氮肥及与两种硝化抑制剂配施均显著降低了CH4的排放量。3种温室气体的综合温室效应在处理间差异显著...     

不同形态氮添加对毛竹林土壤N2O排放的影响

  【目的】  氧化亚氮(N₂O)排放是亚热带地区氮损失的主要途径,我们研究了不同形态含氮化合物对土壤N₂O排放的影响。  【方法】  以毛竹(Phyllostachys edulis)林土壤为研究对象进行了室内培养试验。设置土壤中添加KNO₃、NH₄NO₃、NH₄Cl、KCl处理,以去离子水作为对照(CK),在25oC黑暗条件下培养。在培养0.5 h,1、3、5、7、14、28、60天,测定土壤N₂O排放速率,铵态氮(NH₄+-N)、硝态氮(NO₃?-N)、可溶性有机碳(DOC)和水溶性氮(WSN)含量,采用荧光定量PCR技术测定了土壤氨氧化细菌(ammonia-oxidizing bacteria, AOB)、氨氧化古菌(ammonia-oxidizing archaea, AOA)、nirS、nirK、nosZⅠ、nosZⅡ基因丰度。  【结果】  培养第60天,氮添加与KCl添加处理均显著增加了土壤DOC含量,NH₄NO₃、NH₄Cl处理显著增加了WSN含量,但显著降低了土壤pH。氮添加及KCl添加处理均增加了土壤AOA、AOB、nirK基因丰度,降低了nosZⅠ、nosZⅡ基因丰度。氮添加处理N₂O排放速率均在培养第14天达到峰值,且相较于CK处理均增加了N₂O累积排放量,KNO₃、NH₄NO₃、NH₄Cl和KCl处理累积排放量的增幅分别为524.3%、771.1%、652.7%、98.6%。N₂O排放速率与NO₃?、WSN、nirK基因丰度呈显著正相关,而与pH、nosZⅠ、nosZⅡ基因丰度呈显著负相关。  【结论】  铵态氮添加能显著促进毛竹林土壤N₂O的排放,其效果高于硝态氮,NH₄NO₃作为混合氮,外源性NH₄+-N、NO₃?-N同时输入对土壤N₂O排放的促进作用比单独添加NH₄+-N、NO₃?-N更显著,但并未出现叠加效应。     

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。     

滨海盐渍化土壤中氨氧化微生物丰度和多样性特征

为探究滨海盐土不同盐度梯度下氨氧化微生物的丰度和多样性特征,利用土壤化学和分子生态学手段(定量PCR、T-RFLP)对莱州湾南岸及黄河口4个河口断面(黄河、白浪河、堤河、胶莱河)14个不同盐度(6.4‰～51.1‰)盐渍化土壤样品的氨氧化古菌(AOA)和氨氧化细菌(AOB)的硝化潜势、丰度及多样性进行了分析。结果发现：土壤硝化潜势在高盐度(34.7‰～51.1‰)条件下被显著抑制,主要受土壤盐度、p H和NO₃^–-N水平显著影响;AOA-amoA基因丰度比AOB-amoA高出两个数量级,在中盐度时丰度最高(9.92×10⁶ copies/g土),在高盐度时受到显著抑制(5.28×10⁶ copies/g土,P<0.05);AOB-amoA基因丰度受盐度的影响,低盐度时显著高于中、高盐度条件;然而AOA和AOB的多样性和群落结构受盐度梯度影响不大。相关分析表明,硝化潜势与AOA和AOB丰度均无显著相关性,而与AOA/AOB比值以及AOA的Shannon指数显著负相关。由此可见,滨海盐土中,盐度的...     

长期施肥对土nirS型反硝化细菌的影响及其与N2O排放的关系

土壤反硝化作用是土壤N₂O产生的重要过程,亚硝酸盐还原酶(NIR)催化的亚硝态氮(NO^-₂)还原为一氧化氮(NO)是反硝化作用的关键环节,研究长期施肥对反硝化微生物的影响及其与N₂O排放的关系对于全面理解土壤反硝化过程具有重要意义。基于28年的旱作雨养长期施肥试验,通过常规监测、定量PCR和高通量测序等探讨了长期不同施肥(不施肥CK、偏施肥的单施氮肥N和氮钾配施NK、以及氮磷钾平衡施肥NPK)下■土N₂O排放和nirS反硝化细菌群落特征及两者之间的关系。结果表明:长期化肥施用(N,NK和NPK)均显著提高了N₂O累积排放量,其中平衡施肥(NPK)最高。长期化肥施用对nirS基因丰度和nirS型反硝化细菌的α-多样性无显著影响,但长期平衡施用化肥提高了uncultured_bacterium₂303和Rhodanobacter_sp._D206a的相对丰度,降低了unclass...     

Decreased nitrous oxide emissions associated with functional microbial genes under bio-organic fertilizer application in vegetable fields

Bio-organic fertilizers enriched with plant growth-promoting microbes(PGPMs)have been widely used in crop fields to promote plant growth and maintain soil microbiome functions.However,their potential effects on N₂O emissions are of increasing concern.In this study,an in situ measurement experiment was conducted to investigate the effect of organic fertilizer containing Trichoderma guizhouense(a plant growth-promoting fungus)on soil N₂O emissions from a greenhouse vegetable field.The following four treatments were used:no fertilizer(control),chemical fertilizer(NPK),organic fertilizer derived from cattle manure(O),and organic fertilizer containing T.guizhouense(O+T,referring to bio-organic fertilizer).The abundances of soil N cycling-related functional genes(amoA)from ammonium-oxidizing bacteria(AOB)and archaea(AOA),as well as nirS,nirK,and nosZ,were simultaneously determined using quantitative PCR(qPCR).Compared to the NPK plot,seasonal total N₂O emissions decreased by 11.7%and 18.7%in the O and O+T plots,respectively,which was attributed to lower NH₄⁺-N content and AOB amoA abundance in the O and O+T plots.The nosZ abundance was significantly greater in the O+T plot,whilst the AOB amoA abundance was significantly lower in the O+T plot than in the O plot.Relative to the organic fertilizer,bio-organic fertilizer application tended to decrease N₂O emissions by 7.9%and enhanced vegetable yield,resulting in a significant decrease in yield-scaled N₂O emissions.Overall,the results of this study suggested that,compared to organic and chemical fertilizers,bio-organic fertilizers containing PGPMs could benefit crop yield and mitigate N₂O emissions in vegetable fields.     

生物质炭对土壤N2O消耗的影响及其微生物影响机理

生物质炭在温室气体减排方面具有很大的发展前景,它不仅能实现固碳,对于在大气中停留时间长且增温潜势大的N₂O也能发挥积极作用。本研究采用室内厌氧培养试验,按照生物质炭与土壤质量比（0、1%和5%）加入一定量生物质炭,土壤重量含水率控制在20%。利用Robotized Incubation平台实时检测N₂O和N₂浓度变化,通过测定土壤中反硝化功能基因丰度（nirK、nirS、nosZ）分析生物质炭对N₂O消耗的影响及其微生物方面的影响机理。结果表明：经过20 h厌氧培养后,0生物质炭处理的反硝化功能基因丰度（基因拷贝数·g^-1）分别为6.80×10⁷（nirK）、5.59×10⁸（nirS）和1.22×10⁸（nosZ）。与0生物质炭处理相比,1%生物质炭处理的nirS基因丰度由最初的2.65×10⁸基因拷贝数·g^-1升至7.43×10⁸基因拷贝数·g^-1,nosZ基因丰度则提高了一个数量级,由4.82×10⁷基因拷贝数·g^-1升至1.50×10⁸基因拷贝数·g^-1,然而nirK基因丰度并无明显变化;5%生物质炭处理的反硝化功能基因丰度并未发生显著变化。试验结束时,添加生物质炭处理的N₂/（N₂O+N₂）比值也明显高于0生物质炭处理。相关性分析结果表明,nirS基因丰度和nosZ基因丰度均与N₂O浓度在0.01水平上显著相关。试验末期nirS基因丰度和nosZ基因丰度均随着N₂O浓度的降低而升高。因此在本试验中,添加1%生物质炭可显著提高nirS和nosZ基因型反硝化细菌的丰度,增大N₂/（N₂O+N₂）比值,促进N₂O彻底还原成N₂。生物质炭对于N₂O主要影响机理是增大了可以还原氧化亚氮的细菌活性,促进完全反硝化。     

Simulated animal trampling of a free‐draining stony soil stimulated denitrifier growth and increased nitrous oxide emissions

Winter forage grazing systems in New Zealand cause compaction of soil by grazing animals, especially when the soil is wet. However, there is little information on the effects of animal trampling on denitrifiers in soil, despite their importance for N₂O production. Here, we report a field study of the abundance of the denitrifying genes nirS, nirK, and nosZ and N₂O emissions following the application of dairy cow urine in a free‐draining stony soil. Importantly, we found that simulated animal trampling altered some of the denitrifying microbial communities, thus leading to increased N₂O emissions. Over the 111 day measurement period, the abundance of nitrite (NO₂^?)‐reducing nirS gene copy numbers increased significantly by 87% in the trampled soil with urine (P < 0.01) and increased by 40% in the trampled soil without urine (P < 0.05), but the nirS gene abundance did not change significantly in the nontrampled soil. The abundance of NO₂^? reducing nirK gene copy numbers was not affected by trampling, but increased significantly following urine application. The abundance of N₂O‐reducing nosZ clade I and nosZ clade II gene copy numbers increased significantly in the trampled soil, but did not change significantly in the nontrampled soil. N₂O emissions from the trampled soil were about twice that from the nontrampled soil without urine (1.20 and 0.62 kg N₂O‐N per ha, respectively) and about eight times greater (6.24 kg N₂O‐N per ha) than from nontrampled soil (0.80 kg N₂O‐N per ha) when urine was applied. These results strongly suggest that animal trampling during winter forage grazing can have a major impact on denitrifying communities in soil, which in turn stimulate greater denitrification with increased N₂O emissions.     

Linkage of N2O emissions to the abundance of soil ammonia oxidizers and denitrifiers in purple soil under long-term fertilization

Abstract

Microbial nitrification and denitrification are responsible for the majority of soil nitrous (N₂O) emissions. In this study, N₂O emissions were measured and the abundance of ammonium oxidizers and denitrifiers were quantified in purple soil in a long-term fertilization experiment to explore their relationships. The average N₂O fluxes and abundance of the amoAgene in ammonia-oxidizing bacteria during the observed dry season were highest when treated with mixed nitrogen, phosphorus and potassium fertilizer (NPK) and a single N treatment (N) using NH₄HCO₃as the sole N source; lower values were obtained using organic manure with pig slurry and added NPK at a ratio of 40%:60% (OMNPK),organic manure with pig slurry (OM) and returning crop straw residue plus synthetic NH₄HCO₃fertilizer at a ratio of 15%:85% (SRNPK). The lowest N₂O fluxes were observed in the treatment that used crop straw residue(SR) and in the control with no fertilizer (CK). Soil NH₄⁺provides the substrate for nitrification generating N₂O as a byproduct. The N₂O flux was significantly correlated with the abundance of the amoA gene in ammonia-oxidizing bacteria (r = 0.984, p < 0.001), which was the main driver of nitrification. During the wet season, soil nitrate (NO₃^?) and soil organic matter (SOC) were found positively correlated with N₂O emissions (r = 0.774, p = 0.041 and r = 0.827, p = 0.015, respectively). The nirS gene showed a similar trend with N₂O fluxes. These results show the relationship between the abundance of soil microbes and N₂O emissions and suggest that N₂O emissions during the dry season were due to nitrification, whereas in wet season, denitrification might dominate N₂O emission.     

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.     

Tillage system affects fertilizer-induced nitrous oxide emissions

Since the development of effective N₂O mitigation options is a key challenge for future agricultural practice, we studied the interactive effect of tillage systems on fertilizer-derived N₂O emissions and the abundance of microbial communities involved in N₂O production and reduction. Soil samples from 0–10 cm and 10–20 cm depth of reduced tillage and ploughed plots were incubated with dairy slurry (SL) and manure compost (MC) in comparison with calcium ammonium nitrate (CAN) and an unfertilized control (ZERO) for 42 days. N₂O and CO₂ fluxes, ammonium, nitrate, dissolved organic C, and functional gene abundances (16S rRNA gene, nirK, nirS, nosZ, bacterial and archaeal amoA) were regularly monitored. Averaged across all soil samples, N₂O emissions decreased in the order CAN and SL (CAN?=?748.8?±?206.3, SL?=?489.4?±?107.2 μg kg^?1) followed by MC (284.2?±?67.3 μg kg^?1) and ZERO (29.1?±?5.9 μg kg^?1). Highest cumulative N₂O emissions were found in 10–20 cm of the reduced tilled soil in CAN and SL. N₂O fluxes were assigned to ammonium as source in CAN and SL and correlated positively to bacterial amoA abundances. Additionally, nosZ abundances correlated negatively to N₂O fluxes in the organic fertilizer treatments. Soils showed a gradient in soil organic C, 16S rRNA, nirK, and nosZ with greater amounts in the 0–10 than 10–20 cm layer. Abundances of bacterial and archaeal amoA were higher in reduced tilled soil compared to ploughed soils. The study highlights that tillage system induced biophysicochemical stratification impacts net N₂O emissions within the soil profile according to N and C species added during fertilization.     

Abundance of transcripts of functional gene reflects the inverse relationship between CH<Subscript>4</Subscript> and N<Subscript>2</Subscript>O emissions during mid-season drainage in acidic paddy soil

Agricultural management significantly affects methane (CH₄) and nitrous oxide (N₂O) emissions from paddy fields. However, little is known about the underlying microbiological mechanism. Field experiment was conducted to investigate the effect of the water regime and straw incorporation on CH₄ and N₂O emissions and soil properties. Quantitative PCR was applied to measure the abundance of soil methanogens, methane-oxidising bacteria, nitrifiers, and denitrifiers according to DNA and mRNA expression levels of microbial genes, including mcrA, pmoA, amoA, and nirK/nirS/nosZ. Field trials showed that the CH₄ and N₂O flux rates were negatively correlated with each other, and N₂O emissions were far lower than CH₄ emissions. Drainage and straw incorporation affected functional gene abundance through altered soil environment. The present (DNA-level) gene abundances of amoA, nosZ, and mcrA were higher with straw incorporation than those without straw incorporation, and they were positively correlated with high concentrations of soil exchangeable NH₄⁺ and dissolved organic carbon. The active (mRNA-level) gene abundance of mcrA was lower in the drainage treatment than in continuous flooding, which was negatively correlated with soil redox potential (Eh). The CH₄ flux rate was significantly and positively correlated with active mcrA abundance but negatively correlated with Eh. The N₂O flux rate was significantly and positively correlated with present and active nirS abundance and positively correlated with soil Eh. Thus, we demonstrated that active gene abundance, such as of mcrA for CH₄ and nirS for N₂O, reflects the contradictory relationship between CH₄ and N₂O emissions regulated by soil Eh in acidic paddy soils.     

氮肥水平对稻田细菌群落及N2O排放的影响

作为土壤氮素转化的驱动者,微生物群落结构关系着稻田氮素利用及温室气体N_2O排放等问题。本研究分别基于高通量测序和荧光定量PCR技术,分析了不同氮肥水平[CK(不施氮)、N(施N 180 kg·hm-2)、2/3N(施N 120 kg·hm-2)、1/3N(施N 60 kg·hm-2)]下稻田细菌群落及硝化反硝化关键微生物功能基因丰度的变化。结果显示:氮肥水平提高增加了稻田细菌物种丰富度Chao1指数和群落多样性Shannon指数,改变了细菌群落组成,其中与硝化作用相关的硝化螺菌门Nitrospirae和嗜酸的醋杆菌门Acidobacteria的相对丰度随氮肥水平提高而增加,但甲烷氧化菌Methylosinus的相对丰度随氮肥水平提高而降低。氮肥水平对稻田硝化作用关键微生物氨氧化细菌amo A基因丰度的影响较大,0~5 cm和10~20 cm深度土层中的amo A基因丰度均随氮肥用量增加而提高;反硝化作用关键微生物功能基因nir S、qno B和nos Z的丰度在不施肥处理(CK)中显著低于施肥处理(1/3N、2/3N和N)(P0.05),但1/3N、2/3N和N处理的稻田nir S基因丰度没有明显差异;0~5 cm土层中qno B和nos Z基因丰度存在随氮肥水平提高而增加的趋势,10~20 cm土层中nos Z基因丰度在2/3N和N处理下显著高于1/3N处理(P0.05)。N处理的稻田N_2O排放通量显著高于2/3N及1/3N处理(P0.05),后者又显著高于CK处理(P0.05)。相关分析结果表明稻田N_2O排放通量与0~5 cm土层中硝化螺菌门Nitrospirae相对丰度及10~20 cm土层中amo A基因丰度存在显著相关性(P0.05,n=10)。综上所述,氮肥水平提高增加了稻田细菌群落多样性,促进了稻田N_2O排放,且本研究稻田中硝化作用微生物群落及丰度变化与稻田N_2O排放的关系更为密切。     

The effects of silicon fertilizer on denitrification potential and associated genes abundance in paddy soil

This study evaluated the effect of silicate fertilizer on denitrification and associated gene abundance in a paddy soil. A consecutive trial from 2013 to 2015 was conducted including the following treatments: control (CK), mineral fertilizer (NPK), NPK plus sodium metasilicate (NPK + MSF), and NPK plus slag-based silicate fertilizer (NPK + SSF). Real-time quantitative PCR (qPCR) was used to analyze the abundances of nirS, nirK, and nosZ genes. Potential N₂O emissions and ammonium and nitrate concentrations were related to the nirS and nirK gene abundance. Compared with the NPK treatments, the addition of a Si fertilizer decreased N₂O emission rates and denitrification potential by 32.4–66.6 and 22.0–59.2%, respectively, which were probably related to increased rice productivity, soil Fe availability, and soil N depletion. The abundances of nirS and nirK genes were decreased by 17.7–35.8% and 21.1–43.5% with addition of silicate fertilizers, respectively. Rates of total N₂O and N₂O from denitrification (DeN₂O) emission were positively correlated with the nirS and nirK gene abundance. Nitrate, exchangeable NH₄ ⁺, and Fe concentrations were the main factors regulating the nirS and nirK gene abundance. Silicate fertilization during rice growth may serve as an effective approach to decreasing N₂O emissions.     

Carbon-iron coupling reduced N2O emissions via promoting the conversion to N2 in paddy soils

The influence of redox reactions involving carbon-iron coupling (organic carbon and iron oxides) on nitrous oxide (N₂O) production in paddy soils remains poorly understood. In this study, two microcosm experiments were conducted to investigate the effects of carbon-iron coupling on N₂O emissions, and the underlying mechanisms were verified using quantitative denitrification functional genes (nirS, nirK, nosZI and nosZII) and high-throughput sequencing. The results showed that ferrihydrite (iron) significantly promoted N₂O-N emissions (p < 0.05) after adding ammonium nitrogen, while glucose (carbon) significantly inhibited N₂O-N emissions (p < 0.05). Carbon-iron coupling significantly decreased N₂O-N emissions (p < 0.05) but did not affect soil total nitrogen loss and increased nitrogen (N₂) emissions. After adding high concentrations of acetylene (10% C₂H₂), the N₂O-N emissions from carbon-iron coupling treatment increased significantly from 6.4 to 11.9 mg N kg⁻¹ (p < 0.05), which confirmed that the carbon-iron coupling reduced the N₂O emissions by promoting the conversion of N₂O to N₂. The mechanisms behind carbon-iron coupling promoting complete denitrification and reducing N₂O emissions were attributed to glucose promoting iron reduction and carbon-iron coupling enhancing the abundance of nosZI (42.7%) and nosZII (16.6%).