转基因水稻秸秆还田对土壤硝化反硝化微生物群落的影响

转基因作物可能通过根系分泌物和植株残体组成的改变及外源基因的转移释放令土壤微生物群落产生变化,影响土壤微生物的生态功能。氨氧化细菌和反硝化细菌是驱动土壤硝化和反硝化过程的关键微生物,其群落结构的变化直接关系土壤氮素的转化与利用。本研究利用荧光定量PCR和PCR-DGGE技术分析了转cry1Ac/cpti双价抗虫基因水稻‘Kf8’秸秆还田降解过程中,土壤氨氧化细菌和反硝化细菌群落丰度与组成的变化,探讨转基因水稻是否存在影响稻田土壤氮素转化与N2O排放的可能。结果显示:无论是氨氧化细菌amo A基因还是反硝化细菌nirS基因,其丰度在转基因水稻‘Kf8’与非转基因水稻‘Mh86’的秸秆还田土壤中都没有显著差异;转基因水稻‘Kf8’和非转基因水稻‘Mh86’秸秆还田降解过程中0~10 cm土层中的amo A基因丰度均显著高于10~20 cm及20~30 cm土层(P0.05);各深度土层中的nirS基因丰度均存在随秸秆还田时间延长而增加的趋势。水稻秸秆还田降解过程中,转基因水稻‘Kf8’的土壤氨氧化细菌和反硝化细菌的群落多样性指数及组成,均与非转基因水稻‘Mh86’没有显著差异。相关分析结果表明土壤氨氧化细菌和反硝化细菌群落组成均与水稻秸秆还田时间存在显著相关性(P=0.002),反硝化细菌群落组成还与土层深度显著相关(P=0.024)。本研究表明转cry1Ac/cpti抗虫基因水稻秸秆还田对稻田土壤硝化和反硝化关键微生物群落不会产生明显影响。就土壤微生物群落而言,转cry1Ac/cpti抗虫基因水稻秸秆还田不存在影响土壤氮素转化与N2O排放的可能。     

双季稻田冬闲期土壤细菌数量及结构对施氮的响应

[目的]研究不同施氮水平对湖南双季稻区冬闲期土壤细菌结构与数量的影响,为双季稻区水稻可持续生产提供理论依据。[方法]依托湖南省双季稻区连续8年的定位试验,选取3个氮肥水平处理:CK(不施氮肥)、N1(施N 100 kg/hm^2)、N2(施N 200 kg/hm^2),取稻田冬闲期5—20 cm耕层土样,采用高通量测序和荧光定量PCR方法测定了土壤细菌数量与结构。[结果]与CK相比,N1、N2处理显著增加了双季稻产量,提升了冬闲期土壤总氮、全碳含量,降低了土壤pH、硝态氮及碳氮比(P<0.05)。N1和N2处理的细菌16s rDNA基因拷贝数分别为CK的48.25和40.31倍。施氮显著增加土壤总细菌丰度,土壤细菌丰度与土壤全氮、碳氮比呈显著正相关,与土壤全碳呈显著负相关。施氮改变了冬闲期稻田土壤细菌的多样性及群落结构。与CK相比,N1处理提高了稻田土壤微生物多样性;N2处理显著增加稻田土壤细菌丰富度,但显著降低稻田土壤细菌多样性。此外,3个处理土壤菌群相对丰度最高的是Proteobacteria(变形菌门),达40.16%~51.16%。N2处理的变形菌门相对丰度显著高于CK与N1,N1处理的变形菌门相对丰度显著低于CK。3个处理属水平细菌相对丰度较高的是Anaerolineaceae_uncultured(厌氧绳菌属)和Nitrospira(硝化螺旋菌属),其相对丰度分别为8.6%~14.56%和8.16%~11.46%。Spearman相关性分析显示,11个优势门菌群数量均与土壤化学性质存在显著相关性。稻田11个优势菌群的数量与土壤化学性质显著相关。[结论]湖南双季稻区施氮降低冬闲期稻田土壤pH和C/N比,低施氮水平可增加稻田微生物多样性,高施氮量虽然增加稻田细菌丰富度,但降低了微生物多样性。     

短期培养下抑制剂烯丙基硫脲对土壤硝化作用及微生物的影响

硝化抑制剂烯丙基硫脲（ATU）对土壤硝化作用及温室效应的影响及机理尚不清楚。本研究采集典型旱地土壤,进行21天室内微宇宙培养,探究了氮肥与不同剂量ATU（分别为氮素用量的1%, 5%, 10%, 15%和20%）配施对土壤硝化作用及N2O和CO2排放通量的影响,并通过实时荧光定量PCR和高通量测序16S rRNA基因技术监测硝化微生物群落变化,同时与传统硝化抑制剂双氰胺（DCD）进行了保氮减排效果的对比。结果表明,与未施加氮肥的对照相比（CK）,单施氮肥（N）显著提高了土壤硝化强度并促进了N2O排放。DCD能显著抑制硝态氮和N2O的积累,抑制效率分别为68.6%和93.3%。而低浓度ATU对土壤硝化作用无影响,仅在高浓度具有抑制效应,且抑制效率最高仅为14.7%。所有ATU处理N2O排放量均显著降低,降幅为60.3～68.2%,仍远高于DCD处理。处理间N2O和CO2的综合温室效应强弱顺序为N>ATU+N>DCD+N≈CK,且不同ATU施用量处理之间差异不显著。相关分析发现氨氧化细菌（AOB）,而不是氨氧化古菌（AOA）和全程氨氧化细菌（Comammox）,与土壤硝态氮积累和N2O排放显著正相关,与土壤pH显著负相关。高通量测序结果表明Nitrosovibrio tenuis类型AOB对氮肥诱导的硝化过程起主导作用。除此之外,ATU和DCD还能显著提高Cupriavidus,并降低Patulibacter、Aeromicrobium、Actinomycetospora、Defluviicoccus和Acidipila等微生物属在群落中的相对丰度。该研究为深化土壤碳氮循环理论,合理使用硝化抑制剂以及减缓温室气体排放提供科学依据。     

氮肥施用对西南地区紫色土冬小麦N_2O释放和反硝化作用的影响

N2O是重要的温室气体之一,由此引起的全球变暖和臭氧层破坏是当今重要的环境问题。采用遮光密闭箱和气相色谱法研究了氮肥施用对小麦地N2O释放和反硝化作用的影响。结果表明,小麦生长季节里,高氮、中氮以及不施氮处理N2O平均排放通量分别为2.71、2.42、1.97 gN.hm-.2d-1;尿素、硫酸铵、硝酸钾3种氮肥品种处理下,平均N2O排放通量分别为2.42、2.14、3.13 gN.hm-2.d-1。小麦生长季节里,高氮、中氮以及不施氮处理平均反硝化速率分别为4.91、4.50、1.67 gN.hm-.2d-1;尿素、硫酸铵、硝酸钾3种氮肥品种处理下,平均反硝化速率分别为4.50、3.68、5.29 gN.hm-.2d-1。氮肥施用明显促进了土壤-植物系统中N2O排放通量和反硝化作用,氮肥施用量水平和N2O排放通量、反硝化作用呈正相关。硝酸钾对N2O排放通量和反硝化作用贡献最大,硫酸铵最小。研究还表明,小麦地N2O释放和反硝化作用与季节有一定相关性,温度较高季节排放量及反硝化作用明显,反之则较弱。     

施肥对夏玉米季紫色土N2O排放及反硝化作用的影响

采用原状土柱-乙炔抑制培养法研究了施肥对紫色土玉米生长季土壤N2O排放通量和反硝化作用的影响.结果表明:玉米季施肥显著增加土壤N2O排放和反硝化损失,同时,各施肥处理间N2O排放与反硝化损失量差异显著.猪厩肥、猪厩肥配施氮磷钾肥、氮肥、氮磷钾肥和秸秆配施氮磷钾肥等处理的土壤N,O排放量分别为3.01、2.86、2.51、2.19和1.88 kg hm-2,分别占当季氮肥施用量的1.63％、1.53％、1.30％、1.09％和0.88％,反硝化损失量分别为6.74、6.11、5.23、4.69和4.12 kg hm-2,分别占当季氮肥施用量的3.97％、3.55％、2.97％、2.61％和2.23％,不施肥土壤的N2O排放量和反硝化损失量仅为0.56和0.78 kg hm-2.施肥是紫色土玉米生长前期(2周内)土壤N2O排放和反硝化速率出现高峰的主要驱动因子,土壤铵态氮和硝态氮含量是影响土壤N2O排放、土壤硝化和反硝化作用的限制因子,土壤含水量是重要影响因子,降雨是主要促发因素.土壤N2O排放量与反硝化损失量的比值介于0.45 ～0.72之间,土壤反硝化损失量极显著高于土壤N2O排放量,说明土壤反硝化作用是紫色土玉米生长季氮肥损失的重要途径.     

氮肥对盐渍化枸杞园土壤微生物群落和氨氧化微生物丰度的影响

研究主要分析氮元素对宁夏平罗盐渍化枸杞园土壤中氨氧化微生物(氨氧化细菌和氨氧化古菌)的影响。实验共设八个处理:(1)C(不施氮肥,不施脱硫废弃物,原始荒地);(2)不施氮肥(N0);(3)施氮肥25 kg hm~(-2)(N25);(4)施氮肥50 kg hm~(-2)(N50);(5)施氮肥100kg hm~(-2)(N100);(6)施氮肥200 kg hm~(-2)(N200);(7)施氮肥400 kg hm~(-2)(N400);(8)施氮肥800 kg hm~(-2)(N800),在N0-N800处理施用脱硫废弃物3.72×104kg hm~(-2)。2011年8月采集0~20 cm土样。结果显示:脱硫废弃物和氮肥配合施加对土壤理化性质产生了显著影响;NO_3~--N和NH_4~+-N含量在施氮处理中相对于原始样地和不施氮处理组都有显著的升高,微生物生物量和细菌和氨氧化细菌多样性指数在施氮400 kg hm~(-2)达到最大值,施氮肥400 kg hm~(-2)是促进微生物量和群落多样性增加的最佳施用量。实时荧光定量PCR结果显示:氨氧化细菌(AOB)丰度在施氮肥400 kg hm~(-2)和800 kg hm~(-2)显著高于其它处理,脱硫废弃物和氮肥配合施用对AOB的丰度具有叠加效应。相关性分析表明:NH_4~+-N与总磷脂脂肪酸(PLFA)、细菌PLFA、革兰氏阳性菌(G+)、革兰氏阴性菌(G-)、真菌/细菌(F/B)、微生物碳(MBC)、微生物氮(MBN)及16S r RNA基因拷贝数、AOB的基因拷贝数都显著相关。因此,铵态氮是该地区微生物群落可利用的有效氮素。     

强酸性茶园土壤中添加不同肥料氮后N2O释放量变化

茶园由于长期偏施氮肥，造成土壤酸化现象严重和 N2O 大量排放。本文对强酸性茶园土壤进行不同氮肥处理试验，结果表明， 通过31 d的好气培养，各施肥处理均显著提高N2O排放， 其中施硝酸钾（KNO3）处理平均每天排放的N2O最高，总排放量为对照（CK）的17倍，其次是硝酸铵（NH4NO3）处理， 尿素［CO（NH2）2］和硫酸铵［（NH4）2SO4］处理虽然能增加N2O 排放，但远远小于硝酸钾处理。对各氮肥处理硝化势的测定表明，尿素、 硫酸铵和硝酸铵处理均明显增加土壤硝化活性，而硝酸钾处理硝化势与对照相比显著降低。强酸性茶园土壤中N2O排放的主要来源是反硝化作用。氧化亚氮还原酶（nosZ）的定量PCR 分析表明，硝酸钾处理的nosZ 基因拷贝数与对照相比显著降低（P0.05）。因此，强酸性土壤中N2O还原酶活性被NO3-抑制是导致高N2O排放的重要原因之一。     

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。      

菜地氮肥用量与N2O排放的关系及硝化抑制剂效果

通过连续种植四季蔬菜近一年的大田试验,探究高施氮水平和低氮肥利用率的蔬菜生产系统中,N2O排放量与氮肥施用量之间的定量关系及其机理,并研究硝化抑制剂减少菜地N2O排放的效果.结果表明,在氮肥施用水平为N 0～1 733 kg hm-2a-1间,无论氮肥中是否添加硝化抑制剂,N2O总排放量与氮肥施用量均呈指数函数关系,即氮肥施用量高时,N2O排放率也高.在各氮肥水平处理下,硝化抑制剂均能降低N2O排放,抑制率为8.75％ ～ 25.28％,且这种减排效果随着施氮量增加而增加.在氮肥施用量为N 300或400 kg hm-2季-1时,施用硝化抑制剂减少N2O排放所带来的效益略高于其成本,因此,即使不考虑氮肥利用率的提高等因素,施用硝化抑制剂仍是一种有利的选择.     

施氮水平对稻-稻-紫云英稻田土壤细菌数量及群落结构的影响

通过对稻-稻-紫云英稻田土壤细菌数量和群落结构多样性进行研究,揭示不同施氮水平对稻田土壤微生物的影响,以期为促进稻田土壤养分管理提供依据。以湖南省8年定位田间试验稻田土壤为对象,试验处理以冬闲水稻田不施氮肥为对照(CK),设置冬种紫云英还田条件下水稻季三个施氮肥水平分别为不施氮肥(0 kg N·hm~(-2),CN_0)、施中水平氮肥(100 kg N·hm~(-2),CN_(100))、施高水平氮肥(200 kg N·hm~(-2),CN_(200))。采用荧光定量PCR和Illumina MiSeq高通量测序平台分别研究了不同处理下稻田土壤的总细菌数量和群落结构。结果表明:稻田土壤总细菌数量为1.25×10~8～8.47×10~9拷贝数·g-1干土;处理间物种多样性指数(Shannon和Simpson)和物种丰富度指数(Ace和Chao)均存在显著性差异;16S rRNA基因在门水平分类下3个主要类群是变形菌门(Proteobacteria)、绿弯菌门(Chloroflexi)和硝化螺旋菌门(Nitrospirae),分别占总OTU比例的42.22%～54.54%、8.56%～20.73%、10.98%～15.08%;CN100土壤样品中变形菌门的相对丰度低于CN_(200)、CN_0和CK,绿弯菌门的相对丰度分别为CN_(200)、CN_0和CK的2.26、1.58倍和1.17倍。相关性分析表明,土壤细菌16S rRNA基因拷贝数与铵态氮含量呈显著正相关。影响土壤细菌菌群结构的因子分析结果表明,土壤细菌与土壤pH值、铵态氮、硝态氮存在密切相关性。研究表明,在冬种紫云英还田条件下施加氮肥可以显著改变湖南省双季稻区土壤微生物数量与结构。     

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-...     

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.     

氮肥对稻田土壤反硝化细菌群落结构和丰度的影响

以氮肥田间定位试验为研究对象,利用PCR-DGGE(聚合酶链反应变性梯度凝胶电泳)和荧光定量PCR(real-time PCR)技术,通过对反硝化细菌nirS基因的检测,分析了定位试验第2年稻田反硝化细菌群落结构和丰度的变化。DGGE图谱及依据其条带位置和亮度数字化数值进行的主成分分析(PCA)结果均显示:在氮肥定位试验第2年,与不施肥对照(CK)比较,在水稻各个生育期(分蘖期、齐穗期和成熟期)内,施用氮肥[150kg(N)·hm-2]的稻田根层土或表土中的反硝化细菌群落结构均无明显变化;且稻田根层土或表土中的反硝化细菌群落结构在水稻各个生育期间也均无明显差异。荧光定量PCR结果显示,在水稻生长发育过程中,施用氮肥的稻田根层土或表土中的反硝化细菌nirS基因拷贝数始终显著(P<0.05)高于其对应的不施肥对照。此外,无论施用氮肥与否,根层土中的反硝化细菌nirS基因拷贝数在水稻成熟期时都会显著(P<0.05)降低;但表土中的nirS基因拷贝数在水稻各生育期间无明显变化;且水稻成熟期时施用氮肥和不施肥的稻田表土中nirS基因拷贝数都显著(P<0.05)高于根层土。同时,与对照比较施用氮肥可促进水稻增产44%。研究表明,短期定位试验中施用氮肥能够显著提高稻田土壤反硝化细菌的丰度,但对其群落结构没有明显影响。     

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.     

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.     

Incongruous variation of denitrifying bacterial communities as soil N level rises in Canadian canola fields

Soil N fertilization stimulates the activity of the soil bacterial species specialized in performing the different steps of the denitrification processes. Different responses of these bacterial denitrifiers to soil N management could alter the efficiency of reduction of the greenhouse gas N₂O into N₂ gas in cultivated fields. We used next generation sequencing to show how raising the soil N fertility of Canadian canola fields differentially modifies the diversity and composition of nitrite reductase (nirK and nirS) and nitrous oxide reductase (nosZ) gene-carrying denitrifying bacterial communities, based on a randomized complete blocks field experiment. Raising soil N levels increased up to 60% the ratio of the nirK to nirS genes, the two nitrite reductase coding genes, in the Brown soil and up to 300% in the Black soil, but this ratio was unaffected in the Dark Brown soil. Raising soil N levels also increased the diversity of the bacteria carrying the nitrite reductase gene nirK (Simpson index, P = 0.0417 and Shannon index, 0.0181), and changed the proportions of the six dominant phyla hosting nirK, nirS, and nosZ gene-carrying bacteria. The level of soil copper (Cu) and the abundance of nirK gene, which codes for a Cu-dependent nitrite reductase, were positively related in the Brown (P = 0.0060, R² = 0.48) and Dark Brown (0.0199, R² = 0.59) soils, but not in the Black soil. The level of total diversity of the denitrifying communities tended to remain constant as N fertilization induced shifts in the composition of these denitrifying communities. Together, our results indicate that higher N fertilizer rate increases the potential risk of nitrous oxide (N₂O) emission from canola fields by promoting the proliferation of the mostly adaptive N₂O-producing over the less adaptive N₂O-reducing bacterial community.     

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%).     

Contrasting response of two grassland soils to N addition and moisture levels: N<Subscript>2</Subscript>O emission and functional gene abundance

Purpose

Nitrification and denitrification processes dominate nitrous oxide (N₂O) emission in grassland ecosystems, but their relative contribution as well as the abiotic factors are still not well understood.

Materials and methods

Two grassland soils from Duolun in Inner Mongolia, China, and Canterbury in New Zealand were used to quantitatively compare N₂O production and the abundance of bacterial and archaeal amoA, denitrifying nirK and nirS genes in response to N additions (0 and 100 μg NH₄ ⁺–N g^?1 dry soil) and two soil moisture levels (40 and 80 % water holding capacity) using microcosms.

Results and discussion

Soil moisture rather than N availability significantly increased the nitrification rate in the Duolun soil but not in the Canterbury soil. Moreover, N addition promoted denitrification enzyme activities in the Canterbury soil but not in the Duolun soil. The abundance of bacterial and archaeal amoA genes significantly increased as soil moisture increased in the Duolun soil, whereas in the Canterbury soil, only the abundance of bacterial amoA gene increased. The increase in N₂O flux induced by N addition was significantly greater in the Duolun soil than in the Canterbury soil, suggesting that nitrification may have a dominant role in N₂O emission for the Duolun soil, while denitrification for the Canterbury soil.

Conclusions

Microbial processes controlling N₂O emission differed in grassland soils, thus providing important baseline data in terms of global change.

     

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.