Chronic effects of different fertilization regimes on nirS-type denitrifier communities across the black soil region of Northeast China

Denitrification is one of the major processes causing nitrogen loss from arable soils.This study aimed to investigate the responses of nir S-type denitrifier communities to different chronic fertilization regimes across the black soil region of Northeast China.Soil samples were collected from sites located in the north(NB),middle(MB),and south(SB)of the black soil region of Northeast China,each with four chronic fertilization regimes:no fertilizer(No F),chemical fertilizer(CF),manure(M),and chemical fertilizer plus manure(CFM).Methods of quantitative polymerase chain reaction(q PCR)and Illumina Mi Seq sequencing were applied to assess the abundance and composition of denitrifier communities by targeting the nir S gene.The results showed that the M and CFM regimes significantly increased the abundances of nir S-type denitrifiers compared with No F at the three locations.The majority of nir S sequences were grouped as unclassified denitrifiers,and the different fertilizers induced little variation in the relative abundance of known nir S-type denitrifier taxa.Over 90%of the sequences were shared among the four fertilization regimes at each location,but none of the abundant operational taxonomic units(OTUs)were shared among the three locations.Principal coordinate analysis(PCo A)revealed that the communities of nir S-type denitrifier were separated into three groups that corresponded with their locations.Although similar fertilization regimes did not induce consistent changes in the nir S-type denitrifier communities,soil p H and NO-3-N content simultaneously and significantly influenced the structure of nir S-type denitrifier communities at the three locations.Our results highlight that geographical separation rather than chronic fertilization was the dominant factor determining the nir S-type denitrifier community structures,and similar chronic fertilization regimes did not induce consistent shifts of nir S-type denitrifier communities in the black soils.     

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.     

Temporal shifts in diversity and quantity of nirS and nirK in a rice paddy field soil

Denitrification is an important part of the nitrogen cycle in the environment, and diverse bacteria, archaea, and fungi are known to have denitrifying ability. Rice paddy field soils have been known to have strong denitrifying activity, but the microbes responsible for denitrification in rice paddy field soils are not well known. Present study analyzed the diversity and quantity of the nitrite reductase genes (nirS and nirK) in a rice paddy field soil, sampled four times in one rice-growing season. Clone library analyses suggested that the denitrifier community composition varied over sampling time. Although many clones were distantly related to the known NirS or NirK, some clones were related to the NirS from Burkholderiales and Rhodocyclales bacteria, and some were related to the NirK from Rhizobiales bacteria. These denitrifiers may play an important role in denitrification in the rice paddy field soil. The quantitative PCR results showed that nirK was more abundant than nirS in all soil samples, but the nirK/nirS ratio decreased after water logging. These results suggest that both diversity and quantity changed over time in the rice paddy field soil, in response to the soil condition.     

华北平原水浇玉米-小麦轮作农田氨挥发与反硝化损失

Ammonia (NH3) volatilization, denitriflcation loss, and nitrous oxide (N2O) emission were investigated from an irrigated wheat-maize rotation field on the North China Plain, and the magnitude of gaseous N loss from denitrification and NH3 volatilization was assessed. The micrometeorological gradient diffusion method in conjunction with a Bowen Ratio system was utilized to measure actual NH3 fluxes over a large area, while the acetylene inhibition technique (intact soil cores) was employed for measurement of denitrification losses and N2O emissions. Ammonia volatilization loss was 26.62% of the applied fertilizer nitrogen (N) under maize, while 0.90% and 15.55% were lost from the wheat field at sowing and topdressing, respectively. The differences in NH3 volatilization between different measurement events may be due to differences between the fertilization methods, and to differences in climatic conditions such as soil temperature. Denitrification losses in the fertilized plots were 0.67%-2.87% and 0.31%-0.49% of the applied fertilizer N under maize and wheat after subtracting those of the controls, respectively. Nitrous oxide emissions in the fertilized plots were approximately 0.08%-0.41% and 0.26%-0.34% of the applied fertilizer N over the maize and wheat seasons after subtracting those of the controls, correspondingly. The fertilizer N losses due to NH3 volatilization were markedly higher than those through denitriflcation and nitrous oxide emissions. These results indicated that NH3 volatilization was an important N transformation in the crop-soil system and was likely to be the major cause of low efficiencies with N fertilizer in the study area. Denitriflcation was not a very important pathway of N fertilizer loss, but did result in important evolution of the greenhouse gas N2O and the effect of N2O emitted from agricultural fields on environment should not be overlooked.     

东北黑土nirS型反硝化细菌群落和网络结构对长期施用化肥的响应

【目的】探明长期施用氮磷钾化肥对东北黑土农田土壤nirS型反硝化细菌群落和网络结构的影响,为更加合理的肥料配施提供理论依据。【方法】基于农业农村部黑龙江耕地保育与农业环境科学观测实验站平台,选取不施肥(NoF)、氮肥(N)、磷肥(P)、钾肥(K)、氮钾肥(NK)、氮磷肥(NP)、磷钾肥(PK)、氮磷钾肥(NPK)8个施肥处理,借助荧光定量PCR、Illumina MiSeq高通量测序和分子生态网络技术,分析东北黑土nirS型反硝化细菌丰度、群落及网络结构,及影响反硝化细菌群落变异的主要环境因子。【结果】1)长期施用氮肥均在显著增加nirS基因拷贝数的同时,降低了nirS型反硝化细菌的群落多样性,而磷、钾肥对其影响并不显著。2)Proteobacteria是所有处理中的优势反硝化细菌门,相对丰度为16.96%~27.34%,且氮肥的施用促进了隶属于该菌门中Bradyrhizobium的生长。3)PCoA分析结果显示,8个施肥处理nirS型反硝化细菌群落主要分成施氮肥和不施氮肥两组,说明长期氮肥的施用显著改变了东北黑土反硝化细菌的群落结构。结合Mantel test的结果可知,土壤pH是影响nirS型反硝化细菌群落改变的主要因素。4)分别构建施氮肥和不施氮肥的反硝化细菌分子生态网络,发现施氮肥和不施氮肥网络结构存在很大差异,长期施用氮肥明显简化了nirS型反硝化细菌的网络结构,同时使其网络结构稳定性降低,易受外界环境扰动。【结论】尽管施用化学氮肥有利于土壤养分的增加,但其土壤nirS型反硝化细菌群落及网络结构发生了较大改变,而磷、钾肥的施用对反硝化细菌群落无显著影响。本试验结果为进一步研究东北黑土区农田土壤反硝化微生物对不同施肥管理的响应机制提供了重要科学依据。     

Effect of straw management, tillage timing and timing of fertilizer nitrogen application on the crop utilization of fertilizer and soil nitrogen in an irrigated cereal rotation

In irrigated grain-growing soils on Canada's prairies, straw management can affect nitrogen (N) fertility and long-term soil organic matter reserves. We conducted a 2-year field experiment in southern Alberta, on a Dark Brown Chernozemic Lethbridge loam (Typic Boroll), to determine the effects of straw removal, tillage, and fertilizer timing on crop uptake of soil and fertilizer N. During the study (1991 and 1992), the crop was oat (Avena sativa L.) and wheat (Triticum aestivum L.), respectively, in an experiment that had been in a wheat-wheat-oat-wheat rotation since 1986. Five straw-tillage treatments were: straw-fall plow, straw-pring plow, no straw-fall plow, no straw-spring plow and no straw-direct seeding. Fertilizer N was applied in fall or spring. Ammonium nitrate (5 at.% ¹⁵N) was added at 100 kg N ha⁻¹ in fall 1990 or spring 1991. For oat (1991), plant N derived from soil was higher under fall plow than under spring plow, higher with tillage than direct seeding, and unaffected by straw removal. The plant N derived from fertilizer was not affected by straw removal in fall plow treatments, but under spring plow, it was higher with straw removal. The plant N derived from fertilizer showed a significant straw-tillage × fertilizer timing interaction; with fall incorporated straw, plant N derived from fertilizer was 44.0 kg N ha⁻¹ for spring-applied, and 30.6 kg N ha⁻¹ for fall-applied N, but in other straw-tillage treatments there was no effect of fertilizer timing. Cumulative fertilizer N recovery (plant + soil) over the 2 years averaged 64.2%, and was unaffected by straw-tillage treatment. Fertilizer N recovery, however, was less with fall-applied N (61.3%) than spring applied N (66.8%). At mid-season, fall plow treatments had higher soil inorganic N and inorganic N derived from fertilizer than spring plow treatments, apparently because of less immobilization. The fall plow treatments also retained higher inorganic N after harvest. Straw removal and fertilizer timing did not influence soil inorganic N and soil inorganic N derived from fertilizer. N removal in straw (16 kg N ha⁻¹ yr⁻¹) could deplete soil N in the long-term. Long-term effects of tillage timing on soil N will depend on the relative amount of N lost by leaching with fall plowing and that lost by denitrification under spring plowing. With direct seeding, crop yield and uptake of soil N was less, and N losses by denitrification could be greater. Application of N in spring, rather than fall, should enhance crop N uptake, reducing N losses and enhancing long-term soil organic N.     

Impacts of different N management regimes on nitrifier and denitrifier communities and N cycling in soil microenvironments

Real-time quantitative PCR assays, targeting part of the ammonia monooxygenase (amoA), nitrous oxide reductase (nosZ), and 16S rRNA genes were coupled with ¹⁵N pool dilution techniques to investigate the effects of long-term agricultural management practices on potential gross N mineralization and nitrification rates, as well as ammonia-oxidizing bacteria (AOB), denitrifier, and total bacterial community sizes within different soil microenvironments. Three soil microenvironments [coarse particulate organic matter (cPOM; >250 μm), microaggregate (53-250 μm), and silt-and-clay fraction (<53 μm)] were physically isolated from soil samples collected across the cropping season from conventional, low-input, and organic maize-tomato systems (Zea mays L.-Lycopersicum esculentum L.). We hypothesized that (i) the higher N inputs and soil N content of the organic system foster larger AOB and denitrifier communities than in the conventional and low-input systems, (ii) differences in potential gross N mineralization and nitrification rates across the systems correspond with AOB and denitrifier abundances, and (iii) amoA, nosZ, and 16S rRNA gene abundances are higher in the microaggregates than in the cPOM and silt-and-clay microenvironments. Despite 13 years of different soil management and greater soil C and N content in the organic compared to the conventional and low-input systems, total bacterial communities within the whole soil were similar in size across the three systems (∼5.15 × 10⁸ copies g⁻¹ soil). However, amoA gene densities were ∼2 times higher in the organic (1.75 × 10⁸ copies g⁻¹ soil) than the other systems at the start of the season and nosZ gene abundances were ∼2 times greater in the conventional (7.65 × 10⁷ copies g⁻¹ soil) than in the other systems by the end of the season. Because organic management did not consistently lead to larger AOB and denitrifier communities than the other two systems, our first hypothesis was not corroborated. Our second hypothesis was also not corroborated because canonical correspondence analyses revealed that AOB and denitrifier abundances were decoupled from potential gross N mineralization and nitrification rates and from inorganic N concentrations. Our third hypothesis was supported by the overall larger nitrifier, denitrifier, and total bacterial communities measured in the soil microaggregates compared to the cPOM and silt-and-clay. These results suggest that the microaggregates are microenvironments that preferentially stabilize C, and concomitantly promote the growth of nitrifier and denitrifier communities, thereby serving as potential hotspots for N₂O losses.     

作物茬口对黑土农田土壤反硝化细菌数量及群落结构的影响

【目的】反硝化作用导致农田土壤氮素损失和温室气体N2O的排放。研究不同作物茬口对土壤反硝化细菌群落结构的影响,旨在揭示作物茬口影响N2O排放的相关机制。【方法】定位试验位于黑龙江省海伦市前进乡光荣村(47°23′N,126°51′E),种植方式包括玉米连作(CC)、大豆连作(SS)以及玉米–大豆轮作,每年一季。取样时,轮作体系玉米已倒茬三次、大豆两次。采集CC、SS以及轮作体系中的大豆茬口(SSC)和玉米茬口(CSC)的表层土壤(0—15 cm)样品,利用实时定量PCR (qPCR)和高通量测序技术,分析土壤中的nirS和nirK型反硝化细菌丰度和群落组成。【结果】在4个作物茬口土壤中,CC处理的反硝化速率最高,玉米–大豆轮作体系中SSC和CSC处理的反硝化速率显著高于SS处理。轮作体系两个茬口SSC和CSC处理的nirS和nirK型反硝化细菌基因丰度多显著高于SS处理,而与CC处理多差异不显著。PCoA结果显示,SSC和CSC处理的nirS型反硝化细菌群落间差异显著,而CC和SS处理的nirK型反硝化细菌群落间存在显著差异。RDA分析结果表明,NO₃~–-N和C...     

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。     

长期施肥对稻田土壤微生物群落结构及氮循环功能微生物数量的影响

  【目的】   稻田是陆生生态系统中重要的氮库之一,在氮素生物地球化学循环中具有重要地位。研究不同施肥处理对稻田土壤微生物群落结构及其功能的影响具有重要意义。   【方法】   田间试验位于江苏省金坛市,在取样时试验已进行了6年。施肥处理包括:不施肥对照 (CK)、施化肥 (CF)、化肥+猪粪混施 (CMF)、化肥+秸秆混施 (CSF)。采用高通量测序和定量PCR方法测定稻田土壤微生物群落结构及氮循环相关功能微生物数量。   【结果】   在施用肥料6年后,土壤全碳、可溶性有机碳、全氮、铵态氮和硝态氮含量均不同程度地提高。与CF相比,CSF和CMF处理土壤pH升高,全碳、可溶性有机碳与养分含量升高。CK与施肥处理的土壤细菌群落结构差异明显,不同施肥处理的细菌群落结构之间有明显差别。聚类结果显示,CK与CMF处理细菌群落聚类更接近,CF处理和CSF处理细菌群落结构更为接近;与CK相比,CF、CMF、CSF处理土壤中氨氧化细菌 (AOB) 和铁氨氧化微生物Feammox A6的丰度显著提高,其中Feammox A6分别增长87.6%、158%和157%。冗余分析结果表明,施肥过程及其对土壤化学性质的改变显著影响土壤细菌群落的组成和分布。   【结论】   施肥导致的反应底物 (NH₄+、NO₃–含量) 及土壤理化性质的差异,是土壤微生物群落结构和功能微生物数量响应的主要决定因素。不施肥与化肥配施猪粪的土壤细菌群落聚类更接近,施化肥与化肥配施秸秆的细菌群落结构更为接近。施肥对氨氧化细菌AOA数量影响不明显,但显著提高氨氧化古菌AOB和厌氧铁氨氧化功能微生物Feammox A6的数量,特别是有机肥 (猪粪、秸秆) 提高Feammox A6数量的效果大于化肥。长期单施化肥土壤中厌氧氨氧化细菌丰度显著降低,反硝化功能基因nirK、nosZ丰度显著增高;化肥配施猪粪土壤中的厌氧氨氧化细菌丰度变化不明显,反硝化功能基因narG、nirK、nosZ丰度显著增高;化肥配施秸秆处理厌氧氨氧化细菌丰度变化不明显,反硝化功能基因nirK、nosZ丰度显著增高。     

Soil residual nitrogen under various crop rotations and cultural practices

Crop rotation and cultural practice may influence soil residual N available for environmental loss due to crop N uptake and N immobilization. We evaluated the effects of stacked vs . alternate‐year crop rotations and cultural practices on soil residual N (NH₄‐N and NO₃‐N contents) at the 0–125 cm depth, annualized crop N uptake, and N balance from 2005 to 2011 in the northern Great Plains, USA. Stacked rotations were durum (Triticum turgidum L.)–durum–canola (Brassica napus L.)–pea (Pisum sativum L.) (DDCP) and durum–durum–flax (Linum usitatissimum L.)–pea (DDFP). Alternate‐year rotations were durum–canola–durum–pea (DCDP) and durum–flax–durum–pea (DFDP). Both of these are legume‐based rotations because they contain legume (pea) in the crop rotation. A continuous durum (CD) was also included for comparison. Cultural practices were traditional (conventional tillage, recommended seeding rate, broadcast N fertilization, and reduced stubble height) and improved (no‐tillage, increased seeding rate, banded N fertilization, and increased stubble height) systems. The amount of N fertilizer applied to each crop in the rotation was adjusted to soil NO₃‐N content to a depth of 60 cm observed in the autumn of the previous year. Compared with other crop rotations, annualized crop biomass N was greater with DCDP and DDCP in 2007 and 2009, but was greater with DDFP than DCDP in 2011. Annualized grain N was greater with DCDP than CD, DFDP, and DDFP and greater in the improved than the traditional practice in 2010 and 2011. Soil NH₄‐N content was greater with CD than other crop rotations in the traditional practice at 0–5 cm, but was greater with DDCP than CD and DDFP in the improved practice at 50–88 cm. Soil NO₃‐N content was greater with CD than other crop rotations at 5–10 cm, but was greater with CD and DFDP than DCDP and DDCP at 10–20, 88–125, and 0–125 cm. Nitrate‐N content at 88–125 and 0–125 cm was also greater in the traditional than the improved practice. Nitrogen balance based on the difference between N inputs and outputs was greater with crop rotations than CD. Increased N fertilization rate increased soil residual N with CD, but legume N fixation increased N balance with crop rotations. Legume‐based crop rotations (all rotations except CD) reduced N input and soil residual N available for environmental loss, especially in the improved practice, by increasing crop N uptake and N immobilization compared with non‐legume monocrop.     

旱地土壤中残留肥料氮的动向及作物有效性

氮素是作物生长最重要的必需元素之一。合理施用氮肥能促进作物生长并提高产量,但是,过多施用氮肥则抑制作物生长并导致大量的肥料氮残留在土壤中,这部分氮素不但会引起土壤养分不平衡,而且为生态环境带来潜在威胁,因此,研究残留氮的动向及作物有效性可为合理施用化肥氮、高效利用土壤残留氮素和减少残留氮素的损失提供依据。应用~(15)N示踪技术,通过4年定位试验,研究了黄土高原南部旱地冬小麦/夏玉米轮作过程中土壤残留肥料氮的变化及作物吸收利用。在冬小麦和夏玉米轮作的第一个周期,为了制造高肥料氮残留背景,于冬小麦播种前向微区施入240 kg hm~(-2)的~(15)N标记氮素;在夏玉米拔节期,为了研究氮肥施入对残留肥料氮的影响,设置0和120 kg hm~(-2)两个氮水平,以普通尿素施入微区。在第2至第4个轮作周期内,为了分析残留肥料氮的动向及其对作物的有效性,微区内不施任何肥料。结果发现,冬小麦播种前施用的~(15)N标记氮肥于收获期在0~200 cm土壤剖面中均有残留,但大部分累积在0~40 cm土层中,累积总量达到200.9 kg hm~(-2),占当季施入量的83.7%。在随后的夏玉米生长季残留的肥料氮迅速减少,之后随生长季的后移缓慢减少,然后保持相对稳定。经过4年的冬小麦/夏玉米轮作,0~300 cm土壤剖面仍残留大量的~(15)N肥料,后季不追施氮肥和追施氮肥处理的残留量分别为47.1 kg hm~(-2)和54.0 kg hm~(-2)。可见,有一部分肥料氮被固定在土壤有机质中。作物对残留氮的回收量逐年减少,且因后季追施氮肥与否而异,4年中作物对肥料氮的总利用率不追施氮肥和追施氮肥处理的分别为46.9%和50.4%,其中在第1个轮作周期中,小麦和玉米的总利用率分别41.6%和42.0%,后3年利用率分别仅有5.3%和8.4%;4年中残留~(15)N的损失率分别达38.1%和29.7%,其损失主要发生在第1个轮作周期的夏玉米生长季节。说明,在旱地土壤上,氮肥的残留是不可避免的,残留肥料氮的有效性较低,只有少量被作物逐年吸收,一部分以有机形态残留在土壤剖面中,另一部分发生了无效损失。后季追施氮肥可促进作物对土壤残留肥料氮的吸收且增加肥料氮在土壤中的保留,减少残留肥料氮的无效损失,但是以自身的大量损失为代价的。     

黄土高原南部夏季不施肥种植玉米对旱地土壤残留养分的利用

夏季降水是造成我国西北黄土高原区旱地土壤硝态氮淋溶的主要原因。通过田间长期定位试验研究了冬小麦收获后,不施肥种植夏玉米而利用土壤残留养分阻止硝态氮淋溶的效应。结果表明,小麦播前施氮量增加,夏玉米收获期生物量和子粒产量增加,但磷肥用量增加对其影响不明显。小麦播前施氮量增加,夏玉米氮磷钾累积增加,施磷量增加,氮钾素累积降低,磷素累积无显著变化。土壤剖面含水量随小麦播前施氮量增加而降低,不同施磷量土壤剖面水分累积量的差异显著减少。不施肥种植夏玉米可以有效阻止和减少土壤剖面硝态氮淋溶,但在小麦播前施氮240和320kg·hm^-2时仍有较明显淋溶,其累积峰逐渐向深层土壤转移,造成氮素损失。施磷时,土壤剖面0-220cm硝态氮累积量下降,220cm以下土层变化不明显。     

闽南农业小流域土壤反硝化作用研究

土壤反硝化是流域土壤氮损失的重要途径之一。利用乙炔抑制培养法对五川流域内表层土壤的反硝化进行测定,研究发现,闽南农业小流域土壤具有较强的反硝化作用强度,在种植季节土壤平均反硝化作用强度为N 0.1 kg/(hm2.d),最高达到N 0.6 kg/(hm2.d),其中蔬菜地反硝化作用强于其他土地利用类型。反硝化作用同土壤的含水量、温度以及NO3-含量都存在有正相关关系,温度是流域土壤反硝化作用的最主要影响因子。五川流域土壤经由反硝化作用氮损失量占流域施肥量的16%,稍高于国内其他地区。     

Denitrification Rate and Controlling Factors for Accumulated Nitrate in the Deep Subsoil of Intensive Farmlands: A Case Study in the North China Plain

Denitrification in subsoil(to a depth of 12 m) is an important mechanism to reduce nitrate(NO_3~-) leaching into groundwater.However, regulating mechanisms of subsoil denitrification, especially those in the deep subsoil beneath the crop root zone, have not been well documented. In this study, soil columns of 0–12 m depth were collected from intensively farmed fields in the North China Plain. The fields had received long-term nitrogen(N) fertilizer inputs at 0(N0), 200(N200) and 600(N600) kg N ha~(-1) year~(-1). Main soil properties related to denitrification, i.e., soil water content, NO_3~-, dissolved organic carbon(DOC), soil organic carbon(SOC),pH, denitrifying enzyme activity(DEA), and anaerobic denitrification rate(ADR), were determined. Statistical comparisons among the treatments were performed. The results showed that NO_3~- was more heavily accumulated in the entire soil profile of the N600 treatment, compared to the N0 and N200 treatments. The SOC, DOC, and ADR decreased with increasing soil depth in all treatments,whereas considerable DEA was observed throughout the subsoil. The long-term fertilizer rates affected ADR only in the upper 4 m soil layers. The ADRs in the N200 and N600 treatments were significantly correlated with DOC. Multiple regression analysis indicated that DOC rather than DEA was the key factor regulating denitrification beneath the root zone. Additional research is required to determine if carbon addition into subsoil can be a promising approach to enhance NO_3~- denitrification in the subsoil and consequently to mitigate groundwater NO_3~- contamination in the intensive farmlands.     

低量施氮对小青菜生长和氮素损失的影响

采用田间试验和微区试验相结合,研究了低量施氮对小青菜(Brassica.chinensis)产量、氮肥利用率和氮素损失的影响,其中氮素总损失用15N示踪法测定,氨挥发用通气密闭室法测定,反硝化损失用乙炔抑制-原状土柱培养法测定,不加乙炔测定N2O排放。结果表明,施用氮肥显著增加了小青菜的产量和吸氮量,在75和150kg/hm2氮肥水平下,氮肥利用率分别为46.8%和39.4%。由于试验地土壤pH低(5.38),各处理的氨挥发均很低且差异不大,施用氮肥没有增加氨挥发。试验地土壤反硝化损失和N2O排放量较高,分别为N4.34kg/hm2/sup和N2.65kg/hm2,施用氮肥没有增加反硝化损失和N2O排放,表明氮源不是反硝化作用的限制因子。在N75和150kg/hm2两个施氮水平下,氮素回收率分别为103%和91.3%,并且土壤残留氮主要累积在020cm土层,表明肥料氮损失很少,这与氨挥发、反硝化损失较低的结果相吻合。     

施肥方式对冬小麦—夏玉米轮作土壤N_2O排放的影响

氧化亚氮(N_2O)是一种重要的农田温室气体,本研究利用紫色土长期施肥试验平台,采用静态箱/气相色谱法对紫色土旱作农田冬小麦—夏玉米轮作系统的N_2O排放进行了定位观测(2012年11月至2013年9月),研究单施氮肥(N)、常规氮磷钾肥(NPK)、猪厩肥(OM)、猪厩肥配施氮磷钾肥(OMNPK)和秸秆还田配施氮磷钾肥(ICRNPK)等施肥方式对紫色土N_2O排放特征的影响;不施肥(NF)作为对照计算排放系数,以探寻紫色土地区可操作性强、环境友好的施肥方式。结果表明,所有施肥方式的N_2O排放均呈现双峰排放,峰值出现在施肥初期;玉米季N_2O排放峰值显著高于小麦季(p0.05)。在相同的施氮水平(小麦季130 kg hm~(~(-2)),玉米季150 kg hm~(~(-2)))下,施肥方式对N_2O排放和作物产量均有显著影响(p0.05)。N、OM、NPK、OMNPK和ICRNPK处理的土壤N_2O周年累积排放量分别为1.93、1.96、1.12、1.50和0.79 kg hm~(~(-2)),排放系数分别为0.62%、0.63%、0.33%、0.47%和0.21%,全年作物产量分别为4.35、11.95、8.39、9.77、10.93 t hm~(~(-2))。施用猪厩肥显著增加N_2O排放量,而秸秆还田在保证作物产量的同时显著降低N_2O排放量,可作为紫色土地区环境友好的施肥方式。土壤无机氮(NO_3~--N和NH_4~+-N)是N_2O排放的主要限制因子。因此,在施氮水平相同时,施肥方式对紫色土活性氮含量的影响导致N_2O排放差异显著,是土壤N_2O排放差异的根本原因。土壤孔隙充水率也是影响N_2O排放的重要环境因子,并且其对N_2O排放的影响存在阈值效应。     

Shifts in size,genetic structure and activity of the soil denitrifier community by nematode grazing

Bacterial-feeding nematodes represent an important driver of the soil microbial activity and diversity. This study aimed at characterizing the impact of nematode grazing on a model functional bacterial guild involved in N-cycling, the denitrifiers. Bacterial-feeding nematodes (Cephalobus pseudoparvus) were inoculated into soil microcosms whose indigenous nematofauna had previously been removed. The size, genetic structure and activity of the soil denitrifier community were characterized 15 and 45 days after nematodes inoculation using quantitative PCR of the nirK, nirS and nosZ denitrification genes, fingerprinting of the nirK and nirS genes and denitrification enzyme activity measurements, respectively. A significant impact of C. pseudoparvus was observed on genetic structure of the nirK community, mainly due to shifts in the relative abundance of the dominant populations, but not on the nirS community. The grazing pressure also tended to decrease the density of all denitrification genes as well as that of 16S rRNA genes. Despite being non-significant, the extent of this decline in gene copy numbers ranged between 60 and 80% of the control microcosm genes densities. Finally, compared to non-inoculated microcosms, denitrification activity significantly decreased by 8% in response to the nematodes inoculation. The herewith data showed that predation by a single species of bacterial-feeding nematode can affect the soil denitrifier community.     

黄土丘陵区土壤—作物系统氮素特征与管理

基于长期定位试验,对黄土丘陵区的晋西北河曲县砖窑沟试验区土壤氮素资源与分布,土壤氮素变化和农田氮素平衡进行了分析,并探讨了该区氮素管理策略和施肥方案。结果表明,试验区土壤全氮含量平均为0.4.g/kg,土壤碱解氮为16.76.mg/kg,氮素水平低,供氮能力差。土壤氮素含量呈现耕地草地林地的分布态势。经过连续13年的施肥处理,施有机肥和有机、无机肥配施020cm土层全氮含量显著提高了12.0%～105.6%,2040cm土层全氮含量变化没有显著差异;各施肥处理020cm土壤碱解氮均呈上升趋势,增加了32%～299%,土壤有效氮的增加与连续施入有机肥的量有关。施肥处理的糜子农田的土壤氮素均呈现出盈余状况,马铃薯田则基本保持平衡。因此,在轮作周期中应根据前茬作物的需氮特性和收成情况确定当季作物的氮肥施用量。糜子和马铃薯的施肥应以氮肥和有机肥为主,氮磷比例分别为1∶0.5～0.7和1∶0.7左右为宜。     

Denitrification activity in the root zone of a sludge-amended desert soil

Summary We evaluated potential NO _inf3 ^sup- losses from organic and inorganic N sources applied to improve the growth of cotton (Gossypium hirsutum) on a Pima clay loam soil (Typic Torrifluvent). An initial set of soil cores (April 1989) was collected to a depth of 270 cm from sites in a cotton field previously amended with anaerobically digested sewage sludge or an inorganic N fertilizer. The denitrification potential was estimated in all soil samples by measuring N₂O with gas chromatography. Soils amended with a low or high rate of sludge showed increased denitrification activity over soil samples amended with a low rate or inorganic N fertilizer. All amended samples showed greater denitrification activity than control soils. The denitrification decreased with soil depth in all treatments, and was only evident as deep as 90 cm in the soils treated with the high sludge rate. However, when soils collected from depths greater than 90 cm were amended with a C substrate, significant denitrification activity occurred. These date imply that organisms capable of denitrification were present in all soil samples, even those at depths far beneath the root zone. Hence, denitrification was C-substrate limited. A second series of soil cores taken later in the growing season (July 1989) confirmed these data. Denitrification losses (under laboratory conditions) to a soil depth of 270 cm represented 1–4% of total soil N depending on treatment, when the activity was C-substrate limited. With additional C substrate, the denitrification losses increased to 15–22% of the total soil N.