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。     

三种硝化抑制剂在石灰性土壤中的应用效果比较

在人工气候室内采用25℃黑暗培养法研究双氰胺(DCD)、3,4-二甲基吡唑磷酸(DMPP)及2-氯-6-三氯甲基吡啶(Nitrapyrin)在石灰性土壤中的硝化抑制效果。结果表明:施用DCD、DMPP、Nitrapyrin的土壤NH4+-N含量较单施硫酸铵的土壤(对照)分别提高228.45~244.85 mg/kg(砂土)、209.75~254.79 mg/kg(黏土),NO3--N含量较对照分别降低93.85%~94.99%(砂土)、91.82%~95.38%(黏土)。表观硝化率随培养进程增加缓慢,培养期间只增加了1.28%~2.09%(砂土)、2.72%~8.40%(黏土),而对照增加了86.00%(砂土)、80.89%(黏土)。3种硝化抑制剂均显著抑制了石灰性土壤中硫酸铵水解铵硝化作用的进行,并且在砂土中的硝化抑制率高于黏土,硝化抑制效果最好的为DMPP处理,0.54%Nitrapyrin处理次之但用量最小,0.27%Nitrapyrin和10.8%DCD处理抑制效果相对较弱。     

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。     

硝化抑制剂类型和剂量对不同类型土壤硝化抑制作用机理的研究

氮肥配施硝化抑制剂是提高氮肥利用率、减少活性氮损失和降低环境代价风险的有效措施。为探讨不同硝化抑制剂类型和剂量对不同类型土壤硝化作用的机理,采用室内土壤培养试验,对3种硝化抑制剂[双氰胺(DCD)、3,4-二甲基吡唑磷酸盐(DMPP)和2-氯-6-(三氯甲基)吡啶(NP)]设置不同剂量,研究其对我国不同区域典型土壤(红壤、水稻土、潮土)硝化过程无机氮含量、土壤pH值及土壤表观硝化率变化特征的影响。结果表明,与单施硫酸铵处理相比,3种硝化抑制剂均能抑制水稻土和潮土中铵态氮向硝态氮的转化,且对潮土铵态氮向硝态氮转化抑制效果优于水稻土,而对红壤硝化作用的抑制效果均不明显,3种土壤pH值差异是影响硝化抑制剂作用效果的主因。此外,DCD和NP随着用量的增加,对水稻土和潮土的硝化抑制效果越明显,而DMPP对2种土壤硝化抑制作用无明显的剂量效应。在水稻土中,NP的抑制效果强于DMPP和DCD;在潮土中,DCD的抑制效果优于NP和DMPP,这可能是由于不同硝化抑制剂类型硝化抑制机理性的差异以及其自身特性的差异导致的。综上,针对特定土壤类型筛选适宜的硝化抑制剂类型和用量对农业优化氮肥管理、提高氮肥利用率尤为重要。     

用于黑土的稳定性氯化铵的适宜硝化抑制剂和氮肥增效剂组合

【目的】本文研究添加不同种类硝化抑制剂的高效稳定性氯化铵氮肥在黑土中的施用效果，旨在筛选出适合旱作黑土的高效稳定性氯化铵态氮肥。【方法】在氯化铵中分别添加硝化抑制剂3,4-二甲基吡唑磷酸盐 (DMPP)、双氰胺 (DCD)、2-氯-6-三甲基吡啶 (Nitrapyrin，CP)、氨保护剂 (N-GD) 和1种氮肥增效剂 (HFJ) 及其组合，制成9种稳定性氯化铵氮肥。以不施氮肥 (CK) 和施普通氯化铵 (CK-N) 为对照，以9种稳定性氯化铵为处理进行了等氮量盆栽试验。在玉米苗期、大喇叭口期、灌浆期和成熟期测定了土壤中铵态氮和硝态氮含量，在玉米成熟期测定植株生物量、籽粒产量和氮素含量，计算铵态氮肥的表观硝化率、硝化抑制率、氮肥农学效率、氮肥偏生产力。【结果】1) 与CK-N处理相比，9个处理均显著提高玉米的产量，HFJ的效果均为最显著，可增加玉米籽粒产量3.99倍，提高氮肥吸收利用率4.98倍，显著高于8个硝化抑制剂处理 (P < 0.05)。CP + DMPP和CP + DCD处理提高玉米籽粒产量1.90～2.11倍，两个处理之间无显著差异；CP + DMPP玉米生物量显著高于CP处理，而与DMPP和DCD处理无显著差异；CP + DMPP玉米氮肥吸收利用率显著高于CP和DMPP处理，显著提高3.71倍 (P < 0.05)；2) CP + DMPP和CP + DCD土壤中铵态氮含量提高2.09～2.42倍，且显著高于CP、DMPP和DCD处理 (P < 0.05)，而硝态氮含量和土壤表观硝化率均显著降低24%和66%～68%，与CP和DCD处理存在显著差异 (P < 0.05)；苗期CP + DMPP和CP + DCD硝化抑制率高达23.9%～24.3%，显著高于CP和DCD (P < 0.05)。【结论】在黑土中，氯化铵中添加硝化抑制剂组合的硝化抑制率显著高于添加单一抑制剂，能够有效减缓土壤中铵态氮向硝态氮的转化，减少土壤中氮素损失，降低环境污染。CP + DMPP组合玉米的氮肥吸收利用率显著高于CP + DCD组合。氮肥增效剂HFJ显著增加玉米的氮素吸收量，提高氮肥利用率，从而使玉米获得高产并获得较高的收获指数和经济系数。因此，综合考虑产量和抑制硝化作用等因素，黑土区氯化铵作为玉米生产用氮肥时，建议首选添加氮肥增效剂HFJ来保证作物的高产和氮肥高利用率，也可以添加硝化抑制剂组合CP + DMPP，或者CP + DCD制备稳定性氯化铵来提高氯化铵的增产效果和氮肥利用率，减少氮素损失，降低环境污染。     

稻虾共作模式下稻田土壤氨氧化微生物丰度和群落结构的特征

【目的】研究稻虾共作模式下土壤氨氧化微生物数量、群落多样性及群落结构,深入了解该模式下的土壤微生态环境的演变。【方法】试验点在湖北省荆州市长江大学农学院基地,设置稻虾共作模式 (CR) 与常规中稻种植模式 (MR),借助荧光定量PCR技术与Illumina Miseq高通量测序平台,分析了土壤氨氧化细菌 (AOB) 与古菌 (AOA) 丰度、多样性及群落结构。【结果】与MR模式相比,CR模式显著提高了土壤硝态氮、总碳及总氮含量,对土壤pH、碱解氮及土壤碳氮比无显著影响。CR模式土壤AOA与AOB amoA基因拷贝数为3.13 × 105和7.01 × 105 copies/g干土,MR模式土壤AOA、AOB amoA基因拷贝数为1.41 × 105和3.87 × 105 copies/g干土,两个模式土壤AOB的数量均显著高于AOA,CR模式土壤AOA、AOB的数量均显著高于MR模式 (P < 0.05)。α群落多样性指数表明,相比MR,CR模式显著降低了土壤AOA群落多样性,对AOB群落多样性无显著影响。Venn结果分析,CR模式增加了AOA amoA基因的物种,改变了AOB amoA基因的物种组成,且AOB amoA物种数量下降。在属水平上,norank_c_environmental_samples_p_Thaumarchaeota、unclassified_k_norank_d_Archaea、norank_c_environmental_samples_p_Crenarchaeota、norank_p_environmental_samples_k_norank为AOA的优势类群,相对丰度占AOA amoA基因总序列的99.25%～99.46%,CR模式显著提高了norank_c_environmental_samples_p_Crenarchaeota在AOA群落属水平的相对丰度;unclassified_k_norank_d_Bacteria、norank_f_environmental_samples、norank_o_environmental_samples_c_Betaproteobacteria、unclassified_o_Nitrosomonadales为AOB的优势类群,相对丰度共占97.78%～98.49%,且CR模式显著增加了norank_o_environmental_samples_c_Betaproteobacteria与unclassified_o_Nitrosomonadales在AOB群落属水平的相对丰度。冗余分析 (RDA) 结果显示,土壤基本理化性质对于土壤AOA、AOB群落结构影响有着相似的趋势,其中对AOA、AOB群落结构影响最大的因子是硝态氮,其次分别为总碳、铵态氮、碱解氮、pH。根据RDA投影距离分析,稻虾共作模式对土壤AOA群落结构的影响大于AOB,且MR与稻虾共作模式土壤AOB的群落结构具有一定的相似度。【结论】稻虾共作模式显著降低了AOA群落多样性,而对AOB群落无显著影响;稻虾共作模式显著增加了AOA与AOB的丰度并显著影响了群落结构组成。土壤硝态氮、总碳、铵态氮、碱解氮、pH含量是导致土壤微生物数量、多样性及群落结构变化的主要原因。     

脲酶抑制剂与硝化抑制剂对稻田土壤硝化、反硝化功能菌的影响

[目的]在农业生产中,脲酶抑制剂(urease inhibitor,UI)与硝化抑制剂(nitrification inhibitor,NI)常作为氮肥增效剂来提高肥料利用率。本文研究了在我国南方红壤稻田施用脲酶抑制剂与硝化抑制剂后,土壤中氨氧化细菌(ammonia oxidizing bacteria,AOB)、氨氧化古菌(ammonia-oxidizing archaea,AOA)以及反硝化细菌的丰度以及群落结构的变化特征,旨在揭示抑制剂的作用机理及其对土壤环境的影响。[方法]试验在我国南方红壤稻田进行,共设5个处理:1)不施氮肥(CK);2)尿素(U);3)尿素+脲酶抑制剂(U+UI);4)尿素+硝化抑制剂(U+NI);5)尿素+脲酶抑制剂+硝化抑制剂(U+UI+NI),3次重复。脲酶抑制剂与硝化抑制剂分别为NBPT[N-(n-butyl)thiophosphrictriamide,N-丁基硫代磷酰三胺]和DMPP(3,4-dimethylpyrazole phosphate,3,4-二甲基吡唑磷酸盐)。通过荧光定量PCR(Real-time PCR)研究水稻分蘖期与孕穗期抑制剂对三类微生物标记基因拷贝数的影响,并分析土壤铵态氮、硝态氮与三种菌群丰度的相关性;利用变性梯度凝胶电泳(DenaturingGradient Gel Electrophoresis,DGGE)分析抑制剂对土壤AOB、AOA以及反硝化细菌群落结构的影响,并对优势菌群进行系统发育分析。[结果]1)荧光定量PCR结果表明,施用氮肥对两个时期土壤中AOB的amoA基因与反硝化细菌nirK基因的拷贝数均有显著提高,而对AOA的amoA基因始终没有明显影响;AOB与nirK反硝化细菌的丰度与两个时期的铵态氮含量、分蘖期的硝态氮含量呈极显著正相关,与孕穗期的硝态氮含量相关性不显著;DMPP仅在分蘖期显著减少了AOB的amoA基因拷贝数,表明DMPP主要通过限制AOB的生长来抑制稻田土壤硝化过程;NBPT对三类微生物的丰度无明显影响;2)DGGE图谱表明,在分蘖期与孕穗期,施用氮肥均明显增加了图谱中AOB的条带数,而对AOA却没有明显影响;氮肥明显增加了孕穗期反硝化细菌的条带数;与氮肥的影响相比,抑制剂NBPT与DMPP对AOA、AOB以及反硝化菌的群落结构影响甚微;系统发育分析结果表明,与土壤中AOB的优势菌群序列较为接近的有亚硝化单胞菌和亚硝化螺菌。[结论]在南方红壤稻田中,施入氮肥可显著提高AOB与反硝化细菌的丰度,明显影响两种菌群的群落结构,而AOA较为稳定;NBPT对三类微生物的群落结构丰度无明显影响;硝化抑制剂DMPP可抑制AOB的生长但仅表现在分蘖期,这可能是其缓解硝化反应的主要途径;这也说明二者对土壤生态环境均安全可靠。     

水氮耦合对设施农田土壤自养和异养硝化作用的影响

水氮措施影响设施土壤氮素的转化及硝化微生物活性,但水氮耦合对设施土壤自养和异养硝化作用差异的影响尚不明确。以连续8年设施水氮耦合田间定位试验土壤为研究对象,控制不同土壤田间持水量(WHC)(40%WHC、60%WHC和80% WHC)进行室内微宇宙培养试验,通过添加乙炔抑制剂抑制自养硝化途径,研究水氮耦合对设施土壤自养和异养硝化速率及参与自养硝化的氨氧化微生物的影响,分析氨氧化微生物氨氧化古细菌(AOA)和氨氧化细菌(AOB)对自养硝化作用的贡献。结果表明,水氮耦合下,不同硝化途径NH4+-N、NO^3--N含量以及参与自养硝化的AOA amoA和AOB amoA基因拷贝数均有显著差异。无乙炔培养7 d后,NO^3--N含量显著增加,而NH4+-N含量显著降低,AOA amoA和AOB amoA的基因丰度显著增加。添加乙炔后,NO^3--N、NH4+-N含量基本保持恒定,AOA amoA和AOB amoA基因丰度显著减少。水氮耦合显著影响自养和异养硝化速率,冗余分析(RDA)表明,NH4+-N含量、AOB amoA、NO^3--N-C2H2、AOA amoA可分别解释自养和异养硝化速率变异的68.9%、34.9%、32.8%和24.4%。设施土壤存在自养硝化和异养硝化两种途径,60%～80%WHC各施氮处理均以自养硝化为主,占总硝化速率的65%～86%;仅40%WHC下,氮纯养分量300和525 kg·hm-2处理以异养硝化为主,占总硝化速率的61%～77%。AOB和AOA共同驱动自养硝化,且AOB贡献更大。     

不同硝化抑制剂组合对铵态氮在黑土和褐土中转化的影响

【目的】添加硝化抑制剂和氮肥增效剂是提高氮肥利用率的有效方法。研究不同硝化抑制剂和氮肥增效剂组合对不同性质土壤中铵态氮转化特征的影响,为科学合理选择抑制剂提供理论依据。【方法】供试生化抑制剂包括2-氯-6 (三氯甲基) -吡啶 (Nitrapyrin,CP)、3,4-二甲基吡唑磷酸盐 (DMPP)、1-甲氨甲酰-3-甲基吡唑 (CMP)、3-甲基吡唑 (MP)、2-氨基-4-氯-6-甲基嘧啶 (AM)、N-guard、二氰二胺 (DCD)。供试土壤为黑土和褐土,以氯化铵为氮肥,按照常用量添加各生化抑制剂制备稳定性肥料,用于室内恒温、恒湿土壤培养试验。试验设不施肥 (CK)、氯化铵 (N)、N + CP、N + CP + AM、N + CP + DCD、N + CP + N-guard、N + CP + DMPP、N + CP + CMP、N + CP + MP等9个处理。在培养第1、4、7、11、15、22、30、45、60、75、90、105、120天取土样,测定土壤含水量、土壤NH₄+-N和NO₃–-N含量,并计算硝化抑制率。【结果】在黑土和褐土两种类型土壤中,铵态氮转化特征具有显著差异,在弱酸性黑土中硝化反应速率显著低于碱性褐土。在黑土中,不同硝化抑制剂组合N + CP、N + CP + N-guard、N + CP + DMPP、N + CP + DCD、N + CP + CMP、N + CP + AM、N + CP + MP都表现出较好的硝化抑制效果,可以维持黑土中较高的铵态氮含量超过4个月以上。其中N + CP、N + CP + DCD、N + CP + N-guard处理在120天时,其硝化抑制率为37%～40%。而N + CP + AM、N + CP + MP、N + CP + DMPP为32%～36%,N + CP + CMP为26%。在褐土中,N + CP + DCD组合硝化抑制效果最大,在培养120天,其硝化抑制率为20%;其次是N + CP、N + CP + AM,其硝化抑制率在培养第105天时分别为23%、12%,在培养第90天时分别为63%、60%;N + CP + N-guard、N + CP + DMPP、N + CP + MP、N + CP + CMP在培养第75天时硝化抑制率分别为43%、42%、37%、35%,有效硝化抑制作用时间可维持75天左右。【结论】在黑土和褐土2种不同类型土壤中施用氯化铵氮肥,应添加专一硝化抑制剂或组合制成高效稳定性铵态氮肥。在湿润地区pH较低的酸性土壤上,例如黑土,适宜的硝化抑制剂较多,其中N + CP或N + CP + N-guard、N + CP + DCD组合的硝化抑制效果显著且持续时间长。在干旱半干旱的碱性土壤上,例如褐土,N + CP + DCD组合的硝化抑制效果和持续时间优于其他组合,可用于褐土上施用的高效稳定性氯化铵氮肥的生产。     

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。     

两种农田土壤中的氨氧化细菌和氨氧化古菌对硝化抑制剂DCD和DMPP的响应-盆栽试验

Taking two important agricultural soils with different pH, brown soil （Hap-Udic Luvisol） and cinnamon soil （Hap-Ustic Luvisol）, from Northeast China, a pot culture experiment with spring maize （Zea mays L.） was conducted to study the dynamic changes in the abundance and diversity of soil ammonia-oxidizing bacteria （AOB） and ammonia-oxidizing archaea （AOA） populations during maize growth period in response to the additions of nitrification inhibitors dicyandiamide （DCD） and 3,4-dimethylpyrazole phosphate （DMPP） by the methods of real-time polymerase chain reaction （PCR） assay, PCR-denaturing gradient gel electrophoresis （DGGE）, and construction of clone library targeting the amoA gene. Four treatments were established, i.e., no urea （control）, urea, urea plus DCD, and urea plus DMPP. Both DCD and DMPP inhibited growth of AOB significantly, compared to applying urea alone. Soil bacterial amoA gene copies had a significant positive linear correlation with soil nitrate content, but soil archaeal amoA gene copies did not. In both soils, all AOB sequences fell within Nitrosospira or Nitrosospira-like groups, and all AOA sequences belonged to group 1.1b crenaxchaea. With the application of DCD or DMPP, community composition of AOB and AOA in the two soils had less change except that the AOB community composition in Hap-Udic Luvisol changed at the last two growth stages of maize under the application of DCD. AOB rather than AOA likely dominated soil ammonia oxidation in these two agricultural soils.     

Growth of comammox Nitrospira is inhibited by nitrification inhibitors in agricultural soils

Purpose

The discovery of comammox Nitrospira being capable of complete oxidising ammonia to nitrate radically challenged the conventional concept of two-step nitrification. However, the response of comammox Nitrospira to nitrification inhibitors (NIs) and their role in soil nitrification remain largely unknown, which has hindered our ability to predict the efficiency of NIs in agroecosystems.

Materials and methods

We evaluated the effect of four NIs, 2-chloro-6-(trichloromethyl) pyridine (nitrapyrin), 3,4-dimethylpyrazole phosphate (DMPP), allylthiourea (ATU) and dicyandiamide (DCD) on the growth of comammox Nitrospira, ammonia-oxidising archaea (AOA) and ammonia-oxidising bacteria (AOB) in two pasture and arable soils.

Results and discussion

The amendment of nitrogen fertiliser significantly increased soil nitrate concentrations over time, indicating a sustaining nitrification activity in both soils. The addition of all the four NIs effectively reduced the production of nitrate in both soils, but to varying degrees during incubation. The abundances of comammox Nitrospira clade A were significantly increased by addition of nitrogen fertilisers and significantly impeded by the four NIs in the pasture soil, but their abundances were only remarkably hindered by nitrapyrin in the arable soil. All the four NIs obviously inhibited the AOB abundances in both soils. Except for DMPP, the other three NIs effectively suppressed the AOA abundances in both soils.

Conclusions

We provided new evidence that growth of comammox Nitrospira clade A can be stimulated by nitrogen fertilisers and inhibited by various nitrification inhibitors, suggesting their potential role in nitrification of agricultural soils.

     

亚热带松林中强酸性土壤自养和异养硝化作用的研究

The occurrence of nitrification in some acidic forest soils is still a subject of debate. Identification of main nitrification pathways in acidic forest soils is still largely unknown. Acidic yellow soil (Oxisol) samples were selected to test whether nitrification can occur or not in acidic subtropical pine forest ecosystems. Relative contributions of autotrophs and heterotrophs to nitrification were studied by adding selective nitrification inhibitor nitrapyrin. Soil NH₄⁺-N concentrations decreased, but NO₃^--N concentrations increased significantly for the no-nitrapyrin control during the first week of incubation, indicating that nitrification did occur in the acidic subtropical soil. The calculated net nitrification rate was 0.49 mg N kg^-1 d^-1 for the no-nitrapyrin control during the first week of incubation. Nitrapyrin amendment resulted in a significant reduction of NO₃^--N concentration. Autotrophic nitrification rate averaged 0.28 mg N kg^-1 d^-1 and the heterotrophic nitrification rate was 0.21 mg N kg^-1 d^-1 in the first week. Ammonia-oxidizing bacteria (AOB) abundance increased slightly during incubation, but nitrapyrin amendment significantly decreased AOB amoA gene copy numbers by about 80%. However, the ammonia-oxidizing archaea (AOA) abundance showed significant increases only in the last 2 weeks of incubation and it was also decreased by nitrapyrin amendment. Our results indicated that nitrification did occur in the present acidic subtropical pine forest soil, and autotrophic nitrification was the main nitrification pathway. Both AOA and AOB were the active biotic agents responsible for autotrophic nitrification in the acidic subtropical pine forest soil.     

Nitrapyrin affects the abundance of ammonia oxidizers rather than community structure in a yellow clay paddy soil

Purpose

Yellow clay paddy soil (Oxisols) is a low-yield soil with low nitrogen use efficiency (NUE) in southern China. The nitrification inhibitor nitrapyrin (2-chloro-6- (tricholoromethyl)-pyridine, CP) has been applied to improve NUE and reduce environmental pollution in paddy soil. However, the effects of nitrapyrin combined with nitrogen fertilizers on ammonia oxidizers in yellow clay paddy soil have not been examined.

Materials and methods

A randomized complete block design was set with three treatments: (1) without nitrogen fertilizer (CK), (2) common prilled urea (PU), and (3) prilled urea with nitrapyrin (NPU). Soil samples were collected from three treatments where CK, PU, and NPU had been repeatedly applied over 5 years. Soil samples were analyzed by quantitative PCR and 454 high-throughput pyrosequencing of the amoA gene to investigate the influence of nitrapyrin combined with nitrogen on the abundance and community structure of ammonia oxidizers in yellow clay paddy soil.

Results and discussion

The potential nitrification rate (PNR) of the soil was significantly correlated with the abundances of both ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB). Application of urea significantly stimulated AOA and AOB growth, whereas nitrapyrin exhibited inhibitory effects on AOA. Phylogenetic analysis showed that the most dominant operational taxonomic units (OTUs) of AOA and AOB were affiliated with the Nitrosotalea cluster and Nitrosospira cluster 12, respectively. AOA and AOB community structures were not altered by urea and nitrapyrin application.

Conclusions

Nitrogen fertilization stimulated nitrification and increased the population sizes of AOA and AOB. Nitrapyrin affected the abundance, but not community structure of ammonia oxidizers in yellow clay soil. Our results suggested that nitrapyrin improving NUE and inhibiting PNR was attributable to the inhibition of AOA growth.

     

Can Nitrogen Source and Nitrification Inhibitors Affect In-Season Nitrogen Supply?

ABSTRACT

This study sought to identify whether piggery effluent-derived nitrogen sources can be formulated with urea and nitrification inhibitors to better synchronize nitrogen (N) supply with crop demand than conventional urea fertilizer alone. A 288 pot pasture growth and leaching growth accelerator trial (5 pasture cuts) was completed with a factorial treatment structure of three N sources (2.63 g N [kg soil]^?1 applied as 100% urea-N, 8% struvite-N + 92% urea-N, and 8% piggery pond sludge-N + 92% urea-N), five rates of three nitrification inhibitors (including 3,4-Dimethylpyrazole phosphate, DMPP; limonene+ethanol; and dicyandiamide, DCD), and matrix encapsulated forms of these inhibitors. Applying a combination of piggery sludge with urea increased N uptake during the first 4 weeks of plant growth (by 65%), though total N uptake throughout the trial (22 weeks) did not differ across the N-sources. The microbial community of the soil to which the sludge was added was significantly different from the un-amended soil at the conclusion of the trial. All inhibitor formulations significantly decreased leaching losses of mineral-N relative to the control (by 14 to 61%). The use of DMPP decreased initial nutrient uptake, deferring uptake until later in the experiment. Inhibitor addition resulted in microbial community effects that persisted throughout the trial. The study demonstrated that a piggery-derived N-source and a nitrification inhibitor can be used to manipulate plant N uptake to occur later or earlier in a growing period with equal cumulative uptake, achieving an 11% increase in residual N store, and decreased N leaching losses.     

Responses of ammonia-oxidizing bacteria and archaea to nitrogen fertilization and precipitation increment in a typical temperate steppe in Inner Mongolia

As the first and rate-limiting step of nitrification, ammonia oxidation can be realized either by ammonia-oxidizing bacteria (AOB) or archaea (AOA). However, the key factors driving the abundance, community structure and activity of ammonia oxidizers are still unclear, and the relative importance of AOA and AOB in ammonia oxidation is unresolved. In the present study, we examined the effects of long-term (6 years) nitrogen (N) addition and simulated precipitation increment on the abundance and community composition of AOA and AOB based on a field trial in a typical temperate steppe of northern China. We used combined approaches of quantitative PCR, terminal-restriction fragment length polymorphism (T-RFLP) and clone library analyses of amoA genes. The study objective was to determine (1) AOA and AOB diversity and activity in response to N addition and increased precipitation and (2) the relative contributions of AOA and AOB to soil ammonia oxidation in the typical temperate steppe. The results showed that the potential nitrification rate (PNR) increased with N addition, but decreased with increased precipitation. Both N addition and increased precipitation significantly increased AOB but not AOA abundance, and a significant correlation was only observed between PNR and AOB amoA gene copies. The T-RFLP analysis showed that both N and precipitation were key factors in shaping the composition of AOB, while AOA were only marginally influenced. Phylogenetic analysis indicated that all AOA clones fell within the soil and sediment lineage while all AOB clones fell within the Nitrosospira. The study suggested that AOA and AOB had distinct physiological characteristics and ecological niches. AOB were shown to be more sensitive to N and precipitation than AOA, and the ammonia oxidation process was therefore supposed to be mainly driven by AOB in this temperate steppe.     

The effect of soil pH and dicyandiamide (DCD) on N2O emissions and ammonia oxidiser abundance in a stimulated grazed pasture soil

Purpose

Nitrous oxide (N₂O) is a potent greenhouse gas which is mainly produced from agricultural soils through the processes of nitrification and denitrification. Although denitrification is usually the major process responsible for N₂O emissions, N₂O production from nitrification can increase under some soil conditions. Soil pH can affect N₂O emissions by altering N transformations and microbial communities. Bacterial (AOB) and archaeal (AOA) ammonia oxidisers are important for N₂O production as they carry out the rate-limiting step of the nitrification process.

Material and methods

A field study was conducted to investigate the effect of soil pH changes on N₂O emissions, AOB and AOA community abundance, and the efficacy of a nitrification inhibitor, dicyandiamide (DCD), at reducing N₂O emissions from animal urine applied to soil. The effect of three pH treatments, namely alkaline treatment (CaO/NaOH), acid treatment (HCl) and native (water) and four urine and DCD treatments as control (no urine or DCD), urine-only, DCD-only and urine + DCD were assessed in terms of their effect on N₂O emissions and ammonia oxidiser community growth.

Results and discussion

Results showed that total N₂O emissions were increased when the soil was acidified by the acid treatment. This was probably due to incomplete denitrification caused by the inhibition of the assembly of the N₂O reductase enzyme under acidic conditions. AOB population abundance increased when the pH was increased in the alkaline treatment, particularly when animal urine was applied. In contrast, AOA grew in the acid treatment, once the initial inhibitory effect of the urine had subsided. The addition of DCD decreased total N₂O emissions significantly in the acid treatment and decreased peak N₂O emissions in all pH treatments. DCD also inhibited AOB growth in both the alkaline and native pH treatments and inhibited AOA growth in the acid treatment.

Conclusions

These results show that N₂O emissions increase when soil pH decreases. AOB and AOA prefer different soil pH environments to grow: AOB growth is favoured in an alkaline pH and AOA growth favoured in more acidic soils. DCD was effective in inhibiting AOB and AOA when they were actively growing under the different soil pH conditions.