O3浓度升高对麦田土壤氨氧化细菌、氨氧化古菌和硝化细菌数量的影响

硝化作用在氮循环过程中至关重要,包括氨氧化作用和亚硝酸盐氧化作用,通过氨氧化反应和亚硝酸盐氧化反应将N素转化为植物可利用的NO3-形态。利用开顶式臭氧气室(OTCs,open-top chambers)试验平台,通过大田模拟熏气试验,结合Real-timePCR探讨大气O3浓度升高对麦田土壤氨氧化细菌(AOB)、氨氧化古菌(AOA)及硝化细菌(NOB)数量的影响。结果表明,AOB、AOA和NOB对O3胁迫的反应不一样,AOB基因拷贝数基本上随着O3浓度的升高呈现出降低的趋势,而AOA和NOB基因拷贝数随O3浓度的升高变化不明显。冬小麦拔节期,当O3浓度为40、80、120nmol·mol-1时,20～40cm土层的AOB基因拷贝数分别比对照处理降低39.8%、51.2%和59.4%。AOB和NOB基因拷贝数灌浆期多于收获期,0～10cm土层多于10～20cm。AOA基因拷贝数随季节的变化不大。O3胁迫可通过影响AOB、AOA和NOB的数量和活性来影响土壤的硝化反应,从而影响土壤的氮素循环过程。     

稻田垄作免耕对紫色水稻土硝化微生物的影响

为研究长期垄作免耕对紫色水稻土硝化微生物的影响,以常规平作和冬水平作作为对照处理,采用实时荧光定量PCR对土壤氨氧化细菌(AOB)和氨氧化古菌(AOA)的amoA基因拷贝数进行了测定.结果表明:垄作免耕显著增加土壤有机质和全氮含量.垄作免耕条件下,AOB的amoA基因拷贝数为8.3×105/g干土,比常规平作和冬水平作分别高43%,158%;AOA的amoA基因拷贝数为4.5×107/g干土,比常规平作和冬水平作分别高201%,632%,均具有显著性差异(p0.05),稻田垄作免耕可增加土壤中硝化微生物的数量,对土壤中具有硝化功能的氨氧化细菌和氨氧化古菌具有积极影响.3种耕作方式土壤中AOA amoA基因拷贝数均比AOB amoA基因拷贝数高1~2个数量级,AOA可能在其中起重要作用.     

长期施氮对酸性紫色土氨氧化微生物群落及其硝化作用的影响

目的研究长期施氮对酸性紫色土壤中氨氧化古菌（AOA）和氨氧化细菌（AOB）群落特征的影响,揭示氨氧化微生物群落的驱动因子及其调控硝化作用的微生物学机制。方法依托四川雅安玉米体系施氮长期定位试验（始于2010年）,试验处理包括5个供氮水平,即0（N0）、90（N90）、180（N180）、270（N270）和360（N360）kg N·hm^-2,通过Illumina Miseq高通量测序技术测定AOA和AOB的群落,探究长期施氮对氨氧化微生物群落介导的硝化作用的影响。结果 长期施氮影响AOA和AOB的α-多样性（包括丰富度指数和香农-威纳指数）、群落结构和群落组成。其中,随着施氮量的增加,AOA 丰富度指数无显著变化,香农-威纳指数显著降低,AOB 丰富度指数和香农-威纳指数均显著增加;长期不同供氮水平显著影响AOA和AOB的群落结构,供氮水平的增加显著降低AOB优势类群中 Nitrosospira Cluster 3a.1的相对丰度（P<0.05）,同时显著增加了Cluster 3a.2、Cluster 9和Cluster 1的相对丰度（P<0.05）,而对AOA优势类群无显著影响。土壤pH、全氮（TN）、有机质（SOM）、NH₄⁺-N和NO₃^--N均显著影响AOA和AOB的α-多样性,其中,pH与AOB 丰富度指数和香农-威纳指数呈显著负相关（P<0.01）,而与AOA 香农-威纳指数呈显著正相关,而TN、SOM、NH₄⁺-N和NO₃^--N与AOB 丰富度指数和香农-威纳指数呈显著正相关,与AOA 香农-威纳指数呈显著负相关。同时,pH、TN、NO₃^--N 、SOM 和NH₄⁺-N显著影响AOA和AOB的群落结构（P<0.05）。结构方程模型（SEM）的结果表明,长期施氮通过降低土壤pH、提高TN和NO₃^--N含量、改变AOA和AOB的α-多样性和群落结构,进而提高了土壤的硝化势。结论 长期施氮通过改变酸性紫色土壤pH、TN、NH₄⁺-N和NO₃^--N和氨氧化微生物的α-多样性和群落结构进而影响了硝化势。     

生物炭与有机肥对菜田土壤氨氧化菌丰度的影响

通过土壤培育试验,运用定量PCR技术,研究了添加生物炭和有机肥对土壤氮营养及氨氧化古菌(AOA)和氨氧化细菌(AOB)丰度的影响。结果表明,土壤培育时间、有机肥显著增加硝态氮的含量(P0.001),促进硝化作用;生物炭通过抑制有机氮矿化作用降低硝化作用,106d时生物炭处理的硝态氮、可溶性总氮含量低于没有生物炭的处理(P0.05),并且中和了有机肥对矿化作用的促进效应。随着处理时间的延长,生物炭促进氨氧化菌群的丰度,AOA丰度的提高较AOB显著,且AOA丰度的变化与硝态氮含量的变化呈正相关。因此认为AOA是土壤中氨氧化作用的主要驱动力,且生物炭与有机肥主要对AOA的丰度产生影响。     

长期施用尿素对东北黑土中氨氧化古菌群落的影响

【目的】表征34年施肥条件下东北黑土中氨氧化古菌（AOA）群落特征,明确不同尿素水平对其丰度和群落结构的影响,并与土壤化学性质进行关联分析,探讨引起影响东北黑土中AOA群落变化的主效环境因子,为进一步揭示黑土土壤硝化作用过程和机制,以及改良施肥方式提供依据。【方法】依托黑龙江省农业科学院34年长期定位试验,选取3组不同施肥处理：无尿素组（对照CK和磷钾处理PK）、一倍尿素组（单施一倍氮肥处理N₁,氮磷处理NP,氮磷钾处理NPK和氮钾处理NK）和二倍尿素组（单施二倍氮肥处理N₂）共7个处理的耕层土壤为研究对象,借助454高通量测序和Real-time PCR技术,以AOA Arch-amoA基因为分子标靶,解析不同尿素水平对黑土中AOA群落组成和丰度的影响,并对AOA群落结构与环境因子进行相关性分析。【结果】随着尿素施用量的增加,黑土中AOA 的Arch-amoA拷贝数由2.64×10⁷/g土壤显著降低至8.34×10⁵/g土壤,且黑土pH是引起其数量降低的直接原因;OTU水平上的聚类和非度量多维度分析（NMDS）结果均表明,相同尿素水平的各处理AOA群落结构相似,不同尿素水平处理间的AOA群落差异显著;冗余分析表明,土壤pH、水溶性有机碳和硝态氮浓度是影响土壤AOA群落变化的主效环境因子（P＜0.05）;系统发育分析表明,东北黑土中AOA主要有Nitrososphaera和Nitrosotalea两个属：无尿素组中99.3%和一倍尿素组中90.1%的Arch-amoA序列归类于Nitrososphaera,二倍尿素组中67.9%的Arch-amoA序列归类于Nitrosotalea。【结论】本研究表征了长期不同施肥下黑土中的AOA群落特征,通过与土壤化学性质耦合分析,确定了引起黑土中AOA群落变化的主要因素。长期施用氮肥的黑土中,AOA Arch-amoA拷贝数和群落结构受尿素施用量的影响显著,低倍尿素提高其多样性,而高倍尿素则使之降低,土壤pH、有机碳和硝态氮浓度是导致AOA群落差异主要因素。     

长期咸水滴灌对土壤氨氧化微生物丰度和群落结构的影响

为探讨长期咸水滴灌对棉田土壤氨氧化细菌（AOA）和氨氧化古菌（AOB）的丰度和群落结构的影响,于2018年采集已经过10年咸水滴灌的棉田土壤,通过实时荧光定量PCR和高通量测序技术测定土壤AOA和AOB的丰度和群落结构。试验设3个灌溉水盐度水平：0.35、4.61 dS·m^-1和8.04 dS·m^-1（分别代表淡水、微咸水和咸水）。结果表明：微咸水、咸水灌溉显著降低土壤NO₃^--N含量和潜在硝化势（PNR）,但显著增加土壤盐分和NH₄⁺-N含量。不同处理AOA和AOB的amoA基因拷贝数分别为2.2×10⁶~3.6×10⁶copies·g^-1和1.9×10⁵~3.2×10⁵ copies·g^-1干土;微咸水、咸水处理AOA和AOB amoA基因拷贝数均显著低于淡水处理,且微咸水处理显著降低AOA/AOB。PNR与AOA丰度（P<0.001）和AOB丰度（P<0.001）均呈显著正相关关系。此外,不同灌溉水盐度下AOA群落操作分类单元（OTUs）的数量大于AOB,微咸水、咸水灌溉显著降低AOB群落的OTUs。与淡水处理相比,咸水、微咸水处理显著增加AOA群落的香农指数,咸水处理显著降低AOB群落的香农指数。AOA和AOB群落的优势类群分别为Candidatus Nitrosocaldus和Nitrosospira;咸水、微咸水处理抑制AOA群落的Betaproteobacteria生长,而咸水处理中Candidatus Nitrosocaldus显著高于淡水和微咸水处理。AOB群落中Nitrosomonas的相对丰度随着灌溉水盐度的增加而显著降低。LEf Se分析显示,AOA在咸水灌溉下仅有1个差异物种,而AOB在微咸水灌溉时有5个差异物种。冗余分析结果显示：AOA群落结构的改变与土壤NO₃^--N、pH和盐度的变化密切相关,而AOB群落结构的改变仅与NO₃^--N和pH显著相关。盐分是影响氨氧化微生物生长及群落结构的主导因素,AOA和AOB共同参与土壤硝化作用,淡水、微咸水灌溉条件下AOB可能是硝化作用主导微生物种群,而咸水灌溉条件下AOA可能是主导微生物种群。     

不同类型稻田中全程氨氧化微生物的分异特征

选用微碱性氮贫瘠的上海市崇明岛稻田和微酸性氮丰富的南京市稻田剖面(0～50 cm),比较研究稻田土壤中氨氧化微生物类群丰度的差异及其环境驱动机制,评价其与氨氧化潜力的内在关系。结果表明,崇明稻田的净硝化速率为12.82～22.30 mg/(kg·d),明显高于南京稻田[4.26～7.46 mg/(kg·d)]。崇明稻田土壤剖面的Comammox amoA基因总拷贝数(Clade A与Clade B之和)均值为1 g 8.8×10~6拷贝,是南京稻田的2.4倍,且Clade A与Clade B的比值范围为2.5～12.7,证实了Comammox存在于2种不同类型的稻田土壤中。崇明稻田和南京稻田剖面的氨氧化细菌(AOB)的amoA基因拷贝数均值分别为1 g 3.75×10~8拷贝和1.23×10~8拷贝,氨氧化古菌(AOA)的amoA基因拷贝数均值分别为1 g 2.05×10~7拷贝和0.35×10~7拷贝,这2种菌群基因拷贝数均在10.1～20.0 cm土层达到最高。回归分析发现,2个稻田中氨氧化细菌(AOB)对氨氧化潜力的贡献率达到90%～94%,而Comammox仅为3%左右,表明氨氧化细菌(AOB)在氨氧化过程中发挥主要作用。     

东北沼泽湿地土壤中氨氧化微生物活性和丰度研究

为了研究东北典型沼泽湿地——三江湿地和扎龙湿地表层土壤中氨氧化微生物的活性和丰度,采用氯酸盐抑制亚硝酸盐氧化菌(Nitrite-oxidizing bacteria,NOB)和氨苄青霉素抑制氨氧化细菌(Ammonia-oxidizing bacteria,AOB)以及荧光定量PCR的方法,分析了生长季和非生长季氨氧化微生物的活性和丰度的关系。结果表明:在不同类型的湿地土壤中,pH值显著地影响湿地土壤中氨氧化古菌(Ammonia-oxidizing archaea,AOA)的潜在氨氧化活性(Potential ammonia oxidation activity,PAO)(P0.01)。AOA的丰度值与PAO_(AOA+AOB)呈显著正相关(r=0.96,P0.05),而与PAO_(AOA)不存在相关性(P0.05)。三江湿地的毛苔草和小叶章土壤中的PAO_(AOA+AOB)、PAO_(AOA)、AOA丰度与AOB丰度,并无显著差异。研究表明植被的差异并没有影响土壤中氨氧化微生物的潜在氨氧化活性以及氨氧化微生物的丰度,但在抑制AOB活性之后毛苔草土壤中的PAO出现了显著的下降(P0.05),表明AOB在毛苔草土壤中的氨氧化过程中发挥着重要作用。     

模拟水位下降与刈割对高寒湿地土壤氨氧化与反硝化微生物的影响

湿地土壤是温室气体重要的源和汇,认识湿地生态系统氮循环过程有助于预测氮循环对未来气候变化的响应与反馈机制。为探讨硝化作用和反硝化作用对土壤水位变化和刈割的响应机制,依托于2013年在青藏高原东部若尔盖泥炭地南部湿地设置的野外实验,通过在样地周围挖掘不同深度的排水沟模拟水位下降,结合刈割处理,研究水位下降和刈割对泥炭地土壤氨氧化古菌(Ammonia-oxidizing archaea,AOA)、氨氧化细菌(Ammonia-oxidizing bacteria,AOB)和反硝化细菌(Denitrifying bacteria)丰度的影响。2014年7月取样分析结果表明:水位下降显著降低土壤含水量,水位下降与刈割均显著降低土壤呼吸;氨氧化及反硝化微生物功能基因丰度在各处理间无显著差异,但刈割及其与水位下降的交互作用显著影响AOA-amo A与AOB-amo A基因丰度比。刈割处理显著增加AOB-amo A基因相对丰度,但对AOA-amo A基因丰度无显著影响,揭示AOB可能在湿地土壤硝化过程中占主导地位。土壤nir S基因丰度显著高于nir K基因,表明nir S基因对水位下降及刈割的响应更为敏感。随着土壤水位的下降,刈割促进了由AOB主导的氨氧化过程,而反硝化微生物丰度的增加削减了氨氧化产物硝酸盐的积累,继而降低了土壤硝酸盐含量。     

橘园绿肥种植对土壤氨氧化微生物的影响

本研究以当阳凤凰山长期定位施肥柑橘试验基地为平台,进行了自然生草（NG）、人工种植光叶苕子（LP）以及光叶苕子混播于自然生草（NL）等三种绿肥处理,通过植物群落调查、土壤理化性质测定,以及 qPCR和 Illumina Miseq高通量测序的方法,阐明橘园土壤氨氧化微生物AOA和AOB的功能基因丰度和群落结构对不同绿肥种植处理的响应特征,并探讨植物群落和土壤理化因子对它们的影响。研究结果表明：相比于NG,人工种植绿肥（LP和NL）对AOA和AOB数量的影响不一,LP降低了AOA和AOB的 amoA 基因拷贝数,NL 降低了 AOA-amoA 但增加了 AOB-amoA 基因拷贝数;LP 和 NL 中 AOA 的 Ace 指数和 Chao指数显著高于NG,LP中AOB的Shannon指数显著高于NG,三个处理的β多样性差异显著;三种绿肥种植处理下AOA共获得119个OTU,优势门和属分别是泉古菌门和 norank—Crenarchaeota,其中 norank—Crenarchaeota 在 NG 中的相对丰度显著高于 LP 和 NL;AOB 共获得142 个 OTU,优势门和属分别是变形菌门和亚硝化螺菌属,其中亚硝化螺菌属的相对丰度在不同处理中有显著差异,其 LP 中最高,为45.87%;Mantel检验和冗余-层次分割分析结果表明,氨氧化微生物的丰度、多样性和组成受到绿肥种植后的植物因子（如群落高度、盖度、多样性、地上生物量和地下生物量、根系碳氮含量等）和土壤因子（如pH、土壤含水量、总碳、总氮、铵态氮、硝态氮和速效磷含量等）的共同影响。该研究说明在橘园种植绿肥可改变地上植物群落特征和土壤理化性质,从而使土壤中氨氧化微生物的数量和群落结构发生变化,其中光叶苕子混播于自然生草的生草模式可明显增加土壤氨氧化微生物量,促进土壤的硝化作用和改善果园生态环境,因此,实际应用中可优先考虑此模式。     

Changes in the activities of key enzymes and the abundance of functional genes involved in nitrogen transformation in rice rhizosphere soil under different aerated conditions

Soil microorganisms play important roles in nitrogen transformation.  The aim of this study was to characterize changes in the activity of nitrogen transformation enzymes and the abundance of nitrogen function genes in rhizosphere soil aerated using three different methods (continuous flooding (CF), continuous flooding and aeration (CFA), and alternate wetting and drying (AWD)).  The abundances of amoA ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB), nirS, nirK, and nifH genes, and the activities of urease, protease, ammonia oxidase, nitrate reductase, and nitrite reductase were measured at the tillering (S1), heading (S2), and ripening (S3) stages.  We analyzed the relationships of the aforementioned microbial activity indices, in addition to soil microbial biomass carbon (MBC) and soil microbial biomass nitrogen (MBN), with the concentration of soil nitrate and ammonium nitrogen.  The abundance of nitrogen function genes and the activities of nitrogen invertase in rice rhizosphere soil were higher at S2 compared with S1 and S3 in all treatments.  AWD and CFA increased the abundance of amoA and nifH genes, and the activities of urease, protease, and ammonia oxidase, and decreased the abundance of nirS and nirK genes and the activities of nitrate reductase and nitrite reductase, with the effect of AWD being particularly strong.  During the entire growth period, the mean abundances of the AOA amoA, AOB amoA, and nifH genes were 2.9, 5.8, and 3.0 higher in the AWD treatment than in the CF treatment, respectively, and the activities of urease, protease, and ammonia oxidase were 1.1, 0.5, and 0.7 higher in the AWD treatment than in the CF treatment, respectively.  The abundances of the nirS and nirK genes, and the activities of nitrate reductase and nitrite reductase were 73.6, 84.8, 10.3 and 36.5% lower in the AWD treatment than in the CF treatment, respectively.  The abundances of the AOA amoA, AOB amoA, and nifH genes were significantly and positively correlated with the activities of urease, protease, and ammonia oxidase, and the abundances of the nirS and nirK genes were significantly positively correlated with the activities of nitrate reductase.  All the above indicators were positively correlated with soil MBC and MBN.  In sum, microbial activity related to nitrogen transformation in rice rhizosphere soil was highest at S2.  Aeration can effectively increase the activity of most nitrogen-converting microorganisms and MBN, and thus promote soil nitrogen transformation.      

雷竹林土壤氨氧化微生物对不同肥料的响应

雷竹Phyllostachys violascens林冬季地表覆盖施肥为核心的集约经营技术被广泛推广应用,冬季覆盖以及施肥处理必将对土壤氨氧化微生物群落结构产生影响。为揭示雷竹林覆盖和施肥等集约经营措施对土壤氨氧化微生物的影响,进行了田间试验。设计如下2个试验:试验1为施用等量硝态氮复合肥:m(氮)∶m(五氧化二磷)∶m(氧化钾)=16∶16∶16的对照1(不覆盖)和处理1(覆盖);试验2为覆盖的4个不同肥料处理,分别为对照2(不施肥),处理1,处理2(尿素),处理3(尿素+氯化钾),各处理以含氮量360 kg·hm-2为标准折算成相应的肥料用量,氯化钾与复合肥等钾量折算。应用聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)技术和定量PCR技术分析土壤氨氧化细菌(AOB)和氨氧化古菌(AOA)群落结构多样性和功能基因丰富度。结果表明:不同处理土壤氨氧化细菌和氨氧化古菌有较高比例的共性种类,群落结构尚未发生显著改变;前期未覆盖的对照1与其他前期曾覆盖的所有处理的氨氧化细菌群落结构差异相对较明显,同为前期覆盖的处理3与其他处理又稍有不同;土壤氨氧化细菌多样性为处理1明显高于(P<0.05)处理3;处理3氨氧化细菌和氨氧化古菌的基因的丰度均较高(P<0.05)。综合比较不同处理氨氧化细菌和氨氧化古菌的DGGE条带的多样性以及定量分析得出结论,处理3的氨氧化微生物多样性和功能基因最丰富,处理2最差。建议生产上采用尿素与氯化钾搭配施用,有利于土壤氨氧化微生物的活动和氮素循环利用。     

Abundance and Community Composition of Ammonia-Oxidizers in Paddy Soil at Different Nitrogen Fertilizer Rates

Ammonia oxidation, the first and rate-limiting step of nitrification, is carried out by both ammonia-oxidizing bacteria （AOB） and ammonia-oxidizing archaea （AOA）. However, the relative importance of AOB and AOA to nitrification in terrestrial ecosystems is not well understood. The aim of this study was to investigate the effect of the nitrogen input amount on abundance and community composition of AOB and AOA in red paddy soil. Soil samples of 10-20 cm （root layer soil） and 0-5 cm （surface soil） depths were taken from a red paddy. Rice in the paddy was fertilized with different rates of N as urea of N1 （75 kg N ha＂ yr-1）, N2 （150 kg N ha~ yrl）, N3 （225 kg N ha1 yrl） and CK （without fertilizers） in 2009, 2010 and 2011. Abundance and community composition of ammonia oxidizers was analyzed by real-time PCR and denaturing gradient gel electrophoresis （DGGE） based on amoA （the unit A of ammonia monooxygenase） gene. Archaeal amoA copies in N3 and N2 were significantly （P〈0.05） higher than those in CK and N1 in root layer soil or in surface soil under tillering and heading stages of rice, while the enhancement in bacterial amoA gene copies with increasing of N fertilizer rates only took on in root layer soil. N availability and soil NO3--N content increased but soil NH4＋-N content didn＇t change with increasing of N fertilizer rates. Otherwise, the copy numbers of archaeal amoA gene were higher （P〈0.05） than those of bacterial amoA gene in root lary soil or in surface soil. Redundancy discriminate analysis based on DGGE bands showed that there were no obvious differs in composition of AOA or AOB communities in the field among different N fertilizer rates. Results of this study suggested that the abundance of ammonia-oxidizers had active response to N fertilizer rates and the response of AOA was more obvious than that of AOB. Similarity in the community composition of AOA or AOB among different N fertilizer rates indicate that the community composition of ammonia-oxidizers was relatively stable in the paddy soil at least in short term for three years.     

Reduction of N2O emissions by DMPP depends on the interactions of nitrogen sources (digestate vs. urea) with soil properties

The inhibition of nitrification by mixing nitrification inhibitors (NI) with fertilizers is emerging as an effective method to reduce fertilizer-induced nitrous oxide (N₂O) emissions.  The additive 3,4-dimethylpyrazole phosphate (DMPP) apparently inhibits ammonia oxidizing bacteria (AOB) more than ammonia oxidizing archaea (AOA), which dominate the nitrification in alkaline and acid soil, respectively.  However, the efficacy of DMPP in terms of nitrogen sources interacting with soil properties remains unclear.  We therefore conducted a microcosm experiment using three typical Chinese agricultural soils with contrasting pH values (fluvo-aquic soil, black soil and red soil), which were fertilized with either digestate or urea in conjunction with a range of DMPP concentrations.  In the alkaline fluvo-aquic soil, fertilization with either urea or digestate induced a peak in N₂O emission (60 μg N kg^–1 d^–1) coinciding with the rapid nitrification within 3 d following fertilization.  DMPP almost eliminated this peak in N₂O emission, reducing it by nearly 90%, despite the fact that the nitrification rate was only reduced by 50%.  In the acid black soil, only the digestate induced an N₂O emission that increased gradually, reaching its maximum (20 μg N kg^–1 d^–1) after 5–7 d.  The nitrification rate and N₂O emission were both marginally reduced by DMPP in the black soil, and the N₂O yield (N₂O-N per NO₂^–+NO₃^–-N produced) was exceptionally high at 3.5%, suggesting that the digestate induced heterotrophic denitrification.  In the acid red soil, the N₂O emission spiked in the digestate and urea treatments at 50 and 10 μg N kg^–1 d^–1, respectively, and DMPP reduced the rates substantially by nearly 70%.  Compared with 0.5% DMPP, the higher concentrations of DMPP (1.0 to 1.5%) did not exert a significantly (P<0.05) better inhibition effect on the N₂O emissions in these soils (either with digestate or urea).  This study highlights the importance of matching the nitrogen sources, soil properties and NIs to achieve a high efficiency of N₂O emission reduction.     

Impact of Long-Term Fertilization on Community Structure of Ammonia Oxidizing and Denitrifying Bacteria Based on amoA and nirK Genes in a Rice Paddy from Tai Lake Region,China

Ammonia oxidizing （AOB） and denitrifying bacteria （DNB） play an important role in soil nitrogen transformation in natural and agricultural ecosystems. Effects of long-term fertilization on abundance and community composition of AOB and DNB were studied with targeting ammonia monooxygenase （amoA） and nitrite reductase （nirK） genes using polymerase chain reaction- denaturing gradient gel electrophoresis （PCR-DGGE） and real-time PCR, respectively. A field trial with different fertilization treatments in a rice paddy from Tai Lake region, centre East China was used in this study, including no fertilizer application （NF）, balanced chemical fertilizers （CF）, combined organic/inorganic fertilizer of balanced chemical fertilizers plus pig manure （CFM）, and plus rice straw return （CFS）. The abundances and riehnesses of amoA and nirK were increased in CF, CFM and CFS compared to NF. Principle component analysis of DGGE profiles showed significant difference in nirK and amoA genes composition between organic amended （CFS and CFM） and the non-organic amended （CF and NF） plots. Number of amoA copies was significantly positively correlated with normalized soil nutrient richness （NSNR） of soil organic carbon （SOC） and total nitrogen （T-N）, and that of nirK copies was with NSNR of SOC, T-N plus total phosphorus. Moreover, nitrification potential showed a positive correlation with SOC content, while a significantly lower denitrification potential was found under CFM compared to under CFS. Therefore, SOC accumulation accompanied with soil nutrient richness under long-term balanced and organic/inorganic combined fertilization promoted abundance and diversity of AOB and DNB in the rice paddy.     

农田土壤中氨氧化细菌群落多样性研究进展

氨氧化细菌(AOB)在全球氮循环中具有重要作用,与农田土壤潜在硝化率(PNR)的高低有直接关系。基于长期定位试验氨氧化细菌的研究报道,综述在我国主要农田土壤中氨氧化细菌多样性和丰富度对不同肥料处理、种植方式的响应,旨在为提高土壤硝化能力、促进碳氮良性循环、构建健康土壤提供参考。     

ROOT EXUDATES FROM CANOLA EXHIBIT BIOLOGICAL NITRIFICATION INHIBITION AND ARE EFFECTIVE IN INHIBITING AMMONIA OXIDATION IN SOIL

● First evidence of BNI capacity in canola. ● BNI level was higher in canola cv. Hyola 404RR than in B. humidicola, the BNI positive control. ● BNI in canola may explain increased N immobilization and mineralization rates following a canola crop which may have implications for N management in rotational farming systems that include canola. A range of plant species produce root exudates that inhibit ammonia-oxidizing microorganisms. This biological nitrification inhibition (BNI) capacity can decrease N loss and increase N uptake from the rhizosphere. This study sought evidence for the existence and magnitude of BNI capacity in canola ( Brassica napus). Seedlings of three canola cultivars, Brachiaria humidicola (BNI positive) and wheat ( Triticum aestivum) were grown in a hydroponic system. Root exudates were collected and their inhibition of the ammonia oxidizing bacterium, Nitrosospira multiformis, was tested. Subsequent pot experiments were used to test the inhibition of native nitrifying communities in soil. Root exudates from canola significantly reduced nitrification rates of both N. multiformis cultures and native soil microbial communities. The level of nitrification inhibition across the three cultivars was similar to the well-studied high-BNI species B. humidicola. BNI capacity of canola may have implications for the N dynamics in farming systems and the N uptake efficiency of crops in rotational farming systems. By reducing nitrification rates canola crops may decrease N losses, increase plant N uptake and encourage microbial N immobilization and subsequently increase the pool of organic N that is available for mineralization during the following cereal crops.     

氟磺胺草醚及其降解菌对大豆生长及生物固氮的影响

从生长于氟磺胺草醚污染土壤的大豆根瘤中筛选出的Sinorhizobium sp.W16菌株,能高效降解氟磺胺草醚并能缓解氟磺胺草醚的生物负效应。以大豆（苏C1008）为研究对象,采用盆栽试验,探究Sinorhizobium sp.W16对大豆生长、氮累积量、根瘤固氮酶活性及根际土壤氮循环相关微生物功能基因丰度等的影响。结果表明：氟磺胺草醚的施用量（以有效成分计）超过450 g·hm^-2时显著降低了大豆生物量,抑制了大豆根瘤固氮酶活性和土壤脲酶活性,降低了土壤固氮细菌（nifH）、氨氧化古菌（AOA）、氨氧化细菌（AOB）基因丰度,限制了植物-根际系统的生物固氮及有机氮素转化;接种Sinorhizobium sp.W16降解菌显著提高土壤中氟磺胺草醚的降解率至81.97%,且显著提高了大豆根瘤干质量、根瘤固氮酶活性和土壤nifH基因丰度,增强了大豆的固氮作用,同时刺激了脲酶活性,提升土壤AOA和AOB的基因丰度,增加了土壤有效氮素的供应,从而使大豆植株氮含量提高了15.85%~24.93%。研究表明,氟磺胺草醚的施用抑制了大豆-根际系统的生物固氮作用,但接种Sinorhizobium sp.W16降解菌不仅能有效降低土壤中氟磺胺草醚的残留量、缓解氟磺胺草醚对大豆的持续药害,还增强了植物-根际系统中生物固氮能力、土壤有效氮素供应及大豆的氮素累积,对修复氟磺胺草醚污染土壤、增强大豆固氮具有较好的应用价值和市场前景。