Bacterial diversity in soils subjected to long-term chemical fertilization can be more stably maintained with the addition of livestock manure than wheat straw

Addition of organic matter such as livestock manures and plant residues is a feasible practice to mitigate soil degradation caused by long-term application of chemical fertilizers, and the mitigation is largely mediated though activities of the soil-dwelling microorganisms. However, the roles of different kinds of organic matter in maintaining bacterial community structure have not been assessed in a comparative manner. In this study, 454 pyrosequencing of 16S rRNA gene was employed to compare the bacterial community structure among soils that had been subjected to 30 years of NPK fertilization under six treatment regimes: non-fertilization control, fertilization only, and fertilization combined with the use of pig manure, cow manure or low- and high-level of wheat straws. Consistent with expectation, long-term application of NPK chemical fertilizers caused a significant decrease of bacterial diversity in terms of species richness (i.e. number of unique operational taxonomic units (OTU)), Faith's index of phylogenetic diversity and Chao 1 index. Incorporation of wheat straw into soil produced little effects on bacterial community, whereas addition of either pig manure or cow manure restored bacterial diversity to levels that are comparable to that of the non-fertilization control. Moreover, bacterial abundance determined by quantitative PCR was positively correlated with the nutritional status of the soil (e.g., nitrate, total nitrogen, total carbon, available phosphorus); however, bacterial diversity was predominantly determined by soil pH. Together, our data implicate the role of livestock manures in preventing the loss of bacterial diversity during long-term chemical fertilization, and highlight pH as the major deterministic factor for soil bacterial community structure.     

长期施肥与地膜覆盖对土壤微生物量碳氮的影响

通过田间定位试验研究长期施肥与地膜覆盖对土壤微生物量C、N的影响。结果表明，长期施肥与地膜覆盖提高了土壤微生物量C、N含量，长期施有机肥和有机无机肥配施，土壤微生物量C、N显著高于单施化肥和不施肥。相关分析表明，土壤微生物量C、N与土壤有机C、全N均呈极显著的正相关。土壤微生物量C、N可作为指示土壤肥力的重要指标。本试验土壤微生物量C占有机C的比例平均为9．95％，微生物量N占全N的比例平均为10．78％。     

长期有机与无机肥配施的黄壤稻田土壤细菌群落结构特征

  【目的】  比较长期施用不同肥料黄壤稻田土壤细菌群落结构的差异,剖析不同肥料维持土壤细菌群落多样性的作用及其机理,为农田施肥管理提供理论依据。  【方法】  以农业农村部贵州耕地保育与农业环境科学观测试验站为依托,采用高通量测序技术,分析连续24年不施肥(CK)、全量化肥(NPK)、1/4牛厩肥+3/4化肥(1/4M+3/4NP)、1/2牛厩肥+1/2化肥(1/2M+1/2NP)和全量牛厩肥(M)处理黄壤稻田土壤细菌群落组成及多样性,并揭示其主要环境影响因子。  【结果】  不同施肥处理土壤细菌α多样性指数分析结果显示,长期施用有机肥提高了土壤细菌多样性指数(Shannon)、优势度指数(Simpson)和均匀度指数(Pielou),对丰富度指数(Chao1)影响较小。与CK相比,长期施肥不同程度地提高了变形菌门(Proteobacteria)的相对丰度,化肥的作用更明显。与CK和NPK相比,施用有机肥处理降低了棒状杆菌门(Rokubacteria)和亚硝酸盐氧化菌门(Nitrospinae)相对丰度,提高了拟杆菌门(Bacteroidetes)相对丰度,其他菌门变化不明显(P<0.05)。土壤细菌群落结构主成分分析结果表明,施用有机肥1/4M+3/4NP和1/2M+1/2NP处理土壤环境较为相似,细菌群落组成相似度较高,而CK和NPK处理土壤环境相似,细菌群落组成相似度高。土壤细菌群落结构组成与土壤环境因子冗余分析显示,土壤理化性质对细菌群落结构的影响重要性由大到小依次为全氮、碱解氮、速效钾、pH、有效磷、全磷,其中全氮、碱解氮和速效钾是关键因素。  【结论】  长期施用有机肥能够提高黄壤稻田土壤肥力,改变细菌生长环境,进而改变细菌群落结构组成,提高细菌群落多样性,促进土壤生态系统稳定和健康。     

地膜覆盖对土壤微生物群落结构的影响

采用磷脂脂肪酸(PLFA)法测定了沈阳农业大学棕壤长期定位试验站地膜覆盖条件下土壤微生物磷脂脂肪酸图谱及群落结构。结果表明:在玉米苗期,长期施氮肥处理土壤覆膜后大部分脂肪酸含量都有所提高;施有机肥处理的土壤覆膜后脂肪酸的含量有降低的趋势;有机无机肥配合施用处理的土壤覆膜会增加真菌的含量,但却降低了其他脂肪酸的含量;在不施肥处理的土壤中,覆膜会使一些支链脂肪酸的含量降低。抽雄期,施有机肥的土壤覆膜后除单饱和脂肪酸含量有所下降外,其他脂肪酸都会提高;不施肥土壤覆膜处理双不饱和支链脂肪酸及放线菌标志性脂肪酸10Me18:0的含量会有所提高。成熟期,施氮肥的土壤覆膜处理大部分脂肪酸的含量降低;有机肥处理土壤中各种脂肪酸如:15∶0,a17∶0,i17∶0,i19∶0,18∶0,10Me18∶0含量覆膜高于裸地;有机无机配施处理土壤中17∶0,br17∶0,18∶0,18∶2w6,10Me18∶0含量覆膜处理高于裸地;对照土壤中i15∶0,16∶0,17∶0,a17∶0,18∶0,18∶2w6,19∶0含量覆膜高于裸地。另外,从脂肪酸的变化看出,覆膜处理土壤微生物群落整体结构发生了改变,没有表现出明显的种群优势。但是,尽管土壤的施肥处理不同,覆膜处理土壤微生物群落结构有一致化发展的趋势。     

长期施肥对旱地红壤细菌群落的影响

为探讨长期不同施肥对旱地红壤细菌群落的影响,以中国农业科学院祁阳红壤实验站的冬小麦—夏玉米定位试验为研究对象,选取不施肥(CK)、单施氮肥(N)、施化学氮磷钾肥(NPK)和化学氮磷钾+有机肥配施(NPKM)4个处理,于试验开展25年(2015年)小麦收获后采集各处理0~20 cm的土壤样品,利用Illumina MiSeq高通量测序技术对土壤细菌群落进行测定,并深入揭示影响旱地红壤细菌群落的关键因素。结果表明:(1)长期不同施肥显著改变了旱地红壤的化学性质,N和NPK处理的土壤pH显著降低至4.02和4.15,而NPKM处理的土壤pH显著上升至5.99。NPK和NPKM处理均显著改善土壤肥力,但后者效果明显优于前者,而N处理对土壤肥力的提升效果微弱。(2)长期不同施肥改变了旱地红壤优势菌的相对丰度,非度量多维度分析(NMDS)和相似性分析(ANOSIM)表明不同处理的土壤细菌群落发生显著变化。(3)与CK相比,N处理的4种多样性指数(物种丰富度、Chao1指数、系统发育多样性和香农指数)显著降低了21.4%~49.4%,而NPKM处理显著增加了7.0%~66.9%,NPK处理也会使系统发育多样性和香农指数显著降低10.3%和13.0%。(4)逐步回归分析表明土壤pH是决定优势菌相对丰度及4种多样性指数的首要因素,多元回归树分析(MRT)探明土壤pH共解释了83.1%的细菌群落变异,不同处理间细菌群落转变均由土壤pH驱动。(5)STAMP分析发现,N、NPK和NPKM处理与CK分别有11、14和8个显著差异细菌属。综上所述,长期施肥后旱地红壤细菌群落主要受土壤pH的影响,而土壤肥力的作用相对较弱,长期施用化学氮肥造成的红壤酸化的负面效应已远超肥力改善的正面效应。因此,旱地红壤施肥应以防治土壤酸化为前提,长期化肥有机肥配施是一项适宜的施肥措施。     

Bacterial diversity as affected by application of manure in red soils of subtropical China

A plot experiment was conducted to understand the response of the soil bacterial community to manure application rates and the relationship between the composition of bacterial community and soil chemical properties. The experiment involved gradients of manure combined with chemical fertilizer in red soils from granite, red sandstone and red clay between 2013 and 2015. The soil bacterial community composition was significantly affected by different manure rates. The relative abundances of Burkholderiaceae, Micrococcaceae and Streptomycetaceae were higher at low manure rates (1.75 to 3.5 t·ha^?1·yr.^?1), whereas the relative abundance of Xanthomonadaceae was higher at high manure rates (7 to 28 t·ha^?1·yr.^?1). Manure application increased the bacterial abundance but decreased the diversity when its rates were higher than 7, 14 and 14 t·ha^?1·yr.^?1 in soils from granite, red sandstone and red clay, respectively. Redundancy analysis revealed that soils from different parent materials had different bacterial communities with soil pH and available phosphorus (AP) being determinant factors. The peanut yields exhibited significantly positive correlations with the bacterial diversity in soil, implying the importance of bacterial diversity for soil productivity. Soil AP was correlated with bacterial diversity by parabolic equations and probably AP may be an indicator of declining bacterial diversity at high manure rates. The critical value were 39.71, 65.75 and 90.16 mg·kg^?1 in soils from granite, red sandstone and red clay, respectively. This study suggests the importance of maintaining soil bacterial diversity under moderate and balanced applications of manure.     

Response of the bacterial diversity and soil enzyme activity in particle-size fractions of Mollisol after different fertilization in a long-term experiment

Particle-size soils were fractionated for evaluating changes in the composition of bacterial community and enzyme activity in response to 13 years of fertilization. This study focused on Mollisol and its particle-size fractions of 200–2,000 μm (coarse sand sized), 63 to 200 μm (fine sand sized), 2 to 63 μm (silt sized), and 0.1 to 2 to μm (clay-sized). Long-term chemical fertilization lowered the pH of all particle fractions, whereas organic fertilizer application mitigated soil acidification. Nutrient concentrations depended on both fertilizer treatment and particle fractions and enzymes were unevenly active throughout the soil. Generally, the highest enzyme activities were observed in the silt and clay fractions of control soil and the soil treated with chemical fertilizer (N, P, and K (NPK)) and in the sand-sized fraction of soil treated with manure and chemical fertilizer (MNPK). Except for acid phosphomonoesterase, the other tested enzyme activities in coarse-sized fractions of MNPK soil were significantly higher than those of the control and NPK soils. Fertilization and soil fraction interactively (p?<?0.05) affected the enzyme activity. Denaturing gradient gel electrophoresis analysis showed that the bacterial community structure significantly differed in different particle sizes with a higher bacterial diversity in small-sized than in coarse-sized fractions. Dominant bands were excised and sequenced. We have found the following bacterial groups: Actinobacteria, γ-proteobacteria, and Acidobacteria. In addition, enrichment of organic matter in coarser fractions was related to greater bacterial diversity than any other treatment. Principal component analysis showed a smaller variability among fractions of the organic amended treatment. Redundancy analysis showed that the tested properties significantly affected the composition of bacterial community with the exception of C/N and available P. No significant correlation between enzyme activity and bacterial community composition was detected, whereas positive correlations between other soil properties and enzyme activities were observed to various extents. Probably, enzyme activities might be affected by specific functional bacterial communities rather than by the overall bacterial community. We concluded that the long-term application of organic manures contributed to the increase of soil organic matter content of particles higher than 200 mm, with higher bacterial diversity and increases in most of the enzyme activities.     

西湖平原区连续13年定位施肥对麦、稻产量及土壤肥力的影响

以浙江省水网平原水稻主产区土壤为对象,通过定位试验,研究了连续13年的不同施肥处理对麦稻产量、土壤养分状况和物理性状的影响。结果表明,化肥配施有机肥可显著提高麦、稻产量; 不同施肥处理的长期定位试验土壤有机质含量和全氮均呈上升趋势,增幅依次为: 栏肥+NPK秸秆+NPKNPK秸秆栏肥CK处理; 土壤碱解氮和速效磷也呈增加趋势,以栏肥+NPK处理的增幅最为明显。土壤物理性状的分析表明,长期施肥均能明显增加土壤水稳性团粒含量和土壤孔隙度。经土壤养分平衡分析,栏肥+NPK、秸秆+NPK和NPK处理的氮和磷呈现盈余,秸秆和CK处理氮和磷亏缺; 栏肥+NPK和秸秆+NPK处理钾基本平衡,NPK、秸秆、栏肥和CK处理钾严重亏缺。长期定位试验进一步证明有机肥与氮、磷、钾化肥长期配合施用可实现当地农作物持续稳产,农田施肥管理要注意适当减少氮、磷投入,增加钾肥施用量,保持农田土壤养分平衡。     

化肥配施有机肥对花生根际细菌群落结构及共存网络的影响

为评估化肥配施有机肥对花生根际细菌群落的影响，田间试验条件下研究了不施肥(CK)、施化学氮磷钾肥(NPK)和化学氮磷钾肥+有机肥配施(NPKM)对花生根际细菌群落多样性、结构和共存网络的影响。结果表明：NPKM处理下花生根际土养分状况明显改善，特别是有效磷的含量，较CK和NPK处理分别提高了5.31倍～12.16倍和3.24倍～6.50倍；而NPK处理下花生根际土养分状况并没有明显提升；Source Tracker分析显示，NPKM处理下根际细菌群落中只有2.1%～5.5%的物种来源于有机肥自身，但其花生根际细菌群落多样性在生育期前期要明显高于CK和NPK处理，而CK和NPK处理下根际细菌群落多样性差异不大；限制性主成分分析(CAP)显示，施肥措施和生育期均对根际细菌群落结构产生了显著影响，NPKM处理下花生根际土中富集了属于根瘤菌目(Rhizobiales)、梭菌目(Clostridiales)和芽孢杆菌目(Bacillales)这一类根际促生菌，根际细菌共存网络结构更加复杂，且网络中重要连接点的比例要也明显高于CK和NPK处理网络。可见，化肥配施有机肥有助于构建一个健康稳定的根际细...     

Long-term and legacy effects of manure application on soil microbial community composition

We analyzed soil prokaryotic and fungal community composition in soils with varying histories of cattle manure application. The manure treatments were (i) annual application for 43 years (MF), (ii) annual application for 14 years followed by 29 years without application (MF14), and (iii) annual application for 30 years followed by 13 years without application (MF30). An annual application of chemical nitrogen (N) fertilizer (CNF) and a non-amended control (Con) were also included. Soil prokaryotic evenness and diversity significantly decreased in MF relative to other treatments in fall, but were similar to the other fertilizer treatments in spring and summer. Distinct prokaryotic and fungal community composition was observed in MF compared to other treatments across fall, spring, and summer seasons. The MF treatment significantly increased the relative abundance of Firmicutes, Gammaproteobacteria, and Gemmatimonadetes, but significantly decreased the relative abundance of Acidobacteria. In fall, the soil prokaryotic and fungal community composition with MF30 was significantly different than the other fertilization treatments. Overall, the study showed that annual manure application (MF) led to a different microbial community composition than the other fertilizer treatments. Soil without manure application for 13 years (MF30) had a significantly different microbial community composition from other fertilizer treatments in fall, while the soil without manure application for 29 years (MF14) resembled a microbial community that had never received manure.     

长期施用有机肥潮土细菌的多样性及功能预测

  【目的】  细菌作为土壤中最多的微生物物种,其多样性是土壤质量和土壤健康的标志。施肥不仅为土壤细菌提供了矿质养分,也为细菌提供了不同的碳源。探究长期不同施肥下潮土细菌的多样性及其功能,对于深入理解土壤养分转化的微生物驱动过程及优化施肥管理措施具有重要意义。  【方法】  选取潮土上连续37年不施肥(CK)、单施化学氮肥(N)、单施有机肥(M)、有机肥和化学氮肥配施(MN) 的4个处理,采用高通量测序方法分析土壤细菌丰度(16S rRNA基因拷贝数)、α多样性和β多样性,采用FAPROTAX功能预测的方法分析主要功能种群的丰度在处理间的差异,采用Mantel 检验分析细菌多样性等与产量、土壤养分含量间的相关关系。  【结果】  长期施用有机肥显著提高了土壤细菌丰度,MN和M处理下土壤中的16S rRNA基因拷贝数分别是CK和N处理平均值的11.8和10.7倍(P< 0.05)。土壤细菌多样性指数(Shannon index)和丰富度指数(Chao1 index)均在M处理下最高。潮土中变形菌门、酸杆菌门和放线菌门为优势菌门。与CK相比,施肥处理均提高了潮土拟杆菌门、厚壁菌门和疣微菌门的相对丰度。与N处理相比,MN处理显著提高了潮土变形菌门、厚壁菌门和拟杆菌门的丰度,降低了酸杆菌门的丰度。潮土细菌生态功能以化能异养、有氧化能异养、发酵作用、硝化作用和硝酸盐还原为主,相应种群丰度均在1%以上。MN处理下化能异养功能细菌的丰度较其他处理显著提高2.2%～4.2%。MN和M处理中起发酵作用的种群相对丰度是其他处理的3倍以上。与CK 相比,3个施肥处理均显著增加了光异作用和光异养作用功能种群的相对丰度。土壤细菌β多样性和功能结构均与土壤有机质(SOM)、全氮(TN)、硝态氮(NO₃?-N)含量及作物产量呈显著(P < 0.05)或极显著(P < 0.01)相关。  【结论】  长期施用有机肥提高了潮土中细菌的数量和多样性,提高了土壤中变形菌门、厚壁菌门和拟杆菌门的丰度,降低了酸杆菌门的丰度。有机肥和化肥配施处理中化能异养型细菌丰度较高,可能是施用有机肥后优化土壤养分循环和响应作物产量提高的主要菌群,今后可进一步深入研究其菌群组成和功能调控。     

长期施肥对黑土农田土壤微生物群落的影响

基于中国科学院海伦农业生态试验站长期定位试验区,应用实时荧光定量PCR(Real-time PCR)和变性梯度凝胶电泳(DGGE)技术研究了无施肥(NF)、单施N、P化肥(NP)以及化肥配施有机猪粪肥(NPM)等3种长期施肥措施对黑土区玉米田土壤微生物群落密度和结构的影响.Real-time PCR方法定量NF、NP及NPM措施土壤细菌群落基因组DNA质量分别为381、1 351和1 773 ng g-1干土,真菌群落基因组DNA质量分别113.3、127.3和20.6 ng g-1干土,真菌与细菌的比率分别为0.31、0.09和0.01,NPM措施显著低于另两种施肥方式(p＜0.05).DGGE方法研究表明,NP和NPM措施不能改善土壤细菌和真菌群落的多样性、均匀性及优势菌优势程度;但主成分分析结果显示NP和NPM措施均可改变土壤细菌和真菌群落的构成,且真菌群落的变化更为显著;聚类分析结果显示NP和NPM措施下细菌群落结构较相近,其相似系数为0.89,真菌群落中NP措施与NF措施相近,相似系数为0.63,高于NP与NPM措施的相似系数0.51.上述结果表明有机猪粪肥的长期施用可以显著降低黑土农田土壤真菌与细菌的比率,且明显地改变土壤细菌和真菌群落的结构.     

长期施肥对黄棕壤细菌多样性的影响

  【目的】   基于武汉黄棕壤长期定位试验 (1981—2016年),探究不同施肥措施下土壤肥力演变和土壤微生物多样性变化,为黄棕壤培肥以及农业绿色可持续发展提供依据。   【方法】   本研究采用Illumina MiSeq高通量测序技术和偏最小二乘路径模型 (PLS-PM) 综合分析了不施肥 (CK)、氮磷钾 (NPK)、常量有机肥 (OM)、氮磷钾+常量有机肥 (NPK+OM) 和氮磷钾+高量有机肥 (NPK+OMM) 5种不同施肥方式对黄棕壤理化性质、细菌多样性的影响及其与产量的关系。   【结果】   1) 与CK相比,施有机肥处理显著增加了土壤碱解氮、有效磷、速效钾和有机碳含量,而NPK处理只显著增加了土壤有机碳含量。NPK+OMM处理的土壤有效磷和速效钾含量显著高于OM和NPK+OM处理,3个有机肥处理间的碱解氮、有机碳含量差异不显著。4个施肥处理均显著提高了水稻产量,但处理间差异不显著。2) OM处理土壤微生物多样性最高,而高量粪肥投入的NPK+OMM处理,细菌多样性有下降的趋势,但各施肥处理间细菌多样性差异不显著。3) 长期不同施肥方式影响土壤细菌群落结构,其中NPK+OM和NPK+OMM处理细菌群落结构更接近。OM处理提高了变形菌纲和放线菌纲的相对丰度,降低了绿弯菌纲和硝化螺旋菌纲的相对丰度。NPK处理降低了放线菌纲和硝化螺旋菌纲相对丰度,提高了酸杆菌纲相对丰度。与NPK+OM处理比较,NPK+OMM处理降低了放线菌纲和α-变形菌纲相对丰度,提高了厌氧绳菌纲、绿弯菌纲和硝化螺旋菌纲相对丰度。4) PLS-PM显示土壤有机碳 (SOC)、碱解氮、有效磷和速效钾对细菌群落结构表现出正调控 (路径系数 = 0.36),而pH表现负调控 (路径系数 = ?0.48);但是对细菌多样性的影响都较小;影响产量的理化指标主要是SOC、碱解氮和有效磷。土壤细菌多样性对产量显示正调控 (路径系数 = 0.42)。   【结论】   土壤有机碳 (SOC) 和碱解氮、有效磷、速效钾含量对细菌群落结构有正调控作用,而pH有负调控作用。与NPK处理相比,长期施用常量有机肥 (OM) 处理提高了细菌多样性和水稻产量,而高量有机肥配施氮磷钾肥 (NPK+OMM) 会导致细菌多样性和产量降低。     

Microbial residues were increased by film mulching with manure amendment in a semiarid agroecosystem

Amino sugars, as a kind of microbial residue, are strongly associated with cycling of microbial-derived soil organic matter. However, responses of amino sugars to agricultural practices on the Loess Plateau in North-western China are poorly known. The objective was to evaluate effects of film mulching (no film mulching + NPK fertilizers, CK; film mulching + NPK fertilizers, PF; film mulching + NPK fertilizers + cow manure, FM) on accumulations of amino sugars in this region. FM significantly increased total amino sugar by 190.46 mg kg^?1 in 0–10 cm layer and 214.66 mg kg^?1 in 10–20 cm layer relative to CK, but PF significantly decreased it by 139.28 mg kg^?1 in 0–10 cm layer. Ratios of glucosamine to muramic acid were markedly decreased by 2.50 in 0–10 cm layer and 2.28 in 10–20 cm layer in FM than CK, suggesting a tendency of microbial residues pool shift towards bacterial residues in this agroecosystem. These results indicated film mulching alone was not benefitial to accumulation of amino sugar while organic manure contributed to the build-up of amino sugar partly due to manure contained microbial residues. The different patterns of amino sugars suggested significant changes in the quality of microbial-derived organic matter.     

等碳投入的有机肥和生物炭对红壤微生物多样性和土壤呼吸的影响

施用有机肥是快速培育瘠薄土壤的一个重要措施。针对中亚热带第四纪红黏土发育的红壤旱地,建立了玉米和花生单作系统等碳量投入有机肥和生物炭的田间试验,利用聚合酶链式反应—变性梯度凝胶电泳(polymerase chain reaction-denaturing gradient gel electrophoresis,PCR-DGGE)方法研究了土壤细菌和真菌群落组成和多样性的变化,分析了土壤呼吸速率(CO2通量)的变化及其与微生物多样性的关系。两年的试验表明,不同施肥方式导致微生物群落结构显著分异,施用有机肥和生物炭显著增加了细菌多样性,但施肥第二年真菌多样性有下降趋势。秸秆和猪粪配施显著增加了土壤呼吸速率,土壤呼吸速率与细菌和真菌多样性呈显著正相关,细菌多样性对土壤呼吸的影响(相对贡献率为71%)显著高于真菌(29%)。土壤磷素(全磷和速效磷)含量的变化是驱动红壤微生物多样性变化的主导因素,其对细菌和真菌多样性的相对贡献率分别为44.8%和47.4%。因此,合理配施秸秆和猪粪可以快速提高瘠薄红壤的生物功能。     

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。     

长期不同施肥方式下土壤有机碳的垂直分布及碳储量

为了研究施肥对土壤有机碳含量的影响,在湖北省农业科学院南湖实验站进行了25年不同施肥方式的长期定位试验。研究结果表明:与对照相比,除单施氮肥与单施有机肥外,其他施肥方式均提高了0～20cm土壤有机碳含量与碳储量;与对照及单施化肥相比,有机肥配施化肥均提高了0～20cm及0～100cm土壤有机碳含量与碳储量。单施化肥与单施有机肥对各土层土壤有机碳含量影响较小,且土壤有机碳累积少;而化肥配施有机肥提高了0～20cm与20～40cm土壤有机碳含量与碳储量。除对照及氮磷钾肥配施过量有机肥处理外,其他处理土壤全氮与有机碳含量间具有显著相关性。有机肥与化肥配合施用是提高农田土壤有机碳,增加土壤碳储量的有效方法。     

Long-term fertilization regimes affect bacterial community structure and diversity of an agricultural soil in northern China

Background, Aims, and Scope  Knowledge about shifts of microbial community structure and diversity following different agricultural management practices could improve our understanding of soil processes and thus help us to develop sound management strategies. A long-term fertilization experiment was established in 1989 at Fengqiu (35°00′N, 114°24′E) in northern China. The soil (sandy loam) is classified as aquic inceptisols and has received continuous fertilization treatments since then. The fertilization treatments included control (CK, no fertilizer), chemical fertilizers nitrogen (N) and potassium (K) (NK), phosphorous (P) and K (PK), NP, NPK, organic manure (OM), and half chemical fertilizers NPK plus half organic manure (1/2NPKOM). The objective of this study was to examine if the microbial community structure and diversity were affected by the long-term fertilization regimes. Materials and Methods  Soil samples were collected from the long-term experimental plots with seven treatments and four replications in April 2006. Microbial DNAs were extracted from the soil samples and the 16S rRNA genes were PCR amplified. The PCR products were analyzed by DGGE, cloning and sequencing. The bacterial community structures and diversity were assessed using the DGGE profiles and the clone libraries constructed from the excised DGGE bands. Results  The bacterial community structure of the OM and PK treatments were significantly different from those of all other treatments. The bacterial community structures of the four Ncontaining treatments (NK, NP, NPK and 1/2NPKOM), as well as CK, were more similar to each other. The changes in bacterial community structures of the OM and PK treatments showed higher richness and diversity. Phylogenetic analyses indicated that Proteobacteria (30.5%) was the dominant taxonomic group of the soil, followed by Acidobacteria (15.3%), Gemmatimonadetes (12.7%), etc. Discussion  Irrespective of the two fertilization treatments of OM and PK, the cluster analysis showed that bacterial communities of the remaining five treatments of CK, NK, NP, NPK and 1/2NPKOM seemed to be more similar to each other, which indicated the relatively weak effects of the four N-containing treatments on soil bacterial communities. N fertilizer may be considered as a key factor to counteract the effects of other fertilizers on microbial communities. Conclusions  Our results show that long-term fertilization regimes can affect bacterial community structure and diversity of the agricultural soil. The OM and PK treatments showed a trend towards distinct community structures, higher richness and diversity when compared to the other treatments. Contrasting to the positive effects of OM and PK treatments on the bacterial communities, N fertilizer could be considered as a key factor in the soil to counteract the effects of other fertilizers on soil microbial communities. Recommendations and Perspectives  Because of the extremely high abundance and diversity of microorganisms in soil and the high heterogeneity of the soil, it is necessary to further examine the effects of fertilization regimes on microbial community and diversity in different type soils for comprehensively understanding their effects through the appropriate combination of molecular approaches. ESS-Submission Editor: Chengrong Chen, PhD (c.chen@griffith.edu.au)     

腐熟羊粪有机肥配施无机肥对植烟土壤微生物群落结构和多样性的影响

为了探究腐熟羊粪有机肥与无机肥配施对洛阳烟区植烟土壤微生物群落结构和土壤肥力特性的影响，采用盆栽控制试验和高通量测序技术，研究了不施肥（CK）、100%无机氮肥（T0）、50%羊粪有机肥氮+50%无机氮肥（T50）和100%羊粪有机肥氮（T100）4个处理下植烟土壤细菌和真菌群落结构及多样性的差异，并结合土壤理化性质分析了土壤肥力指标与土壤微生物多样性的关系。结果表明：植烟土壤细菌优势菌门为变形菌门（Proteobacteria）、酸杆菌门（Acidobacteria）和放线菌门（Actinobacteria），土壤真菌优势菌门为子囊菌门（Ascomycota）和担子菌门（Basidiomycota）；T50处理的变形菌门、酸杆菌门和子囊菌门相对丰度最高，但放线菌门和担子菌门相对丰度最低；NMDS分析和相似性分析发现，T50处理的土壤微生物群落结构与其他处理差异显著；Alpha多样性分析也表明，T50处理土壤细菌和真菌群落的丰富度和多样性最高。不同处理土壤理化性质和土壤关键酶活性差异显著，以T50处理土壤养分含量和土壤碳氮代谢酶的活性最高；Pearson相关性分析显示，土壤碱解氮、有效磷、速效钾含量及蛋白酶和脲酶活性与植烟土壤微生物多样的关系最为密切。总之，采用腐熟羊粪有机肥和无机肥配施通过影响植烟土壤细菌和真菌群落结构和多样性，促进了土壤养分的释放和转化，有利于土壤养分的供应和土壤质量的提升，研究结果为洛阳烟区应用羊粪有机肥改良土壤提供了理论依据。     

Different roles of rhizosphere effect and long-term fertilization in the activity and community structure of ammonia oxidizers in a calcareous fluvo-aquic soil

Ammonia oxidation is a critical step in the soil nitrogen (N) cycle and can be affected by the application of mineral fertilizers or organic manure. However, little is known about the rhizosphere effect on the function and structure of ammonia-oxidizing bacterial (AOB) and archaeal (AOA) communities, the most important organisms responsible for ammonia oxidation in agricultural ecosystems. Here, the potential nitrification activity (PNA), population size and composition of AOB and AOA communities in both the rhizosphere and bulk soil from a long-term (31-year) fertilizer field experiment conducted during two seasons (wheat and maize) were investigated using the shaken slurry method, quantitative real-time polymerase chain reaction and denaturing gradient gel electrophoresis. N fertilization greatly enhanced PNA and AOB abundance, while manure application increased AOA abundance. The community structure of AOB exhibited more obvious shifts than that of AOA after long-term fertilization, resulting in more abundant AOB phylotypes similar to Nitrosospira clusters 3 and 4 in the N-fertilized treatments. Moreover, PNA was closely correlated with the abundance and community structure of AOB rather than that of AOA among soils during both seasons, indicating that AOB play an active role in ammonia oxidation. Conversely, the PNA and population sizes of AOB and AOA were typically higher in the rhizosphere than the bulk soil, implying a significant rhizosphere effect on ammonia oxidation. Cluster and redundancy analyses further showed that this rhizosphere effect played a more important role in shaping AOA community structure than long-term fertilization. Overall, the results indicate that AOB rather than AOA functionally dominate ammonia oxidation in the calcareous fluvo-aquic soil, and that rhizosphere effect and fertilization regime play different roles in the activity and community structures of AOB and AOA.