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。     

鸡粪菌渣好氧堆肥过程中氨氧化古菌及氨氧化细菌群落的动态变化

以amo A基因为标记,通过Real-Time PCR和限制性片段长度多态性(Restriction fragment length polymorphism,RFLP)法对鸡粪菌渣好氧堆肥过程中的氨氧化古菌(Ammonia-oxidizing archaea,AOA)和氨氧化细菌(Ammonia-oxidizing bacteria,AOB)进行了丰度及群落结构的分析。结果表明,在堆制初期、好氧发酵高温期及后熟期,AOB的amo A基因丰度均占主导优势,是AOA的38~992倍。进入好氧发酵高温期,AOA amo A基因丰度下降至发酵前的0.9%,AOB下降至17.6%,后熟期AOA与AOB的amo A基因丰度与好氧发酵高温期相当。在上述3个阶段AOA与AOB各自存在一个绝对优势菌群,分别为Cluster 3和Nitrosomonas europaea,其中Cluster 3克隆子数目分别占整个克隆文库的70.73%、54.28%、72.45%,Nitrosomonas europaea克隆子数目分别占整个克隆文库的78.44%、93.20%、94.27%。堆肥3个阶段AOA的多样性指数变化不大,Shannon-Wiener值维持在1.53~1.60,但群落结构发生明显演替,随着堆肥温度升高,堆肥前期的一些菌群(Cluster 4、Cluster 5、Cluster 6)逐渐消失,新的菌群Cluster 1出现并成为堆肥中后期的第二大优势菌群。AOB无论是多样性指数还是群落组成,都发生剧烈的变化。AOB在堆肥前期Shannon-Wiener指数值最大(1.47),种群数最多(6个基因簇,分别为Nitrosomonas europaea Cluster,Nitrosomonas halophila Cluster,Nitrosomonas communis Cluster,Nitrosomonas nitrosa Cluster,Nitrosospira briensis Cluster,Nitrosospira multiformis Cluster);进入高温发酵期,Shannon-Wiener下降至0.45,群落结构单一,只有Nitrosomonas europaea Cluster和Nitrosomonas halophila Cluster;进入后熟期,AOB多样性及种群数得到一定程度的回升。     

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.     

长期施肥对旱地土壤中氨氧化微生物丰度和分布的影响

采用基于氨单加氧酶基因(amoA)的荧光定量PCR技术,以黄土高原旱地土为材料,研究长期施肥对土壤氨氧化细菌和氨氧化古菌丰度的影响,并分析环境因素与氨氧化菌丰度的关系。以不施肥土壤为对照(CK),设置3个施肥处理,分别为单施磷肥(P),氮、磷共施(NP)和氮、磷、有机肥共施(NPM)3个处理。结果表明,不同处理氨氧化菌amoA基因拷贝数为1.326×106～1.886×106 g-1,各处理间氨氧化细菌丰度差异不显著;氨氧化古菌的arch-amoA基因拷贝数为1.329×106～4.510×106 g-1,表现为处理NPM>NP>CK>P,NPM处理为对照的3.314倍,二者呈现显著性差异。采用DCCA法对4个处理进行环境相似度分类,结果显示,P和NPM处理、CK和NP处理分别构成了2个相似类群;4个处理和12个环境因子的关联(CCA)分析表明,不同处理中的氨氧化微生物活跃度以及氨氧化过程强度表现为处理NMP>NP>CK>P;不同环境因子和不同施肥处理生境相似度分布存在不同的关系,其中反映氨氧化过程的硝态氮含量、氨氧化细菌和氨氧化古菌丰度,以及代表微生物生长主要环境因素的pH值、含水量、全氮和有机碳含量与不同施肥处理导致的生境相似度的分布关系最为紧密。     

牛场肥水灌溉对土壤氨氧化微生物的影响

以不同牛场肥水灌溉制度下河北省徐水县梁家营长期定位施肥试验田为研究对象,通过构建氨氧化细菌和氨氧化古菌的amoA 基因克隆文库,利用T-RFLP方法研究了5种施肥处理(CK:不施肥;CF:300 kg N·hm^-2,120 kg P₂O·hm^-2,75 kg K₂O·hm^-2;T4:105 kg N·hm^-2,39 kg P₂O₅·hm^-2;T5:210 kg N·hm^-2,78 kg P₂O₅·hm^-2; T11:317 kg N·hm^-2,117 kg P₂O₅·hm^-2)下土壤中氨氧化细菌及氨氧化古菌的多样性及其群落结构演变。氨氧化细菌T-RFLP结果显示,T5 处理土壤中氨氧化细菌的 Shannon-Wiener指数(H')和 Pielou指数(E)均最高, T11处理最低,表明T5处理增加了土壤中氨氧化细菌的多样性和丰富度,T11处理一定程度上降低了氨氧化细菌的多样性和丰富度。基于amoA基因建立的系统进化树显示,土壤中氨氧化细菌主要属于Nitrosospira 和Nitrosomanas。氨氧化古菌T-RFLP结果显示,T11处理氨氧化古菌Shannon-Wiener指数(H')、Simpson 指数(Ds)和Pielou指数(E)均最高,表明T11处理增加了土壤中氨氧化古菌的多样性和丰富度。氨氧化古菌的系统发育树显示大部分氨氧化古菌amoA序列与 Cluster S 聚为一类,且占主导地位,少数序列为Unclassified-Archaea(未分类古菌)。最大优势菌与来自中国荷斯坦奶牛瘤胃样品的氨氧化古菌Clone属于同种,推测牛场肥水灌溉增加了土壤的氨氧化古菌的群落多样性。     

Soil Organic Carbon,Black Carbon,and Enzyme Activity Under Long-Term Fertilization

The present study aims to understand the effects of long-term fertilization on soil organic carbon (SOC), black carbon (BC), enzyme activity, and the relationships among these parameters. Paddy field was continuously fertilized over 30 yr with nine different fertilizer treatments including N, P, K, NP, NK, NPK, 2NPK (two-fold NPK), NPK+manure (NPKM), and CK (no fertilization), N, 90 kg urea-N ha⁻¹ yr⁻¹; P, 45 kg triple superphosphate-P₂O₅ ha⁻¹ yr⁻¹; K, 75 kg potassium chloride-K₂O ha⁻¹ yr⁻¹; and pig manure, 22 500 kg ha⁻¹ yr⁻¹. Soil samples were collected and determined for SOC, BC content, and enzyme activity. The results showed that the SOC in the NPKM treatment was significantly higher than those in the K, P, and CK treatments. The lowest SOC content was found in the CK treatment. SOC content was similar in the N, NP, NK, NPK, 2NPK, and NPKM treatments. There was no significant difference in BC content among different treatments. The BC-to-SOC ratios (BC/SOC) ranged from 0.50 to 0.63, suggesting that BC might originate from the same source. Regarding enzyme activity, NPK treatment had higher urease activity than NPKM treatment. The urease activity of NPKM treatment was significantly higher than that of 2NPK, NP, N, P, K, CK, and NPKM treatment which produced higher activities of acid phosphatase, catalase, and invertase than all other treatments. Our results indicated that long-term fertilization did not significantly affect BC content. Concurrent application of manure and mineral fertilizers increased SOC content and significantly enhanced soil enzyme activities. Correlation analysis showed that catalase activity was significantly associated with invertase activity, but SOC, BC, and enzyme activity levels were not significantly correlated with one another. No significant correlations were observed between BC and soil enzymes. It is unknown whether soil enzymes play a role in the decomposition of BC.     

双价转基因抗虫棉花对土壤氨氧化细菌和氨氧化古菌群落结构及丰度的影响

采用末端片段多态性分析(T-RFLP)和实时定量PCR(Quantitative real-time PCR,qPCR)方法,研究了不同生长时期双价转基因抗虫棉花和亲本非转基因棉花(对照)土壤氨氧化细菌(Ammonia-oxidizing bacteria,AOB)和氨氧化古菌(Ammonia-oxidizing archaea,AOA)群落结构和丰度的变化,以探讨双价转基因抗虫棉花种植对土壤氨氧化微生物的影响。结果表明,双价转基因抗虫棉花土壤中AOB和AOA优势种群与对照相同,且随生长时期无显著变化,但在各自样品中所占比例不同。不同生长时期双价转基因抗虫棉花土壤AOB的Shannon指数和Evenness指数均与对照无显著差异,AOA的Shannon指数与对照也无显著差异,而苗期时Evenness指数显著低于对照(P0.05),其他3个生长时期差异均不显著。qPCR分析表明,除花铃期双价转基因抗虫棉花土壤AOB丰度高于对照外,其他3个生长时期均低于对照,而AOA丰度在4个生长时期均低于对照。吐絮期时,双价转基因抗虫棉花与对照土壤AOB和AOA丰度均差异显著(P0.05)。双价转基因抗虫棉花对土壤AOB和AOA群落结构组成无显著影响,但降低了AOB和AOA的数量,双价转基因抗虫棉花对土壤微生物的某些类群可能存在潜在影响。     

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。     

Changes in soil organic carbon pools following long-term fertilization under a rain-fed cropping system in the Loess Plateau,China

Understanding the mechanism of soil organic carbon(SOC) sequestration is of paramount importance in sustaining crop productivity and mitigating climate change. Long-term trials were employed to investigate the responses of total SOC and its pools, i.e., mineral-associated OC(MOC), particulate OC(POC, containing Light-POC and Heavy-POC), to fertilization regimes at Yangling(25-year), Tianshui(35-year) and Pingliang(37-year) under a rain-fed cropping system in the Loess Plateau. The fertilization regimes in each trial included three treatments, i.e., control(no nutrient input, CK), chemical fertilizers(CF), and organic manure plus chemical fertilizers(MCF). Relative to the CK, long-term fertilization appreciably increased SOC storage by 134, 89 and 129 kg ha~(–1) yr~(–1) under CF, and 418, 153 and 384 kg ha~(–1) yr~(–1) under MCF in plough layer soils(0–20 cm), respectively, at the Yangling, Tianshui and Pingliang sites. The MOC pools accounted for 72, 67 and 64% of the total SOC at the above three sites with sequestration rates of 76, 57 and 83 kg ha~(–1) yr~(–1) under CF and 238, 118 and 156 kg ha~(–1) yr~(–1) under MCF, respectively. Moreover, the MOC pool displayed a saturation behavior under MCF conditions. The POC accordingly constituted 27, 33 and 36% of SOC, of which Light-POC accounted for 11, 17 and 22% and Heavy-POC for 17, 16 and 15% of SOC, respectively. The sequestration rates of POC were 58, 32 and 46 kg ha~(–1) yr~(–1) under CF, and 181, 90 and 228 kg ha~(–1) yr~(–1) under MCF at the three respective sites, in which Light-POC explained 59, 81 and 72% of POC under CF, and 60, 40 and 69% of POC under MCF, with Heavy-POC accounting for the balance. Compared with CK, the application of CF alone did not affect the proportions of MOC or total POC to SOC, whereas MCF application markedly reduced the proportion of MOC and increased the POC ratio, mainly in the Light-POC pool. The distribution of SOC among different pools was closely related to the distribution and stability of aggregates. The present study confirmed that organic manure amendment not only sequestered more SOC but also significantly altered the composition of SOC, thus improving SOC quality, which is possibly related to the SOC saturation level.     

Emission and Fixation of CO2 from Soil System as Influenced by Long-Term Application of Organic Manure in Paddy Soils

The observations of 25-yr long-term experiment in Zhejiang paddy soils showed that the soil organic matter could increase continuously with applying organic manure, and the increase in rate enhanced along with the application rates of organic manure. By mathematical modeling, the soil organic matter increased by 22 kg when 1 t of fresh FYM was applied. The CO2 emission resulting from the mineralization of soil organic matter increased with the increase in the application rate of the organic manure as well as the increase in the root residues. It is expected that the CO2 emission will be at 10.04-21.61 t ha^-1 yr^-1 when 16.5-49.5 t ha ^-1 yr^-1 of fresh FYM is applied. The soil organic carbon from mineralization and release of applied organic carbon （fresh FYM and root residues） will affect the CO2 concentration in the atmosphere. So, the higher the application rate of organic manure, the more is the fixed organic carbon. The CO2 fixation will be at 1.885-3.463 t ha^-1 yr^-1 when 16.5-49.5 t ha^-1 yr^-1 of fresh FYM is applied. Thus, the CO2 fixation will increase by 46.7 kg by applying 1 t fresh FYM. To apply organic manure continuously in rice fields may reduce the contribution to the increase of CO2 concentration in the atmosphere.     

Yield gap and production constraints of mango (Mangifera indica) cropping systems in Tianyang County,China

Mango is an important cash crop in the tropics and subtropics. Determining the yield gap of mango and production constraints can potentially promote the sustainable development of the mango industry. In this study, boundary line analysis based on survey data from 103 smallholder farmers and a yield gap model were used to determine the yield gap and production constraints in mango plantations in the northern mountain, central valley and southern mountains regions of Tianyang County, Guangxi, China. The results indicated that the yield of mango in three representing regions of Tianyang County,Northern Mountains, Central Valley and Southern Mountains, was 18.3, 17.0 and 15.4 t ha~(–1) yr~(–1), with an explainable yield gap of 10.9, 6.1 and 14.8 t ha~(–1) yr~(–1), respectively. Fertilization management, including fertilizer N, P_2O_5 and K_2O application rates, and planting density were the main limiting factors of mango yield in all three regions. In addition, tree age influenced mango yield in the Northern Mountains(11.1%) and Central Valley(11.7%) regions. Irrigation time influenced mango yield in the Northern Mountains(9.9%) and Southern Mountains(12.2%). Based on a scenario analysis, the predicted yield would increase by up to 50%, and fertilizer N use would be reduced by as much as approximately 20%. An improved understanding of production constraints will aid in the development of management strategy measures to increase mango yield.     

Responses of Ryegrass （Lolium perenne L.） Grown in Mudflats to Sewage Sludge Amendment

Sewage sludge amendment （SSA） is an alternative waste disposal technique and a potential way to increase fertility of mudflats for crop growth. The present study aimed to assess the suitability of SSA by assessing the nitrogen （N） and phosphorous （P） uptakes, heavy metal accumulation, growth, biomass, and yield response of ryegrass （Lolium perenne L.） at 0, 30, 75, 150, and 300 t ha-1 SSA rates at various growth stages. The results showed that the highest biomass ofryegrass at seedling and vegetative stages were at 300 and 150 t ha-1 SSA rate, respectively. The increments of ryegrass yield at reproductive stage at 30, 75, 150, and 300 t ha1 SSA rates were 98.0, 122.6, 88.1, and 61.2%, compared to unamended soil. N and P concentrations in ryegrass increased with increasing SSA rates at all stages except N and P in roots dropped significantly at 300 t ha-1 rate at vegetative stage. The metal concentration for Mn, Cu, Zn, Ni, Cd, Cr, and Pb in shoot of ryegrass at 300 t ha-1 S SA rate increased by 0.63-, 2.34-, 15.02-, 0.97-, 10.00-, 0.01- and 1.13-fold, respectively, compared to unamended soil. However, heavy metal concentrations in shoot of ryegrass were lower than the standard for forage products in China. The study suggested that sewage sludge amendment in mudflat soils might be feasible. However, the impacts of sludge application on edible crop plants and soil environment need further investigations.     

模拟氮沉降对贝加尔针茅草原土壤氮转化微生物的影响

草地土壤是温室气体重要的源和汇,认识草地生态系统氮转化过程有助于预测氮循环对未来氮沉降增加的响应与反馈机制。依托于2010年在内蒙古贝加尔针茅草原设置的长期模拟氮沉降增加的氮添加试验,共设置了8个氮添加水平(0、15、30、50、100、150、200、300 kg N·hm~(-2)·a~(-1))。应用荧光定量PCR方法,研究氮转化功能基因丰度对不同氮添加水平的响应。2015年8月取样分析结果表明:固氮微生物(nifH)基因丰度随着氮添加水平的升高,表现为先升高后降低的趋势。低于200 kg N·hm~(-2)·a~(-1)硝酸铵处理有利于固氮菌生长。低氮添加(N15、N30和N50)对氨氧化细菌(AOB-amoA)和氨氧化古菌(AOA-amoA)基因丰度无显著影响。高氮添加(N100、N150、N200和N300)显著提高了AOB基因丰度,降低了AOA基因丰度。高氮添加(N150、N200和N300)显著降低了nirK基因丰度。随着氮添加量的增加,高氮添加促进了AOB主导的氨氧化过程,而反硝化微生物丰度的减少提高了氨氧化产物硝酸盐的积累,继而提高了土壤硝酸盐含量。     

Phosphorus fertilization alters complexity of paddy soil dissolved organic matter

The structural complexity of soil dissolved organic matter(DOM) may reflect soil biogeochemical processes due to its spectral characteristics. However, the features of DOM structural complexity in paddy soil amended with long-term chemical P fertilization are still unclear, which may limit understanding of nutrient-related soil C cycle. We collected soil samples from field experiments receiving application of 0, 30, 60, and 90 kg P ha~(–1) yr~(–1) to assess the effect of exogenous P on the complexity of soil DOM structure. Three-dimensional excitation-emission matrix fluorescence analysis and enzymatic activity assay were used to determine the features of soil DOM molecular structure and the associated microbial reactions. The results showed that P input increased the biodegradability of DOM, indicating by the increased lower molecular weight components and decreased humic degree in the DOM. P input also reduced the structural complexity of DOM with blue shifts of fluorescent signals. The fluorescence index and β/α index of DOM increased with increasing P application by 4–5% and 3–11%, respectively, while humification index decreased by 8–13%. The P input increased the abundance of bacteria and fungi by 34–167% and 159–964%, respectively, while 29–54% increments were found for the β-1,4-glucosidase activities. These results implicated that P fertilization accelerated the soil DOM cycle, although the structural complexity of DOM declined, which potentially benefits soil C sequestration in paddy fields and may be a C sequestration mechanism in the P-dependent paddy.     

Trends of Yield and Soil Fertility in a Long-Term Wheat-Maize System

The sustainability of the wheat-maize rotation is important to China＇s food security. Intensive cropping without recycling crop residues or other organic inputs results in the loss of soil organic matter （SOM） and nutrients, and is assumed to be non- sustainable. We evaluated the effects of nine different treatments on yields, nitrogen use efficiency, P and K balances, and soil fertility in a wheat-maize rotation system （1991-2010） on silt clay loam in Shaanxi, China. The treatments involved the application of recommended dose of nitrogen （N）, nitrogen and phosphorus （NP）, nitrogen and potassium （NK）, phosphorus and potassium （PK）, combined NPK, wheat or maize straw （S） with NPK （SNPK）, or dairy manure （M） with NPK （M1NPK and M2NPK）, along with an un-treated control treatment （CK）. The mean yields of wheat and maize ranged from 992 and 2 235 kg ha-1 under CK to 5 962 and 6 894 kg ha-1 under M2NPK treatment, respectively. Treatments in which either N or P was omitted （N, NK and PK） gave significantly lower crop yields than those in which both were applied. The crop yields obtained under NP, NPK and SNPK treatments were statistically identical, as were those obtained under SNPK and MNPK. However, M2NPK gave a significant higher wheat yield than NP, and MNPK gave significant higher maize yield than both NP and NPK. Wheat yields increased significantly （by 86 to 155 kg ha-1 yr-1） in treatments where NP was applied, but maize yields did not. In general, the nitrogen use efficiency of wheat was the highest under the NP and NPK treatments; for maize, it was the highest under MNPK treatment. The P balance was highly positive under MNPK treatment, increasing by 136 to 213 kg ha-1 annually. While the K balance was negative in most treatments, ranging from 31 to 217 kg ha^-1 yr^-1, levels of soil available K remained unchanged or increased over the 20 yr. SOM levels increased significantly in all treatments. Overall, the results indicated that combinations of organic manure and inorganic nitrogen, or retuming straw with NP is likely to improve soil fertility, increasing the yields achievable with wheat-maize system in a way which is environmentally and agronomically beneficial on the tested soil.     

Variation of Potential Nitrification and Ammonia-Oxidizing Bacterial Community with Plant-Growing Period in Apple Orchard Soil

In this study, we investigated the potential nitrification and community structure of soil-based ammonia-oxidizing bacteria （AOB） in apple orchard soil during different growth periods and explored the effects of environmental factors on nitrification activity and AOB community composition in the soil of a Hanfu apple orchard, using a culture-dependent technique and denaturing gradient gel electrophoresis （DGGE）. We observed that nitrification activity and AOB abundance were the highest in November, lower in May, and the lowest in July. The results of statistical analysis indicated that total nitrogen （N） content, NH4＋-N content, NO3-N content, and pH showed significant correlations with AOB abundance and nitrification activity in soil. The Shannon-Winner diversity, as well as species richness and evenness indices （determined by PCR-DGGE banding patterns） in soil samples were the highest in September, but the lowest in July, when compared to additional sampled dates. The DGGE fingerprints of soil-based 16S rRNA genes in November were apparently distinct from those observed in May, July, and September, possessing the lowest species richness indices and the highest dominance indices among all four growth periods. Fourteen DGGE bands were excised for sequencing. The resulting analysis indicated that all AOB communities belonged to the 13-Proteobacteria phylum, with the dominant AOB showing high similarity to the Nitrosospira genus. Therefore, soil-based environmental factors, such as pH variation and content of NHa＋-N and NO3--N, can substantially influence the abundance of AOB communities in soil, and play a critical role in soil-based nitrification kinetics.     

Application of Chinese milk vetch affects rice yield and soil productivity in a subtropical double-rice cropping system

Green manure can be used as a substitute for chemical fertilizer without reducing rice yield. We studied the responses of soil fertility and rice yield to different combinations of Chinese milk vetch(CMV; Astragalus sinicus L.) and chemical fertilizer in a subtropical double-rice cropping system. Our goal is to reduce chemical fertilizer use and decrease environmental contamination. Compared with the recommended rate of chemical fertilizer(CF), both early-and late-rice yields in the two treatments supplied with 15 and 22.5 Mg CMV ha–1 plus 60% CF(represented as 60 A and 60 B, respectively) showed no significant differences while the two treatments supplied with 30 and 37.5 Mg CMV ha–1 plus 60% CF(represented as 60 C and 60 D, respectively) showed significantly higher values. The sustainable yield index(SYI) values in the 60 C and 60 D treatments with double-rice croppong system were significantly higher than those in other treatments(P0.05). Early-rice yield showed a significant positive relationship with the Chinese milk vetch incorporation rate. The coefficients increased annually from 2009 to 2013 and then decreased in 2014. Soil organic matter increased over time by the end of the experiment in all of the treatment groups. Soil organic matter in 60 A, 60 B and 60 C showed no significant difference compared with that in CF, while soil organic matter in 60 D was significantly higher than that in CF. The slopes of soil organic matter and total nitrogen over six years were the highest in 60 C and 60 D. The soil total nitrogen content in 60 A, 60 B, 60 C and 60 D was higher than that in CF, but the differences were not significant(P0.05). Therefore, a relatively high Chinese milk vetch incorporation rate(≥30 Mg ha–1) was more effective in improving the productivity and sustainability of paddy soil. The decreased coefficients of early-rice yield and the Chinese milk vetch incorporation rate in 2014 implied that the benefits of soil fertility and rice yield created by Chinese milk vetch input may decline after five years under a continuously high rate of Chinese milk vetch incorporation.     

Effects of reduced nitrogen and suitable soil moisture on wheat(Triticum aestivum L.) rhizosphere soil microbiological,biochemical properties and yield in the Huanghuai Plain,China

Soil management practices affect rhizosphere microorganisms and enzyme activities, which in turn influence soil ecosystem processes. The objective of this study was to explore the effects of different nitrogen application rates on wheat(Triticum aestivum L.) rhizosphere soil microorganisms and enzyme activities, and their temporal variations in relation to soil fertility under supplemental irrigation conditions in a fluvo-aquic region. For this, we established a split-plot experiment for two consecutive years(2014–2015 and 2015–2016) in the field with three levels of soil moisture: water deficit to no irrigation(W1), medium irrigation to(70±5)% of soil relative moisture after jointing stage(W2), and adequate irrigation to(80±5)% of soil relative moisture after jointing stage(W3); and three levels of nitrogen: 0 kg ha~(–1)(N1), 195 kg ha~(–1)(N2) and 270 kg ha~(–1)(N3). Results showed that irrigation and nitrogen application significantly increased rhizosphere microorganisms and enzyme activities. Soil microbiological properties showed different trends in response to N level; the highest values of bacteria, protease, catalase and phosphatase appeared in N2, while the highest levels of actinobacteria, fungi and urease were observed in N3. In addition, these items performed best under medium irrigation(W2) relative to W1 and W3; particularly the maximum microorganism(bacteria, actinobacteria and fungi) amounts appeared at W2, 5.37×10~7 and 6.35×10~7 CFUs g–1 higher than those at W3 in 2014–2015 and 2015–2016, respectively; and these changes were similar in both growing seasons. Microbe-related parameters fluctuated over time but their seasonality did not hamper the irrigation and fertilization-induced effects. Further, the highest grain yields of 13 309.2 and 12 885.7 kg ha~(–1) were both obtained at W2 N2 in 2014–2015 and 2015–2016, respectively. The selected properties, soil microorganisms and enzymes, were significantly correlated with wheat yield and proved to be valuable indicators of soil quality. These results clearly demonstrated that the combined treatment(W2 N2) significantly improved soil microbiological properties, soil fertility and wheat yield on the Huanghuai Plain, China.     

氮素水平对潮土氨氧化微生物和硝化作用的影响

为了明确不同氮素水平对土壤硝化作用和氨氧化微生物的影响,特别是高氮水平下氨氧化细菌(AOB)和氨氧化古菌(AOA)的响应特异性,以潮土为供试土壤,以尿素和硫酸铵作为氮源,设置5个氮素水平(以N计,分别为0、150、300、600、1 200 mg·kg^-1,相应地,记为N0、N150、N300、N600、N1200),进行28 d的微宇宙培养,研究不同氮素水平下尿素和硫酸铵对土壤AOB、AOA丰度和土壤硝化作用的影响。结果表明,添加氮肥显著(P<0.05)降低了土壤pH值,至培养结束时(28 d),土壤pH值随氮素水平增加而降低。培养结束时,各加氮处理的土壤NH⁺₄-N含量都处于较低水平(0.72～2.01 mg·kg^-1)。与N0处理相比,添加尿素或硫酸铵增加了AOB amoA基因拷贝数,且AOB amoA基因拷贝数随氮素水平增加而增加;但施用氮肥对AOA amoA基因拷贝数无显著影响。土壤平均净硝化速率随氮素水平的增加而增加,且同一氮素水平下,尿素处理的土壤平均净硝化速率高于硫酸铵处理。相...