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.     

渭北旱塬长期施肥试验中氨氧化细菌的多样性及群落结构分析

以中国科学院长武黄土高原农业生态试验站23 a长期施肥定位试验的土壤样品为研究对象,通过构建氨氧化细菌的amoA基因克隆文库,采用PCR-RFLP方法分析了裸地（LD）、种植不施肥（CK）、单施氮肥（N）、单施磷肥（P）和氮磷共施（NP）这5个处理条件下土壤中氨氧化细菌的多样性及其群落结构。结果表明,N处理的土壤中氨氧化细菌的Shannon-Wiene（rH′）和Margalef（dM）a指数均最高,其次是CK、NP、P,而LD处理中最低,表明长期单施氮肥后增加了土壤中氨氧化细菌的多样性和丰富度,长期种植作物后也同样会增加土壤中氨氧化细菌的多样性和丰富度,但单施磷肥和氮磷共施后土壤中氨氧化细菌的多样性和丰富度都有所降低。基于amoA基因建立的系统进化树显示,所有来自于各处理条件下土壤中氨氧化细菌的优势种群都是属于Nitrosospira和Nitrosospira-like,与Nitrosospira cluster 3聚为一组,但优势菌种在克隆文库中所占的比例不同,表明不同的施肥处理下土壤中氨氧化细菌的群落结构发生了改变。     

长期定位施肥对中性紫色土硝化作用及氨氧化微生物的影响

为了研究长期不同施肥措施对中性紫色土氨氧化微生物及其硝化作用的影响,以国家紫色土肥力与肥料效益监测基地的中性紫色土为研究对象,进行土壤氨氧化细菌和氨氧化古菌amo A基因的Real-time PCR分析,比较长期不同定位施肥对土壤氨氧化潜势和硝化强度的影响,并分析不同施肥制度对功能微生物丰度与功能的作用。数据显示,土壤中氨氧化古菌amo A基因拷贝数(Log值6.21～7.14)远大于氨氧化细菌(Log值3.65～5.73),相对于对氨氧化细菌丰度的影响,施肥对土壤氨氧化古菌丰度影响较小。施用氮肥与磷肥都显著提高了土壤氨氧化细菌丰度,1.5NPK+M处理氨氧化细菌丰度最高(Log值5.73),有机无机肥配施可以显著提高土壤氨氧化微生物丰度;而含氯化肥的施用在一定程度上降低了土壤氨氧化细菌丰度与硝化细菌生长,与施用不含氯的肥料处理相比,含氯肥料处理的土壤氨氧化细菌丰度与硝化细菌数分别降低了3.74%和88.12%。研究表明,长期施肥能影响中性紫色土中氨氧化细菌的丰度,有机无机肥配施能够提高土壤的氨氧化潜力与土壤的硝化能力。     

长期施肥对氨氧化古菌丰度及群落结构的影响

【目的】氨氧化古菌对土壤氮素转化有着重要的作用。本研究以长期定位施肥黄泥田土壤为研究对象探讨了长期不同施肥模式对土壤氨氧化古菌数量和多样性的影响,为制定合理的施肥制度提供理论基础。【方法】试验在福建省农科院试验站上进行,以30年长期定位施肥的红壤性水稻土为研究对象采用荧光定量PCR和克隆文库技术,研究了长期不同施肥模式对氨氧化古菌(ammonia-oxidizing archaea,AOA)丰度及群落结构的影响。试验设4个处理:1)不施肥(CK);2)单施氮磷钾肥(NPK);3)氮磷钾肥配施牛粪(NPKM);4)氮磷钾肥配施秸秆(NPKS)。小区面积为12 m~2每个处理设3个重复。土样采集时间为2012年10月份(水稻收获后),测定土壤养分和氨氧化古菌的数量及多样性。【结果】1)与CK相比,NPKM和NPKS处理显著增加了土壤有机质含量,NPKM和NPKS处理之间无显著差异。2)与CK相比施肥均能提高土壤全氮含量;NPKM和NPKS处理能够显著提高土壤全磷含量,NPKM处理全磷含量最高;仅NPKS处理能显著增加全钾含量。3)与CK相比,长期施肥均能提高土壤有效氮(AN)、速效磷(AP)和速效钾(AK)的含量,并且NPKM处理AN和AP含量最高,NPKS处理中AK含量最高。4)与CK相比,长期施肥均对土壤pH值无显著影响。5)与CK处理相比,NPKM和NPKS处理的amoA基因拷贝数显著增加增加幅度分别为168.4%和95.7%;单施化肥处理与CK无显著差异。土壤氨氧化古菌数量与土壤有机质含量呈显著正相关,与土壤全磷、有效氮、速效磷和速效钾含量呈极显著正相关。6)长期不同施肥处理影响土壤氨氧化古菌的种群结构,单施化肥增加了土壤AOA的多样性,而化肥配施有机肥则降低了AOA的多样性。7)本试验中得到的土壤氨氧化古菌amoA基因序列均为不可培养的古菌,包括泉古菌(Crenarchaeote)和奇古菌(Thaumarchaeote)。本试验所得氨氧化古菌绝大多数与来自土壤、沉积物的amoA基因克隆非常相似,少数与水体环境相似。【结论】不同培肥模式下,土壤中氨氧化古菌均为不可培养的古菌,包括泉古菌(Crenarchaeota)和奇古菌(Thaumarchaeota),然而水稻土壤养分和氨氧化古菌丰度及群落结构变化显著。单施化肥的作用不明显,有机无机肥配施有利于土壤有机质和养分的积累以及氨氧化古菌的生长增加了氨氧化古菌优势菌群的比例。     

长期施用氮肥和磷肥对渭北旱塬土壤中氨氧化古菌多样性的影响

采用基于氨单加氧酶基因的PCR-RFLP和DNA测序技术,以黄土高原旱地黑垆土为材料,研究长期施用氮肥和磷肥对土壤氨氧化古菌多样性的影响。结果显示,不同施肥处理土壤样品得到的氨氧化古菌的OTU数分别为25（种植不施肥,CK）、21（不施肥不种植,LD）、18（单施氮肥,N）、25（单施磷肥,P）和13（氮、磷共施,NP）。氨氧化古菌的多样性指数H′和优势度指数Ds变化趋势基本相同,分别为P〉CK〉LD〉NP〉N和P〉CK〉NP〉LD〉N;种群丰富度和均匀度指数在不同处理间变化较大,分别为CK〉P〉LD〉N〉NP和P〉NP〉LD〉N〉CK。各处理优势氨氧化古菌绝大部分属于Cluster S,少数属于Cluster M,获得的序列全部属于难培养泉古菌门。不同施肥方式的长期定位试验土壤中氨氧化古菌多样性变化较大,而优势氨氧化古菌系统进化定位没有显著变化。     

氮肥对盐渍化枸杞园土壤微生物群落和氨氧化微生物丰度的影响

研究主要分析氮元素对宁夏平罗盐渍化枸杞园土壤中氨氧化微生物(氨氧化细菌和氨氧化古菌)的影响。实验共设八个处理:(1)C(不施氮肥,不施脱硫废弃物,原始荒地);(2)不施氮肥(N0);(3)施氮肥25 kg hm~(-2)(N25);(4)施氮肥50 kg hm~(-2)(N50);(5)施氮肥100kg hm~(-2)(N100);(6)施氮肥200 kg hm~(-2)(N200);(7)施氮肥400 kg hm~(-2)(N400);(8)施氮肥800 kg hm~(-2)(N800),在N0-N800处理施用脱硫废弃物3.72×104kg hm~(-2)。2011年8月采集0~20 cm土样。结果显示:脱硫废弃物和氮肥配合施加对土壤理化性质产生了显著影响;NO_3~--N和NH_4~+-N含量在施氮处理中相对于原始样地和不施氮处理组都有显著的升高,微生物生物量和细菌和氨氧化细菌多样性指数在施氮400 kg hm~(-2)达到最大值,施氮肥400 kg hm~(-2)是促进微生物量和群落多样性增加的最佳施用量。实时荧光定量PCR结果显示:氨氧化细菌(AOB)丰度在施氮肥400 kg hm~(-2)和800 kg hm~(-2)显著高于其它处理,脱硫废弃物和氮肥配合施用对AOB的丰度具有叠加效应。相关性分析表明:NH_4~+-N与总磷脂脂肪酸(PLFA)、细菌PLFA、革兰氏阳性菌(G+)、革兰氏阴性菌(G-)、真菌/细菌(F/B)、微生物碳(MBC)、微生物氮(MBN)及16S r RNA基因拷贝数、AOB的基因拷贝数都显著相关。因此,铵态氮是该地区微生物群落可利用的有效氮素。     

不同施肥方式下土壤氨氧化细菌的群落特征

为了研究长期定位施肥对棕壤中氨氧化细菌（ammonia-oxidizing bacteria,AOB）种群结构多样性和垂直分布特征的影响,本研究采用化学分析、荧光定量PCR（qPCR）和变性梯度凝胶电泳（PCR-DGGE）技术,针对沈阳农业大学试验区不同施肥方式（不施肥、低量无机氮肥、高量无机氮肥、无机氮肥与有机肥配施）下不同土壤深度（0~20 cm、20~40 cm、40~60 cm）的土壤理化性质、AOB丰度及种群多样性进行分析,比较不同施肥方式对土壤AOB种群的影响。结果显示,与不施肥相比,施肥会降低土壤pH,增加土壤铵态氮（70.5%~939.21%）和硝态氮（253.20%~625.48%）含量。随土壤深度增加,土壤pH升高,铵态氮和硝态氮含量除低量无机氮肥处理外,多呈降低趋势。土壤增施氮肥可提高AOB丰度,降低总细菌丰度。其中,0~20 cm土层中AOB丰度较高,且高量无机氮肥处理的AOB数量最高,为9.65×10⁵拷贝数·g^-1（干土）。DGGE图谱分析显示,不同处理下,AOB群落结构多样性指数存在明显差异（P<0.05）,各多样性指数均在表层（0~20 cm）最高,增施氮肥则显著降低AOB的多样性。聚类分析表明,4个施肥处理中,高量无机氮肥处理聚为一类,其他处理则因土壤深度不同而异;3个土壤深度中,除不施肥处理外,所有施肥处理均表现为0~20 cm、20~40 cm土层发生聚类,40~60 cm则明显与其他两层分开。冗余梯度分析（RDA）显示,硝态氮（P=0.027）是造成影响AOB群落结构差异的主要原因。上述研究结果表明,长期定位施肥土壤AOB的数量和群落结构多样性受施肥方式显著影响,并表现出明显的垂直分布特征。与无机氮肥相比,有机无机配施处理有助于改善土壤pH,维持不同土壤深度下AOB群落结构多样性。     

长期轮作与施肥对土壤主要微生物类群的影响

根据长期田间试验,对施肥、作物、种植方式等对土壤细菌、真菌与放线菌含量的影响进行分析.结果显示施肥,特别是施有机肥明显提高土壤微生物数量,玉米、苜蓿与冬小麦连作不同施肥土壤细菌数量相对休闲地变幅在-197.04×105～+187.07×105个/g土,真菌在-6.08×103～+36.57×103个/g土,放线菌在-47.21×104～+301.4×104个/g土.轮作系统不同施肥处理真菌与放线菌数量高于休闲地.在施氮磷化肥条件下不同作物轮作,苜蓿与糜子对细菌有相对抑制作用,对真菌与放线菌有促进作用,红豆草抑制放线菌促进细菌,豌豆对三类微生物均有抑制作用.     

覆盖作物对猕猴桃园土壤氨氧化微生物丰度和群落结构的影响

[目的]果园行间播种覆盖作物有利于改善土壤理化性状,改变氮循环过程中的氨氧化微生物群落结构.因此,我们分析不同覆盖作物下猕猴桃园土壤氨氧化微生物的群落结构和基因丰度,以探究其与土壤硝化作用的关系.[方法]选择湖北十堰的一个5年生猕猴桃果园进行了种植多年生覆盖作物试验,供试覆盖作物有白三叶草(Trifolium repe...     

华北潮土长期施肥对土壤跳虫群落的影响

通过试验研究了河南封丘潮土长期施肥处理下土壤跳虫的群落结构及其多样性,试验包括氮磷钾(NPK)、氮磷(NP)、氮钾(NK)、磷钾(PK)、有机肥(OM)、1∶1化学氮肥与有机氮(MNPK)和不施肥(CK)7个处理。结果表明,7种施肥试验处理下,平均土壤跳虫密度为30 000头m-2,施用有机肥显著增加了土壤跳虫数量,而磷肥可能是该地区跳虫数量增长的一个限制因素。OM和MNPK处理下土壤跳虫多样性显著高于其他处理,OM和NPK处理具有最高的相似性,其次是MNPK和OM处理之间,而CK处理与其他处理相似性指数较小。不同施肥处理下表现了土壤跳虫类群对环境的选择性,多数跳虫与OM处理成正相关关系,而MNPK处理促进了Isotoma和Xenylla属在数量上的增长,PK处理与Hypogastrura和Sminthurinus属有一定的正相关关系。施肥处理也引起了土壤跳虫的垂直分布差异,MNPK、NPK和OM处理下土壤跳虫趋于表层(0～10cm)分布,而NK和CK处理下趋于下层(10～20cm)分布。土壤跳虫的量化指标(如个体数、类群数和多样性)对施肥处理表现敏感,可作为土壤质量评价的指标之一。     

Long-term fertilization effects on active ammonia oxidizers in an acidic upland soil in China

The effects of long-term fertilization of acidic soils on ammonia-oxidizing archaea (AOA) and bacteria (AOB) communities and its ecological implications remain poorly understood. We chose an acidic upland soil site under long-term (27-year) fertilization to investigate ammonia oxidizer communities under four different regimes: mineral N fertilizer (N), mineral NPK fertilizer (NPK), organic manure (OM) and an unfertilized control (CK). Soil net nitrification rates were significantly higher in OM soils than in CK, N or NPK soils. Quantitative analysis of the distribution of amoA genes by DNA-based stable isotope probing revealed that AOA dominate in CK, N and NPK soils, while AOB dominate in OM soils. Denaturing gradient gel electrophoresis and clone library analyses of amoA genes revealed that Group 1.1a-associated AOA (also referred to as Nitrosotalea) were the most dominant active AOA population (>92%), while Nitrosospira Cluster 3 and Cluster 9 were predominant among active AOB communities. The functional diversity of active ammonia oxidizers in acidic soils is affected by long-term fertilization practices, and the responses of active ammonia oxidizers to mineral fertilizer and organic manure are clearly different. Our results provide strong evidence that AOA are more highly adapted to growth at low pH and low substrate availability than AOB, and they suggest that the niche differentiation and metabolic diversity of ammonia oxidizers in acidic soils are more complex than previously thought.     

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.     

Linkage of N2O emissions to the abundance of soil ammonia oxidizers and denitrifiers in purple soil under long-term fertilization

Abstract

Microbial nitrification and denitrification are responsible for the majority of soil nitrous (N₂O) emissions. In this study, N₂O emissions were measured and the abundance of ammonium oxidizers and denitrifiers were quantified in purple soil in a long-term fertilization experiment to explore their relationships. The average N₂O fluxes and abundance of the amoAgene in ammonia-oxidizing bacteria during the observed dry season were highest when treated with mixed nitrogen, phosphorus and potassium fertilizer (NPK) and a single N treatment (N) using NH₄HCO₃as the sole N source; lower values were obtained using organic manure with pig slurry and added NPK at a ratio of 40%:60% (OMNPK),organic manure with pig slurry (OM) and returning crop straw residue plus synthetic NH₄HCO₃fertilizer at a ratio of 15%:85% (SRNPK). The lowest N₂O fluxes were observed in the treatment that used crop straw residue(SR) and in the control with no fertilizer (CK). Soil NH₄⁺provides the substrate for nitrification generating N₂O as a byproduct. The N₂O flux was significantly correlated with the abundance of the amoA gene in ammonia-oxidizing bacteria (r = 0.984, p < 0.001), which was the main driver of nitrification. During the wet season, soil nitrate (NO₃^?) and soil organic matter (SOC) were found positively correlated with N₂O emissions (r = 0.774, p = 0.041 and r = 0.827, p = 0.015, respectively). The nirS gene showed a similar trend with N₂O fluxes. These results show the relationship between the abundance of soil microbes and N₂O emissions and suggest that N₂O emissions during the dry season were due to nitrification, whereas in wet season, denitrification might dominate N₂O emission.     

Aggregate-related changes in network patterns of nematodes and ammonia oxidizers in an acidic soil

Nitrification plays a central role in global nitrogen cycle, which is affected by biological interaction between soil microfauna and microorganisms. However, the complexity of soil biotic communities made it difficult to reveal organizational principles of the community and the interactions among species. Here, we used the network analysis to decipher the interactions between nematodes and ammonia oxidizers within aggregate fractions under 10-year manure application, and examine their associations with soil variables and potential nitrification activity (PNA). Three aggregate fractions included large macroaggregates (>2000 μm, LA), small macroaggregates (250–2000 μm, SA), and inter-aggregate soil and space (<250 μm, IA). Aggregate factions showed a remarkable effect on association networks of nematodes and ammonia oxidizers. The average connectivity (avgK) and the number of edges in overall networks increased with increasing aggregate sizes, while the average geodesic distance (GD) followed the opposite trend. The LA network could be viewed as a better organized or a better operational soil food web with more functional interrelated members than the SA and IA networks. The modules related to PNA were significantly correlated and clustered together as meta-modules in networks of aggregate fractions. The role-shifts prevailed among the network members such as significant module memberships (MMs) and generalist/specialist operational taxonomic units (OTUs). A half of shared nodes were further identified as shared MMs, dominated by ammonia-oxidizing bacteria (AOB) especially for Nitrosospira cluster 3a and 10. Soil pH could explain partly the shift of module hubs in different networks, while grazing by bacterivores might account for three exclusively connecters related to Nitrososphaera clusters 1.1. The strongly coupled modules correlated positively to pH and total carbon (TC), regardless of aggregate fractions. The network analysis approach provided new insights into potential importance of network interactions between nematodes and ammonia oxidizers in soil nitrogen cycling.     

Long-term greenhouse vegetable cultivation alters the community structures of soil ammonia oxidizers

Journal of Soils and Sediments - Continuous cropping obstacles derived from long-term intensive cultivation has severely affected the healthy development of greenhouse vegetable industry in China....     

长期施肥对土壤肥力的影响

基于长期试验资料,从土壤肥力的角度综述了长期施肥对土壤肥力指标有机质、N素、P素和K素含量,微生物生物量及数量和土壤酶活性的影响,指出长期施用有机肥及有机肥与化肥配施是维持和提高土壤肥力的关键,可促进农田生态系统可持续发展。     

Effect of agricultural land use change on community composition of bacteria and ammonia oxidizers

Purpose

Soil microbial communities can be strongly influenced by agricultural practices, but little is known about bacterial community successions as land use changes. The objective of this study was to determine microbial community shifts following major land use changes in order to improve our understanding of land use impacts on microbial community composition and functions.

Materials and methods

Four agricultural land use patterns were selected for the study, including old rice paddy fields (ORP), Magnolia nursery planting (MNP), short-term vegetable (STV), and long-term vegetable (LTV) cultivation. All four systems are located in the same region with same soil parent material (alluvium), and the MNP, STV, and LTV systems had been converted from ORP for 10, 3, and 30 years, respectively. Soil bacteria and ammonia oxidizer community compositions were analyzed by 454 pyrosequencing and terminal restriction fragment length polymorphism, respectively. Quantitative PCR was used to determine 16S rRNA and amoA gene copy numbers.

Results and discussion

The results showed that when land use was changed from rice paddy to upland systems, the relative abundance of Chloroflexi increased whereas Acidobacteria decreased significantly. While LTV induced significant shifts of bacterial composition, MNP had the highest relative abundance of genera GP1, GP2, and GP3, which were mainly related to the development of soil acidity. The community composition of ammonia-oxidizing bacteria (AOB) but not ammonia-oxidizing archaea was strongly impacted by the agricultural land use patterns, with LTV inducing the growth of a single super predominant AOB group. The land use changes also induced significant shifts in the abundance of 16S rRNA and bacterial amoA genes, but no significant differences in the abundance of archaea amoA was detected among the four land use patterns. Soil total phosphorous, available phosphorous, NO₃ ^?, and soil organic carbon contents and pH were the main determinants in driving the composition of both bacteria and AOB communities.

Conclusions

These results clearly show the significant impact of land use change on soil microbial community composition and abundance and this will have major implications on the microbial ecology and nutrient cycling in these systems, some of which is unknown. Further research should be directed to studying the impacts of these microbial community shifts on nutrient dynamics in these agroecosystems so that improved nutrient management systems can be developed.     

长期施肥对农田黑土微生物活力与群落结构的影响

以黑土长期肥料定位站CK、N、P、NP、MN(有机肥+氮)、MP(有机肥+磷)、MNP(有机肥+氮+磷)等处理为供试土样,研究氮、磷及有机肥不同配施条件下土壤基本理化性质、磷脂脂肪酸(PLFA)、中性磷脂脂肪酸(NLFA)、酸(碱)性磷酸酶、微生物量碳或氮(SMB-C或N)等变化规律。结果表明,26年有机肥-化肥配施明显提高土壤有机质(OM)、总氮(TN)以及速效氮、磷、钾等养分含量,其SMB-C或N、SMB-C/OM比值、磷酸酶活力以及各菌群PLFA含量显著高于CK和各化肥处理。长期N处理微生物活力明显低于CK,而P处理对微生物活力与结构影响最小。NLFA在各菌体相对含量与活体生物标识物变化趋势相反,表明养分充足的有机肥处理中各菌群休眠体或贮存物质积累有限。PLFA、NLFA与NLFA/PLFA主成分分析表明,有机肥与化肥处理微生物群落结构显著不同。个别PLFA或NLFA载荷值分析表明,真菌C18∶2ω6,9受有机肥施用影响显著;而G+菌iC15∶0、iC17∶0、iC16∶0等更易受化肥施用影响。与PLFA或NLFA相比,NLFA/PLFA比值PC分析能更有效地区分不同施肥处理。