长期不同施肥条件下红壤性水稻土微生物群落结构的变化

以位于江西省红壤研究所内长期定位试验的水稻土(始于1981年)为研究对象,运用磷脂脂肪酸(phospholipid fatty acid,PLFA)和BIOLOG分析技术研究了不施肥(CK)、单施化肥(NPK)及有机肥与化肥混施(NPKM)三种施肥方式对土壤微生物群落结构的影响。结果表明:长期施化肥和有机肥与化肥混施处理的PLFA总量均高于未施肥处理,两者分别较未施肥处理高91%和309%;PLFA主成分分析(PCA)显示施肥促进了土壤微生物群落结构的变化,其中NPKM处理增加了革兰氏阴性细菌(G-细菌)、真菌、放线菌和原生动物的数量,NPK处理增加了革兰氏阳性细菌(G+细菌)的数量,不施肥处理较施肥处理提高了真菌/细菌比例,CK和NPK处理的微生物群落结构更为相似;各施肥处理间土壤的AWCD值(平均每孔颜色变化率,average well color development,AWCD)表明,NPKM处理能够促进土壤微生物群落对碳源的利用能力,进而增加土壤中微生物的整体活性,而NPK处理减弱了土壤微生物的活性。代谢功能多样性分析同时表明,NPKM处理增加了微生物群落的多样性,而NPK处理使土壤微生物的多样性降低;土壤PLFA与土壤养分的相关性分析显示,土壤总PLFA量与土壤有机质和全氮呈极显著相关(p0.01),与速效养分相关性不大。     

太湖地区长期不同施肥水稻土N2和CO2固定细菌群落结构的特征与差异

土壤N2固定细菌和CO2固定细菌是土壤碳氮循环重要的微生物群落,不同施肥下土壤有机碳积累与这两类微生物群落结构及活性的关系有助于了解施肥对农田碳氮循环的影响特点.本文研究了太湖地区自1987年开始的水稻土长期施肥试验,选取不施肥(NF)、氮磷钾(NPK)肥(CF)、NPK肥与猪粪配施(CFM)和NPK肥与秸秆还田配施(CFS)小区,采集(0-20 cm)表层样本,以nifH和cbbLR基因分别作为N2和CO2固定细菌的指示基因,用PCR-DGGE和荧光定量PCR的方法研究二者的群落结构,并用平板菌落计数法测定土壤自生固氮细菌数量和用乙炔还原法测定土壤固氮酶活性.结果表明,与CF处理相比,CF、CFM和CFS处理下的自生固氮细菌数量分别提高了58％、66％和106％;CF、CFM和CFS处理下的nifH基因丰度分别提高了213％、1079％和344％.CF与CFM处理的土壤固氮酶活性显著高于NF和CFS处理.因此,施肥提高了cbbLR基因的多样性.与NF处理相比,CF、CFM和CFS处理下的cbbLR基因丰度分别提高了465％、1827％和758％.相关性分析表明,土壤自生固氮菌数量与土壤有机碳含量呈显著正相关,cbbLR和nifH基因丰度均与归一化的土壤养分呈极其显著正相关.研究表明,有机/无机肥配施下土壤养分平衡对维持N2和CO2固定细菌较高的丰度具有重要的作用.     

长期不同施肥对塿土大团聚体中有机碳组分特征的影响

【目的】研究长期施肥对土大团聚体中有机碳组分特征的影响,揭示不同施肥方式下土壤有机碳的固持机制,为合理施肥提供理论依据。【方法】采集土35年长期肥料定位试验不同施肥处理0—10 cm和10—20 cm土样,分析其大团聚体中各组分有机碳含量的变化。试验处理为:不施肥(CK)、单施化肥(NP)、单施有机肥(M)和有机肥配施化肥(MNP)。【结果】与CK相比,长期NP处理对大团聚体中粗颗粒有机碳(cPOC)、细颗粒有机碳(fPOC)、大团聚体中微团聚体内颗粒有机碳(iPOC)以及矿质结合态有机碳(MOC)组分的有机碳(OC)含量均无显著影响;而M处理以及MNP处理可显著提高两土层cPOC和iPOC组分的OC含量以及0—10 cm土层MOC组分的OC含量,其中,cPOC含量增幅分别为174%~338%和215%~245%,iPOC含量增幅分别为127%~241%和106%~130%,MOC含量增幅达28.9%~34.6%。MNP处理显著提高了0—10 cm土层fPOC组分的OC含量,增幅达482.1%。累积碳投入量与大团聚体中各组分的OC含量呈显著线性相关,尤其是iPOC含量,表明长期施肥过程中土有机碳在大团聚体中固存的差异主要受物理保护的颗粒有机碳组分的影响。【结论】关中地区土长期施化肥对大团聚体中各组分OC含量没有显著影响,而长期单施有机肥能进一步增加大团聚体中各组分OC含量,有机肥配施化肥能显著增加团聚体中各组分OC含量,特别是大团聚体中微团聚体内颗粒有机碳组分的含量,进而增加土的有机碳固持。因此,有机肥配施化肥是提高土有机碳含量的有效措施。     

长期不同施肥措施下■土细菌群落结构变化及其主要影响因素

【目的】研究不同施肥处理下土壤细菌群落的特征,为建立促进土壤生态系统稳定和健康的养分管理制度提供依据。【方法】陕西省杨凌示范区"国家黄土肥力与肥料效益监测基地"的28年长期定位试验始于1990年秋,种植制度为冬小麦–夏休闲,无灌溉。本研究选取定位试验中不施肥(CK)、施用氮磷钾肥(N、P_2O_5、K_2O分别为135、108、67.5 kg/hm~2,NPK)和有机无机肥配施(70%N来自牛粪,MNPK) 3个处理。于2018年6月小麦收获后采集0—20 cm耕层土样,测定养分含量、含水量、微生物量碳含量、微生物量氮含量及目标土壤微生物。以1%琼脂糖凝胶电泳法检测土壤中DNA,根据97%相似度对序列进行OTU聚类、α多样性分析(包括Shannon、ACE和Chao1等指数),使用CANOCO 4.5软件对土壤细菌门水平群落结构、细菌相对丰度等与土壤理化性质进行冗余分析。【结果】与CK相比,NPK和MNPK处理显著提高了土壤有机碳、全氮、微生物量碳、微生物量氮、硝态氮和铵态氮含量,显著降低了土壤pH值。不同处理细菌基因拷贝数为每克干土6.69×10~9～16.46×10~9,与CK相比,NPK和MNPK处理细菌数量分别提高了77%和146%。MNPK处理的土壤细菌Shannon多样性指数显著高于CK和NPK处理,而Simpson指数显著低于CK和NPK处理,NPK与CK处理间两个指数无显著差异。3个处理的细菌丰富度指数(Chao1指数和ACE指数)和均匀度指数均没有显著差异。在门水平上,共获得35个细菌类群,其中,放线菌门(Actinobacteria)、变形菌门(Proteobacteria)、酸杆菌门(Acidobacteria)和绿弯菌门(Chloroflexi)为主要优势菌门(相对丰度 10%),占到全部菌门的80.1%～81.7%。与CK相比,MNPK处理显著降低了放线菌门(F=5.845,P 0.05)的相对丰度,增加了拟杆菌门(F=4.461,P 0.05)的相对丰度,3个处理间其他菌门均无显著差异。冗余分析结果显示,CK与NPK、MNPK处理的土壤细菌群落结构具有明显差异,且MNPK处理对土壤细菌群落组成的影响更大。土壤理化性质对细菌菌群影响表现为:土壤硝态氮可溶性有机碳 pH铵态氮有机碳土壤含水量,这些理化因子均是影响微生物生长的关键因子。【结论】关中■土区旱作雨养条件下,化肥平衡施用和有机无机肥配施均显著提高了土壤中细菌数量、多样性和丰富度,有机无机肥配施还改变了细菌群落结构,特别是降低了放线菌门、增加了拟杆菌门的丰度,更有利于土壤生态系统的稳定和健康。     

不同施肥年限对红壤细菌多样性及群落结构演替的影响

为探讨不同年限施肥处理对旱地红壤细菌多样性和群落结构演替的影响,基于中国科学院红壤生态实验站设置的不同秸秆还田方式长期定位试验,采集种植玉米后第1年(Y2011)、第3年(Y2013)和第7年(Y2017)的土样进行分析。试验处理分为不施肥(CK)、化肥(N)和化肥+秸秆猪粪配施(NSM)3个处理,通过高通量测序技术研究旱地红壤细菌多样性和群落结构差异。结果表明:(1)施肥处理显著提高了土壤有机碳、全氮、全磷和有效磷养分含量,NSM处理对土壤肥力的提升效果比N处理好,且随着施肥年限的延长,提升效果越显著;(2)与Y2011相比,Y2013和Y2017下CK、N和NSM处理的细菌α多样性均显著提高,且N和NSM处理细菌多样性显著高于CK处理;(3)主坐标分析和聚类分析表明,土壤细菌群落主要通过施肥年限聚类在一起,但同一施肥年限下不同施肥处理之间细菌群落结构没有显著差异;(4)土壤全磷是驱动细菌多样性和群落结构变异的最关键因子。本研究从细菌多样性增加的角度,为探索有机培肥下红壤肥力提升和生态系统健康管护提供了科学依据。     

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。     

潮土长期不同施氮水平对秸秆降解及其细菌群落结构的影响

农田过量施氮所引发的问题已经引起广泛关注,但长期施氮后土壤无机氮水平状况及其对秸秆降解的作用尚不清楚。本研究以中国科学院封丘农业生态实验站长期（2005～2018年）施氮肥（5个施氮水平：0(N0)、150(N1)、190(N2)、,230(N3)和270 (N4)kg ha-1 yr-1）的潮土为研究对象,开展短期（50天）的秸秆降解-土壤培育实验。培育期间监测秸秆碳的矿化、土壤无机氮（硝态氮和铵态氮）、可溶性有机碳、微生物生物量碳的动态变化,利用高通量测序测定细菌群落结构。结果表明,长期施氮后,土壤无机氮含量和秸秆碳的矿化率随施氮水平的升高而增加。不同长期施氮水平的土壤细菌群落结构呈现显著差异。网络分析揭示：秸秆降解过程中细菌群落内部物种间的共现模式随长期施氮水平发生改变,具体体现为长期高施氮水平下细菌群落彼此间的负相关得到了加强;同时,变形菌主导地位减弱、酸杆菌主导地位增强。综上,土壤无机氮含量、细菌群落结构及物种之间的关系随着长期施氮水平的不同发生了改变。本研究探究了长期不同施氮水平下土壤中无机氮的水平状况、秸秆降解状况以及秸秆降解过程中土壤细菌的生物特性,以期为秸秆还田和科学施氮提供一定的数据支撑和思路启示。     

长期不同施肥下太湖地区黄泥土肥力的变化

对太湖地区典型水稻土———黄泥土进行了12年的长期定位不同施肥试验,并保持化肥的施用水平与本地区常规施用量相当。对12年来的肥力变化与不同施肥小区的肥力差异的分析研究表明,长期单施化肥下土壤耕层中养分都有所下降,且表层土壤养分的下降量不及亚表层土壤,年际间生产力不稳定。化肥配施有机肥(秸秆还田或猪粪),可长期保持土壤中的养分,稳定提高作物生产力;有机无机配合施肥下12年间全碳和全氮分别增加1.1g/kg和3.0g/kg,而全磷提高了0.3g/kg以上。同时也表明,表层土壤全氮的增加明显地促进了有机碳的固定。     

不同施肥水稻土可溶性有机氮组分差异及影响因素

可溶性有机氮(Soluble Organic Nitrogen,SON)在研究土壤供氮能力方面至关重要,而目前关于水田生态系统SON组成及其影响因素尚不清楚。运用红外光谱和氨基酸自动分析仪研究了长期(33年)不同施肥处理对水稻土SON含量、组分的影响及其主控因子。结果表明:单施化肥、化肥+牛粪、化肥+稻草处理土壤SON较不施肥处理提高23.49%、58.70%和106.30%,游离氨基酸氮提高32.27%、84.42%和95.21%,可溶性蛋白氮提高20.83%、70.00%和95.83%;不同施肥处理土壤游离氨基酸组成均以中性氨基酸占优势;土壤可溶性氮官能团组成均具有酰胺类化合物的红外特征吸收峰;RDA结果表明,土壤SON含量和组成差异明显,CK和NPK处理主要受土壤容重影响,而NPKM和NPKS则主要受有机质、全氮、微生物量氮、蛋白酶和谷氨酰胺酶影响。因此,SON含量和组分变化与施肥处理密切相关,长期添加有机肥能够显著增加土壤SON及其组分的含量,提高土壤供氮潜力。     

长期不同施肥和土地利用方式对塿土耕层碳储量的影响

利用土20年长期肥料定位试验研究了不同土地利用方式和施肥对土壤有机碳和无机碳储量变化的影响。试验包括休闲（Fallow, FL）、 撂荒（Setaside, SL）、 不施肥（CK）、 单施氮（N）、 氮钾（NK）、 磷钾（PK）、 氮磷（NP）、 氮磷钾（NPK）、 氮磷钾配合秸秆还田（SNPK）、 氮磷钾配合低量有机肥（M1NPK）和氮磷钾配合高量有机肥（M2NPK）11个处理。结果表明,CK和 FL 处理等质量耕层土壤有机碳储量仍维持在试验前水平,NP和 SL 处理显著提高了耕层土壤有机碳储量,年均增加分别达到 347 kg/hm2 和518 kg/hm2, 此4个处理等质量耕层土壤无机碳储量均较试验开始前（Initial soil, IniS）显著下降,尤其是NP处理显著低于其它3个处理。与IniS和CK相比,除NK处理外的所有施肥处理均显著地提高了等质量耕层土壤有机碳储量,其大小顺序为 M2NPKM1NPKSNPKNPKNPPKN,最大年均增加量为M2NPK 944 kg/hm2,最小为N 127 kg/hm2。施肥处理除PK和M2NPK处理外,其它处理等质量耕层土壤无机碳储量均较试验前明显降低,可能是由于土壤酸化所致。PK和M2NPK处理无机碳储量能够维持不下降,表明土壤无机碳和有机碳在适合条件下可能有某种关系。试验结果还显示,长期试验20年除M1NPK和M2NPK处理外,其它处理耕层土壤容重均明显高于试验开始前,表明等质量土壤碳储量与等深度碳储量相比可以更好地反映土壤碳的变化。     

长期定位施肥下黑土呼吸的变化特征及其影响因素

阐明长期不同施肥下的土壤呼吸特征及其影响机制对黑土区固碳减排研究至关重要。该研究基于1990年开始的国家土壤肥力与肥料效益监测网站-吉林省公主岭市黑土监测基地,选取不施肥(CK)、单施氮磷钾肥(NPK)、无机肥配施低量有机肥(NPKM1)、1.5倍的无机肥配施低量有机肥(1.5(NPKM1))、无机肥配施高量有机肥(NPKM2)和无机肥配施秸秆(NPKS)6个处理,明确了长期不同施肥下土壤总呼吸和异养呼吸的季节变化特征,并分析了土壤温度、水分、微生物量碳氮、铵态氮、硝态氮与土壤呼吸和异养呼吸的关系。结果表明:长期有机无机肥配施可以显著提高土壤有机碳、全氮、土壤速效磷、有效钾的含量和土壤活性有机碳库组分含量(P0.05);与不施肥相比,长期有机无机肥配施和无机配施秸秆处理分别显著增加土壤呼吸及异养呼吸碳累积排放量56.32%~86.54%和70.01%~100.93%;根系呼吸对土壤呼吸的整体贡献为23.68%~34.30%;相关分析表明,土壤呼吸速率和异养呼吸速率与土壤温度极显著正相关(P0.01),与土壤含水率呈显著负相关(P0.01),土壤温度可以分别解释土壤呼吸和异养呼吸变化的42.79%和39.61%;土壤微生物量碳氮、土壤硝态氮均与土壤呼吸速率和异养呼吸速率极显著相关(P0.01),土壤微生物量碳氮、土壤硝态氮可以分别解释土壤呼吸和异养呼吸变化的78.42%和77.18%,58.33%和56.79%,59.29%和59.14%;土壤铵态氮虽然显著影响土壤呼吸速率(P0.05),可以解释土壤呼吸变化的5.56%,但其对异养呼吸速率的影响不显著。综合来看,微生物量碳对土壤呼吸及异养呼吸的影响最大,而土壤含水率(15%)越高则土壤呼吸越弱;无机配施秸秆处理可以提高土壤碳库组分含量,且作物生育期内土壤呼吸及异养呼吸碳累积释放量均低于等氮量下施用有机肥(NPKM1)的处理,为最佳的农田管理措施。     

长期施肥下褐土生产力的演变及其影响因素

【目的】 土壤生产力提升和稳定是农业可持续发展的基础。研究过去30年间常规施肥条件下,褐土生产力的变化,厘清影响褐土生产力水平的主要因素,为褐土的培肥改良和生产力提升提供理论依据。 【方法】 基于全国28个长期定位试验点的褐土定位试验,分析了29年 (1988—2016) 不施肥和常规施肥两个处理的土壤肥力指标,并运用主成分分析方法分析了地力和施肥因素对产量的影响。 【结果】 1) 不施肥处理小麦和玉米产量均为先降低后缓慢升高,均值分别为3175和4056 kg/hm2;常规施肥处理小麦产量逐渐升高,玉米产量趋于平稳,均值分别为6124和7432 kg/hm2;小麦和玉米的增产量先升高后降低,均值分别为2901和3429 kg/hm2。施肥提高了作物产量的可持续性指数 (SYI),小麦和玉米SYI值分别为0.57和0.54,分别提高了54.82%和52.49%;降低了变异系数 (CV),小麦和玉米分别降低了44.70%和40.77%。2) 褐土区小麦季和玉米季地力贡献率分别为53.0%和54.2%,变异系数分别为41.2%和37.6%。肥料总量的农学效率,小麦先升高后降低,均值为6.36 kg/kg,玉米先降低后趋于平稳,均值为13.89 kg/kg。氮肥农学效率随施肥年数增加而降低。3) 主成分分析结果表明,影响小麦产量的5个主成分的累积贡献率为71.729%,影响玉米产量的4个主成分的累积贡献率为67.948%,对于两种作物,有机肥用量的影响最大,其次是土壤全氮和有机质含量。 【结论】 褐土区土壤生产力受有机肥用量和土壤全氮、有机质含量的影响最大,因此褐土区生产力的提高与地力的提升和肥料的科学施用密切相关。地力提升可以采取秸秆还田、施用有机肥等措施;肥料施用需要适当提高磷肥的比例,降低氮肥的比例,同时适当提高有机肥的比例。      

Abundance and community structure of sulfate reducing prokaryotes in a paddy soil of southern China under different fertilization regimes

Flooded rice paddy soils represent a typical anaerobic freshwater habitat of microorganisms. The abundance and community structure of sulfate reducing prokaryotes (SRP) were investigated in order to understand their response to different fertilization practices in rice paddy, including control without fertilizers (CT) and arrangements of different chemical fertilizers of nitrogen (N), phosphorus (P) and potassium (K): N, NP, NK and NPK. The abundance of total bacteria and sulfate reducing prokaryotes of the rice paddy in summer and in winter were quantified by real-time PCR assays based on the 16S rRNA gene and the dissimilatory (bi)sulfite reductase gene (dsrAB) β-subunit. No significant differences in the bacterial and SRP abundance were observed among different fertilization treatments in both winter and summer. The mean copy numbers of bacteria was 7.26 × 10⁹ copies g⁻¹ dry soil in winter and 1.27 × 10¹⁰ copies g⁻¹ dry soil in summer. The average dsrAB gene copy numbers of the SRP was 5.08 × 10⁸ copies g⁻¹ dry soil in winter and 5.92 × 10⁸ copies g⁻¹ dry soil in summer. The dsrAB gene clone libraries of the five fertilization treatments were constructed and their RFLP analysis yielded 22-25 restriction patterns, suggesting a high degree of dsrAB sequence diversity in different fertilization treatments. There was no significant change in the soil SRP community structure among the different fertilization regimes. More than half of the sequences were affiliated with novel branching clusters which were uncultured SRP. Clostridia and Deltaproteobacteria were also found with a high proportion in the clone libraries, while Desulfovibrionaceae was absent. High proportion of novel uncultured SRP implies that they may play important roles in paddy soils and deserve further studies.     

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)     

不同施肥处理对农田土壤微生物区系和功能的影响

本研究采用DGGE和Biolog两种方法, 研究了五种不同施肥处理对土壤微生物区系组成和功能的影响。采用UPGMA对DGGE试验结果进行聚类分析得出,小麦季中不施肥对照（T1）和常规氮磷钾肥（T2）聚为一类,相似性为41%,常规氮磷钾肥+秸秆还田（T4）和常规氮磷钾肥+秸秆还田+秸秆腐熟剂（T5）聚为一类,相似性为52%,而70%常规氮磷钾肥+有机肥（T3）单独聚为一类;玉米季也得到类似的结果,只是T1和T2的相似性为68%,T4和T5相似性达78%,而T3依然归为一类,这说明不同施肥处理间土壤微生物区系存在相似性。经过切胶测序及BLAST比对,发现大多为不可培养细菌,可培养细菌中多为芽孢杆菌属和类芽孢杆菌属。Biolog试验采用主成份分析得出,T1和T2土壤微生物功能类似,T4和T5土壤微生物也具有相似的功能,而T3则单独分为一类。其中,T5和T4的土壤细菌群落对底物碳源的代谢活性最强,T3处理次之,T2和T1最低。通过这两种不同试验方法的分析可以看出,不同施肥处理对土壤微生物区系和功能的影响存在关联性。     

塿土区长期施肥农田土壤的可持续性评价

【目的】施肥是实现作物增产和土壤培肥的主要途径,综合评价长期不同施肥条件下农田土壤的可持续性.可为农田合理施肥和管理提供理论支撑。【方法】依托陕西杨凌"国家黄土肥力与肥料效益监测基地"长期肥料定位试验(23年)采集分析了该试验9个处理[不施肥(CK)、单施氮(N)、氮钾配合(NK)、磷钾配合(PK)、氮磷配合(NP)、氮磷钾配合(NPK)、氮磷钾配合秸秆还田(SNPK)、低量有机肥配合氮磷钾(M_1NPK)和高量有机肥配合氮磷钾(M_2NPK]的土壤养分、微生物特性及作物产量,计算了土壤养分指数、微生物指数、作物指数和可持续性指数.综合评价了长期不同施肥处理对(?)土区农田土壤可持续性的影响。【结果】CK和N处理土壤养分指数分别为0.75和0.87,两者之间差异显著(P0.05)均低于土壤养分指数临界值1.00;其余处理的土壤养分指数在1.12~3.12之间均大于临界值,其中NP和NK差异不显著,SNPK和PK差异不显著,其余各处理间均差异显著(P0.05)。CK、N、NK和PK处理土壤微生物指数在0.74~0.84之间低于土壤微生物指数临界值1.00,其中CK和N差异不显著,NK和PK差异不显著但两组之间差异显著(P0.05);NP、NPK、SNPK、M、NPK和M,NPK处理的在0.98~1.21之间,接近或大于临界值,其中M、NPK和M,NPK差异不显著,NP、NPK和SNPK差异不显著,两组之间差异显著(P0.05)。CK、N、NK和PK的作物指数在0.52~0.60之间均低于作物指数临界值1.00,仅PK和N差异显著(P0.05);氮磷配合的施肥处理(NP、NPK、SNPK、M_1NPK和M_2NPK)在0.98~1.02之间,各处理间差异不显著但均接近或大于临界值1.00。CK、N、NK和PK的可持续性指数分别为0.49、0.54、0.74和0.99均低于可持续性指数临界值1.30,其中CK和N差异不显著;NP和NPK处理的可持续性指数分别为1.18和1.26,两者差异不显著,接近于可持续临界值;有机无机配合的处理(SNPK、M、NPK和M,NPK)的可持续性指数差异显著(P0.05)分别为1.55、2.32和2.94,较临界值高19.5%、78.8%和126.3%。土壤养分指数的变异性(50.2%)大于作物指数(29.1%)和微生物指数(18.4%)的变异性综合计算的可持续性指数的变异最高(62.1%)。【结论】冬小麦一夏玉米轮作条件下,长期不施肥和偏施化肥缕土区农田生产不可持续,化肥平衡施用(NP和NPK)基本可持续,NPK化肥配合秸秆还田(SNPK)和适量有机无机配施(M_1NPK)是实现(?)土可持续利用的施肥模式。     

长期不同施肥黑土细菌和古菌群落结构及主效影响因子分析

【目的】依据黑龙江省农科院34年的长期定位试验,探讨影响东北黑土细菌和古菌区系的主效环境因子。【方法】试验采集不施肥(CK)、施氮磷肥(NP)、施氮钾肥(NK)、施磷钾肥(PK)、施氮磷钾肥(NPK)5个施肥处理的耕层土样借助传统化学分析方法和Illumina Miseq高通量测序技术解析土壤化学性状和细菌与古菌的群落结构特征,并对细菌和古菌群落结构与环境因子进行相关性分析。【结果】1)不同施肥处理土壤细菌和古菌群落结构存在显著差异与CK相比,4种施肥处理均降低了古菌丰富度,NPK处理增加了土壤细菌的丰富度、多样性以及古菌的多样性;变形菌门(Proteobacteria)、酸杆菌门(Acidobacteria)是所有土样中的优势菌群,占细菌与古菌总量的38.2%~42.9%。施肥条件下,放线菌门(Actinobacteria)、浮霉菌门(Planctomycetes)、绿弯菌门(Chloroflexi)、蓝藻门(Cyanobacteria)的相对丰度降低芽单胞菌门(Gemmatimonadetes)、厚壁菌门(Firmicutes)的相对丰度升高而其他菌门的相对丰度对不同施肥处理的响应不同。2)施用氮肥显著改变了土壤化学性状,长期施氮(尿素)处理的土壤全氮、速效氮和有机质含量显著高于不施氮处理而pH则显著降低。3)RDA分析结果显示,pH对土壤细菌和古菌群落结构的影响最大解释了66.5%的变化;其他环境因子则共解释了细菌与古菌群落33.5%的变化各因子的贡献率依次为pH速效钾有效磷有机质。Pearson相关性分析结果也表明,细菌和古菌群落与土壤化学指标间存在密切的相关关系。【结论】解析了长期不同施肥条件下土壤细菌和古菌的群落结构特征,确定了pH是影响土壤细菌和古菌区系的主效环境因子,氮磷钾均衡施肥不仅显著提高了作物产量和丰富了土壤养分含量,同时也增加了土壤细菌的丰富度、多样性以及古菌的多样性,有利于维持良好的土壤生态环境。研究结果对揭示东北黑土肥力演变机制、建立合理的施肥制度具有重要的理论价值。     

Chemical nature of soil organic carbon under different long-term fertilization regimes is coupled with changes in the bacterial community composition in a Calcaric Fluvisol

Fertilization is an important factor influencing the chemical structure of soil organic carbon (SOC) and soil microbial communities; however, whether any connection exists between the two under different fertilization regimes remains unclear. Soils from a 27-year field experiment were used to explore potential associations between SOC functional groups and specific bacterial taxa, using quantitative multiple cross-polarization magic-angle spinning ¹³C nuclear magnetic resonance and 16S rRNA gene sequencing. Treatments included balanced fertilization with organic materials (OM) and with nitrogen (N), phosphorus (P), and potassium (K) mineral fertilizers (NPK); unbalanced fertilization without one of the major elements (NP, PK, or NK); and an unamended control. These treatments were divided into four distinct groups, namely OM, NPK, NP plus PK, and NK plus control, according to their bacterial community composition and SOC chemical structure. Soil total P, available P, and SOC contents were the major determinants of bacterial community composition after long-term fertilization. Compared to NPK, the OM treatment generated a higher aromatic C–O and OCH₃ and lower alkyl C and OCH abundance, which were associated with the enhanced abundance of members of the Acidobacteria subgroups 6 and 5, Cytophagaceae, Chitinophagaceae, and Bacillus sp.; NP plus PK treatments resulted in a higher OCH and lower aromatic C–C abundance, which showed a close association with the enrichment of unclassified Chloracidobacteria, Syntrophobacteraceae, and Anaerolineae and depletion of Bacillales; and NK plus control treatments resulted in a higher abundance of aromatic C–C, which was associated with the enhanced abundance of Bacillales. Our results indicate that different fertilization regimes changed the SOC chemical structure and bacterial community composition in different patterns. The results also suggest that fertilization-induced variations in SOC chemical structure were strongly associated with shifts in specific microbial taxa which, in turn, may be affected by changes in soil properties.     

Effects of different fertilization regimes on nitrogen and phosphorus losses by surface runoff and bacterial community in a vegetable soil

Purpose

Vegetables are major economic crops in China. Their cultivation usually involves high fertilizer application rates leading to significant losses of N and P to the wider environment, resulting in water contamination and low nutrient use efficiency. Hence, it is a matter of urgency to understand the mechanisms and factors that affect N and P losses in vegetable production systems in order to develop optimum fertilization regimes.

Materials and methods

Different fertilization regimes were applied in a long-term chili (Capsicum spp. L.) production soil to study the effects on nitrogen (N) and phosphorus (P) runoff losses, microbial biomass, microbial community, and crop yields. Three fertilization regimes were implemented: control (no fertilizer; CK), farmer’s fertilization practice (FFP), and site-specific nutrient management (SSNM). A fixed collection device was used to quantify the total volume of water output after each precipitation event. All water samples were analyzed for total nitrogen, ammonium nitrogen (NH₄⁺-N), nitrate nitrogen (NO₃^?-N), total phosphorus (TP), and available phosphorus (AP). Soil samples were collected for analysis of the physicochemical properties and for DNA extraction after chili harvest. High-throughput sequencing was used to further investigate the relationship between the microbial community and nutrient losses.

Results and discussion

The SSNM fertilizer regime resulted in a 23.3% yield increase and enhanced agronomic N use efficiency from 11.87 to 15.67% compared with the FFP treatment. Soil available nutrients (i.e., AN and AP) and ATP content increased significantly after SSNM implementation. Under the SSNM regime, N losses decreased by 25.8% compared with FFP but did not lead to significantly different P losses. High-throughput sequencing results showed that each treatment formed a unique microbial community structure. VPA results revealed that the microbial community structure was mainly (50.56%) affected by the interactions between N and P. Mantel results indicated that the soil properties that significantly affected soil microbial community structure followed the order: AP, AK, and salinity.

Conclusions

Our study has demonstrated that SSNM not only generates lower N losses but also provides higher contents of soil available nutrients and plant yield, which were mainly attributed to the multiple top dressings and meeting of the plants’ demand with adequate nutrient supplies. The combined data showed that the microbial community differentiation between the different fertilizer regimes was mainly linked to the interactions between N and P in the soil.

     

设施种植模式对土壤细菌多样性及群落结构的影响

为了研究有机和常规设施种植模式及轮作对土壤细菌多样性和群落结构的影响,本研究采用Illumina平台Hiseq 2500高通量测序技术,于2016年6月(作物处于收获期)对北京市顺义区不同设施种植模式(分别为有机设施种植模式和常规设施种植模式下的叶菜连作、茄果连作和叶茄轮作)下土壤细菌进行16S r RNA测序。测序质控后共获得17 278个操作分类单元(operational taxonomic units,OTUs),共计318 851条有效序列。比较不同种植模式和轮作下土壤细菌多样性、细菌群落结构组成、相对丰度及土壤理化性质与细菌群落多样性关系的差异性。结果表明:土壤微生物群落结构在有机和常规设施种植模式下差异明显,有机设施种植土壤细菌多样性高于常规设施种植;有机设施种植下轮作与连作土壤细菌群落结构表现出明显差异,而常规设施种植下,两者没有明显差异;有机种植模式下,轮作土壤细菌群落多样性高于连作土壤;设施种植土壤细菌群落主要属于鞘氨醇单胞菌属(Sphingomonas,5.05%)和芽孢杆菌属(Bacillus,4.84%),相对丰度大于0.5%的共有14个属。有机设施种植土壤含有较多促进植物生长、有机质分解的细菌,常规设施种植土壤中降解化学杀虫剂、防治土壤病害、促进硝化过程的细菌较多。RDA分析结果显示土壤细菌群落主要受全磷、速效磷、有机质的影响。Tumebacillus、Candidatus Solibacter和Acidothermus都是分解有机质、利用碳源的细菌属,与土壤有机质含量呈正相关关系。由此可见,设施条件下,有机和常规种植土壤微生物群落结构的差异性主要源于肥料使用、有害生物防治措施和管理方式的不同。有机设施种植模式下,轮作更有利于发挥其改良土壤营养循环和防治土壤病虫害的作用。上述结果为在微生物水平上研究设施条件下不同种植模式的土壤生态质量差异提供了参考。