长期不同施肥制度对潮棕壤微生物生物量碳、氮及细菌群落结构的影响

以沈阳生态站长期定位试验为研究平台,采用传统氯仿熏蒸方法和现代PCR-DGGE技术探讨了长期不同施肥制度对土壤微生物生物量碳和氮及细菌群落结构的影响.结果表明:在整个试验期,土壤微生物生物量碳和氮的动态变化趋势基本相同;长期施用有机肥可显著提高土壤有机碳和土壤微生物生物量碳和氮含量,而长期施用化肥明显降低土壤pH,土壤微生物生物量碳和氮含量也显著降低.DGGE图谱表明:不同施肥处理的细菌16S rDNA多数条带分布相同,28条带中有18条为共有条带,说明潮棕壤中细菌类群较稳定,但其数量受到施肥的影响;长期施用有机肥促进潮棕壤细菌群落结构的多样性,而施用化肥处理则降低了其多样性.     

长期施肥对农田黑土r-K策略菌群的影响

直接参与土壤养分代谢周转过程的土壤微生物群落是土壤肥力质量的重要衡量指标。本文研究了长期施肥对农田黑土细菌和真菌r-K策略菌群(生态生理功能)的影响。结果表明:施肥处理菌群生长的均匀度和丰富度有所降低,细菌和真菌生态生理指数(EP)降低幅度分别为0.019～0.106和0.023～0.185。各处理K策略菌数量均大于相应的r策略菌。施肥能增加土壤r策略细菌数量,但不利于r策略真菌生长。与CK相比,r-K策略细菌和K策略真菌数量在单施中量有机肥处理中增幅最大;有机肥与化肥配施处理r-K策略细菌和K策略真菌数量高于单施同种化肥处理。在K策略菌占优势下,施用化肥有利于r策略菌比率提高,而有机肥与化肥配施倾向于使K策略菌比率增加。相关性分析表明,K策略菌群与土壤N素呈极显著正相关(P<0.01),可能是影响土壤N素循环的关键菌群。     

施肥和土壤管理对土壤微生物生物量碳、氮和群落结构的影响

以中国科学院沈阳生态试验站的长期定位试验为平台,研究了不同施肥和土壤管理对潮棕壤微生物生物量碳、氮和群落结构的影响。结果表明,裸地和农田处理的微生物生物量碳、氮较低,但是农田处理下施肥增加了微生物生物量,其中NPK+M效果最明显。DGGE图谱显示,处理间细菌条带分布较相似,其中裸地的细菌多样性最高;长期施肥和土壤管理改变了土壤真菌群落结构,施肥增加了真菌多样性,且有机肥的影响大于化肥;不同处理间氨氧化细菌群落结构差异显著,NPK+M显著增加了氨氧化细菌多样性,且无机肥和有机肥对氨氧化细菌群落影响不同。施肥和土壤管理对细菌影响较小,但显著改变了真菌和氨氧化细菌的群落结构。聚类分析结果显示,土壤管理措施较施肥对细菌、真菌和氨氧化细菌群落的影响更为显著。     

长期施肥对农田黑土微生物群落功能多样性的影响

为了研究长期不同施肥对农田黑土微生物群落功能多样性的影响,采用Biolog ECO微平板培养法,对已经连续施肥35年的农业部哈尔滨黑土生态环境重点野外科学观测试验站4种处理(CK、NPK、M、MNPK)的两个土层(0～20、20～40 cm)微生物群落功能多样性进行研究.结果表明: 在0～20 cm土层,有机无机肥(MNPK)配施能够显著提高土壤微生物对碳源的利用能力以及群落代谢功能的丰富度、多样性和优势度,而在20～40 cm土层则低于单施化肥(NPK)处理,两个土层单施化肥均降低土壤微生物群落代谢均匀度.不同施肥处理土壤微生物对6种碳源的利用率在0～20和20～40 cm两个土层间存在差异,而在每个土层的处理间差异显著(P<0.05),但各处理间的变化规律在两个土层中不一致.典范对应分析(CCA)结果表明,各处理土壤微生物群落代谢功能在0～20和20～40 cm土层间存在差异,且在两个土层的处理间也存在差异,而土壤养分含量对各处理土壤微生物群落代谢功能的影响规律在两个土层中较为相似.长期不同施肥会对耕层及以下土壤微生物群落功能多样性产生影响,在20～40 cm土层中单施化肥对微生物群落功能多样性的影响较大.     

长期施用化肥及秸秆还田对砂姜黑土细菌群落的影响

【目的】在施用化肥的基础上进行秸秆还田是提高砂姜黑土肥力的有效措施,以往的研究只注重秸秆还田对土壤结构、肥力等物理化学性状方面的研究,缺少施肥对砂姜黑土微生物群落影响的研究。本研究以安徽蒙城典型的砂姜黑土为研究对象,以期揭示长期施用化肥和秸秆还田对砂姜黑土细菌群落的影响。【方法】采用454高通量测序对砂姜黑土不同农业施肥措施下的细菌群落进行分析研究,并通过生物信息学的分析方法揭示影响砂姜黑土细菌群落的主要因素。【结果】通过对454高通量测序数据的分析,发现砂姜黑土主要的细菌门类为放线菌、变形菌、酸杆菌、绿弯菌和拟杆菌。长期施用化肥显著提高了砂姜黑土肥力和作物产量,但导致了细菌群落结构的显著变化和多样性的显著降低。秸秆还田有利于土壤肥力的进一步提高,但是并没有缓解长期施用化肥对土壤细菌群落产生的不利影响。分析发现土壤pH的变化是导致土壤细菌群落变异的主要因素。【结论】在施用化肥的基础上进行秸秆还田有利于砂姜黑土肥力的提升,然而并没有缓解由施肥导致的土壤酸化对土壤细菌群落组成和多样性产生的不利影响。这暗示秸秆还田可能并未对砂姜黑土微生物生态产生根本性的有益影响,对于秸秆农田的利用方式还需要进一步研究,以达到农业生产效益和生态效益的并重。     

耕作和施肥对不同深度黑土中细菌群落结构的影响

【目的】为探讨耕作和施加有机肥、化肥对黑土表层(0-30cm)、中层(100-130cm)及深层(200-230cm)细菌群落结构的影响,【方法】应用DGGE技术对相应土层中细菌群落结构进行了解析。【结果】结果表明,与对照相比,耕作和施加有机肥、化肥对表层黑土细菌群落结构影响较大,二者差异度为4%;而对中层和深层细菌群落结构影响较小,二者差异度为2%。对于细菌群落结构的垂向变化,中层和深层中细菌群落结构的相似性远远高于同表层的相似性。【结论】可见,耕作和施加有机肥、化肥仅对黑土表层土壤(0-30cm)具有一定的影响,而对100cm以下土壤细菌群落影响较小,细菌群落随土壤深度不同的垂向变化要远高于土壤管理措施造成的影响。     

连续施肥对农田黑土微生物功能多样性的影响

应用Biolog Eco微平板培养法研究了中国科学院海伦农业生态试验站20余年连续不同施肥处理(对照、氮磷钾、氮磷钾+秸秆、氮磷钾+猪粪1、氮磷钾+猪粪2和氮磷钾+猪粪3)下土壤微生物代谢功能多样性的变化。结果表明:长期施肥条件下,单施化肥和有机肥与化肥配施对土壤细菌、真菌数量影响不明显,但高量有机肥与化肥配施可以显著增加放线菌数量。Biolog结果表明,施用有机肥的平均颜色变化率高于单施化肥和对照。Shannon指数、Simpson指数和McIntosh指数均是有机肥和化肥配施高于化肥单施与对照。主成分分析结果表明,对照、NPK与其他几种处理有较明显的距离,说明有机肥的连年施用导致了土壤微生物代谢功能多样性的变化。     

长期不同施肥下太湖地区黄泥土表土微生物碳氮量及基因多样性变化

农业管理措施影响下土壤微生物群落结构的变化是农业土壤质量研究的前沿问题。运用化学分析方法和 PCR- DGGE技术从土壤微生物碳氮量及基因多样性角度研究了长期不同施肥措施下太湖地区代表性水稻土 -黄泥土的表土微生物活性与分子多样性的变化。结果表明 ,施用化肥以及化肥和有机肥配施在提高土壤有机碳含量的同时 ,不仅提高了水稻土的微生物碳氮量 ,而且改变了微生物的群落结构 ;与长期单施化肥相比 ,长期化肥配施有机肥不仅显著提高了土壤微生物碳氮量 ,而且提高了土壤微生物的分子多样性 ;就土壤的微生物分子群落相似性来说 ,单施化肥下与未施肥下相近 ,而化肥配施秸秆下与化肥配施猪粪下接近 ,说明土壤的有机培肥对土壤微生物群落结构有重要影响。长期单施化肥下水稻产量的年际波动性显著大于化肥配施有机肥下 ,这进一步佐证了化肥配施有机肥显著促进了水稻土的生态系统初级生产力与较高的土壤生态系统稳定性。应用PCR- DGGE技术所揭示的微生物分子群落结构特点可以指示水稻土 10 a尺度的不同农业管理措施下的土壤质量变化     

施用生物质炭和羊粪对宿根连作茶园根际土壤微生物的影响

茶树长期宿根连作会导致土壤酸化严重、土壤营养不平衡、根际土壤微生态结构恶化.研究生物质炭、羊粪对宿根连作茶树生长以及土壤微生物群落结构和功能的影响,探讨其对宿根连作茶树土壤环境的调节效果,可为宿根连作茶园土壤微生态的改善提供理论依据.本研究以宿根连作20年的茶园土壤为对象,利用Biolog技术和磷脂脂肪酸(PLFA)方法,研究施用生物质炭(40 t·hm^-2)和羊粪替代部分化肥对连作茶树产量和品质、土壤化学性质、根际土壤微生物群落功能和结构的影响.结果表明:施用生物质炭、羊粪1年后酸化茶园土壤的p H和土壤养分显著提高,并提高了茶叶产量.与常规施肥相比,施用生物质炭、羊粪替代部分化肥处理显著提高了茶树根际土壤微生物的碳源代谢活性和微生物多样性,对胺类、碳水化合物和聚合物的相对利用有所增加.生物质炭和羊粪处理的根际土壤总PLFA含量分别比常规施肥处理提高了20.9%和47.5%,羊粪处理还显著降低了总饱和/总单一不饱和脂肪酸比例.生物质炭和羊粪可改善茶园土壤酸化状况和土壤肥力,对宿根连作茶树的生长具有促进作用,两种措施均不同程度地增加了土壤微生物的代谢活性和微生物量、提高了多样性指数、改善了微生物群落结构.施用生物质炭和羊粪可作为调节宿根连作茶园根际土壤微生态的有效措施.     

不同富集和分离培养条件下的含酚废水处理生物膜微生物群落结构与活性比较分析

采用Biolog和变性梯度凝胶电泳(DGGE)技术研究了不同苯酚浓度培养对焦化废水处理厂反硝化池生物膜样品中微生物群落结构和代谢类型的影响。DGGE结果表明, 不同浓度苯酚和不同培养方式富集培养后, 细菌16S rDNA的部分条带分布谱形发生改变, 还有部分条带只受到了苯酚浓度变化的影响; 富集培养过程中由于碳源组成相对焦化废水简单, DGGE条带所代表的优势微生物多样性有所降低。Biolog试验结果表明, 生物膜样本的细菌群落代谢能力最强; 低浓度苯酚富集后的样品能利用的底物碳源类型最丰富。对Biolog试验结果的主成分分析显示, 相同浓度苯酚富集培养后的细菌群落代谢功能多样性相似, 但从DGGE结果看出其结构组成产生了变化。富集培养使样品微生物群落的代谢功能发生改变, 低浓度的苯酚富集增加了群落中微生物的代谢类型。而不同条件获得的分离物其苯酚降解能力的初步分析也表明, 富集与分离条件对苯酚降解菌的分离能力和得到的菌株特性具有差别。     

Effects of fertilization on bacterial community structure and function in a black soil of Dehui region estimated by Biolog and PCR-DGGE methods

Soil microbial community structure and function are commonly used as indicators for soil quality and fertility. In this paper, the bacterial community structure and function in a black soil of Dehui region influenced by fertilization were investigated by Biolog and PCR-DGGE (polymerase chain reaction-denaturing gradient gel electrophoresis) methods. Biolog examination showed that substrate richness and catabolic diversities of bacterial communities were the highest in the treatment of farm yard manure, and the lowest in the chemical fertilizer treatment. DGGE fingerprint showed that the majority of bands were similar among all treatments, suggesting that microbial communities with those bands were stable, and not influenced by fertilization. In general, chemical fertilizer decreased the diversity of soil bacterial communities. The PCA (principal component analysis) plots of Biolog and DGGE revealed that the structure and function of bacterial communities were similar in the non-fertilized control and the treatment of farm yard manure alone, which inferred that the application of farm yard manure increased the quantity of soil microbes but had less effect on the changes of community structure. The catabolic function was similar, but the composition structure differed between the treatments of chemical fertilizer alone and combined application of farm yard manure with chemical fertilizer. These results suggest that the use of chemical fertilizer mainly decreased the catabolic activity of the fast growth bacteria or eutrophic bacteria.     

不同施肥制度土壤微生物量碳氮变化及细菌群落16S rDNA V3 片段PCR产物的DGGE分析

应用化学分析和变性梯度凝胶电泳（DGGE）技术分离PCR扩增的16S rDNA的方法,研究了不同施肥制度对土壤微生物量碳、氮变化及微生物多样性的影响。结果表明,连续15a长期试验下,土壤微生物量碳（SMB-C）和微生物量氮（SMB-N）的含量大小均为长期撂荒（CK0）土壤高于农田土壤,而在农田土壤中,长期施肥的处理（NPK、NPKM、NPKSt和NPKF）高于长期不施肥处理（CK）,不同的种植制度中,长期复种轮作（NPKF）高于长期复种连作（NPK）;各处理的SMB-C/SOC（土壤有机碳）和SMB-N/TN（全氮）的比值的变化趋势与SMB-C和SMB-N变化一致;从PCR-DGGE分析,长期氮磷钾化肥配施有机肥（NPKM）处理的微生物量碳、氮的含量最高,微生物丰度最高,细菌物种最多,其次为长期撂荒（CK0）,CK处理细菌物种最少。UPGMC聚类分析表明NPK和NPKF处理细菌的群落结构相似,CK和CK0处理细菌的群落结构相似,而NPKM和NPKSt处理细菌的群落结构相似。     

Effects of Manure Compost Application on Soil Microbial Community Diversity and Soil Microenvironments in a Temperate Cropland in China

The long-term application of excessive chemical fertilizers has resulted in the degeneration of soil quality parameters such as soil microbial biomass, communities, and nutrient content, which in turn affects crop health, productivity, and soil sustainable productivity. The objective of this study was to develop a rapid and efficient solution for rehabilitating degraded cropland soils by precisely quantifying soil quality parameters through the application of manure compost and bacteria fertilizers or its combination during maize growth. We investigated dynamic impacts on soil microbial count, biomass, basal respiration, community structure diversity, and enzyme activity using six different treatments [no fertilizer (CK), N fertilizer (N), N fertilizer + bacterial fertilizer (NB), manure compost (M), manure compost + bacterial fertilizer (MB), and bacterial fertilizer (B)] in the plowed layer (0–20 cm) of potted soil during various maize growth stages in a temperate cropland of eastern China. Denaturing gradient electrophoresis (DGGE) fingerprinting analysis showed that the structure and composition of bacterial and fungi communities in the six fertilizer treatments varied at different levels. The Shannon index of bacterial and fungi communities displayed the highest value in the MB treatments and the lowest in the N treatment at the maize mature stage. Changes in soil microorganism community structure and diversity after different fertilizer treatments resulted in different microbial properties. Adding manure compost significantly increased the amount of cultivable microorganisms and microbial biomass, thus enhancing soil respiration and enzyme activities (p<0.01), whereas N treatment showed the opposite results (p<0.01). However, B and NB treatments minimally increased the amount of cultivable microorganisms and microbial biomass, with no obvious influence on community structure and soil enzymes. Our findings indicate that the application of manure compost plus bacterial fertilizers can immediately improve the microbial community structure and diversity of degraded cropland soils.     

Community structure of ammonia-oxidizing bacteria under long-term application of mineral fertilizer and organic manure in a sandy loam soil

The effects of mineral fertilizer (NPK) and organic manure on the community structure of soil ammonia-oxidizing bacteria (AOB) was investigated in a long-term (16-year) fertilizer experiment. The experiment included seven treatments: organic manure, half organic manure N plus half fertilizer N, fertilizer NPK, fertilizer NP, fertilizer NK, fertilizer PK, and the control (without fertilization). N fertilization greatly increased soil nitrification potential, and mineral N fertilizer had a greater impact than organic manure, while N deficiency treatment (PK) had no significant effect. AOB community structure was analyzed by PCR-denaturing gradient gel electrophoresis (PCR-DGGE) of the amoA gene, which encodes the alpha subunit of ammonia monooxygenase. DGGE profiles showed that the AOB community was more diverse in N-fertilized treatments than in the PK-fertilized treatment or the control, while one dominant band observed in the control could not be detected in any of the fertilized treatments. Phylogenetic analysis showed that the DGGE bands derived from N-fertilized treatments belonged to Nitrosospira cluster 3, indicating that N fertilization resulted in the dominance of Nitrosospira cluster 3 in soil. These results demonstrate that long-term application of N fertilizers could result in increased soil nitrification potential and the AOB community shifts in soil. Our results also showed the different effects of mineral fertilizer N versus organic manure N; the effects of P and K on the soil AOB community; and the importance of balanced fertilization with N, P, and K in promoting nitrification functions in arable soils.     

Community Structure of Ammonia-Oxidizing Bacteria under Long-Term Application of Mineral Fertilizer and Organic Manure in a Sandy Loam Soil

The effects of mineral fertilizer (NPK) and organic manure on the community structure of soil ammonia-oxidizing bacteria (AOB) was investigated in a long-term (16-year) fertilizer experiment. The experiment included seven treatments: organic manure, half organic manure N plus half fertilizer N, fertilizer NPK, fertilizer NP, fertilizer NK, fertilizer PK, and the control (without fertilization). N fertilization greatly increased soil nitrification potential, and mineral N fertilizer had a greater impact than organic manure, while N deficiency treatment (PK) had no significant effect. AOB community structure was analyzed by PCR-denaturing gradient gel electrophoresis (PCR-DGGE) of the amoA gene, which encodes the α subunit of ammonia monooxygenase. DGGE profiles showed that the AOB community was more diverse in N-fertilized treatments than in the PK-fertilized treatment or the control, while one dominant band observed in the control could not be detected in any of the fertilized treatments. Phylogenetic analysis showed that the DGGE bands derived from N-fertilized treatments belonged to Nitrosospira cluster 3, indicating that N fertilization resulted in the dominance of Nitrosospira cluster 3 in soil. These results demonstrate that long-term application of N fertilizers could result in increased soil nitrification potential and the AOB community shifts in soil. Our results also showed the different effects of mineral fertilizer N versus organic manure N; the effects of P and K on the soil AOB community; and the importance of balanced fertilization with N, P, and K in promoting nitrification functions in arable soils.     

长期施用有机无机肥对潮土微生物群落的影响

微生物群落结构是土壤生态系统变化的预警及敏感指标,可用于表征土壤质量及其生态功能变化。本文用磷脂脂肪酸法研究了有机肥和NPK肥料长期施用对华北平原潮土微生物群落结构的影响及其变化特征。结果表明：长期施用有机和无机肥不仅提高了土壤有机碳、全氮、速效磷和速效钾等含量,改善了土壤酸碱度,而且显著增加了土壤微生物生物量,其中以有机肥的效果最为明显,增幅达到15.4%。长期施用肥料有机肥也改变了土壤微生物的群落结构,提高了细菌数量,降低了放线菌含量,而对真菌数量没有明显影响,导致真菌与细菌的比值下降。主成分分析表明,长期施用有机肥的土壤,细菌以含a19:0、br14:0、16:1w5c和17:1w9而真菌以含18:1w10c的微生物为优势种群,NPK处理土壤中细菌以含18:1w7、i19:0、br18:0、16:1w7t和a15:0的微生物为优势种群,CK处理中没有明显的优势种群。     

长期施肥对砂姜黑土丛枝菌根真菌群落的影响

在农业生态系统中,丛枝菌根真菌(AM真菌)与很多作物的根系都存在互惠互利的共生关系,与作物的生长和健康密切相关,同时,这类特殊的真菌群落也会受到施肥等农业措施的影响.本研究依托长期定位试验4个试验处理(不施肥、单施化肥、化肥配施秸秆、化肥配施粪肥),研究砂姜黑土AM真菌群落对不同施肥措施的响应及其影响因素,探索不同处理AM真菌指示种的存在.结果表明: 砂姜黑土中的主要AM真菌类群为原囊霉科、多孢囊霉科、巨孢囊霉科、近明球囊霉科、球囊霉科和类球囊霉科;其中类球囊霉属在化肥和有机物料配施中具有显著指示作用.与对照相比,长期单施化肥显著改变AM真菌群落结构并降低其多样性,配施秸秆处理进一步降低AM真菌群落多样性,而配施粪肥明显缓解因施用化肥而造成的多样性减少现象.检验发现,导致AM真菌群落变化最主要的影响因素是土壤pH和可溶性碳.总之,长期不同有机物料和化肥配施对砂姜黑土AM真菌群落结构和多样性会产生不同影响,其中化肥配施粪肥更有利于土壤AM真菌群落多样性的维持.     

长期施肥条件下黄土高原黑垆土作物产量与土壤碳氮的关系

通过设置在黄土高原黑垆土区的长期定位试验系统,研究了长期施肥条件下作物产量与土壤碳氮的互馈关系.试验设不施肥(CK)、单施氮肥(N)、氮磷配施(NP)、秸秆与氮磷配施(SNP)、施有机肥(M)和有机肥与氮磷配施(MNP)6个处理.结果表明: 与对照相比,长期平衡施用化肥、单施有机肥、化肥与有机肥配合施用和秸秆还田配施化肥显著增加了作物产量及其稳定性, NP、SNP、M、MNP处理玉米和小麦产量分别增加92%、97%、93%、141%和147%、164%、139%、214%.NP处理玉米和小麦年均产量与当地常规施肥作物产量相当且稳定,小麦-玉米轮作体系施肥量为N 90 kg·hm^-2、P₂O₅ 75 kg·hm^-2能够满足作物需要.秸秆还田与隔年施磷相配合的SNP处理与NP处理作物产量相似,且可减少磷肥施用量50%.平衡施用化肥、有机肥、化肥与有机肥配施和秸秆还田配施化肥均可显著增加土壤有机碳含量,而施用化肥对土壤全氮含量影响不明显,综合所有处理,土壤有机碳和全氮含量呈显著正相关.不同处理土壤有机碳固存率在15%～41%.SNP处理土壤有机碳累积投入量增加1 t·hm^-2,土壤有机碳含量增加0.06 g·kg^-1,而CK、N、NP、M和MNP处理的增幅在0.12～0.15 g·kg^-1.玉米和小麦产量都与土壤全氮含量呈显著正相关,玉米产量随土壤有机碳含量的增加而增加,但小麦产量随土壤有机碳含量的增加先快速增加后趋于平稳,拐点出现在6.8 g·kg^-1.长期平衡施用化肥、有机肥、有机肥与化肥配合施用及秸秆还田配施化肥可显著增加黄土高原黑垆土土壤有机碳和全氮含量、作物产量和根茬还田量,根茬还田量的增加又进一步增加了土壤有机碳和全氮含量,形成了相互促进的互馈关系.