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1.
梯度有机负荷下农业废弃物厌氧发酵特性及微生物群落   总被引:4,自引:2,他引:2  
为了解梯度负荷对不同C/N农业废弃物厌氧发酵特性及微生物群落的影响,该试验以猪粪、金针菇菌包、稻秆和甘蔗叶等C/N差异较大的废弃物作为原料,通过逐渐提高全混合厌氧反应器(continuous stirred tank reactor,CSTR)的有机负荷率(organic loading rate,OLR),研究物料在4个OLR(1.11、1.67、2.22和2.78g/(L·d),以可挥发性固体计)下的产甲烷特性和微生物群落结构变化.结果表明:4种物料的日甲烷产量均随OLR递增而增加,但单位物料甲烷产率与微生物菌群结构则因物料C/N差异而表现出不同的变化趋势.其中:猪粪的日甲烷产量最高,细菌和古菌的菌群结构与对照组相比变化不大;多样性指数先增后减,甲烷产率也在OLR达到1.67g/(L·d)后逐渐下降.金针菇菌包甲烷产率相对稳定,细菌和古菌的多样性指数随OLR递增而增长.稻秆和甘蔗叶同属于碳质量分数高的秸秆类物料,二者的细菌和古菌的菌群结构变化明显;但在OLR达到2.78g/(L·d)时,稻秆受系统酸化(VFA/TIC>0.8)影响,甲烷产率下降明显,细菌和古菌的多样性指数也出现下降;而甘蔗叶则因VFA/TIC<0.8,其甲烷产率在发酵过程中未出现明显下降.此外,不同C/N物料对优势菌的形成存在影响.其中:Methanolobus zinderi为猪粪物料特有的优势古菌;Proteiniphilum acetatigenes,Acetivibrio cellulolyticus为金针菇菌包、甘蔗叶特有的优势细菌;Methanospirillum hungatei为稻秆物料独有的优势古菌.  相似文献   

2.
基于PCR-DGGE技术研究芦苇秸秆氢气-甲烷厌氧联产过程中,细菌微生物群落结构特征和演替规律。结果表明,厌氧联产过程中细菌群落结构分布存在明显的阶段性差异。产氢阶段初期,细菌群落相似性较小,随着厌氧联产的进行,细菌种类逐渐增多并在产氢高峰期保持稳定,群落相似性较高,戴斯系数(Cs)为83.6%(第12、24小时,泳道H3和H4)。产甲烷高峰期Cs值达到87.4%(第210、258小时,泳道M6,M7),群落结构稳定,产甲烷末期Cs值降低至51.5%(第210、432小时,泳道M6,M9)。序列分析表明,Enterobacter aerogenes产气肠杆菌是具有高效产氢潜力的兼性厌氧细菌,Sedimentibacter产氢产乙酸菌是产氢阶段的优势微生物。Clostridium thermocellum嗜热纤维素菌是厌氧联产过程的优势微生物,具有降解纤维素功能,对芦苇秸秆的能源化利用起到重要作用。  相似文献   

3.
为了探究双污泥系统下反硝化除磷-诱导磷结晶工艺中硝化池内微生物特性,该文利用原位荧光杂交(fluorescence in situ hybridization,FISH)技术、电子显微镜扫描(scanning electron micrograph,SEM)方法和Image-Pro Plus(IPP)软件考察了该工艺中硝化细菌的种群结构、形态和硝化污泥微观三维结构图。结果表明:该工艺中硝化池内氨氧化细菌(ammonia-oxidizing bacteria,AOB)数量要多于亚硝酸盐氧化细菌(nitrite-oxidizing bacteria,NOB),占总细菌的比例分别为46.2%,28.5%,且AOB处于污泥颗粒外层而NOB处于污泥颗粒内层,可能由于NOB利用AOB的代谢产物所致;工艺中硝化细菌多以球形或短杆菌为主,NOB生长时多以几个细胞形成小团聚体,而AOB生长时则形成大的团聚体;通过硝化污泥微观三维结构发现,污泥外层呈密实状而内层较疏松且有空洞存在,可能由于污泥外层和内层微生物的丰度差异、营养物质和溶解氧的浓度差异所致。此外,与传统的单污泥污水处理工艺相比,双污泥工艺明显地增强了硝化细菌的生长和富集能力。  相似文献   

4.
基于高通量测序技术的樱桃根际细菌群落研究   总被引:2,自引:0,他引:2  
本研究基于Illumina Hiseq测序平台和16S r DNA扩增测序技术,对三年生砂蜜豆/马哈利樱桃5个生长发育期(萌芽期、展叶期、新梢迅速生长期、新梢停长期及落叶期)根际土壤细菌群落分别进行了基因序列测定,分析了不同生长发育期根际细菌种群结构的变化。结果表明变形菌门(Proteobacteria)、酸杆菌门(Acidobacteria)、厚壁菌门(Firmicute)以及放线菌门(Actinobacteria)细菌在所有样品中均为优势菌群,检出率高达80%以上。在樱桃生长过程中,不同生长发育期根际细菌群落结构不同,萌芽期种群类别最少,然后逐渐增加,到新稍迅速生长期达最多,多样性最丰富,进而又逐渐减少。对样品序列进行聚类分析,5个样品归属于两大类群,萌芽期、展叶期及新稍迅速生长期的根际土壤样品具有更加相似的细菌群落结构;而停长期和落叶期样品中细菌组成相似。研究还发现绿弯菌门细菌是新稍迅速生长期和其他样品之间具有统计学差异的物种。由此可见,砂蜜豆/马哈利樱桃生长发育期对根际土壤细菌群落具有一定的影响。  相似文献   

5.
以两只750ml UASB反应器分别作为酸化相和甲烷相进行农业废弃物厌氧处理的研究,两相中,都培养出了颗粒污泥。酸化相有机负荷达240kgCOD/m~3[bed]·d,HRT=1.6h,酸化率66%,COD去除率4%~8%,甲烷相有机负荷达88kgCOD/m~3[bed]·d,HRT=3.7h,COD去除率大于85%。两相COD总去除率大于90%,总有机负荷60kgCOD/m~3[bed]·d。并结合扫描电镜照片,对两相颗粒污泥的形成,菌体分布规律等进行了探讨。  相似文献   

6.
  【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。  相似文献   

7.
甲烷营养菌(methanotrophs)是一类以CH4为唯一碳源和能源的细菌,广泛分布在水稻土、森林土、苔原土、泥炭地、海洋与湖泊底泥、堆肥、垃圾填埋场及地下水等环境中,并作为大气甲烷(CH4)唯一的生物汇(库),在全球温室效应研究中备受关注。目前,关于土壤甲烷营养菌的研究主要包括菌株的多样性、生态分布以及环境因素对微生物氧化CH4过程的影响。本文从甲烷营养菌的分类入手,概述稻田土壤CH4的氧化与释放、旱地土壤CH4的氧化以及影响土壤CH4氧化的因素等方面的研究进展,同时介绍了土壤甲烷营养菌研究领域的几种主要的分子研究技术,以期为甲烷营养菌相关的研究提供参考。  相似文献   

8.
以两只750mlUASB反应器分别作为酸化相和甲烷相进行农业废弃物厌氧处理的研究,两相中,都培养出了颗粒污泥。酸化相有机负荷达240kgCOD/m3[bed]·d,HRT=1.6h,酸化率66%,COD去除率4%~8%,甲烷相有机负荷达88kgCOD/m3[bed]·d,HRT=3.7h,COD去除率大于85%。两相COD总去除率大于90%,总有机负荷60kgCOD/m3[bed]·d。并结合扫描电镜照片,对两相颗粒污泥的形成,菌体分布规律等进行了探讨。  相似文献   

9.
亚硝酸盐型甲烷厌氧氧化(nitrite-dependent anaerobic methane oxidation, n-damo)是控制稻田甲烷排放的一种新途径,但有关其在稻田甲烷氧化中的贡献及其对大气CO2升高的响应尚不清楚。本文依托开顶式气室组成的CO2浓度自动调控平台,在环境 CO2浓度(CK)基础上,设置了CO2缓增处理(EC:每年增加 40 μL·L-1, 至采样时CO2浓度为CK+160 μL·L-1)。采用稳定性同位素示踪、定量PCR和高通量测序等手段,分析了不同CO2处理下水稻关键生育期(分蘖期、拔节期和开花期)稻田n-damo活性及其功能微生物Candidatus Methylomirabilis oxyfera (M. oxyfera)-like细菌的丰度、多样性和群落组成。结果发现,EC处理在一定程度上刺激了土壤中n-damo活性以及M. oxyfera-like细菌丰度,且在拔节期达到了显著性水平。EC处理还使M. oxyfera-like细菌的群落组成发生了显著改变,并影响了其多样性。EC处理下土壤可溶性有机碳含量和无机氮含量的改变很可能是导致n-damo活性及M. oxyfera-like细菌群落结构发生变化的重要原因。综上,稻田n-damo过程对大气CO2浓度缓增具有正响应,表明了其在未来气候变化背景下对稻田甲烷减排的积极作用。  相似文献   

10.
长期不同施肥处理对栗褐土可培养微生物数量的影响   总被引:1,自引:0,他引:1  
在24年长期定位试验的基础上,以选择性培养基为基础研究长期定位施肥对土壤微生物种群及数量的影响,同时测定了土壤速效养分的含量,探讨了微生物与土壤速效养分的相关关系。结果表明,栗褐土微生物以细菌为主体,约占微生物总数的92.72%~95.91%;施用有机肥可以显著提高土壤微生物的数量,处理M1N细菌数量最多,显著高于对照;处理M1NP放线菌数量最多,处理M2NP真菌数量最多;施用有机肥可以显著提高土壤解磷细菌数量;细菌、真菌、放线菌和无机磷细菌与碱解氮、速效磷和速效钾相关性均达显著或极显著水平。  相似文献   

11.
The community structure of methanogenic archaea is relatively stable,i.e.,it is sustained at a high abundance with minimal changes in composition,in paddy field soils irrespective of submergence and drainage.In contrast,the abundance in non-methanogenic oxic soils is much lower than that in paddy field soils.This study aimed to describe methanogenic archaeal community development following the long-term submergence of non-methanogenic oxic upland field soils in pot and field experiments.In the pot experiment,a soil sample obtained from an upland field was incubated under submerged conditions for 275 d.Soil samples periodically collected were subjected to culture-dependent most probable number(MPN)enumeration,polymerase chain reaction-denaturing gradient gel electrophoresis(PCR-DGGE)analysis of archaeal 16 S r RNA gene,and quantitative PCR analysis of the methyl-coenzyme M reductase alpha subunit gene(mcr A)of methanogenic archaea.The abundance of methanogenic archaea increased from 102 to 103 cells g-1 dry soil and 104 to 107 copies of mcr A gene g-1 dry soil after submergence.Although no methanogenic archaeon was detected prior to incubation by the DGGE analysis,members from Methanocellales,Methanosarcinaceae,and Methanosaetaceae proliferated in the soils,and the community structure was relatively stable once established.In the field experiment,the number of viable methanogenic archaea in a rice paddy field converted from meadow(reclaimed paddy field)was monitored by MPN enumeration over five annual cycles of field operations.Viability was also determined simultaneously in a paddy field where the plow layer soil from a farmer’s paddy field was dressed onto the meadow(dressed paddy field)and an upland crop field converted from the meadow(reclaimed upland field).The number of viable methanogenic archaea in the reclaimed paddy field was below the detection limit before the first cultivation of rice and in the reclaimed upland field.Then,the number gradually increased over five years and finally reached 103–104 cells g-1 dry soil,which was comparable to that in the dressed paddy field.These findings showed that the low abundance of autochthonous methanogenic archaea in the non-methanogenic oxic upland field soils steadily proliferated,and the community structure was developed following repeated and long-term submergence.These results suggest that habitats suitable for methanogenic archaea were established in soil following repeated and long-term submergence.  相似文献   

12.
巢湖十五里河不同河段沉积物古菌群落结构特征研究   总被引:2,自引:0,他引:2  
[目的]为了探究城市河流沉积物古菌群落结构特征及其环境指示意义,[方法]应用高通量测序技术获取巢湖十五里河不同河段表层沉积物古菌群落组成,用冗余分析(RDA)和相关性分析等方法探究古菌群落结构特征与沉积物环境因子间的联系.[结果]十五里河表层沉积物古菌的优势菌门和次优势菌门分别是广古菌和奇古菌与深古菌,其相对丰度分别为...  相似文献   

13.
Paddy field is a major emission source of methane. Methane is the terminal product of anaerobic decomposition of organic matter and generated by methanogenic archaea under flooded conditions in paddy fields. This study aimed to reveal the effect of winter flooding on methanogenic archaeal community structure in paddy fields of Andosols under organic farming. Soil samples were collected from experimental paddy fields in the Field Science Center, Tohoku University, for two years. They were under flooding conditions during winter with organic farming, under non-flooding conditions during winter with organic farming and under non-flooding conditions during winter with conventional farming (non-organic farming). Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) analysis of methanogenic archaeal 16S rRNA gene revealed that the DGGE patterns were nearly the same irrespective of the treatment and sampling times. Twenty-three bands were observed from each treatment and 4, 13 and 6 sequences were closely related to Methanomicrobiales, Methanosarcinales and Methanocellales, respectively. Real-time quantitative PCR analysis indicated that the abundance of methanogenic archaeal 16S rRNA gene and mcrA gene, encoding α subunit of methyl-coenzyme M reductase, was not significantly different among the paddy fields. This study first revealed a methanogenic archaeal community in an Andosol paddy field and showed that the community was not affected by winter flooding under organic farming.  相似文献   

14.
15.
Community structure of methanogenic archaea in paddy field soil under double cropping (rice [Oryza sativa L.] and wheat [Triticum aestivum L.]) was studied by the denaturing gradient gel electrophoresis (DGGE) method. Soil samples under flooded and upland conditions were collected 7 and 6 times, respectively, from two paddy fields throughout a year, and two primer sets, 0357F-GC/0691R and newly designed 1106F-GC/1378R, were used for DGGE analysis. The 25 and 29 different bands were observed on the DGGE gels with the primers 0357F-GC/0691R and 1106F-GC/1378R, respectively. DGGE band patterns of the methanogenic archaeal community were stable throughout a year including the cultivation periods of rice under flooded conditions and of wheat under upland conditions. Cluster analysis and principal component analysis suggested that the difference in the soil type (sampling region) largely influenced the community structures of methanogenic archaea in paddy field soil, while the effects of sampling period and different fertilizer treatments on them were small. Most of the sequences obtained from the DGGE bands were closely related to Methanomicrobiales, Methanosarcinaceae, Methanosaetaceae and Rice cluster-I.  相似文献   

16.
Addition of rice straw, which is a common practice in rice agriculture, generally results in enhanced production and emission of the greenhouse gas methane (CH4). However, it is unclear whether straw addition affects only the activity or also the composition of the methanogenic microbial community. It is also unclear to what extent methanogenic archaea would be able to proliferate in the soil. Anoxic slurries of Italian rice‐field soil produced CH4 after a lag, during which ferric iron and sulfate were reduced. Addition of rice straw slightly decreased this lag and greatly enhanced the subsequent production of CH4. At the same time, addition of rice straw enhanced the intermediate production of H2 and acetate that served as the methanogenic substrates. Compared with the unamended control, the addition of rice straw resulted in an increased concentration of phospholipid fatty acids in the soil. Quantitative ‘real‐time’ PCR targeting the 16S rRNA gene also showed increased copy numbers of both Bacteria and Archaea in the straw‐amended soil at the end of the experiment. The composition of the archaeal community was followed over time by terminal restriction length polymorphism (T‐RFLP) analysis of the archaeal 16S rRNA genes extracted from straw‐amended soil and the control. Rice Cluster‐I (RC‐I) methanogens and Methanosarcinaceae were the most abundant methanogenic populations, followed by Methanobacteriales, Methanomicrobiales and Methanosaetaceae. Addition of rice straw resulted in a relative increase of Methanosarcinaceae and Methanobacteriales and a relative decrease of RC‐I methanogens and Methanomicrobiales. Our results revealed a dynamic methanogenic community in anoxic rice‐field soil and showed that addition of organic matter selectively enhanced the growth of particular methanogenic populations, which were apparently better adapted to the presence of straw than the others. The extent of archaeal growth was consistent with that expected theoretically from the ambient Gibbs free energies of hydrogenotrophic and acetoclastic methanogenesis.  相似文献   

17.
A comprehensive comparison about microbial community (bacterial, archaeal and fungal) response to different tillage managements in Northern China remain little studied, in this study we compared no-tillage (NT) versus conventional tillage (CT) management on topsoil microbial community diversity and composition in field experiment. We found that NT practice significantly increased the soil moisture content (SMC), bulk density, stocks of soil organic carbon (SOC), total nitrogen (TN), and microbial biomass carbon and nitrogen (P < 0.05). Moreover, higher levels of bacterial and archaeal alpha diversity were observed in NT relative to CT while unexpectedly, there was no significant difference found in fungal diversity between two treatments. The most pronounced shifts in the composition of the different microbial groups were found for the archaeal community, which followed by bacterial and fungal. NT practice markedly enhanced abundances of Proteobacteria (belongs to bacteria) phyla, Thaumarchaeota phyla (belongs to archaea) and Glomeromycota phyla (belongs to fungi). Redundancy analysis revealed that the factor that most closely correlated with bacterial, archaeal and fungal composition were SMC, TN and SOC, respectively. Considering NT enhanced both microbial composition and C storage in topsoil, we suggest that NT offers significant promise to improve topsoil health in this region.  相似文献   

18.
间隙灌溉和控释肥施用对稻田土壤产甲烷微生物的影响   总被引:1,自引:0,他引:1  
纪洋  于海洋  Conrad Ralf  徐华 《土壤》2017,49(6):1132-1139
间隙灌溉和控释肥施用影响稻田CH_4的产生和排放,然而其微生物机理尚不清楚。本研究通过采集稻季田间原位试验新鲜土样,采用核酸定量技术(qPCR)和末端限制性片段长度多态性(T-RFLP)技术,研究间隙灌溉和控释肥施用对稻田土壤产甲烷微生物群落丰度和结构的影响。结果表明,稻季CH_4排放量与古菌、产甲烷菌(mcr A基因)和甲烷氧化菌(pmo A基因)数量均呈极显著正相关关系(P0.01),而与细菌数量无显著相关性。间隙灌溉显著影响产甲烷菌和甲烷氧化菌数量的季节变化,其中烤田抑制产甲烷菌生长,而对甲烷氧化菌数量没有显著影响。与尿素相比,施用控释肥增加了稻田土壤细菌、古菌和产甲烷菌数量,降低了甲烷氧化菌数量。土壤古菌群落的优势T-RFs片段为184bp和391bp,其中184bp片段的相对丰度随着间隙灌溉的进行由45%~55%降低到23%~30%;而391bp片段则相反,其相对丰度由12%~18%增加到23%~26%。典型相关性分析(CCA)表明间隙灌溉显著影响土壤古菌群落结构(P0.001),而控释肥施用对土壤古菌群落结构没有明显影响。  相似文献   

19.
Flooded paddy fields are the major anthropogenic sources of methane (CH4) emission, and organic materials of rice plant origin were estimated to be important as its source. This study used rice (Oryza sativa L. cv, Yukihikari) callus cells as a model material for slough-off root cap cells, and carbon-13 (13C)-labelled callus cells were subjected to decomposition in aerobic and anaerobic soil microcosms for 56 days. DNA was extracted from a soil incubated with carbon-12 (12C)- and 13C-callus cells and subjected to buoyant density gradient centrifugation to identify methanogenic archaeal species that assimilated carbon from the callus cells. 13C-labelled 16S rRNA gene (16S rDNA) fragments from methanogenic archaea were not polymerase chain reaction (PCR)-amplified in heavy fractions under aerobic soil conditions, while they were successfully done from day 3 onwards under anaerobic soil conditions. Eighty-four denaturing gradient gel electrophoresis (DGGE) bands in heavy fractions were sequenced, revealing that they were members of Methanosarcina spp. (20 clones), Methanosaeta spp. (18 clones), Methanocella spp. (25 clones), Methanomicrobiales (10 clones), Methanobacterium spp. (7 clones) and Cluster ZC-I (2 clones). They included hydrogenotrophic and acetoclastic methanogens and were phylogenetically different from those residing in rice roots and, presumably, from those assimilating root exudate and mucilage-derived carbon. This study indicates that carbon of slough-off root cap cells propagates specific methanogenic species in rice rhizosphere under anaerobic soil conditions and thus augments the diversity of the total rhizospheric methanogenic community.  相似文献   

20.
Diversity of methanogenic archaeal communities in Japanese paddy field ecosystem was evaluated by the denaturing gradient gel electrophoresis (DGGE) after PCR amplification of the 16S rRNA genes (16S rDNAs), sequencing analysis and data evaluation by principal component analysis. Data were obtained from samples collected from the plowed soil layer, rice roots, rice straws incorporated in soil, plant residues (mixture of weeds, rice litters, rice roots, and rice stubbles) in soil, and composing rice straw. The number of bands of DGGE profiles ranged from 12 to 26 with the highest numbers in rice roots and rice straws incorporated in soil. However, the diversity indices based on both the numbers and intensity of bands indicated that the community of the plowed soil layer was the most diverse, even, and stable. Sequencing of the main DGGE bands showed the presence of Methanomicrobiales, Methanosarcinales, Methanobacteriaceae, and Methanocellales. The plowed soil layer included all phylogenetic groups of the methanogenic archaea of the other studied habitats, with prevalence of the members of Methanomicrobiales and Methanocellales. The phylogenetic diversity was compared with that of paddy soils collected in Italy, China, and the Philippines and that of 12 anaerobic environments (fen, waste, coast, permafrost, natural gas field, bovine rumen, riparian soil, termite, ciliate endosymboints, lake sediment, landfill, and seep rumen). The phylogenetic diversity was more similar among paddy soils than with the other anaerobic environments. Probably, the methanogenic archaeal communities of the paddy field soils were characterized by indigenous members and some of the members of the community of the plowed soil layer colonized rice roots, rice straws, and plant residues.  相似文献   

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